1. Introduction:
3. Lotus Activities: reports
and abstracts:
4. Request for wild - colected Lotus material. D.A. Jones.
Purpose: The Lotus Newsletter
consists of informal communications of research information on
Lotus. 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.
This is the 23 rd year of publication for
the Lotus Newsletter. Now is the
time to consider contributing to the 24 rd volume of the
Lotus Newsletter. Contributions generally
are compiled without editing.
1. Prepare your contribution using any Macintosh or IBM (MS-DOS) 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.
2. Send your contributions by December 31, 1993 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
The expense of publishing the Lotus Newsletter has been partially covered by unrestricted research support. This issue of the Lotus Newsletter is provided to you without charge. I will continue to strive for financial support of the Lotus Newsletter to provide you with an unencumbered communication resource.
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 research and management on the many species of Lotus.
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.
Your suggestions are helpful, and I will strive to incorporate them to make this publication more useful. As you compile your data for analysis please think about making a contribution to the Lotus Newsletter. Use the Lotus Newsletter as a resource for communication: it is published to aid you (and me), the international researchers of Lotus.
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 receiving the Lotus Newsletter have them submit a questionnaire and they'll be added to the mailing list.
Please note the instructions for submitting your contribution by computer disk or e-mail.
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.
The illustration on the cover is of a Lotus spp. L. graciously provided by Ana Arambarri (Argentina) . The illustration of L. unifoliatus Benth. (syn. L. purshianus) is the third in a series of illustrations that started with L. edulis in Volume 23.
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Checkmark all categories that apply
to your area of Lotus research:
| O Genetics | O Breeding | O Taxonomy | O Physiology |
| O Pathology | O Ecology | O Biology | O Forage |
| O Utilization | O Germplasm | O Tissue culture | O Biotechnology |
| O Entomology | O Seed | O Reclamation | O Other (please indicate) |
List the Lotus
species you study: _______________________________
Give a brief description of your research _________________________
___________________________________________________________
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Send or FAX your completed questionnaire to:
P. R. Beuselinck, USDA-ARS
University of Missouri
207 Waters Hall
Columbia, MO 65211 USA
FAX 573-882-1467
Arrangements have been made to hold this symposium at the Missouri Botanical Gardens in St. Louis, Missouri. Meeting dates are 2224 March 1994. The Lotus Symposium is cosponsored by the Missouri Botanical Society, the University of Missouri, and the Agricultural Research ServiceUSDA.
Objective of the Lotus Symposium will be to cover the many aspects of this genus including breeding, genetics, taxonomy, management, utilization or physiology, from applied agronomy to genetic manipulation. Secondary objective is to publish the proceedings in a manner which can be used as the standard reference for Lotus.. Plenary speakers are planned, and voluntary papers are encouraged.
A mailing will be made to all recipients of the Lotus Newsletter with details of the program, accommodations, and instructions for participation.
Further information can be obtained by writing P. R. Beuselinck,
Plant Genetic Research Unit, University of Missouri, 207 Waters
Hall, Columbia, MO 65211, USA, or by FAX (3148821467), or EMail
(AGROBELL@MIZZOU1.MISSOURIEDU).
Second Conference on Forage Quality, Evaluation, and Use. 1315 April 1994. Lincoln, Nebraska. Contact Dr. Lowell Moser (402) 4721558 for further details.
B. G. Cameron and N. Prakash
Department of Botany, University of New England, Armidale, N.S.W.,
2351. Australia.
Lotus cruentus Court, the redflowered trefoil, is
one of two indigenous species of Lotus in Australia. It
is a decumbent or ascending perennial herb found in all mainland
states in a diverse range of soil types and plant communities,
varying from flood and sand plains to rocky hillsides (Cunningham,
Mulham, Milthorpe and Leigh, 1981).
The leaves consist of five narrow obovate to broadly cuneate leaflets.
The flowers are red, pink or white, 68mm long and are borne
in leaf axils in groups of 13 (Norris and Harden, 1991).The
calyx lobes are acute, almost equal, and with long silky hairs
(Fig 1 A). Unlike in many
other papilionoid genera, the wing petals have no sculpturing
but have a distinct pocket in the upper basal region. The pod
is brown, hairless and curves upwards (Fig 1A).
