LOTUS NEWSLETTER

1996 Volume 27

Editor: P.R. BEUSELINCK USDA-Agricultural Research Service

CONTENTS

1. Introduction: instructions to contributors notes from the editor

2. Lotus Activities: reports and abstracts

3. Recent Lotus Literature


Lotus Newsletter Survey

Please answer the following questions to help update the mailing list and provide information about your research interests.

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P. R. Beuselinck, USDA-ARS

University of Missouri

207 Waters Hall

Columbia, MO 65211 USA

FAX 573-882-1467


LOTUS NEWSLETTER, No. 27 1996

Purpose: The Lotus Newsletter consists of informal communications of research information on Lotus spp. Reports of any phase of research on Lotus breeding, genetics, taxonomy, management, utilization or physiology are welcome. Your biographic sketches and information about your research objectives, approaches, and progress including titles of your publications are encouraged. Seed requests and news items are accepted.

INSTRUCTIONS TO CONTRIBUTORS

This is the 27th year of publication for the Lotus Newsletter. Now is the time to consider contributing to the 28th volume of the Lotus Newsletter. Contributions generally are compiled without editing.

IF YOU USE A WORD PROCESSOR

1. Prepare your contribution using any IBM (MS-DOS) or Macintosh word processing program. Then you have two options:

a. submit the file on 3.5 " (90 mm) disk accompanied by a printed copy of the contribution. Identify which program you used. OR

b. submit the file to my e-mail address (pbeuselinck@plantsci.missouri.edu) and send me a hardcopy by FAX to 573-882-1467, or by regular mail.

IF YOU DO NOT HAVE ACCESS TO A COMPUTER OR E-MAIL

1. Single space typewritten text on white 8.5" x 11" (21.5 cm x 27.5 cm) paper. Double space between paragraphs and tables. Do not number pages.

2. Leave a minimum of 3/4" (1.9 cm) on the left and bottom of each page.

3. On the first page of each contribution, indicate state or country, title of report, and name(s) of contributor(s).

4. Send your contributions by December 31, 1996 to:

Lotus Newsletter

Dr. P. R. Beuselinck, USDA-ARS

Plant Genetics Research Unit

207 Waters Hall

University of Missouri

Columbia, MO 65211 U.S.A.

E-mail: pbeuselinck@plantsci.missouri.edu

FAX 573-882-1467


COST of the Lotus Newsletter

The expense of publishing the Lotus Newsletter has been partially covered by unrestricted research support. I will continue to strive for financial support of the Lotus Newsletter to provide you with an unencumbered communication resource. Please note my comments about an internet version of the Lotus Newsletter below.


THANKS AGAIN!

Many thanks to you who respond to my requests for information about your Lotus research. Your contributions to the Lotus Newsletter help generate a better perspective of the international research and management on the many species of Lotus.


LIBRARY REQUESTS

Requests for distribution the Lotus Newsletter to university or research libraries are accepted. If you have a library that needs a copy for your research group or center please notify me. Please note my comments about an internet version of the Lotus Newsletter below.


BACK ISSUES

There is a limited supply of back issues available. Supplies of most volumes have been depleted, but requests will be handled on a first-come first-served basis.


INTERNET AVAILABILITY

This was the last issue of the Lotus Newsletter to be printed in a hardcopy form. The cost of printing and mailing has become quite expensive. You should be able to go the Lotus Newsletter homepage and print a copy for yourself or you simply access the homepage and read the volume of your choice without filling your bookshelf. My goal is to work my way back through all the volumes until they are all in electronic format.

From the homepage you should be able to access the current volume or past volumes. The current volume for the year will be a compilation of all Lotus news and articles received between January 1 through December 31.

The internet address for the Lotus Newsletter is: http://www.plantsci.missouri.edu/lnl


QUESTIONNAIRE

If you have not filled-out and sent in a questionnaire in the last two years please complete one. If you know of others interested in the Lotus Newsletter have them submit a questionnaire and they'll be added to the mailing list.


SPECIAL THANKS

The illustration on the cover is of Lotus conjugatus L. has again been graciously provided by Ana Arambarri (Argentina) . The Lotus illustration is the 4th in a series of illustrations that started with Volume 23.


EVALUATION OF VARIETIES AND METHODS OF ESTABLISHING BIRDSFOOT TREFOIL INTO IRRIGATED MOUNTAIN MEADOWS

Joe Brummer and Don Rill
Colorado State University, Mountain Meadow Research Center
P.O. Box 598, Gunnison, CO 81230

Preliminary Report
March 12, 1996

Justification

Birdsfoot trefoil (Lotus corniculatus) possesses characteristics that should make it well suited for use in high elevation mountain meadows. However, its potential benefits in that environment have not been demonstrated through research. Therefore, very few producers ever consider birdsfoot trefoil when selecting a legume for seeding.

Most producers cut their high elevation meadows (6,000 - 10,000 ft.) once for hay and then graze any regrowth off in the fall. Birdsfoot trefoil fits this production scheme because first cutting yields are high (i.e. comparable to alfalfa) and the regrowth can be safely grazed due to birdsfoot trefoil's non-bloating characteristic. Birdsfoot trefoil also maintains good forage quality over an extended period of time because of its indeterminate growth habit and better leaf retention capabilities compared to many other legumes. This trait has positive implications for producers that cannot realistically harvest all their hay at peak quality due to constraints such as time, labor, or weather.

The addition of legumes to grass dominated meadows can increase both the quantity and quality of forage produced. Legumes such as red clover (Trifolium pratense), alsike clover (Trifolium hybridum), and alfalfa (Medicago sativa) have been successfully interseeded into mountain meadows in the past. Birdsfoot trefoil seedlings lack vigor and do not compete well with existing vegetation which often makes establishment difficult. The objectives of this study were to evaluate varieties of birdsfoot trefoil potentially adapted for use in mountain meadows and to determine the impact of different seedbed preparations on establishment success of the varieties.

Study Area

This study was established on the Trampe Ranch approximately 6.5 miles north of Gunnison, Colorado at 7800 ft. elevation. The soil is classified as a Fola cobbly sandy loam (loamy-skeletal, mixed Borollic Camborthid). Species present at the site include: common meadow foxtail (Alopecurus pratensis), timothy (Phleum pratense), smooth brome (Bromus inermis), Kentucky bluegrass (Poa pratensis), red clover, alsike clover, dandelion (Taraxacum officinale), herbaceous cinquefoil (Potentilla pulcherrima), and alfalfa. The alfalfa was from an old interseeding and consisted of widely scattered plants. Annual precipitation is evenly distributed and averages 10.5 inches at Gunnison. Snowfall averages 54 inches per year. Meadows used for forage production are flood irrigated. The average frost-free growing season is 63 days, however, freezing temperatures can occur on any day of the year.

Methods and Materials

Birdsfoot trefoil was seeded on May 24, 1994 at the rate of 6 lbs pure live seed/ac. Varieties evaluated were: 'Carroll', 'Norcen', 'Leo', 'Tretana', and 'Empire'. An entry of 'Empire' that was coated with clay containing the appropriate Rhizobium bacteria for root nodulation and the fungicide Apron® was also evaluated and was designated as 'Empire-C'. All other varieties were inoculated just prior to seeding with standard inoculant (Nitragin Company). Plots were seeded with a John Deere Powr-till® drill (Model 1550, 8 ft.). An unseeded plot was left as a control.

The different varieties were seeded into 3 types of seedbeds. The first consisted of seeding directly into the existing vegetation with no effort to suppress competition. The existing vegetation was approximately 4-6 inches tall at the time of seeding. This treatment was considered the control. The second seedbed treatment consisted of suppressing the existing vegetation with Roundup® sprayed 2 weeks prior to seeding at the rate of 1.5 qts/ac. Although Roundup® is supposed to kill all plants that it comes in contact with, it generally only suppresses the more vigorous perennial grasses allowing the seedlings to become established. The third seedbed treatment consisted of tilling with a tractor-mounted rototiller to a depth of about 1 inch. The objective of this treatment was to eliminate or reduce shallow-rooted species such as Kentucky bluegrass and set back the deeper-rooted, desirable perennial grasses giving the seedlings time to become established.

Experimental design was a split-block with 3 replications. Plot size for each variety/seeding method combination was 10' x 24'. Each block measured 72' x 80'. The entire study area was fertilized with 80 lbs/ac of P2O5 just prior to seeding using triple superphosphate (0-45-0).

The study area was flood irrigated under the management of the landowner. Harvest of plots was timed to coincide with the landowner's normal haying which occurred on August 3 in 1995. Plots were harvested with a New Holland mower/conditioner (9 ft., 3 in. header) which left the forage in a windrow. The center 20 ft. of each windrow was collected and weighed to estimate production. Two grab samples were collected per plot. The first was used to determine moisture content and overall hay quality. The second was separated into components of alfalfa, birdsfoot trefoil, clover, forbs, and grass to determine hay composition. Forage quality of individual components was also analyzed. Crude protein and invitro dry matter digestibility were analyzed as measures of forage quality. Production and quality estimates will be gathered for at least one more year. Persistence will be monitored over time.

Results

Interseeding birdsfoot trefoil into existing mountain meadow vegetation increased total forage yield an average of 650 lbs/ac compared to the unseeded control (Table 1). The variety 'Empire-C' that was established using coated seed was the only one that did not significantly increase total yield above the control. For some unknown reason, the seed coating inhibited establishment of this variety instead of enhancing it. The 'Empire-C' variety contributed only 4% to total production while the other varieties contributed between 15 and 22%. The relationship between the amount of birdsfoot trefoil in the hay and total yield was strong (r = 0.97). Plots seeded to the 'Norcen' variety produced the highest total yield at just over 2 tons/ac and had the highest average contribution from birdsfoot trefoil at 22%.

Averaged across varieties, birdsfoot trefoil increased crude protein content of the hay 2.3 percentage points compared to the control (Table 2). Crude protein content of the hay was lowest from plots seeded with the 'Empire-C' variety and highest from plots seeded with either the 'Leo' or 'Norcen' varieties. As with production, this trend was related to the amount of birdsfoot trefoil in the hay (r = 0.94). Digestibility of the hay was not affected by any of the birdsfoot trefoil varieties (Table 2).

Total forage yield was not affected by either the spray or tillage seedbed preparations (Table 3). Contribution of birdsfoot trefoil to total yield was 3 times higher from plots sprayed with Roundup® and about 2 times higher from tilled plots compared to the control. This result emphasizes the need to suppress the existing vegetation in some manner prior to seeding to improve establishment success of birdsfoot trefoil. However, there were some tradeoffs associated with the different suppression methods evaluated in this study. The tillage treatment increased grass composition compared to the spray and control treatments and decreased clover composition compared to the control (Table 3). The spray treatment also decreased clover composition compared to the control.

The changes in hay composition (Table 3) due to interseeding and suppression of the existing vegetation affected both crude protein content and digestibility of the hay (Table 4). An increase in grass and decrease in clover composition led to a reduction in both crude protein content and digestibility of the hay produced from the tillage plots compared to the control. Digestibility of hay from the sprayed plots was reduced about 3 percentage points compared to the control, possibly in response to reduced clover composition. Crude protein content of hay from the sprayed plots was not affected. The contribution of crude protein from birdsfoot trefoil probably offset any loss associated with decreased clover composition.

No significant differences in crude protein content or digestibility were measured among the birdsfoot trefoil varieties (Table 5). Crude protein content of birdsfoot trefoil averaged 17.3% which was comparable to clover (Table 6). These 2 components were higher in crude protein compared to alfalfa, grass, or forbs. Birdsfoot trefoil was also the most digestible (67.2%) compared to the other hay components (Table 6). Crude protein content and digestibility of alfalfa were lower than expected because of the large, robust growth form of the plants that were present. As density of alfalfa declines over time, the remaining plants compensate by producing stemmier growth that is generally higher in fiber (i.e. less digestible) and lower in crude protein.

Summary

Birdsfoot trefoil that was interseeded into a mountain meadow increased both the quantity and quality of forage produced in the first production year. Differences among the varieties tested were generally small. Suppression of the existing vegetation improved establishment success. Plots sprayed with Roundup® prior to seeding had the highest contribution of birdsfoot trefoil to total production. Although these results from the first production year appear promising, persistence of the different varieties needs to be documented before birdsfoot trefoil can be commonly recommended for seeding in mountain meadows.


Table 1. Effect of interseeding various varieties of birdsfoot trefoil on production and composition of mountain meadow hay. Samples were taken August 3, 1995.
Variety1
Production
Grass
Birdsfoot Trefoil
Alfalfa
Clover
Forbs
(lb/ac)
---------------------------------%---------------------------------
Control
3050 a2
76 a
0 a
5 a
15 a
4 a
Carroll
3710 bc
62 a
15 bc
5 a
13 a
5 a
Empire
3700 bc
58 a
18 c
6 a
14 a
4 a
Empire-C
3310 ab
65 a
4 ab
10 a
15 a
6 a
Leo
3700 bc
54 a
19 c
12 a
13 a
2 a
Norcen
4010 c
55 a
22 c
7 a
13 a
3 a
Tretana
3750 bc
52 a
17 c
5 a
15 a
11 a

1Variety means were averaged across seeding methods.
2Means within columns followed by the same letter are not significantly different (P>0.05).


Table 2. Effect of interseeding various varieties of birdsfoot trefoil on crude protein concentration and invitro dry matter digestibility of mountain meadow hay. Samples were taken August 3, 1995.
Variety1
Crude Protein
Dry Matter Digestibility
------------------------------%----------------------------------
Control
7.4 a2
62.7 a
Carroll
9.4 bc
62.8 a
Empire
9.9 bc
62.3 a
Empire-C
8.7 ab
62.7 a
Leo
10.5 c
60.3 a
Norcen
10.2 bc
61.5 a
Tretana
9.5 bc
61.1

1Variety means were averaged across seeding methods.
2Means within columns followed by the same letter are not significantly different (P>0.05).

Table 3. Effect of method of seeding (i.e. vegetation suppression method) birdsfoot trefoil on production and composition of mountain meadow hay. Samples were taken August 3, 1995.
Seeding Method1
Production
Grass
Birdsfoot Trefoil
Alfalfa
Clover
Forbs
(lb/ac)
-----------------------------------%----------------------------------
Control
3660 a2
58 a
7 a
9 a
19 b
7 a
Spray
3630 a
57 a
21 c
6 a
13 a
3 a
Till
3520 a
67 b
13 b
5 a
10 a
5 a

1Seeding method means were averaged across birdsfoot trefoil varieties.
2Means within columns followed by the same letter are not significantly different (P>0.05).


Table 4. Effect of method of seeding (i.e. vegetation suppression method) birdsfoot trefoil on crude protein concentration and invitro dry matter digestibility of mountain meadow hay. Samples were taken August 3, 1995.
Seeding Method1
Crude Protein
Dry Matter Digestibility
----------------------------%---------------------------------
Control
9.9 b2
65.4 c
Spray
9.8 b
62.6 b
Till
8.4 a
57.8 a

1Seeding method means were averaged across birdsfoot trefoil varieties.

2Means within columns followed by the same letter are not significantly different (P>0.05).


Table 5. Crude protein concentration and invitro dry matter digestibility of various birdsfoot trefoil varieties that were interseeded into a mountain meadow. Samples were taken August 3, 1995.
Variety1
Crude Protein
Dry Matter Digestibility
-------------------------------%--------------------------------
Carroll
16.9 a2
65.0 a
Empire
17.4 a
67.4 a
Empire-C
15.9 a
66.7 a
Leo
18.4 a
69.6 a
Norcen
17.4 a
66.2 a
Tretana
16.7 a
67.6 a

1Varieties were composited across seeding methods.

2Means within columns followed by the same letter are not significantly different (P>0.05).


Table 6. Crude protein concentration and invitro dry matter digestibility of various components of mountain meadow hay following interseeding of birdsfoot trefoil. Samples were taken August 3, 1995.
Component1
Crude Protein
Dry Matter Digestibility
------------------------------%--------------------------------
Alfalfa
14.1 b2
56.7 a
Birdsfoot Trefoil
17.3 c
67.2 d
Clover
16.8 c
63.8 c
Forbs
7.6 a
61.8 bc
Grass
6.7 a
59.5 b

1Varieties were composited or averaged across seeding methods.
2Means within columns followed by the same letter are not significantly different (P>0.05).

SEED QUALITY IN LOTUS CORNICULATUS IN RELATION TO POD MATURITY AND HARVEST TREATMENT

Wilson Hugo Ettlin1 and Giles Laverack2
1
INIA Experimental station La Estanzuela, Ruta 50 km 12 Colonia, Uruguay cc. 39.173

2Institute of Ecology and Resource Management
University of Edinburgh, School of Agriculture
West Mains Road, Edinburgh EH9 3JG. Scotland, U.K.

ABSTRACT

Seed development and maturation at different stages of pod development were studied Lotus corniculatus. Pods were classified into 4 different stages of development using morphological characters such as colour and size. The effect on seed quality of three different harvesting procedures at each stage of development was studied. The number of normal seedlings and hard seed increased with stage of development. Direct combining resulted in the lowest germination potential whilst highest germination potential was obtained after threshing seed from dried plants. Collecting pods and drying them artificially showed post harvest maturation mainly by means of hardseedness.

INTRODUCTION

The genus Lotus belongs to the subfamily Papilionatae (Wheeler and Hill, 1957). It consists of a diverse group of annual and perennial species widely distributed throughout the world (Callen, 1959).Lotus corniculatus, a perennial forage legume is utilised in several countries in different parts of the world (Seaney and Henson, 1970: Robinson, 1934: Orsi, 1953: Panikar, 1949: Gardner and Elliot (1945). This species has advantages due to its forage quality and ability to tolerate low phosphorous levels in soil and drought (Formoso, 1983). However lack of sufficient seed yield has prevented its spread (Anderson, 1955: McGraw and Beuselinck, 1983: Buckovic, 1952: Metcalfe et al, 1957). The gap between the theoretical and actual seed yield is enormous (Seaney and Henson, 1970). Lack of proper guides to determine the best moment for harvesting have been mentioned by Pieroni (1992) and this situation may lead to loss of seeds because of dehiscence of pods or because of damage to seed if harvest is too early. Several authors have studied the development of seeds of forage legume from defined points early in the process of maturation (Pieroni, 1992: Anderson, 1955: Hare and Lucas, 1984 ). The variability of environment in different parts of the world, seasonal variation in weather conditions and differences in populations mean that it is difficult to make generalisations and practical recommendations for crop management from these studies as the timing of maturation may change. The objectives of the present work were to relate the physiological development of seeds and seed quality to recognisable morphological stages of pod development and to relate this to harvesting treatment.

MATERIALS AND METHODS:

Field experiments were carried out at Terrington St. Clements, Norfolk, England (52o 45' N: 0o15' W) during July and August 1995. Laboratory tests were carried out at The Official Seed Testing Station for Scotland, East Craigs, Edinburgh. A 0.8 hectare crop of Lotus corniculatus was sown in 1991, at a seed rate of 3.5 kg of seed per hectare at 45 cm between furrows. Paraquat® and Linacil® were applied during spring for weed control. The crop was grown from wild populations collected in the British isles to produce seed for natural habitat reconstruction. Experiments on reserve accumulation: Pods were collected at each stage of development in five replications taken from five random sites marked in the field. The stages of development were defined as follows:1- Purple-green pod colour, shorter than 2 cm.2- Green pod colour , longer than 2 cm and thicker than 1 mm, green seed.3- Yellow pod, turning to light brown in colour, green- light brown seed.4- Dry, brown pod colour, brown seed. After collection, pods were put in air-water proof plastic bags, and stored at -20 oC prior to laboratory studies. Laboratory studies involved opening the pods and removing four sets of 50 seeds from each replicate at each stage of development. The sub samples were weighed and dried in oven following ISTA rules for moisture content (Seed Science and Technology, 1993) . The following indices were calculated for each stage of development: -1000 seed weight at harvest moisture content;- Moisture content at harvest;- 1000 seed weight of dried seed.

Experiments on post harvest maturation: After pollination and during the different stages of seed development, five replicates of 200 pods were harvested from each of the sites described above in three different ways as follows: Treatment 1 (t1)- Direct harvest and threshing with a commercial combine, a HEGE 125b, with the cylinder speed set at 1043 rpm and with cylinder-concave set as small as possible (less than 0.5 cm). Treatment 2 (t2)- Picking pods from the plants, drying and threshing them after drying as in t1. Treatment 3 (t3)- Cutting entire plants and drying them. Seeds were threshed after drying as above. These different harvesting methods were applied at all the stages of development described above, with the exception of the first one, where the seeds were not physically big enough to be threshed by a combine. For all the treatments, seed, pods or plants were dried on trays inside a green house for at least one week. Plant material in the trays was covered with paper to avoid direct exposure to sun. The average temperature reached was 25oC with a maximum of 38oC and a minimum of 12oC. The relative humidity of the air varied between 40% and 60% Germination tests: Germination tests were carried out according to ISTA rules (Seed Science and Technology, 1993). Four replications of 100 seeds were counted for each stage of development and harvesting treatment. The seed was set to germinate on top of paper (Whatman 181) at 20/30oC. The first count was made at 7 days and the second at 12 days. Germinated, hard, abnormal, dead and fresh seeds were recorded.

RESULTS

All results were subject to analysis of variance using the GENSTAT program.

Reserve accumulation: Results are shown in table 1.

Table 1: Development of moisture content, dry weight and fresh seed weight in Lotus corniculatus.
stage of
moisture
1000 seed
1000 seed
development
content %
dry weight (gr.)
fresh weight (gr.)
1
78.40
0.03
0.15
2
76.20
0.47
1.98
3
59.15
1.56
3.82
4
17.70
1.63
1.98

Water content showed a decline through the stages of development as shown in Table 1. The maximum fresh seed weight was reached at stage 3, which represents a light yellow-brown pod. Dry seed weight also changed drastically from stage 2 to stage 3, going form 0.47 to 1.56 gr. per 1000 seeds.

Post harvest maturation.

Germination tests: The results from the germination test were classified into the following categories:

a) Normal seedling after 12 days.
b) Hard seeds: after 12 days at 20/30oC.
c) Normal seedlings plus abnormal seedlings
d)Total germination: This is the germination value of a seed lot as assessed in an official germination test and includes normal seedlings plus hard and fresh seeds. The results can be seen in Figures 1 and 2 as the addition of normal, hard and fresh seeds.

A significant difference in amount of normal seedling and in amount of hard seeds with P< 0.001 was found between stages of development and harvesting treatment. Total germination value showed significant differences between harvesting treatments, stages of development and the interaction between treatments and stages (P< 0.01).Normal plus abnormal seedling were scored to find any possible differential mechanical damage in any treatment. No difference was found neither in harvesting treatment nor stage of development for both crops. The overall amount of abnormal seed recorded was 4.4% The proportion of normal seedling, fresh seeds, hard seeds and total germination are presented in Figures 1 and 2 according to stage of development and harvesting treatment respectively.

Figure 1: Development of normal seedling, hard seed, fresh seed and total germination (addition) at different stages of development in Lotus corniculatus. Standard error bars are displayed for total germination value.

Figure 2: Amount of normal seedling, hard seed, fresh seed and total germination (addition) at different harvesting treatment in Lotus corniculatus. Standard error bars are displayed for total germination value.

Table 2: Total germination value in Lotus corniculatus at different combinations of harvesting treatment and stage of development. LSD = 6.5 for P=0.05
stage of develop
Harvest treatment
T 1
T 2
T 3
2
0.25
3.40
11.85
3
13.65
88.85
70.95
4
81.90
89.70
88.65

A significant effect of the harvesting treatment was found (P< 0.01). As shown in Figure 2, the direct combining treatment presented the lowest germination rate, and both harvesting treatment 2 and 3 presented the highest quality without significant difference between them. The harvesting treatment 3 showed some pods attacked by a weevil that was identified as Apion loti. It was present in plants picked from the field and attacked some pods during the drying process decreasing the final quality of seed. That could be the reason of a lower trend in the germination of seed from dried plants. The higher germination rate in harvesting treatment 3 is mainly due to a higher amount of hard seeds (Figure 2), while in the case of harvesting treatment 2 it is due to higher number of normal seedling. A similar trend in development of germination through the stages of pod development was found. Seed collected from green pods showed an extremely low germination rate of 5.1% , rising to 57.8% in seed from light yellow-brown pods and up to 86.7 % in seed from brown pods. A significant difference was also found in the interaction between harvesting treatment and stage of development (P< 0.01). As shown in table 2 seed did not tolerate direct combining during stage 3 but germination increased dramatically at the same stage when seed were dried in pods or with the entire plant. It also shows that after this stage and treatment the germination value seems to reach a maximum between 80% and 90%.

DISCUSSION

According to Bewley and Black (1994) seed grows from a single fertilised egg to a multicellular embryo by cell division and differentiation. These events are marked by a period of rapid increase in whole seed fresh weight and water content. A period of rapid gain in dry weight then follows as a result of the synthesis and deposition of stored reserves. The whole seed fresh weight then remains relatively stable, although the seed loses water as this is displaced by the accumulation of insoluble reserves within the cells of storage tissues. The evolution of water content shown in Table 1 is as expected in seeds during accumulation of reserves. It seems that the very early stages of embryo development, where amount of water content increases has been missed, which means it happens before the first stage of development studied. Bewley and Black (1994) also described that the decline in water content of a developing seed slows as it approaches its maximum dry weight. Finally as the seed undergoes maturation drying and approaches the quiescent stage, when it may be shed from the plant, there is a period of fresh weight loss accompanied by a rapid decline in whole seed water content. The fact that there is a sharp drop in water content going from 59% to 17% between the last two stages seems to indicate that the desiccation occurs in Lotus corniculatus at the transition from light yellow to brown colour. Winch and MacDonald (1961) reported similar trend for pods, in this case pod moisture content fell from 65% to 25% when changing to light brown colour. A slightly higher moisture content was reported by Anderson (1955) who stated that seed become morphologically mature at the time pods have light brown colour, having a moisture content of 35% to 40%.However, Seaney and Henson (1970) suggest that seed become physiologically mature slightly before pods change colour from green to light brown, which corresponds in this case to the transition from stage 2 to stage 3. This might indicate that seeds increase in fresh and dry matter content even after physiological maturation as will be discussed ahead. Hare and Lucas (1984) defined the development stages in Lotus as: a) growing stage, b) food reserve accumulation stage and c) the ripening stage, counting days from pollination to classify these stages. In this experiment the evolution of dry matter content and water content seems to indicate that the transition from stage 2 (green pods) to stage 3 (light yellow-brown) could be the beginning of the food reserve accumulation stage, meanwhile the transition from stage 3 to stage 4 (brown colour) could be the ripening one.

Post harvest maturation. As explained in results he higher germination rate in the harvesting treatment 3 is mainly due to a higher amount of hard seeds (Figure 2), while in the case of harvesting treatment 2 it is due to higher number of normal seedling. These results also indicates that some post harvest maturation occurs in a different way according to the drying environment. Bewley and Black (1994) state that hardseedness is a common feature in the Leguminosae family and is a kind of dormancy in which water entry is the limiting factor for germination. They also state that the testa is generally responsible for impeding water uptake through different structures like the waxy cuticle, suberin or the thick-walled palisade cells. Which of these anatomical structures are able to be changed during the last stages of development or during different conditions of drying has not been studied in this paper. It could be worth more research in the matter since the understanding of this maturation process can have implications in commercial seed production. The trend in total germination value found at different stages of development is as expected. However, Anderson (1955) working in Iowa found higher germination rate during the green pod stage, reaching 90 % with the sum of normal and hard seeds. It is not known whether these differences in physiological maturity are due to a genetic effect or due to a year difference. The small drop in germination value in dried plants during the stage of light yellow-brown pods could be explained by the Apion loti attack. Hare and Lucas (1984) stated that Lotus pedunculatus reached seed maturity at the time pods were light-brown yellowish colour, at this stage seed had 65% moisture content. This stage correspond with stage 3 in this experiment in which maximum germination was still not reached even if seed had the same moisture content. The results of the interaction between harvesting treatment and stage of development also agree with Hare and Lucas (1984) who recommended, as mentioned above, that mowing should start at light brown colour stage. The differences shown in table 2, could be important for commercial seed production and opens a field for more research. If technology allowed a mechanised way of picking pods from the field as early as the light-yellow brown pod stage and drying artificially, harvesting could start in an earlier stage of maturation, before dehiscence in field. If dehiscence happens in-building it will not mean seed loses. During seed drying in the harvesting treatment 2 and 3 no dehiscence of pods was noticed in all the stages of development, but as mentioned above, temperature reached an average of 25oC with variation in relative humidity between 40% and 60%. Metcalfe et al. (1957) mentioned that in the Midwest and Northeast U.S.A. seed losses from pod shattering can be high when relative humidity drops below 40%. Similarly, Phillips and Keim (1968) found 35% of relative humidity as the point where pods start a rapid dehiscence. Under different climatic situations it is not sure that mowing the crop and allowing it to dry in the field in an earlier stage would have not dehiscence of pods.

BIBLIOGRAPHY:

Anderson S.R. (1955) Development of pods and seed of birdsfoot trefoil (Lotus corniculatus L.) as related to viability and to seed yield. Agronomy Journal 47: 483-487.

Bewley, J.D and Black, M. (1994) Seeds, Physiology of development and germination. Second edition. Plenum press, New York and London.

Buckovic, R.G. (1952). MSc. Thesis. Oregon State College, Corvallis, Oregon.

Callen, E.O. (1959) Studies of the genus Lotus. Canadian Journal of Botany 37: 157-165.

Formoso, F. (1983). Effect of defoliation in Lotus corniculatus. Lotus Newsletter 14: 3.

Gardner, C.A. and Elliot, H.G. (1945) West Australia Dep. Agr. J. 22: 358-360.

Hare, M.D. and Lucas, R.J. (1984). `Grassland Maku lotus (Lotus pedunculatus L.) seed production. Journal of Applied Seed Production 2: 58-64.

McGraw, R.L. and Beuselinck, P.R. (1983) Growth and seed yield characteristics of birdsfoot trefoil. Agronomy Journal 75: 443-446.

Metcalfe, D.S., Johnson, I.J. and Shaw, R.H. (1957). The relationship between pod dehiscence, relative humidity and moisture equilibrium in birdsfoot trefoil (Lotus corniculatus L.). Agronomy Journal 49: 130-134.

Orsi, S. (1953). Ann. Ente. Consor Interprov. Tosc. Sementa IV, 30.

Panikar, M.R.(1949). Indian Farming 10: 444-447.

Phillips, R.L. and Keim, W.F. (1968). Seed pod dehiscence in Lotus and interspecific hybridisation involving L. corniculatus L. Crop Science 8: 18-21.

Pieroni, S.J. (1992) Factors affecting seed yield and quality in temperate forage legumes. MSc Thesis. The Edinburgh School of Agriculture. University of Edinburgh.

Robinson, D.H. (1934) Birdsfoot trefoil in Agriculture. Empire J. Exp. Agr. 2: 274-283.

Seaney, R.R. and Henson, P.R. (1970). Birdsfoot trefoil. Advances in Agronomy 22: 119-157.

Seed Science and Technology (1993). V 21 Supplement Rules. International Rules for Seed testing.

Wheeler, W.A. and Hill, D.D. (1957). Seed formation and germination. In Grassland seeds. Eds. Wheeler, W.A. and Hill, D.D. London, D. Van Nostrand Company, Inc. pp 12-34.

Winch, J.E. and MacDonald, H.A. (1961). Flower pod and seed development relative to the timing of the seed harvest of Viking birdsfoot trefoil (Lotus corniculatus L.) Canadian Journal of Plant Science 41: 523-532.


ERRATA

CORRECTIONS SHOULD BE NOTED IN THE TWO FOLLOWING PAPERS

William F. Grant

Department of Plant Science
P.O. Box 4000
McGill University, Macdonald Campus
Ste. Anne de Bellevue, Que., Canada H9X 3V9

1. Error in title of paper:
"A chromosome atlas and interspecific-intergenic index for Lotus and Tetragonolobus (Fabaceae). Can. J. Bot. 73: 1787-1809, 1995.

The correct title is:
A chromosome atlas and interspecific-intergeneric index for Lotus and Tetragonolobus (Fabaceae).

2. Error in Table 1 in the list of species studied:
"Genome relationships among Lotus species based on random amplified polymorphic DNA (RAPD)" by L. P. Campos, J. V. Raelson and W. F. Grant. Theor. Appl. Genet. 88: 417-422, 1994.

Table 1 lists the species and their chromosome numbers studied. Accession B-557 listed as L. japonicus, 2n = 12, is incorrect. Accession B-557 is L. corniculatus and the chromosome number is 2n = 24.


REDUCTION IN BASIC CHROMOSOME NUMBER IN THE GENUS LOTUS (FABACEAE)

William F. Grant

Department of Plant Science
P.O. Box 4000
McGill University, Macdonald Campus
Ste. Anne de Bellevue, Que., Canada H9X 3V9

The greatest number of Lotus species are those with a basic chromosome number of 7 (x = 7). Evolutionarily, these species are considered to be the oldest. This group is followed by those species which have a basic chromosome number of x = 6. Only a single species has a basic chromosome number of 5 (L. glareosus). Chromosome evolution in the genus is considered to be occurring by means of a reduction in the basic number from x = 7 to x = 6 to x = 5, in a descending aneuploid series from an eight-chromosomed ancestor of the tribe Galegeae (Senn, 1938). In the new world only a reduction from x = 7 to x = 6 has taken place to date (Grant 1995). The question arises as to how this reduction in basic number came about?

Chromosome measurements were made representing three European species. A drawing of the chromosomes for Lotus strictus (previously unpublished) was used for the 7-chromosome species. Idiograms of the species L. glacialis (2n = 12) and L. glareosus (2n = 10) are given in the paper of Fernandes and Queiros (1980) and chromosome measurements were made from these idiograms for these two species. From the measurements for all three species, idiograms were drawn using the chromosome analysis package CHROMPAC III (Green et al. 1984).

The data from the chromosome analyses are given in Table I. The idiograms in Figures 1 and 2 are photocopied from Fernandes and Queiros (1980). Figs. 3-5 represent the idiograms based on the detailed measurements. Satellites were not included in the measurements of chromosome length.

Karyotype measurements for Lotus strictus, L. glacialis and L. glareosus
Set
Average
Percent
% Length
Centromere*
length
long arm
short arm
ratio
type
index
L. strictus
1
23.0
22.90
17.83
5.07
3.52
ST
0.22
2
19.5
18.80
13.02
5.78
2.25
SM
0.31
3
17.0
16.38
9.16
7.22
1.27
M
0.44
4
14.5
13.98
8.20
5.78
1.42
M
0.41
5
11.0
10.60
7.22
3.38
2.14
SM
0.32
6
10.0
9.64
5.30
4.34
1.22
M
0.45
7
8.0
7.72
3.86
3.86
1.00
M
0.50
Total length of chromosomes = 53.90 µm
L. glacialis
1
16.5
24.78
18.03
6.75
2.67
SM
0.27
2
13.5
20.28
11.27
9.01
1.25
M
0.44
3
11.5
17.26
11.27
5.99
1.88
SM
0.35
4
9.1
13.66
6.90
6.76
1.02
M
0.50
5
9.0
13.52
6.76
6.76
1.00
M
0.50
6
7.0
10.52
5.26
5.26
1.00
M
0.50
Total length of chromosomes = 34.60 µm
L. glareosus
1
18.5
29.36
22.22
7.14
3.11
ST
0.24
2
15.5
24.60
19.84
4.76
4.17
ST
0.19
3
10.0
15.88
10.33
5.55
1.86
SM
0.35
4
10.0
15.88
7.94
7.94
1.00
M
0.50
5
9.0
14.28
7.14
7.14
1.00
M
0.50
Total length of chromosomes = 24.38 µm

*Ratio: Long arm (p) divided by short arm (q)
Type: M, chromosome with centromere in median position; SM, submedian; ST, subterminal
Index: p/p+q

Fernandes and Queiros (1980) have shown 2 pairs of satellited chromosomes on the short arms in both L. glacialis (chromosomes B and D) and L. glareosus (chromosomes B and C) (Figs. 1-2). Although chromosome B is the longest chromosome in L. glacialis, the authors have drawn this chromosome as the second chromosome of the complement (reason unknown) rather than the first. In contrast, no satellited chromosomes were observed in any of the metaphases examined for L. strictus (Fig. 3).

The computer program drew the chromosomes in descending length. Chromosome B in Fig. 1 becomes chromosome 1 in Fig. 4. As may be seen from Figs. 3-5, chromosome 1, for all three species, is morphologically similar with submedian to subterminal centromeres. Fernandes and Queiros (1980) pointed out the similarity of chromosomes C and D between L. glacialis and L. glareosus in Figs. 1 and 2. They explained the reduction in chromosome number from n = 6 to n = 5 on non-separation of the chromatids during anaphase as they observed metaphase plates in root tip cells of L. glacialis (2n = 12) with 13, 11, and 10 chromosomes, in addition to the normal complement of 12. One restitution cell would lack chromosome F (Fig. 1) giving rise to L. glareosus (Fig. 2) lacking this chromosome. They considered chromosome F to be largely heterochromatic and therefore dispensable which lead to the development of a species with a reduced chromosome number (L. glareosus). They also suggested that L. glareosus was derived from L. glacialis by chromosome rearrangements, but the actual mechanism and chromosomes involved were not specified.

Chromosomes 2 and 3 in L. strictus are also very similar to those of chromosomes 3 and 4 in both L. glacialis and L. glareosus. A translocation between chromosomes 4 and 5 in L. strictus would give a largely median chromosome as is chromosome 5 in L. glacialis. The loss of chromosome 7 in L. strictus by being either genetically inert or through missegregation in mitosis would reduce the number to 2n = 12. Similarly, a translocation between chromosomes 1 and 2 in L. glacialis could produce chromosome 2 in L. glareosus . With the loss of chromosome 5, L. glareosus would result with the reduced chromosome number. By the classical translocation route, a reduction in basic chromosome number could have taken place as hypothesized above. However, observations in which chromosome irregularities occur in both root tips of somatic (Fernandes and Queiros 1980; O'Donoughue and Grant 1989), and meiotic cells (Somaroo and Grant 1972) and in Lotus hybrids (O'Donoughue and Grant 1988) with the subsequent development of plants with stable chromosome numbers would be an alternate explanation for the initiation of plants with a reduction in basic chromosome number.

Fernandes, A. and Queiros, M. 1980. Sur l'occurrence de la pseudo- reduction somatique chez Lotus glacialis (Boiss.) Pau. Bol. Soc. Brot., Ser. 2, 54: 133-152.

Grant, W. F. 1995. A chromosome atlas and interspecific - intergeneric index for Lotus and Tetragonolobus (Fabaceae). Can. J. Bot. 73: 1787-1809.

Green, D. M., Myers, P. Z. and Renya, D. L. 1984. CHROMPAC III: an improved package for microcomputer-assisted analysis of karyotypes. J. Hered. 75: 143.

O'Donoughue, L. S. and Grant, W. F. l988. New sources of indehiscence for birdsfoot trefoil (Lotus corniculatus, Fabaceae) produced by interspecific hybridization. Genome, 30: 459- 468.

O'Donoughue, L. S. and Grant, W. F. 1989. Chromosomal variation in Lotus alpinus (Fabaceae). Plant Species Biol. 4: 117-122.

Senn, H. A., 1938. Chromosome numbers in the Leguminosae. Bibliograph. Genet. 12: 175-336.

Somaroo, B. H. and Grant, W. F. 1972. Chromosome differentiation in diploid species of Lotus (Leguminosae). Theor. Appl. Genet. 42: 34-40.


COROLLA, FRUIT AND SEED CHARACTERISTICS IN LOTUS SALSUGINOSUS GREENE VAR. BREVIVEXILLUS OTTLEY AND L. STRIGOSUS (NUTT.) GREENE VAR. TOMENTELLUS ISELY (FABACEAE -- LOTEAE)

Ana M. Arambarri

Area de Botánica, Jardín Botánico y Arboretum "C. Spegazzini," Facultad de Ciencias Agrarias y Forestales, UNLP, Calle 60 y 118, C. C. 31

1900 La Plata, Argentina

INTRODUCTION

In the New World, the genus Lotus L. comprises approximately 40 species distributed from British Columbia to Mexico in North America. Only one species is endemic to Chile in South America. L. salsuginosus Greene and L. salsuginosus var. brevivexillus Ottley; L. strigosus (Nuttall) Greene, L. strigosus var. hirtellus (Greene) Ottley, and L. strigosus var. tomentellus (Greene) Isely, are found in California, Lower California and Mexico, and in the islands west of Lower California.

Nuttall (1838) treated L. strigosus as belonging to genus Hosackia Bentham ex Lindley. Greene (1890), examined and included Nuttall's species into the genus Lotus L. He named L. tomentellus to one species from Lower California. Ottley (1923) considered L. salsuginosus and L. strigosus into the subgenus Acmispon Rafinesque. Later, Ottley (1944) segregated the species belonging to Acmispon into two sections. The section Microlotus Benth. (6 species) containing L. salsuginosus and its variety, and the section Simpeteria Ottley (12 species) containing L. strigosus, L. strigosus var. hirtellus, and L. tomentellus. Isely (1981) reduced L. tomentellus to a variety of L. strigosus.

Until present, the standard with blade attenuated into the claw, the wings asymmetrically positioned and longer than the keel, and the stigma penicillate, are the characters used to separate the section Simpeteria from the section Microlotus. However, these characters and those (e. g., density of the pubescence, presence and shape of the bract, standard shapes) used to separate L. salsuginosus and L. strigosus are not enough to delimitate L. salsuginosus var. brevivexillus and L. strigosus var. tomentellus. These are two varieties of different species frequently confused.

The purpose of this article is to emphasize the most useful characters, and to include news, to delimitate L. salsuginosus var. brevivexillus and L. strigosus var. tomentellus.

MATERIALS AND METHODS

Specimens of herbarium, fruit and seed samples surveyed, were from BRY, MEXU, NSMC, SD, US. The acronyms for the herbaria are given according to Holmgren et al. (1990). Some samples were from Desert Legume Program, Tucson (UAz). Morphological and anatomical characters were examined with a stereoscopic microscope (at 40 x or less) and an optical microscope; both equipped with a camera lucid. The testa topographic features were analysed on whole seeds and portions of them mounted on brass-stubs, sputter-coated with gold-palladium in a JFC-1100 and viewed at 15 kv with a Jeol JSM-T100 scanning electron microscope (SEM). The topographic features were described according to Lersten (1981). Drawings of the characters were prepared by the author.

L. salsuginosus (Greene) Greene

L. salsuginosus Greene var. brevivexillus Ottley

Lotus strigosus (Nutt.) Greene var. tomentellus Isely

Comment

(*) specimens mounted on the same sheet.

(**) the specimens on the sheet correspond to L. strigosus var. tomentelllus, whereas the material into the envelope attached, contains a mixture of the specimen with L. salsuginosus var. brevivexillus.

Results and Discussion

Table 1. Corolla, legume and seed characteristics to distinguish L. salsuginosus Greene var. brevivexillus Ottley and L. strigosus (Nutt.) Greene var. tomentellus Isely.
L. salsuginosus var. brevivexillus L. strigosus var. tomentellus
COROLLA(Fig. 1, A-C-E) (Fig. 1, B-D-F)
Standard venationThe veins show a curved course in the base of the blade. The veins show a ± straight course in the base of the blade.
WingThe auricle is always exposed The auricle is obscured by the upper margin folded downwards.
KeelThe upper margin straight. The apex obtuse. The upper margin convex or slightly so. The apex acute.
LEGUME(Fig. 1, G) (Fig. 1, H)
The hard and persistent base of the style always recurved The hard and persistent base of the style always short and straight.