Redflowered trefoil is known to cause poisoning in livestock,
mainly in sheep, during drought (McBarron, 1978). The toxic chemical
is HCN which is prevalent in young green leaves (Everist, 1974).
The toxicity is still high even at the podded stage (McBarron,
1978).
The stigma of L. cruentus consists of short, denselypacked
papillae (Fig. 1B). The surfaces
of the papillae are coated giving a convoluted, furry appearance.
The coating is no doubt very important for the recognition and
stimulation of pollen to germinate.
The pollen grain (Fig. 1C)
is slightly oblong to spheroidal in shape and shows three furrows
(colpi) that are fairly narrow with undifferentiated margins.
The surface (exine) is generally smooth with a scattering of fine
perforations. The aperture of the pollen is quite distinct. These
features
appear to be common to other species of the genus (Ferguson and
Skvarla, 1981).
The ovule (Fig. 2A)
has two coverings (called integuments) which are continuous except
at the tip where a zigzag micropyle allows access for the
pollen tube to enter. An unusual feature of the inner integument
is the presence of an endothelium which nourishes the embryo sac.
Another unusual feature of the ovule is the presence of large,
glandular cells on the placenta in the region of the funicle constituting
an obturator that assists the pollen tube in its growth towards
the ovule. Following a developmental pattern common to most angiosperms
(Polygonum type), a female gametophyte (also called an embryo
sac) contain" eight nuclei organized into seven cells is
formed inside the ovule.
Upon fertilization, an embryo is formed. Figure 2B shows
longitudinal section through an immature seed. Part of the nourishing
tissue the endosperm) has solidified along the periphery while
the bulk of the endosperm remains liquid with a large number of
free nuclei.
Cellular details of the seed coat (testa) are shown in Figure 2C.
The characteristic macrosclereid (or malpighian) layer is prominent
on the outside with a layer of osteosclereids lying underneath.
Starch grains are abundant in the seed coat.
References
Cunningham, G.M., Mulham, W.E., Milthorpe, P.L. and Leigh, J.H.
(1981). Plants of Western New South Wales. N.S.W. Govt. Printer,
Sydney.
Everist, S.L. (1974). Poisonous Plants of Australia. Angus &
Robertson, Sydney.
Ferguson, I.K. and Skvarla, J.J. (1981). Pollen morphology of
the subfamily Papilionoideae (Leguminosae). In Polhill, R.M. &
Raven, P.H. (eds) Advances in Legume Systematics. Part 2. Royal
Botanic Gardens, Kew.
McBarron, E.J. (1978). Poisonous Plants of Western New South Wales.
N.S.W. Dept. of Agriculture, Sydney.
Norris, E.H. and Harden, G.J. (1991). Lotus. In Harden, G.J.(ed.).
Flora of New South Wales. Vol. 2. NSW University Press, Kensington.
Figure Legends
Figure 1. Lotus cruentus.
A. Bud, flower and fruit. Bar = lcm.
B. S.E.M. of the stigmatic surface showing short, densely packed
rough papillae. Bar = 10mm. C. S.E.M. of a pollen grain showing
3 colpi and a fairly smooth, slightly perforate surface. Bar =
10mm.
Figure 2. Lotus cruentus..
A. L.s. ovule showing obturator cells (ob) at the base
of the funicle (fu), outer integument (oi), inner integument
(ii) with a welldeveloped endothelium (et) and enclosing
a zigzag micropyle (mi). The mature embryo sac ( es)
consists of an egg cell (eg) flanked by 2 synergic
cells (sy), 2 polar nuclei (pn) in a central cell and 3
antipodal cells (an). Bar = 20mm.
B. L.s. of an immature seed showing young embryo (em), solid
and liquid endosperm (en) and a layer of macrosclereids
(ms) in the testa. Bar = 5mm. C. T.s. of seed coat showing
macrosclereid (or malpighian) layer (ms), the osteosclereid
layer (os) containing starch (st) and the solid
endosperm (en) lying underneath. Bar = 50mm.