SEED(Fig. 2, A-a) (Fig. 2, B-b)(Fig. 2, C) (Fig. 2, D)
Seed shapeglobose globosegloboselaterally compressed
Seed shape in outline (Face view) ovate-roundquadrangular to round circularcircular, rarely quadrangular
Testa surface(At 40 x or less)smooth smooth or near sorugose sinuate rugose
Radicular lobeinconspicuous discerniblediscernible prominent
Lenssmall mound small moundconspicuousconspicuous
Hilum elevationsuperficial superficialsuperficial sunken in lateral notch
Hilar rimconspicuously discoloured concolouredconcoloured concoloured
Micropylebifurcate bifurcatebifurcate deltoid
Testa topography, on mid-facepapillose (small papillaes) reticulatereticulate sinuate or non sinuate, with mounds papillose sinuate, with or without very small mounds

Acknowledgments

I wish thank to the staff of the institutions who supplied the specimens and fruit-seed samples studied. My special thank to Mario Sousa S. and Maru García Peńa who borrowed most of the specimens studied. Thank Liliana Katinas for providing a number of suggestions that helped to improve earlier manuscripts and Rafael Urrejola for technical assistance using SEM.

REFERENCES

Greene, E. L. 1890. Enumeration of the North American Loti. Pittonia 2: 133-150.

Holmgren, P. K. , N. H. Holmgren and L. C. Barnett. 1990. Index Herbariorum. P. 1. The Herbaria of the World. [Regnum Veg. 120]. NY Bot. Gard., Bronx.

Isely , D. 1981. Leguminosae of the United States. III. Subfamily Papilionoideae: Tribes Sophoreae, Podalyrieae, Loteae. Mem. NY Bot. Gard. 29(3): 124-264.

Lersten, N. R. 1981. Testa Topography in Leguminosae, Subfamily Papilionoideae. Proc. Iowa Acad. Sci. 88(4): 180-191.

Ottley, A. M. 1923. A revision of the Californian species of Lotus. Univ. California Publ. Bot. 10: 189-305.

Ottley, A. M. 1944. The American Loti with special consideration of a proposed new section, Simpeteria. Brittonia 5(2): 81-123.


ASYMMETRIC SOMATIC CELL HYBRID BETWEEN LEGUMINOUS SPECIES OF BIRDSFOOT TREFOIL AND ALFALFA

Kaimori,N. and M. Niizeki
Laboratory of Bioscience and Biotechnology
Faculty of Agriculture, Hirosaki University
Hirosaki, Aomori-ken 036, Japan

In our laboratory, all combinations of somatic cell hybrids have been carried out among a graminous plant, rice (Oryza sativa L.) and three leguminous plants, soybean (Glycine max (L.) MERR.), birdsfoot trefoil (Lotus corniculatus L.) and alfalfa (Medicago sativa L.). We report here asymmetric somatic hybrid of birdsfoot trefoil and alfalfa.

Materials and Methods

The calli of birdsfoot trefoil cv. Viking and alfalfa cv. Rangelander were used for the isolation of protoplasts. The enzyme solution for the protoplast isolation of both birdsfoot trefoil and alfalfa was constituted with 4% Cellulase Onozuka RS, 1% Macerozyme R-10, 0.2% Pectolyase Y-23 and 0.6 or 0.7 M mannitol. The division of birdsfoot trefoil protoplasts was inhibited by the treatment of 10 mM iodoacetamid (IOA). The protoplasts of alfalfa were irradiated by 40 kR of X-rays. The fused protoplasts were then cultured in the KM8p medium (Kao and Michayluk, 1975) by the agarose-beeds and nurse culture methods. The isozymes of obtained calli and regenerated shoots were examined on aminopeptidase (AMP), esterase (EST), glutamate dehydrogenase (GDH) and catalase (CAT).

Results and Discussion

The 155 callus lines were obtained from the asymmetrc somatic cell fusion of birdsfoot trefoil and alfalfa. Among these calli, 17 callus lines were identified as somatic cell hybrids by the analyses of isozymes (Table 1). After one month of subculture, most of the hybrid calli, however, altered the isozyme banding patterns to those of birdsfoot trefoil on the medium of Murashige and Skoog (1962) (MS medium) with 4 mg/l 1-naphthaleneacetic acid (NAA) and 2.5 mg/l kinetin or with 1.5 mg/l indole-3-acetic acid (IAA) and 1.5 mg/l 6-benzyladenine (BA). This result means that most of the alfalfa chromosomes irradiated by X-rays disappeared during the subcultures. Niizeki et al. (1989) reported that the shoot regeneration did not occur from the symmetric somatic hybrid calli of birdsfoot trefoil and alfalfa. On the other hand, in this study shoot regeneration occurred from the asymmetric somatic cell hybrid calli. All isozyme banding patterns of these shoots investigated so far, however, were the same as those of birdsfoot trefoil. These results show that it is possible to regenerate the shoots from the cybrid calli derived from the asymmetric hybrid in which donor protoplasts irradiated by X-rays, while it seems to be impossible to regenerate the shoots from the symmtrical somatic hybrids which have both parent chromosomes. This phenomenon was also observed in the somatic cell hybrid of birdsfoot trefoil and soybean (Niizeki et al. 1990, 1994).


Table 1. Isozyme analyses of asymmetrical somatic cell hybrids of alfalfa and birdsfoot trefoil.
Total colonies obtained by protoplast fusion = 155.

Number of obtained hybrid clones = 17.

Isozyme pattern A*
Isozyme pattern B
Isozyme pattern C
Hybrid clone Plant regeneration
AMP
EST
GDH
CAT
AMP
EST
GDH
CAT
AMP
EST
GDH
CAT
I-1
-
A+B
-
-
A
B
B
B
B
-
-
-
-
I-3
-
A+B
-
-
A
B
B
B
B
-
-
-
-
I-7
+
A
-
-
B
B
B
B
B
B
B
B
B
I-16
-
A+B
-
-
A
-
-
-
-
-
-
-
-
I-20
-
B
B
-
A
B
B
B
B
-
-
-
-
I-27
+
A+B
B
-
A+B
A+B
B
B
B
-
-
-
-
I-29
+
A+B
A+B
-
A+B
B
B
B
B
B
B
B
B
I-31
-
A+B
A+B
-
A+B
B
B
B
B
-
-
-
-
I-78
-
A+B
A+B
B
A+B
-
-
-
-
-
-
-
-
I-82
-
A+B
A
A
A+B
-
-
-
-
-
-
-
-
II-14
-
A+B
B
B
A+B
-
-
-
-
-
-
-
-
II-34
+
A+B
B
B
A+B
B
B
B
B
-
-
-
-
II-44
-
A
A+B
A+B
A+B
-
-
-
-
-
-
-
-
II-45
+
A+B
A
A+B
A
B
B
B
B
B
B
B
B
II-46
-
A
A
A+B
A
-
-
-
-
-
-
-
-
II-47
-
A+B
-
A
B
B
B
B
B
-
-
-
-
II-63
+
A+B
A+B
B
A+B
B
B
B
B
-
-
-
-

* : +, plant regeneration; -, no plant regeneration.
Isozyme pattern A: Investigate the calli cultured for one month; Isozyme pattern B: Investigate the calli cultured for two months; Isozyme pattern C: Investigate the regenerated plant.
A: Isozyme band pattern of alfalfa; B: Isozyme band pattern of birdsfoot trefoil; A+B: Isozyme band pattern of both parents.


References

Kao, K.M. and M.R.Michayluk 1975. Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media. Planta 126: 105-110.

Murashige, T. and F. Skoog 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant 15: 473-497.

Niizeki, M. and K. Saito 1989. Callus formation from protoplast fusion between leguminous species of Medicago sativa L. and Lotus corniculatus L. Japan. J. Breed. 39: 373-377.

Niizeki, M., K. Kihara and K-N. Cai 1994. Somatic hybridization between birdsfoot trefoil (Lotus corniculatus L.) and soybean (Glycine max L.). In Biotechnology in Agriculture and Forestry, Vol.27 "Somatic Hybridization in Crop Improvement I", Y.P.S. Bajaj (ed), Springer-Verlag pp132-144.

Niizeki, M., K. Cai, M. Kihara, S. Nakajo and T. Harada 1990. Somatic cell hybrids between birdsfoot trefoil and soybean. Lotus Newslett. 21: 14-17.


Lotus tenuis WALD. ET KIT. (FABACEAE) : AN ARCHITECTURAL ANALYSIS

G. Mónica Tourn and G. Germán Roitman
Cátedra de Botánica Agrícola
Facultad de Agronomía. Universidad de Buenos Aires
Av. San Martín 4453. 1417
Buenos Aires. Argentina.

Internet: gmtourn@criba.edu.ar & rqroit@criba.edu.ar

A morphological and architectural study was made on an herbaceous species of Fabaceae, Lotus tenuis Walds. et Kit (narrow birds-foot trefoil), a perennial pasture legume.

This species was introduced into South America (in Argentina, in 1930), from Europe, and became established and naturalized on the Flooding Pampa (Burkart, 1952). Agronomists are selecting various commercial varieties for best performance an alternative to Medicago sativa (lucerne grass) affected by flooding, a very common disturbance in this region (León and Oesterheld, 1982).

This research represents a basic study of the morphology and growth habit of Lotus tenuis, that is essential for a correct grazing management.

The study was conducted during 1993-94, seeds sown in May 17, at experimental fields of the Faculty of Agronomy, Buenos Aires, Argentine,(34ş 35'S, 58ş 29'W), with seeds from a cultivar provided by AgroVerónica, Verónica, Buenos Aires Province. Twenty isolated plants were sampled to determined the architectural unit before nature defoliation.

The architectural unit of any plant are the different (morphological and functional) axes categories (Barthélémy, 1991). These axes are formed by shoot-units (Bell, 1994), each derived from one apical meristem.

The architectural unit is determined by two shoot categories: vegetative and reproductive (flowering branches). The leaves have spiral arrangement and have a single axillary bud with 2 prophylls (&) each with a bud, which can develop vegetative or reproductive branches. The first branches (prb) develop from the prophyllar buds of the axillary complex. Sometimes only one of the prophyllar bud develops, whilst the another and the axillary bud, remain dormant.

The vegetative shoots are basitonic. All vegetative axes -main axis (ma) and branches (b)- are monopodial with continuous growth . Shoot units initially develop horizontally, then change growth direction and procede to develop vertically (Fig. 1 A-D). The proximal nodes of the shoot units have the capacity to produce adventitious roots (ar), this represents a plagiotropic form during establishment (Fig. 1 C-D). The converse reorientation occurs once for the initial shoot unit of many herbaceous plants (Bell, 1994).

The apical meristematic activity is limited, when the growing season finishes in summer after flowering (Fig 1 C & D), all the apical meristems die (md).

During the first year the plant develops the crown, formed by a group of closed branches arising at the basal nodes of the main axis, first and second order branches, near the soil (Fig. 1 B-D).

The reproductive branching pattern is acrotonic (Fig. 1 C). The flowering branches (fb) arise from the axillary buds of the upper nodes of all vegetative axes.

The architecture (growth-form, habit) is determined by one orthotropic, sympodial axis (ma), with limited growth; orthotropic basitonic vegetative branches with a similar pattern (reiterations) occur (Fig. 1 D).

The plant repeats its elementary architecture (the architectural unit, building by shoot units).

At the end of growing season, when the shoots units of the previous year die, the proximal nodes lateral buds of the plagiotropic phase of the branches (Fig. 1 D) give rise new reiterations -proleptic, partial and adaptive- (nr). The old dead stems, of previous year, with adventitious roots, remain as stolons or rhizomes. These regrowth shoots may be vulnerable to damage by too-intensive grazing. The importance of the plagiotropic development of the basal shoots is crucial to long lived axillary meristems.

The independence of the different modules from the main axes may develop, as an important way of propagation, in the second or third period of growth, although it's not clear, the importance of this strategy, during the establishment.

The architectural model responds to Tomlinson's. Shoot units initially develop horizontally, then change growth direction, in established phase as in the Tomlinson's model (Hallé, et al., 1978).

References.

Barthélémy D. 1991. Levels of organization and repetition phenomena. Acta Biotheoretica 39: 309-323.

Bell AD. 1994. A Summary of the Branching Process in Plants. In Ingram, D.S. & Hudson, A. (eds.), Shape and Form in Plants and Fungi. Linn. Soc. Symposium Series 16, pp. 119-142. London: Academic Press.

Burkart A. 1952. Las Leguminosas argentinas silvestres y cultivadas.Ed. ACME, Bs. As. 2da. Edición. Pp.590.

Hallé F, Oldeman RAA & Tomlinson PB. 1978. Tropical trees and forests. Springer, Berlín. Pp. 441.

León RJC y Oesterheld M. 1982. Envejecimiento de pasturas implantadas en el Norte de la Depresión del Salado, un enfoque sucesional. Rev.Fac. Agron. Bs. As. 3: 41-49.


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Lotus tenuis A-E; diagrammatic representation of 5 growing stages: A.- First stage: The main axis bears at the cotyledonary node, one shoot with a prophyllar branch, at each cotyledon axil; all the axes show continuous growth; B.-Second stage : Main axis with basitonic continuous branches from cotyledons to 4-node. The prophyllar branches are larger than cotyledonary shoots; C.- Third stage: Adult phase previous to reproductive stage with complete branching pattern (second and third order branches); D.- Fourth stage: Adult phase with flowering branches; E.- Last stage: Apical meristem of all shoots die. The reiterations (broken line) arise from the crown.

Arrows indicate that stems are still growing. In C all the axes with curved arrows indicate a larger growth than the main axis. In B & C Stars and asterisks represented second order branches. Cotyledons are represented schematically and are not to scale; normal foliage leaves are not represented. In D, little stars represent second order flowering branch.

Abbreviations: ar: adventitious roots; b1, b2, b3 and b4: first node axillary branch to four node branch; c: cotyledon; cs: cotyledonary shoot; fb: flowering branches, ma: main axis; md: meristem death; prb: prophyllar branch; re: reiterations.

Arrows indicate that stems are still growing. Stars represent second order flowering branch. Cotyledons are represented schematically and are not to scale; normal foliage leaves are not represented.


Current Literature

Isolation, chemical structures and biological activity of the lipo-chitin oligosaccharide nodulation signals from Rhizobium etli. Cardenas, L.; Dominguez, J.; Quinto, C.; Lopez-Lara, I. M.; Lugtenberg, B. J. J.; Spaink, H. P.; Rademaker, G. J.; Haverkamp, J.; Thomas-Oates, J. E. Departamento de Biologia Molecular de Plantas, Instituto de Biotecnologia, Universidad Nacional Autonoma de Mexico, Mexico. Plant Molecular Biology vol. 29 (3): p.453-464. Publication Year: 1995

Rhizobium etli is a microsymbiont of plants of the genus Phaseolus. Using MS the lipo-chitin oligosaccharides (LCOs) that are produced by R. etli strain CE3 were identified. They are N-acetylglucosamine pentasaccharides of which the nonreducing residue is n-methylated and n-acylated with cis-vaccenic acid (C18:1) or stearic acid (C18:0) and carries a carbamoyl group at C4. The reducing residue is substituted at the C6 position with O-acetylfucose. Analysis of their biological activity on the host plant Phaseolus vulgaris shows that these LCOs can elicit the formation of nodule primordia which develop to the stage where vascular bundles are formed. The formation of complete nodule structures, including an organized vascular tissue, is never observed. Considering the close resemblance of the R. etli LCO structure of those of R. loti, the ability of R. etli strains to nodulate various Lotus species and of R. loti to nodulate P.vulgaris was tested. The results show that R. etli is able to nodulate Lotus plants. However, several Lotus species are only nodulated when an additional flavonoid independent transcription activator (FITA) nodD gene is provided. Phaseolus plants can also be nodulated by R. loti bacteria, but only when the bacteria contain a FITA nodD gene. Apparently, the type of nod gene inducers secreted by the plants is the major basis for the separation of Phaseolus and Lotus into different cross inoculation groups.

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A chromosome atlas and interspecific-intergenic index for Lotus and Tetragonolobus (Fabaceae). Grant, W. F. Department of Plant Science, P.O. Box 4000, McGill University, Macdonald Campus, Ste Anne de Bellevue, QC H9X 3V9, Canada. Canadian Journal of Botany vol. 73 (11): p.1787-1809 Publication Year: 1995

Basic chromosome numbers in Lotus are x = 5, 6 and 7. It is considered that evolution has proceeded in the genus by means of a descending aneuploid series from an 8-chromosomed ancestor. Chromosome numbers for species of Tetragonolobus are based on x = 7. Somatic chromosome markers are reported for 108 species and 38 varieties. Chromosome numbers are reported for the first time for 5 species (L. hamatus, L. haydonii, L. hintoniorum, L. mearnsii and L. utahensis; all 2n = 14), 6 varieties (L. argophyllus var. argenteus, L. dendroideus var. traskiae, L. heermanii var. orbicularis, L. junceus var. biolettii, L. strigosus var. hirtellus, L. strigosus var. tomentellus; all 2n = 14) and L. uliginosus subsp. vestitus (2n = 12). Natural diploid, tetraploid and hexaploid plants are reported for L. alpinus. Several species are reported as possessing B chromosomes. Mixoploidy is reported to occur in L. alpinus, L. glacialis and L. glareosus. In addition, chromosome numbers are given for plants regenerated from calluses grown in tissue culture having heteroploidy, euploidy and mixoploidy. Root nodules are reported with tetraploid and octoploid cells in addition to the normal number of chromosomes. Trisomic series have been partially developed in L. tenuis and L. uliginosus. Polytene chromosomes were observed in suspensor cells of three species of Lotus. Feulgen cytophotometric measurements, to determine the DNA nuclear content, were made for 16 species of Lotus and one species of Tetragonolobus. The majority of the studies in Lotus concerned the economic species L. corniculatus, L. tenuis and L. uliginosus. Interspecific hybridization was carried out in different combinations between diploids, autoploids and amphidiploids. Intergeneric hybrids were attempted by somatic hybridization, protoplast fusion and assymetric hybridization between Lotus and other species (Glycine max, Medicago sativa, Oryza sativa).

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Effect of inoculation and nitrate on nitrate reductase activity and acetylene reduction activity in Lotus sp.- Rhizobium loti symbiosis. Diaz, P.; Borsani, O.; Monza, J. Departamento de Bioquimica, Facultad de Agronomia, Av. E. Garzon 780 C.P. 12900, Montevideo, Uruguay. Symbiosis (Rehovot) vol. 19 (1): p.53-63. Publication Year: 1995

The effects of nitrate and the Rhizobium strain used in the inoculation on acetylene reduction activity (ARA), nitrate reductase activity (NRA) and nitrate concentration in tissues of two Lotus species inoculated with different Rhizobium loti strains were examined. ARA in the symbiosis Lotus corniculatus-R. loti T1 strain was inhibited 90% by nitrate compared with 36% in the L. tenuis-R. loti Y3 symbiosis. NRA in leaves and nitrate concentration in stems were modified by inoculation. L. tenuis and L. corniculatus nodulated by R. loti T1 strain had lower nitrate concentration in the stem than when the same plants were nodulated by R. loti Y3 and U226 strains and the non-nodulated plants. Inoculation increased the NRA expression in leaf tissues of Lotus spp. nodulated by R. loti T1 strain and the nitrate concentration was similar to that found in non-nodulated plants.

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Forage halophytes in the Mediterranean basin. Le Houerou, H. N. 327 rue A. L. De Jussieu, 34090 Montpellier, France. Book Title: Halophytes and biosaline agriculture. p.115-136. Publication Year: 1995. Editors: Choukr-Allah, R.; Malcolm, C. V.; Hamdy, A. Publisher: Marcel Dekker Inc. New York, USA ISBN: 0-8247-9664-0

It was shown that out of 700 Mediterranean halophyte species, 27.5% were from the Chenopodiaceae, 15% from the Poaceae, 6% from the Asteraceae and 5% from the Caryophyllaceae, Fabaceae and Zygophyllaceae; 70% were perennial and 30% annual or biennial. The most common tree species was Tamarix, the most common shrubs Salicornia, Salsola, Suaeda and Atriplex, the most common perennials Aeluropus, Sporobolus, Puccinellia and the most common annual species Hordeum maritimum, Lepturus cylindricus, Frankenia and Melilotus. The halophytic vegetation was described and aspects of the palatability (ratio of feed intake:fodder on offer) and preference; primary productivity (aboveground biomass of Atriplex halimus was 10-15 t DM/ha and annual productivity 2-5 t DM/ha); grazing and nutritive value (digestible DM 40-70%, digestible OM 50% and CP content 10-12%); and cultivation (Atriplex nummularia, A. semibaccata and Myoporum insulare grown in Australia and A. canescens and A. lentiformis in the USA) were discussed. Many salt tolerant fodder species used in land reclamation with or without irrigation using saline water (Festuca arundinacea, Sporobolus, Trifolium fragiferum, Lotus corniculatus, Lolium rigidum, Hordeum vulgare, Melilotus alba, M. italica and Medicago spp.) gave DM yields of 5-20 and 2-10 t/ha, respectively.

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Comparison of conventional and alternative nursery weed management strategies. Calkins, J. B.; Swanson, B. T. Dept. Hort. Sci., Univ. Minn., St. Paul, MN 55108, USA. Weed Technology vol. 9 (4): p.761-767. Publication Year: 1995 ISSN: 0890-037X

In a field plot at the University of Minnesota, St. Paul, soil cultivation (3 to 5 times/yr) and herbicide management (oxadiazon, 3.92 kg ai/ha), agricultural standards for reducing weed competition, were compared to 3 alternative nursery field management systems regarding weed suppression: 'Norcen' bird's-foot trefoil (Lotus corniculatus) companion crop, 'Wheeler' winter rye cover crop/mulch and grass sod (80% 'Eton' perennial ryegrass (Lolium perenne) and 20% 'Ruby' red fescue (Festuca rubra)). Field management treatment had a significant effect on observed weed populations. Weed densities were also subject to yearly variations caused by climate and endogenous weed life cycles. Herbicide management (oxadiazon) consistently provided the best control of undesired vegetation (0.3 weeds/m2) followed by the grass sod (0.7 weeds/m2), Wheeler rye cover crop/mulch (1.7 weeds/m2), Norcen bird's-foot trefoil companion crop (8.6 weeds/m2), and cultivated (55.7 weeds/m2) treatments. Although the grass sod treatment provided excellent control of undesired vegetation, as an alternative to cultivation and herbicide use, it proved to be excessively competitive with the nursery crop. The bird's-foot trefoil treatment quickly became infested with broadleaf weeds the eradication of which proved difficult. The Wheeler winter rye cover crop/mulch field management system provided acceptable weed control combined with other beneficial effects on the plant/soil environment. Results support the effectiveness of Wheeler winter rye and perhaps other allelopathic cover crop/mulch systems in controlling undesired vegetation in horticultural field production systems.

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Meiosis and seed production in plants of different populations of birdsfoot trefoil (Lotus corniculatus L.). Nikolaichuk, V. I. Uzhgorod University, Uzhgorod, Ukraine. Tsitologiya i Genetika vol. 29 (4): p.61-66. Publication Year: 1995 ISSN: 0564-3783

PMC meiosis was studied in 3 natural populations and 4 varieties. The frequency of disturbances at different stages of meiosis was low, not exceeding 12.5% at the tetrad stage in the variety Smolens'kii. Plants of this variety had the lowest pollen fertility and, accordingly, the lowest seed set.

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Rhizomatous Lotus corniculatus L.: I. Taxonomic and cytological study. Beuselinck, P. R.; Li, B.; Steiner, J. J. USDA-ARS, Plant Genetics Research Unit, Columbia, MO 65211, USA. Crop Science vol. 36 (1): p.179-185. Publication Year: 1996 ISSN: 0011-183X

Wild, putative L. corniculatus accessions from Morocco differ distinctly from domesticated L. corniculatus in that they produce rhizomes. Rhizomes may be useful in increasing stand persistence of domesticated L. corniculatus. The objective of this study was to identify to the species level the wild Lotus accessions from Morocco (G31272, G31273, G31276, G31298 and G31317) that present rhizomatous growth. Comparative morphological and cytological analyses, and intercrossing with L. corniculatus, were used in this study. Six morphological characters that distinguish L. corniculatus and L. uliginosus (the only species in the genus Lotus recognized as producing rhizomes) were used to categorize taxonomically the Moroccan accessions. The Moroccan accessions were morphologically similar to L. corniculatus with the exception of the rhizome trait. The accessions were tetraploid with 2n = 4x = 24 somatic chromosomes. Karyotype analyses detected more similarities than differences between the Moroccan accessions and Norcen. Progeny were produced from intercrosses between L. corniculatus cultivars Norcen and AU Dewey and the Moroccan accessions. Hybridity of progeny was verified by morphological and RAPD analyses. A sample of reciprocal F1 progeny were tetraploid, fertile and consistent in their production of rhizomes. A combination of morphological, chromosome, and karyotype analyses confirmed accessions G31272, G31273, G31276, G31298, and G31317 as L. corniculatus. This is the first description of L. corniculatus with rhizomes.

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Proanthocyanidins from Lotus corniculatus. Foo, L. Y.; Newman, R.; Waghorn, G.; McNabb, W. C.; Ulyatt, M. J. NZ Institute for Industrial Resesarch and Development, P.O. Box 31-310, Lower Hutt, New Zealand. Phytochemistry vol. 41 (2): p.617-624. Publication Year: 1996 ISSN: 0031-9422

The chemical structure of the purified proanthocyanidin polymers of Lotus corniculatus was analysed by 13C NMR and by mild acid catalysed degradation in the presence of excess of phloroglucinol. The NMR data showed that the polymer was partially glycosidated with a number average MW in the range 1800-2100 (6 to 7 flavanoid units). The products from phloroglucinol scission reaction indicated the extender flavan units to consist mostly of epicatechin (67%) and epigallocatechin (30%), with minor amounts of catechin and epiafzelechin units, which were linked together predominantly by C-4/C-8 interflavanoid bonds. The polymer chains were terminated mostly by catechin (83%) and, to a lesser extent, by epicatechin (16%).

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Taproot tip penetration and young seedling growth in relation to root-hair adhesion among surface-sown legume species. Morita, O.; Iwabuchi, K.; Goto, M.; Ehara, H. Faculty of Bioresources, Mie University, Tsu, Mie 514, Japan. Nippon Sochi Gakkaishi = Journal of Japanese Society of Grassland Science vol. 40 (4): p.437-442. Publication Year: 1995 ISSN: 0447-5933

Seeds of white clover (Trifolium repens), alsike clover (T. hybridum), red clover (T. pratense), crimson clover (T. incarnatum), birdsfoot trefoil (Lotus corniculatus), alfalfa (Medicago sativa) and common vetch (Vicia sativa) were surface-sown on yellow soil under controlled conditions with 25 deg C and 100% RH. Penetration percentage of taproot tips, mean days for taproot penetration, and hypocotyl and root growth of three adhesion types in germination behaviour were examined for five days to decide the adhering function of taproot hairs. The three adhesion types in legumes were contact-adhesion, partial-adhesion and non-adhesion, depending on the degree of adhesion in the taproot hairs. Ninety percent of taproot tips of the contact-adhesion type in white clover, alsike clover, birdsfoot trefoil and alfalfa penetrated into the soil, being significantly higher than that of partial-adhesion type. The same tendency was observed in red clover, crimson clover and common vetch. The mean time for taproot tip penetration of the contact-adhesion type was shorter than that of the partial-adhesion type in all species examined. Length and proportion of taproot exposed on the soil surface were shorter in the contact-adhesion type than in the partial-adhesion type. The hypocotyl-root ratio in length was highest in the contact-adhesion type, followed by partial-adhesion type and non-adhesion types. Therefore, good adhesion of taproot hairs to the surface enhanced the hypocotyl growth as compared with taproot growth, and poor adhesion of taproot hairs enhanced taproot growth. It is suggested that the contact-adhesion type of germination behaviour would provide good establishment compared with partial-adhesion and non-adhesion types.

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Effect of adhesion of taproot hairs to the soil surface on germination behaviour in surface-sown legume species. Morita, O.; Iwabuchi, K.; Goto, M.; Ehara, H. Faculty of Bioresources, Mie University, Tsu, Mie 514, Japan. Nippon Sochi Gakkaishi = Journal of Japanese Society of Grassland Science vol. 40 (4): p.429-436. Publication Year: 1995 ISSN: 0447-5933

White clover (Trifolium repens), alsike clover (T. hybridum), red clover (T. pratense), crimson clover (T. incarnatum), birdsfoot trefoil (Lotus corniculatus), alfalfa (Medicago sativa), and common vetch (Vicia sativa) were sown on the surface of yellow paddy field soil under controlled conditions at 25 deg C and 100% RH. Some morphological characteristics of the taproot in relation to germination behaviour were measured. Elongating taproots pushed the seed backwards and grew without penetration of their tips into the soil. After root hairs developing on taproots adhered to the surface, anchoring the seedlings, root tips penetrated into the soil. Germination behaviour was classified into three types: (i) taproot hairs that developed initially adhered to the soil surface, thus taproots grew in contact with the soil surface and root tips penetrated into the soil (contact-adhesion type); (ii) taproot hairs adhered partially to the soil surface, a part of the taproot was not in contact with the surface, but root tips penetrated (partial-adhesion type); and (iii) taproot hairs failed to adhere to the surface, taproots were not in contact with the soil, and failed to penetrate into the soil (non-adhesion type). White clover and birdsfoot trefoil had a higher percentage of the contact-adhesion type in germination. Alsike clover, red clover, crimson clover and alfalfa had higher percentages of the partial-adhesion type, and common vetch had a higher percentage of the non-adhesion type. The percentage of contact-adhesion type seedlings was related to the ratio between length of root hair and root diameter in the legume species. The adhering area that was bound on the soil surface by taproot hairs was significantly larger in the contact-adhesion type than in the partial-adhesion type. It is suggested that an adhering function of taproot hairs in surface-sown legume seedlings may secure good penetration of root tips before establishment can be achieved.

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Novel and complex chromosomal arrangement of Rhizobium loti nodulation genes. Scott, D. B.; Young, C. A.; Collins-Emerson, J. M.; Terzaghi, E. A.; Rockman, E. S.; Lewis, P. E.; Pankhurst, C. E. Molecular Genetics Unit, Department of Microbiology and Genetics, Massey University, Palmerston North, New Zealand. Molecular Plant-Microbe Interactions vol. 9 (3): p.187-197. Publication Year: 1996 ISSN: 0894-0282

A mutational and structural analysis of Rhizobium loti nodulation genes in strs NZP2037 and NZP2213 was carried out. Unlike the case with other Rhizobium strs examined to date, nodB was found on an operon separate from nodACIJ. Sequence analysis of the nodACIJ and nodB operon regions confirm that R. loti common nod genes have a gene organization different from that of other Rhizobium spp. At least 4 copies of nodD-like sequences were identified in R. loti. The complete nucleotide sequence of one of these, nodD3, was determined. A new host-specific nod gene, nolL, was identified adjacent to nodD3. NolL shares homology NodX and other O-acetyl transferases. Mutational analysis of the nod regions of strains NZP2037 and NZP2213 showed that nodD3, nodI, nodJ, and nolL were all essential for R. loti strs to effectively nodulate the extended host Lotus pedunculatus, but were not necessary for effective nodulation of the less restrictive host, Lotus corniculatus. Both nodD3 and nolL were essential for R. loti strs to nodulate Leucaena leucocephala.

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Evaluation and characterization of Lotus corniculatus accessions in spaced and dense stand conditions. (Valutazione e caratterizzazione di accessioni di Lotus corniculatus a piante spaziate e a semina fitta.) Negri, V.; Francia, U.; Veronesi, F. Istituto di Miglioramento Genetico Vegetale, Universita degli Studi, Perugia, Italy. Rivista di Agronomia vol. 29 (2): p.160-164. Publication Year: 1995 ISSN: 0035-6034

In forage plants, single plant evaluation is often reported to be of little interest in predicting sward performance. With the aim of assessing the reliability of spaced plant evaluation in screening birdsfoot trefoil germplasm for breeding purposes, a spaced plant trial and a dense stand trial were set up. Nine natural populations were studied in both trials. The following agronomic, physiological and qualitative characters were recorded in 1993: winter growth, spring regrowth, first flower date, average stem length, total dry matter yield, crude protein, crude fibre, ash, in vitro dry matter digestibility, milk feeding units and meat feeding units. Polcanto, Macerata and Canetra were the most promising natural populations. The data collected for each character and for each population in the spaced plant and dense stand trials showed substantial agreement. When screening birdsfoot trefoil accessions from sites with different ecological conditions, evaluation of spaced plants appears to be reliable in predicting sward performance.

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Use of herbicides for the chemical protection of non-traditional forage crop seed stands. (Vyuziti herbicidu v chemicke ochrane semenarskych porostu netradicnich picnin.) Tesar, O. Vyzkumny Ustav Picninarsky, 664 41 Troubsko, Czech Republic. Scientific Studies - Research Institute for Fodder Plants (No. 13): p.91-104. Publication Year: 1995

One pre-sowing and 14 post-em. herbicides were compared in seed crops of the annual forage legumes Melilotus albus, Trigonella coerulea, Tetragonolobus purpureus, Lotus ornithopodioides and Malva verticillata in 1993 and 1994. The most effective combinations were imazethapyr-ammonium + pyridate + rapeseed oil in T. coerulea and M. albus with yield increases, no phytotoxicity and 95.7-100% control of Amaranthus sp. and 67.8-71.5% control of Chenopodium sp. In M. verticillata, metamitron + triclopyr, metamitron + phenmedipham or desmedipham, and metamitron + fluazifop-P-butyl or haloxyfop gave 72.1-90.7% weed control without phytotoxicity but crop yields were unaffected. T. purpureus and L. ornithopodioides trials were unsatisfactory.

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Seed multiplication of alpine grasses and herbs. (Saatgutvermehrung alpiner Graser und Krauter.) Krautzer, B. Bundesanstalt fur Alpenlandische Landwirtschaft Gumpenstein, 8952 Irdning, Austria. Rasen-Turf-Gazon vol. 26 (4): p.123-131. Publication Year: 1995 ISSN: 0341-9789

Short descriptions are given of 7 grasses, 4 legumes and 6 forbs regarded as promising for use in mixtures for resowing damaged areas at high altitude and suitable for commercial seed production at lower altitude. Information is presented on seed characteristics, 1000-seed weight, germination and seed quality. The latter was comparable with that of commercial seed of lowland species. Seed yields of Festuca nigrescens and Poa alpina decreased with increasing altitude of origin, but adaptation to high altitude may be more important than yield. In Trifolium pratense subsp. nivale the disease susceptibility of some strains meant that their yield was not related to the altitude of origin. Average seed yields of the grasses (F. nigrescens, F. pseudodura, F. supina (F. airoides), F. violacea, Phleum alpinum, P. hirsutum and Poa alpina) were 100-500 kg/ha and those of the legumes (Anthyllis vulneraria, Lotus corniculatus, Trifolium badium and T. pratense subsp. nivale) were 30-350 kg/ha. The highest yielding forb was Dianthus superbus (average 578 kg/ha); the other species (Arnica montana, Crepis aurea, Leontodon hispidus, Melandrium rubrum and Plantago montana) yielded 100-300 kg/ha. Potential yields were considerably higher. Trials of mixtures and sowing techniques for specific areas are continuing.

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Steps toward defining the role of lectins in nodule development in legumes. Hirsch, A. M.; Brill, L. M.; Lim, P. O.; Scambray, J.; Rhijn, P. van Department of Molecular, Cell and Developmental Biology, 405 Hilgard Avenue, University of California, Los Angeles, CA 90095-1606, USA. Conference Title: 10th International congress on nitrogen fixation, St. Petersburg, Russia, 28 May-3 June 1995. Symbiosis (Rehovot) vol. 19 (2/3): p.155-173. Publication Year: 1995 ISSN: 0334-5114

A number of plant and bacterial genes are specifically induced upon the interaction between rhizobia and plants during the onset of nodulation. However, many other genes such as those encoding lectin, a protein which has been hypothesized as being involved in host recognition by rhizobia, are not specifically induced in response either to rhizobial inoculation or to Nod factor treatment. Exactly how lectin is involved in recognition between rhizobia and legume is still not well understood. To this end, two lucerne lectin genes were isolated and characterized from a lucerne genomic library. Lectin mRNA-antisense lucerne plants were generated and the early stages of development of transgenic plants were studied. Differences were seen from the control plants with respect to the number of plants formed per callus. Also, the soyabean lectin gene was transferred into lucerne as well as Lotus corniculatus, and the response of the transgenic plants to inoculation with Bradyrhizobium japonicum, the usual symbiont for soyabean, was examined.

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Seed yield, size and indeterminacy in diverse accessions of Lotus corniculatus L. grown in the UK. Bullard, M. J.; Crawford, T. J Department of Biology, University of York, Heslington, York YO1 5DD, UK. Plant Varieties & Seeds vol. 9 (1): p.21-28. Publication Year: 1996 ISSN: 0952-3863

Eighteen accessions of the forage legume Lotus corniculatus were grown as spaced plants. The period of mature seed production, number of seed pods, seed yield and seed size of these accessions were measured. All accessions produced mature seed over a period of at least 8 weeks. The pattern of production varied greatly between accessions and included unimodal, bimodal and irregular forms. A common date for peak of seed pod production could not be ascertained. There was no indication that any variety or UK ruderal alien exhibited a more contracted period of pod production that could be exploited in a plant breeding programme. The 100-seed weights of the different accessions varied two-fold (between 116.2 and 239.7 mg); ecotypes were generally heavier than varieties or alien accessions. The Norwegian ecotype from Nigardelen had heavier seed than any other accession, and as seedling vigour is positively correlated with seed size in L. corniculatus, it may offer potential for improvement of seed size in commercial material by interaccessional crossing.

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The influence of diet on growth and secretion behaviour of myrmecophilous Polyommatus icarus caterpillars (Lepidoptera: Lycaenidae). Burghardt, F.; Fiedler, K. Lehrstuhl Zoologie II, Theodor-Boveri-Biozentrum der Universitat, Am Hubland, D-97074 Wurzburg, Germany. Ecological Entomology vol. 21 (1): p.1-8. Publication Year: 1996 ISSN: 0307-6946

Larvae of the facultatively myrmecophilus lycaenid Polyommatus icarus were reared on inflorescences, or foliage, of 4 natural hostplant species and on an artificial diet to study dietary effects on larval growth and secretory capacity. Larvae achieved highest weights and relative growth rates when fed flowers of Medicago sativa (lucerne), Lotus corniculatus or Melilotus officinalis. Larvae reared on Coronilla varia (flowers and leaves), foliage of M. sativa and on the artificial diet pupated at lower weights and achieved lower growth rates. In standardized experiments with the ant species Lasius flavus, secretion rates from the dorsal nectar organ (DNO) were 2 times higher among flower-fed larvae than among foliage-fed siblings or larvae on the artificial diet. Larvae reared on C. varia flowers were superior to all other food treatments with respect to secretion rates. High water content of larval diet, as in flowers appears to be important for lycaenid larvae to achieve high secretion rates, whereas the correlation between myrmecophily and nutrient availability, as evidenced by growth rates, was less pronounced. Using experimental data on larval growth and secretion rates, the lifetime volume of secretions from the DNO was estimated to range from 2 to 5 micro l in most food treatments. Only on C. varia flowers (5.5-8.7 micro l) and on M. sativa leaves (0.0-1.1 micro l) did the larvae deviate in their absolute investment in myrmecophily. The estimated lifetime investment accounted for 1.6-5.5% of prepupal fresh weight in all food treatments except on C. varia flowers (7.8-12.3%).

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How much fertilizer will a species-rich chalk grassland in the Swabian Alb stand? (Wieviel Dungung 'vertragt' ein artenreicher Kalkmagerrasen der Schwabischen Alb?) Briemle, G. Staatliche Lehr- und Versuchsanstalt fur Viehhaltung und Grunlandwirtschaft (LVA), Atzenbergerweg 99, 88326 Aulendorf, Germany. Kongressband 1995 Garmisch-Partenkirchen. Vortrage zum Generalthema des 107. VDLUFA-Kongresses vom 18.-23.9.1994 in Garmisch-Partenkirchen: Grunland als Produktionsstandort und Landschaftselement. Conference Title: Kongressband 1995 Garmisch-Partenkirchen. Vortrage zum Generalthema des 107. VDLUFA-Kongresses vom 18.-23.9.1994 in Garmisch-Partenkirchen: Grunland als Produktionsstandort und Landschaftselement. p.631-634 Publication Year: 1995. Publisher: VDLUFA-Verlag Darmstadt, Germany ISBN: 3-922712-55-X

In 1983-93 dry chalk grassland at 780 m alt. dominated by Bromus erectus, Sanguisorba minor, Lotus corniculatus and Trifolium montanum and containing many species of gentians and orchids was mown in July each year and given 20 kg N + 20 kg P + 32 kg K/ha, half these rates, 10 kg P + 16 kg K or no fertilizer, was mulched or the succession was left undisturbed. Mineral fertilizer application or mulching had little effect on the number of species present, which remained at 60-70/25 m2. The number of species fell by about 19% on the other plots. The herbage cut in July contained 10.8% CP and consisted of 44% grasses, 8% legumes and 48% other forbs. Average DM yields ranged from 2.3 t/ha with no fertilizer to 3.1 t with the higher NPK rate. The hay was highly acceptable to growing cattle and gave liveweight gains of 990 g/d.

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Drought stress in seedlings: lipid metabolism and lipid peroxidation during recovery from drought in Lotus corniculatus and Cerastium fontanum. Olsson, M.; Nilsson, K.; Liljenberg, C.; Hendry, G. A. F. Dept. of Plant Physiology, Univ. of Goteborg, Carl Skottsbergs gata 22, S-413 19 Goteborg, Sweden. Physiologia Plantarum vol. 96 (4): p.577-584. Publication Year: 1996 ISSN: 0031-9317

Seedling survival of drought during the first days following germination can be one of the most critical factors in successful establishment of the species. 72-h-old seedlings of Cerastium fontanum and Lotus corniculatus were exposed to severe desiccation for 36 h and the recovery of the whole plants monitored over the subsequent 17 d. The analysis exposed the very different responses to water stress in the two species. The effects of water stress on the less drought tolerant L. corniculatus within the first 5 to 10 d after drought treatment included a 2-fold larger loss in dry weight than in C. fontanum and a 2-fold rise in lipid peroxidation (thiobarbituric acid-reactive substances), in triacylglycerols and in free fatty acids. The ratio of monogalactosyl diacylglycerols/digalactosyl diacylglycerols (MGDG/DGDG) declined 3-fold, while the proportion of MGDG was some 12-fold lower. In contrast, in the relatively drought-tolerant seedlings of C. fontanum no changes of this order were recorded in the days immediately following rehydration. The relationship between these changes in lipid metabolism, lipid peroxidation and drought-induced damage in terms of growth is discussed in the context of seedling recovery and survival in the two species.

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Cotyledon and leaf development associated with seedling vigor of six forage legumes. Hur, S. N.; Nelson, C. J. Department of Animal Science, Chonbuk National University, Chonju 560-756, Korea Republic. Journal of the Korean Society of Grassland Science vol. 15 (1): p.19-23. Publication Year: 1995 ISSN: 1013-9354

Seeds of Medicago sativa, Trifolium hybridum, T. pratense, T. repens, Lotus corniculatus and Lespedeza striata were sown in a silt loam, peat and sand mixture in pots and kept in growth chambers. The cotyledons expanded rapidly in the 1st week after emergence then slowly up to the 3rd week. Maximum cotyledon areas of T. pratense, M. sativa, T. hybridum, T. repens and L. corniculatus were 9.5, 6.5, 6.4, 6.3 and 4.5-fold larger, respectively, than the mean area at emergence, whereas that of L. striata was only 2.9-fold the size. Larger seeds produced greater cotyledon areas and the rate of cotyledon expansion was positively correlated with seedling vigour. The development of leaf area followed a similar relationship. Early onset of leaf production contributed, with early leaf production, to vigorous seedling growth.