Introduction
Lotus pedunculatus cv. Grasslands Maku has been sown widely
on acid infertile soils in coastal regions of eastern Australia
(Harris et. al 1992). Maku lotus is a rhizomatous perennial
legume that persists vegetatively; however, seedling recruitment
may occur following drought or flood if seed is present in the
soil. To determine the size of soil seed banks in farmers fields
in eastern Australia a survey was conducted in 1991.
Methods
Soil samples were taken from 57 paddocks from locations ranging
from Gympie, Queensland (26°10'S) to Bairnsdale, Victoria
(37°51'S). All paddocks had been sown to Maku lotus prior
to 1988 and had been allowed to flower at some stage since that
time. At each site twentyfive 7 cm diameter cores were taken
to a depth of 5 cm. Samples were processed by the method of Jones
and Bunch (1988) and Maku lotus and white clover seeds/m2 were
related to site characters. Site characters used were rainfall,
latitude, aspect, soil type, soil pH, fertilizer history, year
sown, percentage cover of lotus, other species present, paddock
size, type of livestock enterprise, stocking rate and stocking
method.
Results
Lotus seed banks ranged in size from O to 6,621 seedstm2 with
a mean of 662 (s.e. 184.5). White clover seed banks ranged in
size from O to 6,789 seeds/m2 with a mean of 956 (s.e. 171.4).
The size of the lotus seed bank was positively correlated with
latitude (r=0.33; P<0.05) and percentage cover of lotus (r=0.37;
P<0.01) and was independent of other site characters. Samples
collected from low latitude but high altitude at Glen Innes (29042'S;
1000 m) had relatively high soil seed reserves (18902626
seeds/m2) compared with lower altitude sites at the same latitude.
Discussion
Daylength has already been shown to influence flowering in L.
pedunculatus (Force and Thomas 1966) and minimum daylength
requirements may not be met at low latitude sites. The large seed
banks at Glen Innes suggest that a vernalisation requirement may
also need to be met. The size of the seed bank reflects a number
of events, to fully understand the mechanisms involved,
observations need to be made of flower number, yield and yield
components and losses of seed from the seed bank. The effect of
insect predators needs to be monitored throughout these stages.
Whatever the mechanism, seed banks can contribute to the persistence
of Maku lotus in farmers fields, particularly at high altitude
and at sites further south than Taree (340S). The size of lotus
seed banks were comparable to that for white clover. A Maku lotus
seed bank of 600 seeds/m2 is equivalent to 2.7 kg/ha; this should
be sufficient seed for a sward to reestablish following
a drought or flood provided the seed was of reasonable quality
(e.g. 30% germinable).
Acknowledgments
The authors wish to thank Dr J. Ayres, Messrs G. Bunch, R Campbell,
P. Dann, H. Kemp, D. McCoy, P. Midson, A. Rumbel and Ms R. Beardsell
for supplying the samples. The research was funded by the Australian
Dairy Research and Development Corporation.
References
Harris, C A., Blumenthal M.J. and Scott, J.M. 1992. Survey of
use and management of Lotus pedunculatus cv. 'Grasslands
Maku' Proc. 6th Aust. Agron. Conf., Armidale. p. 545.
Jones, R.M. and Bunch, G.A. 1988. A Guide to Sampling and Measuring
the Seed Content of Pasture Soils and Animal Faeces. CSIRO
Trop. Agron. Tech Mern No. 59.
Forde, B.J. and Thomas, R.G. 1966. Flowering in Lotus pedunculatus
Cav. 1. Effects of photoperiod. N.Z. J. Bot. 4,147152.
W.M. Kelman l, M.J. Blumenthal 2 and C.A. Harris2
1 CSIRO, Division of Plant Industry, GPO Box 1600, Canberra, ACT
2601, Australia. 2 NSW Agriculture, Pasture Research Unit, P.O
Box 63, Berry, NSW 2535, Australia.
There is a need to diversify the legume content of pastures in
southern Australia to exploit situations where the more commonly
used species are absent or unproductive. In dairying and beefproducing
regions of southeastern Australia Lotus has proved
a valuable forage for acid and infertile soils (Harris et al.,
1992). A single cultivar (L. pedunculatus cv. Grasslands
Maku) is presently utilized. A wider evaluation of germplasm has
the potential of identifying accessions that can contribute to
the development of cultivars of Lotus with improved establishment,
seed production and forage quality.