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Cotyledon development and seedling growth of Lotus corniculatus, Lotus tenuis and their reciprocal hybrids. Hur, S. N.; Beuselinck, P. R.; Nelson, C. J. Department of Animal Science, Chonbuk National University, Chonju 560-756, Korea. Journal of the Korean Society of Grassland Science vol. 15 (1): p.13-18. Publication Year: 1995 ISSN: 1013-9354

Seeds of Lotus corniculatus, L. tenuis and their reciprocal hybrids were sown in pots in a silt loam, peat and sand mixture. Cotyledon size was measured from seedling emergence to cotyledon senescence at 2-day intervals. Seedlings were harvested 3 weeks after emergence to measure net photosynthetic rate, dark respiration rate and seedling dry weight. Cotyledons expanded rapidly during the 1st week, then only slightly until maximum size was reached 3 weeks after emergence. Cotyledon area at emergence was positively correlated with seed size but after emergence there was a divergence in cotyledon expansion rate, with seed size alone not a reliable indicator of seedling vigour. Rate of leaf area development was a determinant of seedling growth. Some of the hybrids showed increased seedling vigour compared with the parent species.

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Element concentration cadasters of halophytic plant communities in Hungary. Kovacs, M.; Penksza, K.; Turcsanyi, G.; Kaszab, L.; Otvos, E. Agricultural University, Department of Botany and Plant Physiology, H-2103 Godollo, Hungary. Acta Botanica Hungarica vol. 39 (1/2): p.3-16. Publication Year: 1995 ISSN: 0236-6495

At Kiskunsag National Park species from the Lepidio-Camphorosmetum annuae and Agrosti-Caricetum distantis plant communities growing on solonchak soils were assessed and at Hortobagy National Park species from the Camphorosmetum annuae and Artemisio-Festucetum pseudovinae communities occurring on the solonchak-solonetz soils were assessed. Soil samples from both sites were analysed. Foliage of some of the halophytes (e.g. Camphorosma annua, Lepidium cartilagineum, Lotus tenuis and Scorzonera parviflora) contained high quantities of heavy metals. Monocots (including Puccinellia distans and Festuca pseudovina) were characterized by a smaller total content of detectable elements as well as a smaller element concentration range (0.1-1000 micro g/g) than the dicots. Tables are presented of soil element contents and the element contents of the leaves and other plant organs analysed.

Isolation, chemical structures and biological activity of the lipo-chitin oligosaccharide nodulation signals from Rhizobium etli. Cardenas, L; Dominguez, J.; Quinto, C.; Lopez-Lara, I.M.; Lugtenberg, B.J.J.; Spaink, H.P.; Rademaker, G.J.; Haverkamp, J.; Thomas-Oates, J.E. . Universidad Nacional Autonoma de Mexico. Plant molecular biology. 1995. vol. 29 (3), p. 453-464.

Rhizobium etli is a microsymbiont of plants of the genus Phaseolus. Using mass spectrometry we have identified the lipo-chitin oligosaccharides (LCOs) that are produced by R. etli strain CE3. They are N-acetylglucosamine pentasaccharides of which the non-reducing residue is N-methylated and N-acylated with cis-vaccenic acid (C18:1) or stearic acid (C18:0) and carries a carbamoyl group at C4. The reducing residue is substituted at the C6 position with O-acetylfucose. Analysis of their biological activity on the host plant Phaseolus vulgaris shows that these LCOs can elicit the formation of nodule primordia which develop to the stage where vascular bundles are formed. The formation of complete nodule structures, including an organized vascular tissue, is never observed. Considering the very close resemblance of the R. etli LCO structures to those of R. loti (I. M. Lopez-Lara, J. D. J. van den Berg, J. E. Thomas Oates, J. Glushka, B. J. J. Lugtenberg, H. P. Spaink, Mol Microbiol 15: 627-638, 1995) we tested the ability of R. etli strains to nodulate various Lotus species and of R. loti to nodulate P. vulgaris. The results show that R. etli is indeed able to nodulate Lotus plants. However, several Lotus species are only nodulated when an additional flavonoid independent transcription activator (FITA) nodD gene is provided. Phaseolus plants can also be nodulated by R. loti bacteria, but only when the bacteria contain a FITA nods gene. Apparently, the type of nod gene inducers secreted by the plants is the major basis for the separation of Phaseolus and Lotus into different cross inoculation groups.

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Relative effectiveness of phosphorus on narrow-leaf and broadleaf birdsfoot trefoil growth and the effect of added phosphorus on vesicular arbuscular mycorrhizal infection. Mendoza, R.; Gigli, S. Centro de Ecofisiologia Vegetal, Buenos Aires, Argentina.. Journal of plant nutrition. 1995. vol. 18 (7), p. 1483-1494.

The relative effectiveness of phosphorus (P) applications on growth and the effect of added P on the extent of infection of roots of narrowleaf birdsfoot trefoil (Lotus tenuis) and of broadleaf birdsfoot trefoil (Lotus corniculatus) by an indigenous VAM fungi (Glomus sp.) was studied on a P-deficient soil (Typic Natraquoll). In terms of rate of increase of shoot growth per unit of added P, broadleaf was more efficient than narrowleaf birdsfoot trefoil but they did not differ in the relative effectiveness of P for growth. For the two Lotus species, when increasing the level of added P there was an initial increase in the percentage of root length infected, and then with further additions, there was a consistent decrease of the infection. For narrowleaf, the maximum percentage of root length infected was when plants reached 11% of their maximum shoot growth. Whereas for broadfleaf, the maximum percentage of root infected was when plants reached the 66% of their maximum shoot growth. Despite differences in both, the shape of the response curve of shoot growth to P and the extent of infection between d species, they did not differ in their ability to utilize the P that had reacted with the soil for a period of time.

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Evaluation of a phytotoxin(s) from Pseudomonas syringae for weed control in cranberries. Norman, M.A.; Patten, K.D.; Gurusiddaiah, S. Washington State University, Long Beach, WA. HortScience. 1994. vol. 29 (12), p. 1475-1477.

Three indicator species [rye (Secale cereale L.), radish (Raphanus sativus L.), and alfalfa (Medicago sativa L.)] and nonrooted cuttings of 'Stevens' cranberry (Vaccinium macrocarpon Ait.) vines were grown in pots to establish the dose response levels for a sand-applied phytotoxin(s) from a crude extract of Pseudomonas syringae (strain 3366) culture. At 114 ppm [milligrams phytotoxin(s)/kilograms sand], the material was noninhibitory, whereas 1140 ppm reduced root and shoot growth significantly in all four species. In subsequent experiments, a 10-ppm dose controlled corn spurry (Spergula arvensis L.) and fireweed (Epilobium angustifolium L.) seedlings, while 103 ppm reduced root or shoot growth of cuttings of the perennial weeds birdsfoot trefoil (Lotus corniculatus L.) and silverleaf (Potentilla pacifica Howell). Root and shoot growth of partially rooted 'McFarlin' cranberry vines was reduced at 103 and 563 ppm, respectively. The phytotoxin(s) could potentially control germinating annual weeds in newly established 'Stevens' cranberry bogs.

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Novel and complex chromosomal arrangement of Rhizobium loti nodulation genes. Scott, D.B.; Young, C.A.; Collins-Emerson, J.M.; Terzaghi, E.A.; Rockman, E.S.; Lewis, P.E.; Pankhurst, C.E. Massey University, Palmerston North, New Zealand. Molecular plant-microbe interactions. 1996. vol. 9 (3), p.187-197.

A mutational and structural analysis of Rhizobium loti nodulation genes in strains NZP2037 and NZP2213 was carried out. Unlike the case with other Rhizobium strains examined to date, nodB was found on an operon separate from nodACIJ. Sequence analysis of the nodACIJ and nodB operon regions confirm that R. loti common nod genes have a gene organization different from that of other Rhizobium spp. At least 4 copies of nodD-like sequences were identified in R. loti. The complete nucleotide sequence of one of these, nodD3, was determined. A new host-specific nod gene, nolL, was identified adjacent to nodD3. NolL shares homology with NodX and other O-acetyl transferases. Mutational analysis of the nod regions of strains NZP2037 and NZP2213 showed that nodD3, nodI, nodJ, and nolL were all essential for R. loti strains to effectively nodulate the extended host Lotus pedunculatus, but were not necessary for effective nodulation of the less restrictive host, Lotus corniculatus. Both nodD3 and nolL were essential for R. loti strains to nodulate Leucaena leucocephala.

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Molecular cloning and characterisation of asparagine synthetase from Lotus japonicus: dynamics of asparagine synthesis in N-sufficient conditions. Waterhouse, R. N.; Smyth, A. J.; Massonneau, A.; Prosser, I. M.; Clarkson, D. T. IACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, Long Ashton, Bristol BS19 9AF, UK. Plant Molecular Biology vol. 30 (5): p.883-897. Publication Year: 1996.

Two cDNA clones, LJAS1 and LJAS2, encoding different asparagine synthetases (AS) (aspartate-ammonia ligase) have been identified and sequenced and their expression in Lotus japonicus characterized. Analysis of predicted amino acid sequences indicated a high level of identity with other plant AS sequences. No other AS genes were detected in the L. japonicus genome. LJAS1 gene expression was found to be root-enhanced and lower levels of transcript were also identified in photosynthetic tissues. In contrast, LJAS2 gene expression was root-specific. These patterns of AS gene expression are different from those seen in pea. AS gene expression was monitored throughout a 16 h light/8 h dark day, under nitrate-sufficient conditions. Neither transcript showed the dark-enhanced accumulation patterns previously reported for other plant AS genes. To evaluate AS activity, the molecular dynamics of asparagine synthesis were examined in vivo using 15N-ammonium labelling. A constant rate of asparagine synthesis in the roots was observed. Asparagine was the most predominant amino-component of the xylem sap and became labelled at a slightly slower rate than the asparagine in the roots, indicating that most root asparagine was located in a cytoplasmic 'transport' pool rather than in a vacuolar 'storage' pool. The steady-state mRNA levels and the 15N-labelling data suggest that light regulation of AS gene expression is not a factor controlling N-assimilation in L. japonicus roots during stable growth in N-sufficient conditions. Nucleotide sequence data have been submitted to the EMBL/GenBank/DDBJ databases under the accession numbers X89409 (LJAS1) and X89410 (LJAS2).

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Registration of GIFU B-129-S9 Lotus japonicus germplasm. Stougaard, J.; Beuselinck, P. R. Department of Molecular Biology, Aarhus University, Gustav Wiedsvej 10, DK-8000 Aarhus C, Denmark. Crop Science vol. 36 (2): p.476. Publication Year: 1996

GIFU B129-S9 represents an S9 inbred derived from the GIFU accession B129, the original source of which was collected near Gifu, Japan. The growth form of mature GIFU B129-S9 plants is intermediate, with multiple branches up to 30 cm long. Flowering is indeterminate. Plants are cross-fertile, making classical genetic studies possible. Fast-growing strains of Rhizobium loti and some slow-growing strains of Bradyrhizobium spp. will nodulate GIFU B129-S9, but R. loti strain NZP 2235 is recommended for fast and effective nodulation.

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Proposed ecological descriptors and their utility for plant germplasm collections. Steiner, J. J.; Greene, S. L. National Forage Seed Production Research Center, USDA-ARS, 3450 SW Campus Way, Corvallis, OR 97331, USA. Crop Science vol. 36 (2): p.439-451. Publication Year: 1996.

Plant germplasm collections are important repositories of genes that ensure potentially useful sources of genetic variation are preserved for future uses. Detailed information about genetic differences among individuals or groups of accessions can be helpful for management and utilization of germplasm collections. At present a description of the collection site is required when collecting accessions, but the kinds and amount of information recorded that describe the natural environmental features of the collection site may vary greatly depending on the collector. This paper discusses the need for providing standardized and detailed ecological descriptors for collected accessions that can be entered into the US National Plant Germplasm System (NPGS) Germplasm Resources Information Network (GRIN) database and provides a set of necessary tools to accomplish this. Proposed in this paper are (i) a flexible set of ecological descriptors that capture the most important ecological features of the collection site and its environment so that a standardized description can be given of the natural conditions where accessions are found, (ii) a revised accession data collection form that can be used to describe the natural features of the collection site, and (iii) a method to retroclassify poorly documented accessions that do not have detailed descriptions of the collection site. The widely adapted perennial forage legume birdsfoot trefoil (Lotus corniculatus) is used to demonstrate the principles presented in this paper.

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Rhizomatous Lotus corniculatus L.: II. Morphology and anatomy of rhizomes. Li, B.; Beuselinck, P. R. Department of Agronomy, University of Missouri, Columbia, MO, 65211, USA. Crop Science vol. 36 (2): p.407-411. Publication Year: 1996.

Birdsfoot trefoil (Lotus corniculatus) is a valuable forage legume. Rhizomatous L. corniculatus germplasm from Morocco was recently discovered (accessions G31272, G31273, G31276, G31298 and G31317). Increased persistence may result from the rhizomatous habit, because new plants vegetatively generated by rhizomes could replace diseased or dead plants. Rhizome morphology in L. corniculatus from Morocco is unique because taxonomic descriptions for L. corniculatus do not include the presence of rhizomes. The morphology and anatomy of subterranean lateral shoots (rhizomes) and aerial shoots (stems) of these L. corniculatus accessions were described and compared. Rhizomes of Moroccan L. corniculatus are initiated from axillary buds on basal portions of shoots. The rhizome consists of nodes and internodes, with buds, scale leaves, and adventitious roots occurring at each node. Anatomically, the rhizome of L. corniculatus has a large cortex, small pith, and six to seven primary vascular bundles. No endodermis exists in the cortex, and parenchyma cells contain starch. In contrast, the aerial shoot of L. corniculatus has a small cortex, large pith, and nine to 10 primary vascular bundles. An endodermoid layer is peripheral to the concentric ring of vascular bundles of the aerial shoot, and parenchyma cells in the cortex contain chloroplasts. Morphological and anatomical differences between L. corniculatus rhizomes and aerial shoots are distinguishable.

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A simple technique for direct transformation and regeneration of the diploid legume species Lotus japonicus. Oger, P.; Petit, A.; Dessaux, Y. Institut des Sciences Vegetales, CNRS, Batiment 23, Avenue de la Terrasse, F-91198, Gif-sur-Yvette, France. Plant Science (Limerick) vol. 116 (2): p.159-168. Publication Year: 1996.

The diploid legume species Lotus japonicus was transformed using a disarmed Agrobacterium strain. By inoculating the cotyledon attachment site, the regenerative capability of the plant allowed spontaneously regenerated transformed shoots. Some of these shoots were propagated as transgenic plants and later transferred to the glasshouse to produce seeds. Seeds were germinated and yielded non-chimaeric, transgenic stably modified Lotus plants.

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A system for tissue-specific copper-controllable gene expression in transgenic plants: nodule-specific antisense of aspartate aminotransferase-P2. Mett, V. L.; Podivinsky, E.; Tennant, A. M.; Lochhead, L. P.; Jones, W.T.; Reynolds, P. H. S. Plant Improvement Division, Horticulture and Food Research Institute of New Zealand, Private Bag 11030, Palmerston North, New Zealand. Transgenic Research vol. 5 (2): p.105-113. Publication Year: 1996.

A vector system, based on copper controllable gene expression, has been developed to give control over place as well as time of expression of an introduced gene. This system consists of two elements: (1) the yeast ace1 gene encoding a metallo-regulatory transcription factor, ACE1, under control of either an organ-specific or a constitutive promoter; and (2) a gene of interest under control of a chimaeric promoter consisting of the 46 bp TATA fragment of the CaMV 35S RNA promoter linked to four repeats of the ACE1 binding site. The functioning of the system in an organ-specific manner was tested in nodulated Lotus corniculatus plants which consisted of non-transformed shoots plus transformed hairy root tissue 'wild-type tops/transgenic roots'. After addition of copper ions to the plant nutrient solution, beta -glucuronidase (GUS) expression was visualized either specifically in nodules or in both roots and nodules when the ace1 gene was placed under control of the nod45 promoter or the CaMV 35S RNA promoter, respectively. The nodule-specific system was used to express antisense constructs of aspartate aminotransferase-P2 in transgenic Lotus corniculatus plants. When expression was induced by the addition of copper ions to the plant nutrient solution aspartate aminotransferase-P2 activity declined dramatically, and a decrease of up to 90% was observed in nodule asparagine concentration.

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Seasonality of mycorrhizae in coastal sand dunes of Baja California. Siguenza, C.; Espejel, I.; Allen, E. B. Department of Botany and Plant Sciences, University of California, Riverside, CA 92521-0124, USA. Mycorrhiza vol. 6 (2): p.151-157. Publication Year: 1996.

Populations of arbuscular mycorrhizal fungi were estimated from spores associated with 7 plant species in coastal dunes at El Socorro, near Ensenada, Baja California, Mexico, during 6 months in 1992. The seasonal patterns of percentage root colonization were also described in the same species during the wet season (Jan.-Mar.) and the dry season (Apr.-Jul.). Comparisons were made between the pioneer species (Abronia maritima) in the mobile dunes and 6 species (Abronia umbellata, Atriplex julacea, Camissonia californica, Haplopappus venetus, Helianthus niveus and Lotus spp.) in the fixed dunes. Mycorrhizal colonization in Abronia maritima was slight (<1%) and few spores (<1/g soil) were observed. All species in the fixed dune formed mycorrhizae with up to 80% colonization in early summer, and no more than 4 spores/g soil by late summer. The highest percentages of total colonization and abundance of spores did not coincide temporally for any of the 7 species, but the percentages were higher in summer than in spring. Arbuscules were more abundant when the soil was moist, and vesicles more abundant when it was dry.

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Selection for biological nitrogen fixation and nitrogen utilization in birdsfoot trefoil. Ehlke, N. J.; Vellekson, D. J.; Lyman, B. E. Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN 55108, USA. Crop Science vol. 36 (1): p.104-109. Publication Year: 1996.

Birdsfoot trefoil (Lotus corniculatus) is a perennial forage legume which has a lower rate of biological nitrogen fixation (BNF) than alfalfa (Medicago sativa). Studies were conducted to evaluate the gain from two cycles of recurrent selection in the glasshouse in three germplasms of birdsfoot trefoil (Norcen, AU-Dewey and MU81) using traits associated with improved BNF or N utilization. For improved BNF, selection was based on morphological traits correlated with the acetylene reduction assay (ARA) in a nil-N soil environment. For improved N utilization, selection was based on increased top dry weight production in a N-rich soil environment. The 12 selected populations and the three parental populations were evaluated in two growth chambers in a completely randomized design with four replicates. The chambers were watered three times per week with 100 ml of a modified Hoagland's solution containing either 0.0 (nil-N environment) or 10.7 (N-rich environment) mM N. At 8 and 14 weeks after seeding, the plants were individually scored for nodule mass and fibrous roots, top dry weight, ARA rate, and N yield. Correlated trait selection for improved BNF, as measured by the ARA, produced populations with larger plants, more fibrous roots, and greater nodule mass resulting in a 56% increase in ARA in the nil-N environment. In the N-rich environment, the populations selected for improved BNF showed a significant increase in nodule mass and a 79% increase in ARA rate. Selection for improved N utilization resulted in larger plants with more fibrous roots, larger nodule mass, higher N yield and a 77% increase in ARA in the N-rich environment. In the nil-N environment, the populations selected for improved N utilization had larger root systems, slighter better nodulation, and produced larger plants which resulted in a 58% increase in ARA rate. Correlated trait selection for traits which support BNF or N utilization could significantly improve the performance of birdsfoot trefoil in environments with fluctuating N availability.

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Estimation of the crude protein content in grass and legume populations of the spontaneous flora using a chemical colorimetric micromethod. Original Title: Estimarea continutului de proteina bruta din populatiile de graminee si leguminoase din flora spontana prin micrometode chimice colorimetrice. Drochioiu, G.; Avramiuc, M.; Risca, I. M. Banca de Gene, Suceava, Romania. Cercetari Agronomice in Moldova vol. 28 (1/2): p.45-50. Publication Year: 1995.

The crude protein contents of 18 populations of 9 grass spp., 18 populations of 7 Trifolium spp. and 13 populations of 7 other legume spp. were estimated by the coloration of free amino acids with a reagent containing ninhydrin. The intensity of the coloration was proportional to their concentration and indirectly to that of the protein content. This micromethod was rapid and precise. Crude protein contents ranged from 9.84% in the Zamostea population of Lolium perenne to 19.4% in the Poiana Stampei 2 population of Festuca rubra, from 13.32% in the Ponoare population of Trifolium pannonicum to 25.69% in the Danila population of T. repens and from 14.18% in the Ilisesti population of Lotus corniculatus to 32.08% in the Danila population of Vicia cracca.

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Purification and characterization of vegetative storage proteins from alfalfa (Medicago sativa L.) taproots. Cunningham, S. M.; Volenec, J. J. Department of Agronomy, Purdue University, West Lafayette, IN 47907-1150, USA. Journal of Plant Physiology vol. 147 (6): p.625-632. Publication Year: 1996.

Alfalfa (Medicago sativa) accumulates C and N reserves in taproots and utilizes these reserves for shoot growth in spring and for shoot regrowth after defoliation. Three proteins are very abundant in taproots and undergo a cyclic pattern of utilization during early shoot growth followed by reaccumulation during late shoot development. These putative vegetative storage proteins were purified and characterized from alfalfa taproots. The proteins were purified using organic-solvent and ionic-precipitation techniques, gel filtration, and affinity chromatography. Polyclonal antibodies were raised against the purified proteins, and electrophoresis and immunoblotting were utilized to determine protein distribution and relative abundance. These proteins are present in high concentrations in alfalfa taproots, but were not found in seeds, nodules, leaves, or stems of alfalfa. Taproots of all perennial Medicago species examined contained these proteins, whereas roots of annual Medicago species had very low to undetectable amounts of these proteins. Taproots of other forage legume species (Lotus, Melilotus, and Trifolium) did not contain proteins that cross-reacted with antibodies raised against the three alfalfa taproot proteins. The three proteins are glycosylated, and have epitopes in common. The amino acids asparagine and aspartate make up 15 mole percent of the three alfalfa taproot proteins. These proteins possess features consistent with their role being vegetative storage proteins.