A collection of 40 accessions in each of Lotus pedunculatus
and L. corniculatus is under evaluation at two sites
in the dairying region of New South Wales: a coastal site near
Nowra (34°54'S, AAR 1200 mm., aft. 8m) and a more elevated
site at Kangaloon, near Bowral (34°28'S, AAR l500mm., aft.
900m). The accessions were planted in a randomized complete block
design with two replications. As part of a larger plant character
assessment, dry matter production was measured at the end of the
first summer and autumn periods of growth.
The mean level of productivity of Lotus at Kangaloon was
greater than at Nowra (P<0.001), this difference being associated
with the higher rainfall at Kangaloon during this period. Lotus
pedunculatus showed a more uniform level of production over
the two cutting periods than L. corniculatus (Species x
season interaction significant at P=0.001), in which species 72%
and 65% of summer growth was achieved in the autumn at Kangaloon
and Nowra, respectively (Table 1). Within L.
pedunculatus, the New Zealand bred material (Maku, G4703 and
G4704) and a group of Portuguese accessions (including cv.Sharnae)
were the most productive (Fig. 1).
A breeding program based on crosses between these types has been
started. In L. corniculatus a group of 610 entries,
mostly tall hay types developed in North America, were identified
as the most productive (Fig. 1),
while important autumn activity appears to reside in Vega and
some Spanish accessions. A polycross nursery has been established
in an attempt to recombine the productivity of the hay types with
more prostrate, winter active germplasm.
Reference:
Harris C.A., Blumenthal M.J. and Scott J.M. 1992. Survey of use and management of Lotus
pedunculatus cv. Grasslands Maku in eastern Australia Proceedings of 6th Australian
Agronomy Conference, Armidale. p.595.
Table 1: Mean drymatter production (g/plot) over 40 accessions in Lotus pedunculatus and r. corniculatus at two sites in NSW, Australia.
| Species | ||||
| L. pedunculatus | ||||
| L. corniculatus | ||||
W. F. Grant, R. B. McDougall1 and B. Coulman
Department of Plant Science, P.O. Box 4000, Macdona/d Campus
of McGill University, Ste. Anne de Bellevue, Quebec H9X 3V9, Canada
and 15928 99th Street, Edmonton, Alberta TOE 3N9, Canada
A birdsfoot trefoil plant with only a single inflorescence per
individual branch, and in which the inflorescence was sessile,
has been found in a singlespaced field planting of birdsfoot
trefoil. Crosses are planned to obtain inheritance data.
Background of the source of the putative mutant
During the fall of 1990 and the spring of 1991 crosses were made
between four cultivars of birdsfoot trefoil, namely, Cree, Leo,
Upstart, and Viking, with the goal of eventually obtaining a cultivar
with high quality herbage. Seed was germinated in the Spring of
1991 and the resulting seedlings were planted in individual small
pots. Approximately 150 seedlings were obtained and these were
planted in the field during the summer of 1991. In September,
65 of the best plants (growth habit diameter, height, seed
yield) were selected from the field and potted in the greenhouse.
In January, 1992, the plants were brought into flower through
the use of mercury lights which provided a daylength of approximately
17 hours. From February to April, 1992, leaf cutter bees were
hatched and a few were placed in with the flowering plants approximately
every week. Seed was collected from the plants and germinated.
In MayJune, 1992, approximately 640 individual seedlings
were space planted in the field. In September, 1992, seed was
harvested individually from each plant and during the collecting
period, the sessile inflorescent plant was observed.
Character of the putative mutant
It is a relatively large plant with stem lengths up to 70 centimeters.
The stems arise from a very stout crown and are semierect.
The plant possesses only a single inflorescence on each individual
branch (Fig. 1).
In typical birdsfoot trefoil plants, more than one inflorescence
per branch is characteristic. In the putative mutant plant, the
inflorescence is close to sessile and unlike typical trefoil plants
the inflorescence arises only from the first node. The umbels
are 2 to 7flowered which is characteristic of normal birdsfoot
trefoil plants. The plant possesses axillary peduncles which are
1.0 to 2.0 mm in length in contrast to typical birdsfoot trefoil
plants in which the axillary peduncles are from 3 to 10 cm long.