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Immunogold localization of nodule-enhanced phosphoenolpyruvate carboxylase in alfalfa. Robinson, D. L.; Pathirana, S. M.; Gantt, J. S.; Vance, C. P. Department of Biology, Bellarmine College, Louisville, KY 40205-0167, USA. Plant, Cell and Environment vol. 19 (5): p.602-608. Publication Year: 1996.

Phosphoenolpyruvate carboxylase (PEPC) plays a vital role in providing carbon for the synthesis of malate and aspartate in Medicago sativa root nodules. To investigate the relationship between the cellular location of PEPC and root nodule metabolism, enzyme localization was evaluated by immunogold cytochemistry using M. sativa nodule PEPC antibodies. Gold labelling patterns in effective nodules indicated that PEPC is a cytosolic enzyme and was distributed relatively equally in both the infected and uninfected cells of the nodule symbiotic zone. Labelling was also detected in the pericycle cells of the nodule vascular system and within inner cortical cells. When Lotus corniculatus was transformed with a chimeric gene consisting of the 5'-upstream region of the PEPC gene fused to beta -glucuronidase (GUS), subsequent GUS staining in the nodules was consistent with immunogold localization patterns. It is suggested that because of the occurrence of PEPC in both infected and uninfected cells of the symbiotic zone and the reduced amounts observed in ineffective nodules, PEPC plays a direct role in supporting N2 fixation. PEPC localization in uninfected, interstitial cells of the symbiotic zone indicated that these cells may also have a role in nodule carbon metabolism. Moreover, it is concluded that the association of PEPC with the nodule vascular system implies a role for the enzyme in the transport of assimilates to and from the shoot.

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Rhizomatous Lotus corniculatus L. I. Taxonomic and cytological study. Beuselinck, P.R.; Li, B.; Steiner, J.J. USDA, ARS, Plant Genetic Research Unit, Columbia, MO. Crop science. v. 36 (1), p. 179-185. Publication Year: 1996.

Wild, putative Lotus corniculatus L. accessions from Morocco differ distinctly from domesticated L. corniculatus in that they produce rhizomes. Rhizomes may be useful in increasing stand persistence of domesticated L. corniculatus. The objective of this study was to identify to species the wild Lotus accessions from Morocco (G31272, C31273, G31276, G31298, and G31317) that present rhizomatous growth. Comparative morphological and cytological analyses and intercrossing with L. corniculatus were used in this study. Six morphological characters that distinguish L. corniculatus and L. uliginosus Schkuhr. (the only species in the genus Lotus recognized as producing rhizomes) were used to categorize taxonomically the Moroccan accessions. The Moroccan accessions were morphologically similar to L. corniculatus with the exception of the rhizome trait. The accessions were tetraploid with 2n = 4X = 24 somatic chromosomes. Karyotype analyses detected more similarities than differences between the Moroccan accessions and Norcen. Progeny were produced from intercrosses between L. corniculatus cvs. Norcen and AU Dewey and the Moroccan accessions. Hybridity of progeny was verified by morphological and random amplified polymorphic DNA (RAPD) analyses. A sample of reciprocal F1 progeny were tetraploid, fertile, and consistent in their production of rhizomes. A combination of morphological, chromosome, and karyotype analyses confirmed accessions G31272, G31273, G31276, G31298, and G31317 as L. corniculatus. This is the first description of L. corniculatus with rhizomes.

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Frost-seeding legumes into established switchgrass: establishment, density, persistence, and sward composition. Gettle, R.M.; George, J.R.; Blanchet, K.M.; Buxton, D.R.; Moore, K.J. USDA-ARS, Field Crops Res. Unit, Ames, IA. Agronomy Journal. v. 88 (1), p. 98-103. Publication Year: 1996.

Legumes incorporated into established switchgrass (Panicum virgatum L.) could provide symbiotic N, improve herbage quality, and extend the grazing season. Almost no information exists for legume renovation of switchgrass by frost-seeding. The objective of this study was to identify cool-season legumes that can be established into switchgrass by frostseeding, yet not provide excessive competition to the associated grass. Six legumes [biennial 'Polara' white-flowered sweetclover (Melilotus alba Medik.), biennial 'Madrid' yellow-flowered sweetclover (Melilotus officinalis Lam.),'Norcen' birdsfoot trefoil (Lotus corniculatus L.), 'Redland II red clover (Trifolium pratense L.), 'Apollo Supreme' alfalfa (Medicago sativa L.), and a 50:50 mixture of Redland II red clover and Norcen birdsfoot trefoil] were inoculated with rhizobia and frost-seeded into established 'Cave-in-Rock' switchgrass in mid-March of 1991 and 1992. These mixtures were compared with switchgrass fertilized at four N levels (0, 60, 120, and 240 kg ha-1). Successful legume establishment was observed, with an average of 24 and 25% of viable seed producing seedlings by June for 1991 and 1992 seedings, respectively. Mean legume density was 160 and 170 plants m-2 by June of the seeding year (YR 1) and 95 and 55 plants m-2 by June of the second year (YR 2) for 1991 and 1992 seedings, respectively. Legume renovation of switchgrass stands did not affect grass stem density by July YR 1. Mean forage stand composition (based on stem counts) was 87, 67, and 45% legume in June, July, and August YR 2, respectively, for 1991 seedings, and 54, 29, and 23% legume during the same time periods for 1992 seedings. Legumes did not seriously reduce switchgrass stem density, although red clover, birdsfoot trefoil, and their mixture were more competitive than others. Legumes can be successfully introduced into established Cave-in-Rock switchgrass by frost-seeding and they will persist for at least 2 yr with favorable weather.

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Selection of cold-tolerant plants for growth in soils contaminated with organics. Rogers, H. B.; Beyrouty, C. A.; Nichols, T. D.; Wolf, D. C.; Reynolds, C. M. University of Arkansas, Fayetteville, AR, USA. Journal of Soil Contamination vol. 5 (2): p.171-186 Publication Year: 1996.

A mixture of organic chemicals (MOC) containing equal molar amounts of benzoic acid, hexadecane, 2,2-dimethyl 4,n-propyl-benzene, phenanthrene, pyrene, and either cycloheptane or cis-decahydronaphthalene (cis-decalin) was applied to soil at rates of 0 to 8000 mg/kg. In a plant-screening experiment, growth responses of four legumes (lucerne (Medicago sativa), Trifolium pratense, Trifolium repens, and Lotus corniculatus) and five non-legume species (alpine bluegrass (Poa alpina), Deschampsia beringensis, Phalaris arundinacea, Elytrigia repens (Elymus repens) and Artemisia tilesii) were determined at 10 and 25 deg C. The MOC applied at 2000 mg/kg reduced the growth of several species without resulting in significant seedling death. At 10 deg C, the growth of alpine bluegrass in the 1000 and 2000 mg/kg treatments of soil increased by more than 185%. In a plant growth response experiment, alpine bluegrass and lucerne were grown in soil that had been contaminated at rates of 0 and 2000 mg/kg. at 14 weeks, the shoot and root dry weights of lucerne were 97% lower than in the contaminated soil, while the shoot dry weight, root dry weight, and root length of alpine bluegrass were 135, 235, and 268% higher, respectively. Except for pyrene, <23% of the compounds comprising the MOC remained in the soil after 4 weeks and <5% after 14 weeks. The disappearance of the MOC was not significantly influenced by the presence of lucerne or alpine bluegrass.

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Perennial legumes for the high rainfall zone of eastern Australia. 1. Evaluation in single rows and selection of Caucasian clover (Trifolium ambiguum M. Bieb.). Hill, M. J.; Mulcahy, C.; Rapp, G. G. CSIRO Division of Animal Production, Pastoral Research Laboratory, Armidale, NSW 2350, Australia. Australian Journal of Experimental Agriculture vol. 36 (2): p.151-163 Publication Year: 1996.

A range of perennial legume species, including Trifolium ambiguum, T. medium, Astragalus cicer, Coronilla varia, Lotus corniculatus, L. pedunculatus (L. uliginosus) and Lespedeza cuneata, were evaluated in glasshouse and field trials in 1989-92 at Armidale, New South Wales. Hexaploid germplasm of T. ambiguum was subjected to selection for increased seedling vigour, early biomass production and profuse flowering in glasshouse screening and spaced plant field nurseries. On an acid, solodic soil, T. ambiguum and T. medium grew modestly, Lotus spp. grew vigorously initially but died out within 3 years, A. cicer and C. varia failed to persist beyond 1 year, but isolated plants of L. cuneata thrived. On an acid, free-draining, red-basalt soil, all accessions grew vigorously until early 1991. During the succeeding drought, Lotus died out, A. cicer thinned out on the acid soil, but T. ambiguum, T. medium and C. varia remained persistent and productive. A large nursery of A. cicer accessions on the same soil limed to 5 t/ha before planting remained vigorous to the present. Superior hexaploid material of T. ambiguum selected from spaced plants originated chiefly from CPI43909 and the ARS-2678 germplasm collection from Utah, USA.

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The effect of plant growth regulators on growth, morphology and condensed tannin accumulation in transformed root cultures of Lotus corniculatus. Robbins, M. P.; Evans, T. E.; Morris, P. Cell Manipulation Group, Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, Dyfed SY23 3EB, UK. Plant Cell, Tissue and Organ Culture vol. 44 (3): p.219-227 Publication Year: 1996.

The effects were studied of four different classes of plant growth regulators on root morphology, patterns of growth and condensed tannin accumulation in transgenic root cultures of Lotus corniculatus (bird's-foot trefoil). Growth of transformed roots in 2,4-D resulted in decreased tannin levels relative to controls at concentrations of 10-6 M and above, while gibberellic acid (GA3) inhibited tannin accumulation at concentrations of 10-7 M and above. Benzyladenine (BA) had little effect at low concentrations (10-7 M and below) but resulted in an increase in tannin levels at 10-6 M. Abscisic acid had little effect on levels of condensed tannins at any of the concentrations used. Experiments involving growth regulator addition and medium transfer demonstrated that 2,4-D inhibition of tannin accumulation could be reversed by GA3 and BA, while GA3 down-regulation could only be reversed by the addition of 2,4-D. Although 2,4-D inhibited tannin accumulation, addition of 2,4-D to root cultures grown for 14 or 28 days in the absence of plant growth regulators stimulated both growth and tannin biosynthesis. Characteristic alterations in root morphologies accompanied growth regulator-mediated modulation of tannin biosynthesis. Growth in 2,4-D resulted in partially de-differentiated root cultures while growth in GA3 produced roots with an elongated phenotype. Restoration of tannin biosynthesis in 2,4-D-treated roots was accompanied by root re-differentiation and the production of new lateral roots.

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Strategy for catch crop development. I. Hypothetical ideotype and screening of species. Karlsson-Strese, E. M.; Umaerus, M.; Rydberg, I. Julita gard och museer, S-640 25 Julita, Sweden. Acta Agriculturae Scandinavica. Section B, Soil and Plant Science vol. 46 (2): p.106-111 Publication Year: 1996.

Hypothetical criteria were defined for a catch crop insown in spring cereals with the main purpose of reducing leaching of nitrogen. The criteria were applied in the screening of a large variable plant material from three groups: Poaceae, Fabaceae and species from other families. Variation in the characters of interest between and within species was recorded in spaced field plantings. Perennial species adapted to the Nordic climate possessed many of the characteristics expected to be desirable. Accessions, expected to have a potential as catch crops were identified in all three groups. The group "other families" is of great interest. The most interesting species for further experiments were Cichorium intybus, biennial species in Brassicaceae, Lotus corniculatus, Trifolium fragiferum and Lolium spp.

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Yield response of legume seedlings to simulated insect cotyledon damage and nutrient availability. Ferguson, C. M.; Shand, J. A.; Evans, A. A. AgResearch, Invermay Agricultural Centre, Private Bag 500034, Mosgiel, New Zealand. Proceedings of the Forty Eighth New Zealand Plant Protection Conference, Angus Inn, Hastings, New Zealand, August 8-10, 1995. Conference Title: Proceedings of the Forty Eighth New Zealand Plant Protection Conference, Angus Inn, Hastings, New Zealand, August 8-10, 1995. p.213-218 Publication Year: 1995.

Seedlings of 9 legume species (Trifolium repens, T. ambiguum, T. hybridum, T. pratense, Lotus corniculatus, L. pedunculatus (L. uliginosus), Coronilla varia, Lupinus alpinus and lucerne) in pots were subjected to simulated insect defoliation by removing 0, 0.5, 1, 1.5 or 2 cotyledons. Foliage and root yields were measured after 8 weeks. The reaction of T. repens, T. ambiguum, and lucerne to the same levels of defoliation was further investigated with 3 balanced nutrient levels (high, medium or low) but without nitrogen. Foliage and root production and the amount of nitrogen fixed were measured 8 weeks after defoliation. Growth of all legume species decreased as simulated damage increased but some evidence suggested that the response varied between species. No association between simulated damage and nitrogen fixation was demonstrated but nutrient availability was shown to be of greater importance to plant growth than damage to cotyledons.

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Influence of lime and calcium:magnesium ratio on alfalfa and birdsfoot trefoil yields. Reid, W. S. Department of Crop and Atmopsheric Sciences, 803 Bradfield Hall, College of Agriculture and Life Sciences, Cornell University, Ithaca, NY 14853, USA. Communications in Soil Science and Plant Analysis vol. 27 (5-8): p.1885-1900 Publication Year: 1996.

In a field experiment in 1974, the effects of lime and Ca:Mg ratio in the lime on Medicago sativa and Lotus corniculatus DM yields and botanical composition of hay were examined. 0, 6.7, 13.4, 20.2 or 33.6 t lime/ha was applied to an acid (pH 4.7) Mardin silt loam soil (coarse-loamy, mixed, mesic typic Fragiochrepts). Using 4 commercial available lime sources and 2 mixtures of these lime sources, 6 Ca:Mg ratios were established for each lime rate. Calcium and/or magnesium sulfate were applied to plots that received no lime to obtain Ca:Mg ratios similar to those on the limed plots. The initial Morgan's solution extractable Ca was 450 mg/kg and Mg was 25 mg/kg. A 5 x 5 Latin Square design with 5 lime rates and 5 replications was used with the 6 Ca:Mg ratios as lime subplots. To improve mixing, portions of each lime treatment were applied before and after ploughing in 1974 and after ploughing in Apr. of 1975. M. sativa and L. corniculatus were sown to half of each plot. The plots were cut for hay 2 or 3 times each year for 5 years. The M. sativa hay yield and legume component were significantly increased by the first two lime rates (6.7 and 13.4 t/ha); however, the L. corniculatus hay and legume component were significantly increased only by the first lime rate. Maximum yields for the legume component of both crops occurred at a soil pH of 6.5. The Ca:Mg ratios resulting from adding limes with different Ca and Mg contents had no significant effects on the total DM yields, percent legume in hay or M. sativa and L. corniculatus yields. It was concluded that it is difficult to justify the use of Ca and Mg base saturation ratios for lime recommendations, or for determining the Mg concentration needed with lime. Soil samples taken in autumn of 1976 had mean soil pH values of 4.6, 5.7, 6.1, 6.4 and 6.7 for the stated lime rates, respectively.

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Effects of nitrogen supply, variety and Rhizobium inoculation on growth and nitrogen fixation of birdsfoot trefoil. Chun, W. B.; Hoveland, C. S. College of Agriculture, Chonnam National University, Kwangju 500-757, Korea Republic. Korean Journal of Animal Science vol. 37 (2): p.193-202 Publication Year: 1995.

Shoot, root and nodule DW and N-fixing activity were higher in Lotus corniculatus cv. AU Dewey than cv. Fergus. N fertilizer application increased shoot and root DW but decreased N-fixing activity and nodule DW. N-fixing activity increased up to 21 weeks after sowing and decreased thereafter. Rhizobium inoculation only had an effect without N fertilizer application. N contents of shoots and roots were not affected by N fertilizer or Rhizobium inoculation.

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Methods of establishing Lotus corniculatus in native pastures of the region of Campos Gerais, Parana. Original Title: Metodos de implantacao do cornichao (Lotus corniculatus L.) em pastagens nativas de Regiao dos Carupos Gerais do Parana. Postiglioni, S. R. Instituto Agronomico do Parana, 84001-970 Ponta Grossa, PR, Brazil. Boletim Tecnico - Instituto Agronomico do Parana (No. 46): 17 pp. Publication Year: 1995.

In field trials in Ponta Grossa in 1988-91, L. tenuis cv. Sao Gabriel was sown into natural grassland by broadcasting after passage of a heavy or light harrow or sown with no previous cultivation. The pasture was given 3 t/ha dolomitic limestone and 300 kg 10:30:10 NPK/ha before sowing the legume. Although average DM yield of L. corniculatus was highest (1.51 t/ha/year) following direct sowing, differences were not significant and total annual forage DM yield ranged from 3.71 to 4.57 t/ha following light and heavy harrowing, respectively. Sowing method had no significant effect on forage CP content (10.4-11.4%) or in vitro DM digestibility (50.2-51.9%).

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Ballast plants and grain mill plants which appeared after soil was dug out. Original Title: Ballastplanter og molleplanter som spirte etter graving i jorden. 1994. Blyttia vol. 52 (4): p.149-153.

Anagallis arvensis, Coronopus didymus, Euphorbia peplus and Mercurialis annua are recorded from the seed bank of Norwegian habitats. Coronopus didymus, Erucastrum gallicum, Salvia verticillata, Sisymbrium loeselii, Lotus angustissimus, Silene noctiflora and Hyoscyamus niger are recorded from the seed bank around Norwegian grain mill areas.

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Plant regeneration and genetic transformation of Lotus angustissimus. Nenz, E.; Pupilli, F.; Paolocci, F.; Damiani, F.; Cenci, C. A.; Arcioni, S. 1996. Plant Cell, Tissue and Organ Culture vol. 45 (2): p.145-152.

Culture conditions were established for callus induction and growth from different explants of L. angustissimus. Calluses were obtained from hypocotyls, leaves, stems, cotyledons and roots cultured on media containing 2,4-D or alpha -NAA with kinetin, N6- DELTA 2-isopentenyladenine or benzyladenine (BA) in different combinations and concentrations. Only those calluses induced in the presence of alpha -NAA with BA or kinetin produced shoots. Calluses induced from hypocotyl explants were the most efficient in regeneration of shoots. Transformation with an Agrobacterium rhizogenes binary vector carrying the plasmid pBI 121.1 is reported. The percentage of cotransformation was estimated by testing GUS activity in hairy roots. The integration of Ri T-DNA and the NPTII gene in transformed plants was confirmed by molecular analyses and in vitro culture of transgenic tissues in the presence of kanamycin.

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Productivity of Lotus corniculatus L. (bird's-foot trefoil) in the UK when grown under low-input conditions as spaced plants, monoculture swards or mixed swards. Bullard, M. J.; Crawford, T. J. Grass and Forage Science vol. 50 (4): p.439-446: 1995.

The productivity of L. corniculatus was assessed under low-input conditions at 3 sites in the north of England. An initial study in 1989 and 1990 compared 14 accessions of L. corniculatus with Trifolium repens and T. pratense grown as spaced plants at 2 sites with contrasting soil conditions. One site represented good agricultural land, the other poor-quality, marginal land. Yields of all species and accessions were significantly lower on the marginal site. Seasonal yields (averaged over sites) of L. corniculatus cultivars averaged 52 g DM/plant and 170 g in 1989 and 1990 respectively. General trends over both seasons were that T. repens and T. pratense yields were high but some L. corniculatus cultivars (Empire, Franco, Leo) and naturalized UK 'aliens' were similar to at least one of the Trifolium spp. in each year. Yields from L. corniculatus ecotypes from the UK and Norway were very low in comparison with the cultivars. A larger-scale study of 18 L. corniculatus, 2 L. uliginosus, 3 T. repens and 2 T. pratense accessions was undertaken on an upland, calcareous site in 1990. Legumes were assessed in 2 m x 2 m swards either grown alone or in a mixture with grass. Establishment and productivity of L. uliginosus and T. pratense accessions were poor, with seasonal yields no greater that 50 g DM/m2. When grown alone, L. corniculatus yields were higher than those of both Trifolium spp., and averaged 1080 g/m2. In mixed swards, L. corniculatus constituted a higher proportion of the total DM yield (26% and 62% at first and second cuts, respectively) than the other legume species. It is concluded from these studies that L. corniculatus is suited to marginal or free-draining calcareous soils under low-input conditions, and consequently has more potential for use in the UK than has previously been reported.

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Effects of nitrogen supply on growth and nitrogen fixation of birdsfoot trefoil during primary growth. Chun, W. B.; Hoveland, C. S. Proceedings of the 7th AAAP Animal Science Congress, Bali, Indonesia, 11-16 July, 1994. Proceedings of the 7th AAAP Animal Science Congress, Bali, Indonesia, 11-16 July, 1994.

In a greenhouse pot experiment, birdsfoot trefoil (Lotus corniculatus) cv. AU Dewey and Fergus seeds were inoculated with Rhizobium and seedlings were given half-strength Hoagland solution with or without supplemental N. Root and shoot samples were assessed at 15-24 weeks after sowing. Shoot DW was higher in AU Dewey than Fergus at all assessments. Supplemental N increased shoot DW and decreased N fixation.

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Seasonal changes in nonstructural carbohydrates, protein, and macronutrients in roots of alfalfa, red clover, sweetclover, and birdsfoot trefoil. Li Rong; Volenec, J. J.; Joern, B. C.; Cunningham, S. M. Crop Science vol. 36 (3): p.617-623. 1996.