Bracts are present subtending the flower as on typical plants.
As the sessile inflorescent plant was not observed until the plant
had completed flowering, the flower color is not know. It might
be assumed to be yellow as the plant did not appear to be different
from other birdsfoot trefoil plants while in the flowering stage.
Sessile Inflorescence Mutations
No mutant plant possessing a sessile inflorescence is believed
to have been found previously, and such a mutant does not appear
in the list of Lotus mutants given in Beuselinck and Mosjidis
(1991).
Crosses are planned to obtain inheritance data.
Acknowledgment
Financial support from the Natural Sciences and Engineering Research
Council of Canada for a Cooperative Research and Development Grant
to W. F. Grant for studies in Birdsfoot Trefoil is gratefully
acknowledged.
Reference
Beuselinck, P. R. and J. A. Mosjidis. 1991. Genetic nomenclature
in clovers and specialpurpose legumes: III. Lotus, Lespedeza,
Kummerowia, and Vicia sop. Crop Sci. 31: 871874.
El genero Dorycnium propuesto inicialmente por Tournefort
(1700), fue aceptado en principio por Linneo (1735, 1737), pero
posteriormente (1753) no lo acepta y lo sinonimiza con Lotus.
Miller en 1754 publica validamente el genero.
Desde este momento haste la actualidad el genero ha pasado una serie de vicisitudes, pues unos autores lo reconocen como tal mientras que otros lo incluyen en Lotus, e incluso Reichenbach (1832) con alguna de sus especies crea el genero Bonjeanea. Esta situaci6n queda resumida en el siguiente cuadro:
| Autor | ||||
| Tournefort | ||||
| Linneo | ||||
| Linneo | ||||
| Linneo | ||||
| Miller | ||||
| Miller | ||||
| Villars | ||||
| Villars | ||||
| Willdenow | ||||
| Seringe | ||||
| De Candolle | ||||
| Reichenbach | ||||
| Webb & Berthelot 1842 | ||||
| Bentham & Hooker 1865 | ||||
| Boissier | ||||
| Willkomm | ||||
| Taubert | ||||
| Rikli | ||||
| Rikli | ||||
| Coste | ||||
| Bonnier | ||||
| Gams | ||||
| Hutchinson | ||||
| SchulzeMenz | ||||
| Ball | ||||
| Demiriz | ||||
| ,Celebioglu | ||||
| Polhill |
+ = Lo admite como genero
o = No habia sido descrito
= No lo admite como genero
* = Lo admite como seccion
? = no lo trata en su obra
Como indica Polhill (1981), los caracteres usados a los largo
de la historia, pare separar estos dos generos e incluso otros
muy relacionados como Tetragonolobus, Hosackia, Heinekenia,
Vermiinuc, etc, no hen sido muy consistentes, y de ahf su
complejo tratamiento generico.
En las Islas Canarias se encuentran tres especies endemicas de
Dorycuu~n (Lotus) que estan englobadas en un seccion de
Dorycnium (Rikli, 1900) o en un subgenero de Lotus (Gillett,
1959).
Dos especies Dorycnium broussonetcc y Dorycnium spectabile
fueron descritas por Seringe in De Candolle (1825)
como Lotus broussonetii y Lotus spectabilis respectivamente
y la tercera Dorycnium eriophthalmum fue descrita como
tal por Webb & Berthelot (1842), aunque estos autores en una
table publicada con anterioridad lo hicieron como Lotus eriophthalmus.
Rikli (1900, 1901) crea la seccion canaria en el genero
L)orycnium pare incluir a estos tres taxones, y asf hen
sido considerados por los estudiosos de la flora canaria, haste
que Gillett (1959) op ine, al compararlas con l as es pec ies
arb ustivas africanas de Lotus, que estas tres es pecies
estan mas relacionadas con Lotus que con Dorycnium,
y pare ellas propone el subgenero canaria de Lotus.
Nosotros despues de estudiar estas especies canaries, y basandonos
en los estudios de Rikli (1901), Brand (1898), Gillett (1959),
Grant & Sidhu (1967), Ball (1968), Ortega (1979), Monod (1980)
y en nuestras propias observaciones, y siempre que el genero Dorycautm
siga siendo aceptado por .05 botanicos, creemos que estos
taxones endemicos canarios deben de mantenerse en este genero
como una seccidn endemica de el.
Secci6n CANARU Rikli, Ber. Schweiz. Bot. Ges. 10: 15 (1900) Syn.
Loms L. subgen. Canaria (Rikli) Gillett, Kew Bull.
13(3): 364 (1959)
DESCRIPCION
Los caracteres diagndsticos mas destacables de la secci6n consideramos que son los siguientes:
Nano o microfaner6fitos. Fol~olos grandes, con raquis de mas de
1 cm de largo. Pedunculos mas cortos que las hojas. Flores vistosas
de mas de 10 mm, de color blanco, crema o violaceo. Petalos muy
largamente unguiculados, con la una mas large que el tubo del
caliz. Estandarte algo mas corto o ligeramente mayor que los demas
petalos. Alas con el apice no soldado. Quilla ligeramente rostrada,
con el apice mas oscuro.
DISTRIBUCION
Islas Canarias: Gran Canaria, Tenerife, La Gomera, El Hierro y
La Palma. Las tres especies son muy escasas y locales,
viven entre los 160 y 900 m s.m., en la zone de transici6n entre
los pisos bioclimaticos Infra y Termocanario. Mapa n. 1.
ESPECIES
Dorycnium enopAthalmum (Webb & Berth.) Webb
& Berth., Phyt. Canar. 3(2): 88 (1842)
Basiom Lotus eriophthalmus Webb & Berth., Phyt.
Canar. tab.59 (1837); text. explic. tab., op. cit.: 88 (1842),
pro syn.
Distribucion
Dorycnium eriophthalmum es el que tiene una mas amplia
distribuci6n, se encuentra en las isles de Tenerife, La
Palma, La Gomera, El Hierro y posiblemente en Gran Canaria, se
conoce un pliego de Sventenius pare esta isle. Dorycnium broussonetii
(Choisy ex Ser.) Webb & Berth., Phyt. Canar. 3(2): 90
(1842). Basidn. Lotus broussonetii Choisy
ex Ser. in DC., Prodr. 2: 211 (1825) Syn. Dorycnium torulosum
Presl, Symb. Bot. I: 19, tab. XI (1832) Lotus polycephalos
Brouss. ex Rikli, Bot. Jahrb. 31: 324 (1901), pro syn.
Distribucidn
Seringe (1825) cite Tenerife (Isles Canarias) o Mogador (W de
Africa) como lugares de origen de la especie. La cite se establece
en base al itinerario de recoleccion de Broussonet y ha de interpretarse
como orientativa pare aquellas plantas recolectadas por Broussonet
en que existi6 perdida o trasvase de etiquetas. S610 es citada
pare ambas localidades por autores que transcriber la cite de
Seringe. Ningun autor que conozcamos menciona haberla recolectado
en Mogador, por lo que pensamos que alli nunca existio.
Dorycnium broussonetii, solo crece silvestre en dos de
las siete Islas Canarias: Tenerife y Gran Canaria.
Dorycnium spectabile (Choisy ex Ser.) Webb & Berth., Phyt. Canar. 3(2): 89 (1842).
Basion. Lotus spectabilis Choisy ex Ser. in
DC. Prodr. 2: 211 (1825)
Distribuci6n
Para esta especie vale e l mismo comentario real izado en el cap
itulo de distribucion de Dorycnium broussonetii (Choisy
ex Ser.) Webb & Berth.
Dorycnium spectabile, se ha encontrado haste ahora, solamente
en la isle de Tenerife. Tiene una distribucion muy limitada y
por el momento, tanto por testimonios bibliograficos, por exsiccata
de herbarios, asi como por nuestras recolecciones, solamente se
encuentra en una region muy restringida de la isle, en Guimar,
y aqui solo en dos localidades mas o menos proximal, Bco. de Badajoz
(La Ladera) y Bco. del Agua. Y es una especie que se considera
en peligro de extinci6n (Categoria E de la IUCEN).