Root total non-structural carbohydrate (TNC) has been studied, but little is known about seasonal trends in proteins and other macronutrients in legume roots. Studies were conducted to determine how winter hardening and resumption of growth in spring influence (i) concentrations of sugars and starch, and activities of amylases; and (ii) protein concentration and composition, and abundance of N, P, and K. Alfalfa (lucerne) (Medicago sativa), red clover (Trifolium pratense), birdsfoot trefoil (Lotus corniculatus), and biennial sweetclover (Melilotus officinalis) were seeded in rows in the field in May and roots were sampled at approximately monthly intervals in autumn and biweekly in spring. All species accumulated high root TNC and starch concentrations in autumn. Starch and TNC concentrations declined throughout winter, but increased in roots of alfalfa and red clover in May. Starch accumulation and degradation patterns in spring were correlated with root endoamylase activity. Root sugar concentrations increased three-fold from October to November. Root N and soluble protein concentrations increased 50% between September and December, and declined between March and May. Specific proteins accumulated to high concentrations in alfalfa roots during autumn. Root P increased 50% between October and April and was associated with an increase in root phytate. Proteins, N, P and phytate accumulated during winter hardening in autumn in roots of these forage legume species that, in conjunction with TNC, may supply nutrients to regrowing shoots in spring.

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Establishing multi-cut swards. Yushchak, V. S. Kormoproizvodstvo (No. 3): p.29-33: 1994.

Experiments on renovation of degraded grassland on a 15 deg NW-facing slope at 560 m alt. in the Carpathians in 1985-92 are reported. A mixture of Trifolium pratense, Lotus corniculatus, Festuca pratensis, F. arundinacea, Arrhenatherum elatius, Lolium multiflorum and Phleum pratense was successfully established under a companion crop of peas + oats for green forage. In 1986-90, an early maturing mixture of T. pratense, Dactylis glomerata, A. elatius and L. multiflorum, an intermediate maturing mixture of T. pratense, F. arundinacea, F. pratensis and L. multiflorum and a late mixture of T. pratense, F. pratensis, P. pratense and L. multiflorum were sown at 10, 15 or 20 million seeds/ha. Average hay yields of the 3 mixtures were similar at a given sowing rate and increased from 7.30-7.80 t/ha at the lowest sowing rate to 9.92-10.58 t at the highest. Denser sowing tended to increase the proportion of legumes and decrease that of forbs. The effects of delay in cutting on yield and chemical composition of these mixtures in 1988-92 are reported. Combined broadcast and row sowing gave the best results. Mixtures including T. pratense and L. corniculatus were high yielding but the legumes did not persist well. In 1986-89, applying 60 kg K2O/ha increased the hay yield of a grass-legume mixture from 4.05 to 5.17 t/ha and further addition of 45-120 kg P2O5/ha increased yield to 6.22-9.04 t. In 1986-92, hay yield and percentage leaf increased with increasing N rates in the range 120-180 kg/ha. The effect of N varied considerably depending on weather conditions.

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Unusual branching in the seedlings of Lotus japonicus-gibberellins reveal the nitrogen-sensitive cell divisions within the pericycle on roots. Kawaguchi, M.; Imaizumi-Anraku, H.; Fukai, S.; Syono, K. Japan.Plant and Cell Physiology vol. 37 (4): p.461-470: 1996.

The effects of several plant growth regulators on the induction of nodule-like structures on roots of Lotus japonicus were investigated. Addition of gibberellin A3 (GA3) at concentrations between 10-7and 10-4 M resulted in the formation of nodule-like structures on roots when seedlings were plated on nitrogen-free Fahraeus agar medium. GA4 also induced such outgrowths but was less active than GA3. Application of an inhibitor of auxin transport, N-(1-naphthyl)-phthalamic acid (NPA) and of kinetin, which have been reported to induce pseudonodules in other legumes, had no effect on L. japonicus. Microscopic observations showed that GA3-induced nodule-like structures were caused by cell divisions within the pericycle on the roots. In addition, the outgrowths elicited by GA3 could be completely suppressed by the addition of 15 mM potassium nitrate or ammonium nitrate. These results show that the pericycle cells of the roots of L. japonicus are specifically sensitive to gibberellins and that potential for cell division might be modulated by nitrogen compounds. The effects of ancymidol and uniconazole, two synthetic plant-growth retardants, were also investigated. Both compounds at 3 x 10-5 M significantly increased the number of stunted lateral roots. The unusual branching could not be counteracted by the exogenous addition of GA3 but by 10-6 M brassinolide. The physiological role of brassinolide in the initiation of lateral roots is discussed.

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Allelopathic interactions between herbaceous species and trees grown in topsoil and spoil media. Larson, M. M.; Patel, S. H.; Vimmerstedt, J. P. Journal of Sustainable Forestry vol. 3 (1): p.39-52: 1995.

Dried foliage of several herbaceous species was mixed with either vermiculite or 2 mine soils (topsoil and spoil) in 1-litre containers planted with black locust (Robinia pseudoacacia), white ash (Fraxinus americana) or sweetgum (Liquidambar styraciflua) seedlings in a greenhouse. Germinated seed of northern red oak (Quercus rubra) was planted in containers previously sown with the herbaceous species. In vermiculite, black locust growth was inhibited by additions of 12 g of litter of Medicago sativa, Lotus corniculatus, Festuca arundinacea and Poa pratensis, but stimulated by Trifolium pratense litter. White ash growth was strongly inhibited by M. sativa and L. corniculatus litter added to vermiculite at 12 and 18 g. In topsoil, sweetgum was strongly inhibited by M. sativa litter. Both sweetgum and red oak grew poorly in spoil, but sown Lolium perenne, Dactylis glomerata and F. arundinacea grew well. Some aspects of red oak growth were inhibited in containers sown with M. sativa, L. perenne, D. glomerata, T. pratense or F. arundinacea. Isolation and identification of allelopathic compounds was not attempted, but these results suggest that there may be important allelopathic interactions between herbaceous species and trees planted on mined land.

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Nodule development in legumes--the early stages involvement of early nodulins, lectins, and other proteins. Hirsch, A.M.; Fang, Y.; Brill, L.M.; Wycoff, K.L.; Niner, B.M.; Brandt, J.P.; Van Rhijn, P. 1995. In: Nitrogen fixation fundamentals and applications. Proceedings of the 10th International Congress on Nitrogen Fixation, St. Petersburg, Russia, May 28-June 3, p. 299-304.

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Nod metabolites and other compounds excreted by Rhizobium spp. Martinez, E.; Laeremans, T.; Poupot, R.; Rogel, M.A.; Lopez, L.; Garcia, F.; Vanderleyden, J.; Prome, J.C.; Lara, F. In: Nitrogen fixation fundamentals and applications proceedings of the 10th International Congress on Nitrogen Fixation, St. Petersburg, Russia, May 28-June 3, 1995, p. 281-286.

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Influence of lime and calcium magnesium ratio on alfalfa and birdsfoot trefoil yields. Reid, W.S. Communications in soil science and plant analysis. 1996. v. 27 (5/8), p. 1885-1900.

A study was established in 1974 to examine the effects of lime and calcium-magnesium (CaMg) ratio in the limes on alfalfa (Medicago sativa) and birdsfoot trefoil (Lotus corniculatus L.) hay yields and botanical composition of the hay. Lime was applied at rates of 0, 6.7, 13.4, 20.2, and 33.6 Mg/ha to an acid (pH 4.7) Mardin silt loam soil (coarse-loamy, mixed, mesic typic Fragiochrepts). Using four commercially available lime sources and two mixtures of these lime sources, six CaMg ratios were established for each lime rate. Calcium and/or magnesium sulfate were applied to plots that received no lime to obtain CaMg ratios similar to those on the limed plots. The initial Morgan's solution extractable Ca was 450 mg/kg and Mg was 25 mg/kg. A 5 X 5 Latin Square design with five lime rates and five replications was used with the six CaMg ratios as lime subplots. To improve mixing, portions of each lime treatment were applied before and after plowing in 1974 and after plowing in April of 1975. Alfalfa and birdsfoot trefoil were planted to one-half of each plot. The hay was harvested two or three times each year for five years. The alfalfa hay yield and legume component were significantly increased by the first two increments of lime (6.7 and 13.4 Mg/ha); however, the birdsfoot trefoil hay and legume component were significantly increased only by the first increment of lime. Maximum yields for the legume component of both crops occurred at a soil pH of 6.5. The CaMg ratios resulting from adding limes with different Ca and Mg contents had no significant effects on the total dry matter hay yields, percent legume in hay or alfalfa and birdsfoot trefoil yields. It is difficult to justify the use of Ca and Mg base saturation ratios for lime recommendations, or for determining the Mg concentration needed within the lime. Soil samples taken in fall of 1976 had mean soil pH values of 4.6, 5.7, 6.1, 6.4, and 6.7 for the five lime rates.

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Rep-PCR mediated genomic fingerprinting of rhizobia and computer-assisted phylogenetic pattern analysis. Schneider, M.; Bruijn, F. J. de. World Journal of Microbiology & Biotechnology vol. 12 (2) p.163-174. 1996.

A rapid and reproducible method for genomic fingerprinting of rhizobia and other soil microbes that interact with plants is described. The method utilises oligonucleotide primers that correspond to conserved motifs in naturally occurring interspersed repetitive DNA elements in bacteria (rep-elements), and the polymerase chain reaction (rep-PCR). The rep-PCR amplifies inter-element genomic DNA fragments of characteristic lengths thereby generating a genomic fingerprint which resembles UPC bar code patterns, and as such enable bacteria to be identified at the sub-species and strain level, as well as for phylogenetic analyses. Therefore a highly characteristic and reproducible rep-PCR generated genomic fingerprints can be obtained not only from purified genomic DNA, but also directly from rhizobial cells obtained from liquid culture, plate colonies, and root nodule tissue. The effect of cell growth phase, serial subculturing and other parameters on the reproducibility of the rep-PCR fingerprinting protocol were assessed. Moreover, the results of mixing experiments designed to determine if individual genomic fingerprints can be recognized in mixtures of strains was attempted. A review is also provided on the use of computer-based fragment detection and phylogenetic analysis packages to analyse rep-PCR generated genomic fingerprints of a collection of Rhizobium loti and Bradyrhizobium strains nodulating different Lotus spp.

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Differential expression of the Sesbania rostrata leghemoglobin glb3 gene promoter in transgenic legume and non-legume plants. Szczyglowski, K.; Potter, T.; Stoltzfus, J.; Fujimoto, S. Y.; Bruijn, F. J. de. Plant Molecular Biology vol. 31 (4) p.931-935. 1996.

The involvement of the Sesbania rostrata glb3 gene promoter NICE (nodule-infected cell expression) element in root-enhanced expression of the 5'-Srglb3-uidA-3'nos chimaeric gene was investigated in transgenic Nicotiana tabacum plants. The full-length wild-type Srglb3 promoter directed root meristem-enhanced expression in transgenic tobacco plants. The expression pattern of nine selected Srglb3 promoter mutations in the NICE element was examined in transgenic tobacco plants and compared with the pattern observed in nodules of transgenic Lotus corniculatus plants. The results suggest that the highly conserved motifs in the NICE element play an important role in expression in roots of non-legume plants.

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Cytology of 2n pollen formation and pollen morphology in diploid Lotus tenuis (Fabaceae). Rim YongWoo; Beuselinck, P. R. American Journal of Botany vol. 83 (8) p.1057-1062. 1996.

Two genotypes of Lotus tenuis PI204882, a diploid (2n = 2x = 12), were identified as producing 2n pollen (maximum = 6%). Research was undertaken to determine the mechanism(s) of 2n pollen formation in the L. tenuis genotypes, and to morphologically describe n and 2n pollen using light and scanning electron microscopy. Meiotic studies revealed that 2n pollen resulted from tripolar spindles during anaphase II of microsporogenesis. The 2n pollen germinated well, although abnormal pollen tubes were observed. The genetic constitution of 2n pollen resulting from tripolar spindles is equivalent to first division restitution of meiosis. Fresh and air-dried pollen samples exhibited differences in size and shape under light and scanning electron microscopy. The size of diploid (2n) pollen was larger than that of haploid (1n) pollen. Normal haploid (1n) pollen was globose-prolate in shape, while diploid (2n) pollen was tetrahedral.

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Evaluation of Astragalus L. and Lotus L. on two acid soil sites on the southern tablelands of New South Wales. Kelman, W. M. Australian Plant Introduction Review vol. 26 p.17-38. 1996.

A more diverse range of perennial pasture legumes can be used to fill niches not well exploited by commonly used species in southern Australia. Some 162 accessions of Astragalus and 118 accessions of Lotus were evaluated for seasonal productivity and persistence on a limed (pH 5.2 in CaCl2) and an unlimed (pH 4.3 in CaCl2) site on Ginninderra Experiment Station, ACT. A further 59 accessions of L. corniculatus were evaluated on an acid site (pH 4.07 in CaCl2) at Boundary Creek, NSW. The Astragalus species were persistent but unproductive on the unlimed site. On the limed site some accessions of A. cicer and A. glycyphyllos were as productive as white clover (Trifolium repens) and strawberry clover (T. fragiferum) cultivars in the summer. However, lack of cold season growth activity limits the direct use of these accessions in these environments. L. pedunculatus (L. uliginosus) was more tolerant of the unlimed soil site than the other Lotus species and white clover but autumn and spring growth scores were lower than those of the clover species. Improved seed production and lowered condensed tannin levels were identified as breeding objectives in L. pedunculatus. Two L. tenuis accessions had better autumn growth scores than the other Lotus species and a role in saline soils is suggested for this species. The annual L. suaveolens accessions produced vigorous seedling populations on both sites and it is speculated that interspecific hybridization with the perennial species might confer better autumn seedling establishment on the progeny. At Boundary Creek some prostrate forms of L. corniculatus from Spain had high herbage yields but unusually high condensed tannin concentrations. The prostrate forms need further evaluation under grazing by sheep.

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Biochemical study of accessions of birdsfoot trefoil (Lotus corniculatus L.) from the breeding point of view. Kyuchukova, A. Rasteniev"dni Nauki vol. 32 (6) p.135-137. 1995.

Analysis of fodder from the first and second cuts of 15 local and introduced varieties in 1991 showed that crude protein content ranged from 19.43% to 21.53% in the first cut and from 18.89% to 22.15% in the second. The concentration of condensed tannins was 1.32-1.64%. This did not have an adverse effect on dry matter digestibility. Some varieties appeared promising for use in breeding for fodder quality.

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Study of Osmia coerulescens L. (Hymenoptera, Megachilidae) - a pollinator for lucerne. II. Pollinating and building activity. Dochkova, B. Rasteniev"dni Nauki vol. 32 (6) p.64-66. 1995.

Studies during 1988-91 in the Pleven area of Bulgaria indicated that only the females of this solitary bee species engage in nest building and pollinating activity. They opened and pollinated 87.5% of visited Medicago sativa flowers and 98.4 to 100% of visited flowers of Lotus corniculatus and Onobrychis sativa (O. viciifolia). Osmia coerulescens (O. caerulescens) settled readily into artificial nests made of wild cane or of wood and appeared suitable for limited open pollination of lucerne breeding material.

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Application of high molecular weight DNA cloning in legume nodulation gene analysis. Gresshoff, P. M.; Funke, R.; Caetano-Anolles, G.; Prabhu, R.; Pillai, S. ; Kolchinsky, A.; Landau-Ellis, D. Induced mutations and molecular techniques for crop improvement. Proceedings: Induced mutations and molecular techniques for crop improvement. Proceedings, Vienna, Austria, 19-23 June 1995.p.275-285. 1995.

High molecular weight (HMW) DNA was isolated from Glycine max and the model legume Lotus japonicus for legume genome analysis. The primary objectives were the gene regions that control nodulation, early plant-microbe interaction and cell division responses. HMW DNA was separated by pulse field gel electrophoresis and analysed with closely linked RFLP markers co-hybridized with clones, permitting estimation of the physical distances related to recombination frequency. In the distal region of molecular linkage group H containing one of the genes controlling nodule number autoregulation and symbiotic nitrate tolerance (i.e. the nts gene), 1 cM was equivalent to less than 500 kb. Partially digested EcoRI soyabean and L. japonicus HMW DNAs were cloned into pYAC4. Stable yeast artificial chromosomes (YACs) carrying up to 960 kb DNA were generated. The average insert size was 200 kb. Hybridization with total genomic soyabean DNA revealed YACs with different amounts of repeated DNA sequences. Mapping of the end clones demonstrated whether the YACs were chimaeric. YACs of different complexity were used for chromosome identification using degenerate primer PCR and fluorescent in situ hybridization. This approach is a fast alternative to testing for YAC chimaerism. Single arbitrary and structured mini-hairpin primers were used to amplify and DNA fingerprint the YACs, providing a means of identifying the additional markers needed for contig construction. HMW DNA was cloned into the F plasmid bacterial artificial chromosome (BAC) vector. The YACs and BACs were also constructed with DNA from L. japonicus.

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Soil loss control and soil properties improvement based on cropping systems with cover plants and hedgerows in Reunion Island. Perret, S.; Michellon, R.; Tassin, J. Proceedings of the third congress of the European Society for Agronomy, Padova University, Abano-Padova, Italy, 18-22 September 1994. p.736-737. 1994.

Water runoff and erosion were studied on severely and moderately degraded bare andisol soils under 25 years of Pelargonium monoculture, a 6-year rotation including Pelargonium and food crops, and Pelargonium with cover crops of Pennisetum clandestinum or Lotus uliginosus. The use of a cover crop significantly decreased soil erosion. Soil permeability increased under cover crops, especially where Calliandra calothyrsus was used as a hedgerow on plot boundaries. Soil fauna, organic matter turnover and mineralization all increased below the cover crops. Crop rotations alone could not restore soil properties.

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Herbage production from swards containing a range of grass, forb and clover species and under extensive management. Fisher, G. E. J.; Baker, L. J.; Tiley, G. E. D. Grass and Forage Science vol. 51 (1) p.58-72. 1996.

Three small plot experiments were conducted to investigate the effects of species of grass and forbs, defoliation regime, inclusion of white clover (Trifolium repens) and forb blend on the herbage DM yield, botanical composition and mineral content of swards managed with zero fertilizer inputs. The results of all three experiments were characterized by decline in herbage production and large variations in treatment effects over the harvest period. When sown singly with a standard grass mix the species that competed well with grasses and produced annual forb herbage yields greater than 2.0 t DM ha-1 were black knapweed (Centaurea nigra), oxeye daisy (Leucanthemum vulgare), ribwort plantain (Plantago lanceolata), burnet (Sanguisorba officinalis), birdsfoot trefoil (Lotus corniculatus), chicory (Cichorium intybus), kidney vetch (Anthyllis vulneraria), red clover (T. pratense) and white clover. When sown singly with a standard forb mix, grass species significantly affected the annual yield of total, grass and forb herbage. The species that most suppressed the yield of forbs were common bent (Agrostis tenuis (A. capillaris)), Yorkshire fog (Holcus lanatus) and perennial ryegrass (Lolium perenne). Those that allowed for the highest yield of forbs were rough meadow grass (Poa trivialis), sweet vernal grass (Anthoxanthum odoratum) and crested dogstail (Cynosurus cristatus). Averaged over the three harvest years, defoliation regime did not significantly affect herbage production, but the inclusion of white clover in mixtures increased the yield of grasses. The use of rosette-type forb blends increased forb yield, compared with erect-type blends. The effects of treatments on herbage N and mineral contents and yields were inconsistent. However, there was some evidence to support the view that the presence of forb species in swards can result in greater contents of minerals in herbage, compared with grass-only swards.

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Contributions to the improvement of degraded permanent pasture lands from the Moldavian forest steppe, by fertilizer applications, oversowing and radical repairs. Original Title Contributii la imbunatatirea pajistilor permanente degradate din silvostepa Moldovei prin fertilizare, suprainsamantare si refacere radicala.Iacob, T.; Dumitrescu, N.; Vintu, V.; Samuil, C. Cercetari Agronomice in Moldova vol. 28 (3/4) p.127-132. 1995.

In 1988-93, 0-100 kg N + 0-75 kg P2O5 + 0-100 kg K2O + 0-40 t FYM/ha was applied to a degraded Festuca valesiaca pasture. DM yields ranged from 2.9 t/ha with no fertilizers to 5.2 t with 100 kg N + 50 kg P2O5 + 50 kg K2O. In 1986-90 on a permanent F. valesiaca pasture with 10-42 deg slope, 0-150 kg N/ha was applied. DM yields increased with decrease in slope and increase in N rate and ranged from 2.1 t/ha in the unfertilized control on a 40-42 deg slope to 6.1 t with 150 kg N on a 10-15 deg slope. In 1988-93 a pasture was prepared using a rotary sower, chisel plough or disc harrow and sown with Bromus inermis + Onobrychis viciifolia alone or with Dactylis glomerata and Lotus corniculatus and 20 t FYM alone or with 50 kg N + 25 kg P2O5, or 40 t FYM alone or with 100 kg N + 50 kg P2O5/ha were applied. DM yield increases of 72-190% were due to fertilizer applications, 65-131% were due to oversowing + fertilizers and 116-226% were due to resowing + fertilizers and ranged from 1.9 t/ha in the untreated control to 6.2 t with the 2 species mixture + discing + the higher NP rate.

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Frost-seeding legumes into established switchgrass forage yield and botanical composition of the stratified canopy. Gettle, R. M.; George, J. R.; Blanchet, K. M.; Buxton, D. R.; Moore, K. J. Agronomy Journal vol. 88 (4) p.555-560. 1996.

A field study was conducted on a Webster-Nicollet soil (fine-loamy, mixed, mesic, Typic Haplaquoll-Aquic Hapludoll) at Ames, Iowa, to compare herbage DM yield and botanical composition of legume-renovated switchgrass (Panicum virgatum) with that of N-fertilized switchgrass. Five inoculated forage legumes (Melilotus alba, M. officinalis, Lotus corniculatus, Medicago sativa or Trifolium pratense) and a legume mixture (T. pratense and L. corniculatus) were frost-seeded into an excellent stand of established switchgrass cv. Cave-in-Rock in mid-March of 1991 and 1992. Grass-legume DM yield was compared with N application (0, 60, 120 or 240 kg ha-1) in the sowing year (YR 1) and the second year (YR 2). Harvested samples were separated into grass and legume components, and further divided into upper and basal canopy. Nitrogen application provided the greatest upper and basal canopy yields in both June and July of YR 1. By June YR 2, legume renovation increased upper-canopy yields more than 240 kg N. Upper-canopy YR 2 yields of grass-legume mixtures sown in 1991 remained greater than for 240 kg N in July, and were similar to 120 kg N in August. During a wetter and cooler than normal summer, YR 2 yields of upper-canopy grass-legume mixtures sown in 1992 were similar to 60 and 120 kg N by July, and similar to both unfertilized and N-fertilized switchgrass by August. It is concluded that yields of legume-renovated switchgrass were generally greater than for mid to high levels of N application during YR 2. Because of relatively low herbage production during YR 1, it is suggested that producers should consider frost-seeding legumes into only part of existing switchgrass pastures, while applying N to the remaining non-renovated pasture to maintain high forage supply and pasture productivity.