REFERENCL\S BIBLIOGRAfICAS
BALL, P.W. 1968 a. Dorycnium Mill. In: T.G.TUTIN & AL. (eds.), Flora Europaea 2: 172173. Cambridge University Press.
. 1968 b. Lotus L. In: T.G.TUIIN & AL. (eds.), Flora Europaea 2: 173176. Cambridge University Press.
BRAND, A. 1898. Monographie der Gattung Lotus. Bot. Jahrb. 25: 166232.
GILLETT, J.B. 1959. Lotus in Africa south of the Sahara (excludind the Cape Verde islands and Socotra) and its distintion from Dorycnium. Kew Bull. 13(3): 361381.
GRANT, W.F. & B.S. SIDHU. 1967. Basic chromosome number, Cyanogenetic glucoside variation, and geographic distribution of Lotus species. Can. J. Bot. 45: 639647.
LINNEO, C. 1735. Systema naturae. ed. I. Leiden.
. 1737. Genera Plantarum. ed. I. Leiden.
. 1753. Species Plantarum. ed. I. Tomo II. Stockholm.
MILLER, P. 1754. The gardener's dictiorzary Abridged. ed. 4'. London.
MONOD, TH. 1980. Contribution a ['etude des Lotus (Papilionaceae ouestsahariens et macaronesiens. Adansonia, ser. 2, 19 (4): 367 402.
ORTEGA, J. 1979. Citogenetica del genero Lotus en macaronesia. m. Variaci6n en el contenido de gluc6sidos cianogeneticos en Lotus de las isles Canarias y Madeira. Bot. Macar. 5: 919.
POLHILL R.M. 1981 a. Tribe Loteae DC (1825). In: R.M.POLHILL & P.H.RAVEN (eds.), Advances in Legume Systematics, Part 1: 371375. Royal Botanic Gardens, Kew.
REICHENBACH, H.G.L. 18301832. Flora Germanica excursoria, 2. Lipsiae.
RIKLI, M. 1900. Die Scheizerischen Dorycnium. Ber. Scheiz. Bot. Ges. 10: 10 44.
. 1901. Die Gattung Dorycnium. Bot. Jahrb. 31: 314404.
SERINGE, N.C. 1825. Lotus etDorycnium, In: A P. DE CANDOLLE, Prodromus Systematis naturalis regni vegetabilis 2: 208215. Paris.
TOURNEFORT, J.P. 1700. Institutiones rei herbariae. I.
WEBB, P.B. & S. BERTHELOT. 1842. Histoire Naturelle
des Iles Canaries. Phytographia Canariensis 3(2): 4448;
8091; tabs. 49, 57, 58, 59. Paris.
Niizeki, M., S. Nakajo,and T. Harada Laboratory of Plant Breeding, Faculty of Agriculture, University of Hirosaki, Hirosaki,
Aomoriken 036, Japan.
Asymmetric somatic hybrid call), which have only the nuclei of
birdsfoot trefoil (Lotus corniculatus L.), were produced
by protoplast fusion between rice (Oryza sativa L) and
birdsfoot trefoil, and analyzed for their mitochondrial DNAs (mtDNAs)
and chloroplast DNAs (cpDNAs). In the hybrid call), novel mtDNA
fragments were detected in Southern blots. This result shows that
some kind of alteration such as intergenomic and/or intragenomic
recombinations of mtDNA occurred in the hybrid call). On the other
hand, the cpDNA fragment patterns of all hybrid callus lines observed
by Southern analysis were found to be identical with those of
birdsfoot trefoil. Therefore, it is suggested that the cpDNAs
of these hybrid calli sorted out unidirectionally. Interestingly,
some regenerated plants from the hybrid calli were tolerant of
low temperatures and low sunlight intensity.
Introduction
The hybridization of distantly related species by protoplast fusion
has been a practical tool for removing the barriers of incompatibility
in sexual crossing of agriculturally important plant species.