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Characterization of rhizobia nodulating Lotus subbiflorus from Uruguayan soils. Irisarri, P.; Milnitsky, F.; Monza, J.; Bedmar, E. J. Plant and Soil vol. 180 (1) p.39-47 1996.

The generation times, acid production, carbon utilization, immunological properties, plasmid content, protein profile and symbiotic properties of 15 isolates of rhizobia nodulating Lotus subbiflorus (isolated from 8 representative soils in Uruguay) were studied. Rhizobium loti strain NZP2037 and Bradyrhizobium sp. (Lotus) strain NZP2309 were used as reference strains for fast- and slow-growing rhizobia, respectively. Isolates Ls11, Ls22, Ls31, Ls42, Ls71, Ls81, Ls82 and Ls552 were obtained from the Florida region, while Ls3, Ls4, Ls5, Ls7, LsS3, Ls1B3 and Ls1B4 were from Durazno. The seeds of L. subbiflorus cv. Rincon, L. pedunculatus (L. uliginosus), L. tenuis and L. corniculatus were surface sterilised, inoculated with rhizobium isolates and pot-grown under greenhouse conditions. Based on specific growth rates, carbon source utilization and acid production, 13 out of the 15 isolates could be assigned to the slow-growing group of rhizobia (bradyrhizobia). Using antisera against the whole cells of 3 isolates, the 15 isolates were separated into 3 groups. Only the slow-growing isolate Ls4 and the fast-growing Ls5 and Ls552 lacked cross-reactivity with any of the sera tested. Electrophoretic mobilities of whole cell protein from 7 out of the 8 isolates included in the serogroup represented by strain Ls31 were identical. Similarly, isolates Ls1B3 and Ls 1B4 (both in serogroup Ls1B3), exhibited the same pattern of cell proteins. In contrast, isolates Ls3 and Ls7 (belonging to serogroup Ls7), differed in protein profile. Plant growth experiments revealed that all of the isolates effectively nodulated L. subbiflorus and L. pedunculatus, but were unable to form effective nodules on L. tenuis and L. corniculatus. All isolates exhibited similar effectiveness in symbiosis with L. subbiflorus, except isolate Ls7, which resulted in significantly higher plant dry weight.

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Seasonal changes in nonstructural carbohydrates, protein, and macronutrients in roots of alfalfa, red clover, sweetclover, and birdsfoot trefoil. Li, R.; Volenec, J.J.; Joern, B.C.; Cunningham, S.M. Crop Science. v. 36 (3), p. 617-623. 1996.

Root total nonstructural carbohydrate (TNC) has been studied, but little is known about seasonal trends in proteins and other macronutrients in legume roots. Our objectives were to determine how winter hardening and resumption of growth in spring influence (i)concentrations of sugars and starch, and activities of amylases; and (ii)protein concentration and composition, and abundance of N, P, and K. Alfalfa (Medicago sativa L.), red clover (Trifolium pratense L.),birdsfoot trefoil (Lotus corniculatus L.), and biennial sweetclover (Melilotus officinalis L.) were seeded in rows in the field in May and roots were sampled at approximately monthly intervals in autumn and biweekly in spring. All species accumulated high root TNC and starch concentrations in autumn. Starch and TNC concentrations declined throughout winter, but increased in roots of alfalfa and red clover in May. Starch accumulation and degradation patterns in spring were correlated with root endoamylase activity. Root sugar concentrations increased threefold from October to November. Root N and soluble protein concentrations increased 50% between September and December, and declined between March and May. Specific proteins accumulated to high concentrations in alfalfa roots during autumn. Root P increased 50%between October and April and was associated with an increase in root phytate. Proteins, N, P, and phytate accumulated during winter hardening in autumn in roots of these forage legume species that, in conjunction with TNC, may supply nutrients to regrowing shoots in spring.

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Effect of condensed tannins in Lotus corniculatus upon lactation performance in ewes. Wang, Y.,Douglas, G.B.; Waghorn, G.C.; Barry, T.N.; Foote, A.G. Journal of agricultural science. v. 126 (pt.3), p.353-362. 1996.

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Recent data on bird's foot trefoil growth with different seed dosages, herbicide treatments and cutting phenophase. Nagy, L. Irrigated Farming. p.169-170. 1995

Following on from earlier experiments to investigate forage, seed yield and weeding facilities, this short paper outlines tests carried out on yields, number of plants per meter, weeding and also the germination capacity of bird's foot trefoil (Lotus corniculatus). These have shown that higher seeding rates resulted in higher seed yields, higher plant density, lower weed numbers and similar germination compared to lower seeding rates. Crop growth was affected by different herbicide treatments, e.g. the effect of imazethapyr on seed yield and weeds was beneficial, but tests also showed that cutting just before flowering had much better effect on seed yield and germination than cutting after flowering.

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Experiences of usage of imidazoline derivatives in bird's foot trefoil (Lotus corniculatus ssp. tenuifolius) stands. Nagy, L. Irrigation Res. Inst., Szarvas, Hungary. Irrigated Farming. p.163-168. 1995.

Post-em. Pivot 100 LC (with imazethapyr as the a.i.) was used at 0.8 litres/ha after establishment of bird's foot trefoil. This treatment achieved an approximate 30% decrease of gross weed weight by harvesting time. Monocotyledonous weeds decreased more significantly than dicotyledonous ones. Amounts of Chenopodium album, Sonchus oleraceus, Hibiscus trionum and Cirsium arvense increased slightly in plots treated with Pivot 100 LC. The fodder yield of bird's foot trefoil was increased by 75-135% as a result of the treatment. Pivot 100 LC and Scepter 150 LC (imazaquin (ammonium salt)) were more selective when applied at the end of winter than at 5-7 cm plant height. It was concluded that the selectivity of the two herbicides with respect to pioneer bird's foot trefoil is favourable.

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Improving the acid-soil tolerance of Medicago sativa by asymmetric somatic hybridisation. Stoutjesdijk, P.A.; Larkin, P.J.; Sale, P.W.G. Plant-soil interactions at low pH: principles and management. Proceedings of the Third International Symposium, Brisbane, Queensland, Australia, 12-16 September 1993. p.455-457. 1995 Developments in Plant and Soil Sciences Vol. 64 Editors: Date, R.A.; Grundon, N.J.; Rayment, G.E.; Probert, M.E. Kluwer Academic Publishers, Dordrecht, Netherlands.

A protoplast electrofusion method for producing asymmetric somatic hybrid plants which are essentially Medicago sativa with small additions of donor Lotus pedunculatus (L. uliginosus) chromatin is described. Differential aluminium tolerance by M. sativa and L. pedunculatus was observed in vitro in an aluminium containing medium. Hybrid colonies were screened in the same medium for differences in growth rate. Approximately 900 of 4600 somatic hybrid colonies placed on this medium showed growth rates and morphology indicative of transfer of aluminium tolerance from Lotus to M. sativa. Regenerated plants from the selected colonies are being tested for elevated tolerance of aluminium.

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Chemical content of Al-tolerant and Al-susceptible breeding lines of white clover. Caradus, J.R.; Mackay, A.D. Plant-soil interactions at low pH: principles and management. Proceedings of the Third International Symposium, Brisbane, Queensland, Australia, 12-16 September 1993. p.439-445. 1995. Developments in Plant and Soil Sciences Vol. 64. (Eds.) Date, R.A.; Grundon, N.J.; Rayment, G.E.; Probert, M.E. Kluwer Academic Publishers Dordrecht, Netherlands.

Experimental breeding lines were developed from crosses between genotypes selected within white clover (Trifolium repens) cv. Grasslands Huia for either tolerance or susceptibility to aluminium (Al). Using a diallel design, 3 tolerant (T) and 3 susceptible (S) genotypes were crossed and reciprocals bulked. The chemical content of these breeding lines was compared with that of Lotus pedunculatus (L. uliginosus) cv. Maku grown in soil with or without 400 micro g Al/g soil. The Al-tolerant white clover genotypes had lower concentrations of Al and Fe in their shoots but higher concentrations of Cu, K and P than those of Al-susceptible plants. There was a significant trend of increasing percentage change in shoot concentration of Al, Ca, Fe, S and Sr, and decreasing change of Cu, K and P from T x T through T x S to S x S progenies due to added Al. Added Al decreased monovalent cation (K) uptake relative to divalent cation (Ca and Mg) uptake, but this effect was much less for Al-tolerant selections than Al-susceptible selections. It is proposed that the exclusion of Al from roots is a plausible reason for the continued K uptake by Al-tolerant compared with Al-susceptible white clover.

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The effect of aluminium on the growth of a range of temperate legume species and cultivars: a summary of results. Wheeler, D.M.; Dodd, M.B. Plant-soil interactions at low pH: principles and management. Proceedings of the Third International Symposium, Brisbane, Queensland, Australia, 12-16 September 1993. p.439-445. 1995. Developments in Plant and Soil Sciences Vol. 64. (Eds.) Date, R.A.; Grundon, N.J.; Rayment, G.E.; Probert, M.E. Kluwer Academic Publishers Dordrecht, Netherlands.

The aluminium (Al) tolerance of 35 temperate legumes (136 accessions) was determined in 74 experiments over a 2-year period in a low ionic strength (2730 micro M) solution culture. For each accession, the relationship between solution Al3+ activity ( micro M) and relative root yields was determined using polynomial exponential splines, and the Al3+ activity associated with a 50% reduction in root yield (AlRY50) was calculated. All species tested in the genera Adesmia, Medicago and Melilotus tended to be very sensitive to aluminium (AlRY50 <1). Dorycnium pentaphyllum was sensitive, whereas the AlRY50 of the D. hirsutum lines ranged from 1.4 to 2.4 (mean 1.9). In Lotus and Trifolium, the order of tolerance for each species (based on the AlRY50 mean over lines) was L. tenuis, T. resupinatum (AlRY50 <0.5) < T. balansae, T. fragiferum, T. hybridum and T. pratense (AlRY50 0.5-1) < L. angustissimus, L. corniculatus, L. subbiflorus, T. ambiguum, T. dubium, T. glomeratum (AlRY50 1-1.5) < T. amabile, T. repens, T. semipilosum, T. striatum, T. tumens (AlRY50 1.5-2) < T. medium, T. polymorphum (AlRY50 2-3) < T. subterraneum (AlRY50 3-4) < L. pedunculatus (L. uliginosus) (AlRY50 >4). Significant differences in Al tolerance between the lines were noted for (with the range in AlRY50 in parenthesis) T. amabile (0.1-2.6), T. ambiguum (0.8-1.6), T. medium (1.5-4.1), T. repens (0.6-3.1), T. polymorphum (1.5-2.6), T. semipilosum (1.2-2.0) and T. subterraneum (1.8-5.9).

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Genetic transformation of Lotus corniculatus with Agrobacterium tumefaciens and the analysis of the inheritance of transgenes in the T1 generation. Webb, K.J.; Gibbs, M.J.; Mizen, S.; Skot, L.; Gatehouse, J.A. Transgenic Research vol. 5 (5): p.303-312. 1996.

The herbage legume, Lotus corniculatus (bird's-foot trefoil), was transformed using the disarmed Agrobacterium tumefaciens strain LBA4404 (pAL4404) carrying a binary construct, pJit73. This plasmid carries two antibiotic resistance genes, aphIV and nptII encoding resistance to hygromycin and kanamycin respectively, and the easily detectable reporter gene, uidA encoding the enzyme beta -glucuronidase (GUS). Transgenic plants were regenerated from two separate co-cultivations of leaves with A. tumefaciens either with or without an acetosyringone pretreatment. A total of 110 putative transformants were regenerated, 52 (47%) of which grew on selection media containing hygromycin. Twenty-five plants were analysed further for morphological variation and presence of transgenes and were used to study the inheritance of expression of the transgenes in the T1 generation. Expression patterns of the transgenes in the T1 progeny generated from these 25 plants differed. In the majority of plant lines aphIV and uidA transgenes segregated together, but the apparent number of copies of the transgenes varied. No expression of either transgene was detected in the progeny from three plants, while the progeny from six other plants were resistant to hygromycin but had no GUS expression. Progeny of all of the remaining 16 plants had GUS activity. For three plants, inheritance data were consistent with more than one dose of uidA and aphIV; another two plants yielded data expected for exactly one dose of both transgenes. In the progeny of the remaining 11 plants, the percentage of seedlings expressing both uidA and aphIV was lower than expected.

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Elicitor mediated phytoalexin biosynthesis in transgenic root cultures of Lotus corniculatus. Morris, P.; Thomas, B.; Robbins, M.P. International symposium on natural phenols in plant resistance, Volume I, 13-17 Sep., 1993, Weihenstephan, Germany. Acta Horticulturae (No. 381): p.210-213.: 1994.

Agrobacterium rhizogenes-transformed root cultures of L. corniculatus responded to both biotic and abiotic elicitors resulting in the de novo synthesis of the typical Lotus isoflavan phytoalexins vestitol and sativan, which accumulated in both tissue and culture medium. Down regulation of constitutive tannin biosynthesis by auxin resulted in accumulation of chalcones and superinduction of phytoalexins on elicitation, whereas down regulation of tannins with an antisense DFR gene did not result in chalcone accumulation or in modified phytoalexin accumulation. The elicitation of Lotus root cultures with GSH also caused changes in root tip morphology due to differential injury to root meristem cells.

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An 'antisense' strategy for the genetic manipulation of condensed tannin and isoflavonoid phytoalexin accumulation in transgenic Lotus corniculatus L. Colliver, S. P.; Robbins, M. P.; Morris, P. International symposium on natural phenols in plant resistance, Volume I, 13-17 Sep., 1993, Weihenstephan, Germany. Acta Horticulturae (No. 381): p.148-151. 1994.

An antisense chalcone synthase gene construct made using the cDNA for chalcone synthase from Phaseolus vulgaris was introduced into the genome of 3 clonal genotypes of L. corniculatus via Agrobacterium rhizogenes. After initial screening, 30 antisense (AS) and 15 control (C) transformation events were analysed and regenerative ability was found to be reduced in AS root cultures. AS compared with C root cultures appeared to have a higher growth rate which was related to the ability to accumulate tannin.

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Effect of nitrogen fertilizer application on birdsfoot trefoil grown in the foothill region of central North Bulgaria. Aleksandrova, B. Rasteniev"dni Nauki vol. 32 (5): p.193-195. 1995.

In a field trial on light grey pseudopodzolic soil at 385 m alt. in 1989-92, Lotus corniculatus was given no fertilizer or 80, 160 or 240 kg N/ha alone or with 80 kg P2O5 + 80 kg K2O, or these P and K rates without N. Average annual DM yields were 7.78 t/ha with no fertilizer, 8.65 t with PK alone, 8.77-9.53 t with increasing rates of N without PK and 9.04-10.35 t with PK and increasing N rates. The percentage of L. corniculatus in the sward varied considerably between cuts and years; by 1992 it was only about 25% at the highest N rate, compared with about 65% with no fertilizer, and remained over 40% with PK + 80 kg N.

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Yield and forage quality of birdsfoot trefoil grown alone and in mixtures with cocksfoot. Aleksandrova, B. Rasteniev"dni Nauki vol. 32 (5): p.182-184. 1995.

In a trial on light grey pseudopodzolic soil in 1988-90, Lotus corniculatus and Dactylis glomerata were grown separately or in 75/25, 50/50 or 25/75% mixtures and cut for hay at the beginning of flowering of L. corniculatus and heading of D. glomerata. In the 1st year the 50/50% mixture gave the highest DM yield (4.74 t/ha); the other variants yielded up to 7.5% less. The stands nominally containing 100 or 75% L. corniculatus had >50% by weight of forbs in the 1st year, but this later decreased somewhat. In the 2nd year L. corniculatus sown alone gave the lowest yield (14.9 t/ha) and its 75/25% mixture with D. glomerata gave the highest yield (17.83 t), followed by the 25/75% mixture which yielded 17.48 t. In 1990, DM yields ranged from 9.18 t in D. glomerata sown alone (then about 25% forbs) to 11.60 t in L. corniculatus sown alone (then about 45% forbs). As the percentage of D. glomerata increased, CP content and CP yield and Ca content decreased and CF and ash contents increased.

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Study of the influence of foliar nutrition with boron on seed productivity of birdsfoot trefoil. Kyuchukova, A.; Radeva, V. Rasteniev"dni Nauki vol. 32 (5): p.172-174. 1995.

In a field trial on leached chernozem containing 0.05 g B/100 g soil, Lotus corniculatus was sprayed with 250 g B/ha at flowering and seed filling or with 500 g B at one or both of these stages. Average seed yield in 1990-91 increased from 335 and 353 kg/ha in dry and wet controls, respectively, to 424-434 kg with 2 B applications and 389-400 kg with one application. Two applications of 250 g B were most profitable. Numbers of pods/stem and seeds/pod were highest with 2 applications of 500 g B/ha; 1000-seed weight was highest with 1 or 2 applications of 500 g.

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Sowing method for smooth bromegrass cv. Nika for seed production. Tomov, P. Rasteniev"dni Nauki vol. 32 (5): p.141-143. 1995.

In trials on leached chernozem in 1989-92, Bromus inermis cv. Nika was sown in rows 30 or 60 cm apart or in bands 45 cm apart, alone or alternating with rows of Lotus corniculatus or Medicago sativa, or in rows 15 cm apart. alone or mixed with each legume. No seed was harvested in the 1st year. Seed yields in 1990-92 were highest in pure stands with inter-row cultivation and averaged 1040, 986, 991 and 831 kg/ha at spacings of 60, 45, 30 and 15 cm, respectively. At a given spacing, growing B. inermis in alternate rows with L. corniculatus and M. sativa reduced its seed yield by about 10-15 and 50%, respectively. In mixed rows, seed yields were reduced by about 30-40%. Because of the cost, growing seed crops in rows 60 cm apart, with inter-row cultivation, is only recommended for production of elite or super-elite seed. For mass seed production, the use of pure stands in rows 30 cm apart is recommended. Forage yields were highest with alternate or mixed rows of M. sativa and hay yields from pure stands of B. inermis.

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Interrelations between the species in perennial grass mixtures used as pastures. Videva, M. Rasteniev"dni Nauki vol. 32 (5): p.122-125. 1995.

In a trial on cinnamon meadow soil in 1987-91, Lotus corniculatus, Festuca arundinacea and Dactylis glomerata were grown in pure stands or in 30/70% mixtures of L. corniculatus with each of the grasses. Lolium perenne, alone or with L. corniculatus, was grown in 1988-91. Total annual DM yield from 4 cuts ranged from 4.74 t/ha in L. perenne (184 d) to 8.22 t in L. corniculatus (145 d). The mixtures yielded 7.03-7.49 t. The rate of development and dynamics of DM accumulation of D. glomerata made it most compatible with L. corniculatus. F. arundinacea inhibited the growth of L. corniculatus in the 1st year.

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Effect of mineral fertilizer application on the yield and botanical composition of meadow grasses and grass mixtures in the Troyan region. Lingorski, V.; Totev, T.; Mukhovski, Ts.; Tankov, K.; Churkova, B.; Georgieva, D.; Belperchinov, K. Rasteniev"dni Nauki vol. 32 (5): p.100-103. 1995.

In field trials on infertile acid soil in 1989-92, Lotus corniculatus, Trifolium pratense, T. repens, Dactylis glomerata, Festuca rubra and Lolium perenne were grown in pure stands or in mixtures for grazing or hay, and were given 80, 160 or 240 kg N/ha alone or with 80 kg P2O5 + 80 kg K2O/ha. The legumes and the pasture mixture showed little response to N. Yields of D. glomerata, F. rubra and L. perenne were increased by 5-90, 44-84 and 66-134%, respectively. The highest yield of 13 t DM/ha was obtained from D. glomerata with 240 kg N + PK. Applying PK alone had little effect on the grasses, but increased T. repens yield by 41%. Effects on botanical composition are reported; N stimulated growth of grasses and weeds, while PK promoted legumes and suppressed weeds.

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Effect of temperature and moisture potential on germination and emergence in Lotus sp. Blumenthal, M.J.; Aston, S.C.; Pearson, C.J. Australian Journal of Agricultural Research vol. 47 (7): p.1119-1130. 1996.

Two controlled cabinet experiments compared the germination of 6 Lotus genotypes over a range of temperatures (0-40 deg C) and moisture potentials (0 to -1.2 MPa). In addition, a field experiment conducted at Berry, New South Wales (NSW), compared the emergence of 5 Lotus genotypes over 18 sowing dates at a range of soil temperature and moisture potentials. Lotus corniculatus cv. Grasslands Goldie and L. uliginosus cv. Grasslands Maku, G4703, G4704 and Sharnae were included in both the controlled cabinet and field experiments, while Lotus suaveolens CPI 35903 was included in the controlled cabinet experiments only and Trifolium repens cv. Haifa was included as a control in the field study. Of the commercially available Lotus cultivars, Grasslands Goldie provided the greatest flexibility in terms of sowing date in coastal NSW and could be successfully sown even in late autumn and winter as it exhibited the lowest base temperature for germination and field emergence. Cultivars G4703, Sharnae and G4704 had a lower base moisture potential for germination than the other 3 Lotus genotypes, but this was not translated in to a higher percentage seedling emergence in the field. L. suaveolens had an extremely low base temperature for germination (1 deg C) compared with Grasslands Maku (6.4 deg C) and it is suggested that hybridization of this species with both L. corniculatus and L. uliginosus may result in germplasm capable of germinating better at low temperature and able to maintain the perenniality, rhizomatous habit and the condensed tannin content of L. uliginosus.