The cell division of somatic hybrids originating from fused protoplasts
has been observed in several combinations of plant species belonging
to different families (Gleba and Sytnic 1984). Callus formation
has also been reported in somatic hybrids between the species
of different families (Kao 1977, Niizeki et al. 1985, Sala et
al. 1985). By irradiation of the protoplasts of one parent, or
by spontaneous chromosome elimination of one parent, Smith et
al. (1989) and Niizeki et al. (1989) obtained asymmetric somatic
hybrid plants between the species of different families. In such
hybrid calli and plants, new and sexually impossible genetic combinations
can be constructed in the nucleous, chloroplast and mitochondrial
genomes. The hybrid calli and plants may also give new data concerning
the mechanisms of various cellular activities and morphologies
which are not observed in the sexual hybrid cells.
Recently, several researchers have reported successful chloroplast
transformation in higher plants (De Block et al. 1985, Cornelissen
et al. 1987, Haring and De Block 1990, Svab et al. 1990, Staub
and Maliga 1992). However, there are two difficulties that need
to be overcome in order to bring about a stable chloroplast transformation.
The difficulties arise from a number of plastic genomes in higher
plant cells, and the doublelayered envelope of organelle.
Therefore, somatic hybridization will play an important role in
the improvement of cytoplasmic genomes.
Materials and Methods
The plants used in this study are rice (Oryza sativa L.),
strain A58 and birdsfoot trefoil (Lotus corniculatus
L.), cv. Viking. The calli of rice were induced from the scutellums
of seeds on the NS medium (Nurashige and Skoog 1962) with 2 mg/1
2,4dichlorophenoxyacetic acid. The calli of birdsfoot trefoil
were induced from the hypocotyls of young seedlings on the MS
medium with 4 mg/1 1naphthaleneacetic acid and 2.5 mg/1
kinetin. About 12 g of
fresh calli were used for the isolation of protoplasts. An enzyme
solution containing 4% Cellulase Onozuka RS and 1% Macerozyme
R10 was used for the rice calli and a solution of 4% Cellulase
Onozuka RS, 1% MacerozymeR10 and 0.2% Pectolyase Y23 was used
for the birdsfoot trefoil call). The mixed protoplasts of the
two species were treated with polyethylene glycol(PEG) by applying
a somewhat modified method of Melchers et al. (1978). The modified
method has been reported by Niizeki et al. (198577 Selection systems
of hybrid calli wi11 be described in the results and discussion.
NtDNA and a mixture of nuclear DNA and cpDNA were isolated from
23 g of calli of somatic hybrids and their parents by a
somewhat modified method of Milligan (1989). After the first centrifugation
of ground calli with a cold isolation buffer (1.25 M NaCl, 50
mM Tris pH 8.0, 5 mM EDTA, 0.1% BSA, 0.1% 2mercaptethanol)
mtDNA could be obtained from the pellet, and the mixture of nuclear
DNA and cpDNA could be isolated from the aqueous phase. For Southern
hybridization, mitochondrial genes and the PstI and BamHI fragments
of cpDNA were used as probes. Probe labelling, Southern hybridization
and signal detection were performed by using the ECL (enhanced
chemiluminescence) gene detection system (Amersham, UK).
Results and Discussion
When the rice protoplasts were fused with 15 mM IOAtreated
protoplasts of birdsfoot trefoil, and were cultured using the
agarosebead method in the modified KM8P medium (Kao and
Michayluk 1975) containing benzyladenin instead of zeatin and
without coconut water, only hybrid protoplasts initiated cell
division and formed colonies. In this case, rice protoplasts could
not give rise to cell division in the modified KM8P medium. The
hybrid colonies had the flavonoid pigment of rice, strain A58,
or were a brownish yellow in contrast to the green of birdsfoot
trefoil colonies. Also, malformed shoots were derived from the
early subcultured calli and nearnormal shoots of birdsfoot
trefoil from advanced subcultured call). The microscopic observation
of colony cells showed that most of the cells have the chromosomes
of birdsfoot trefoil and a small number of rice chromosomes. Therefore,
the regenerated malformed shoots might presumably be caused by
the presence of a few rice chromosomes. After a considerable number
of subcultures, reduction of the abnormality might be due to the
disappearance of the rice chromosomes retained. In mtDNA of 6
hybrid callus lines digested by 6 restriction enzymes, some novel
fragments were detected by the Southern analysis(Table 1).
This result shows that some modification occurred in the mtDNA
of hybrid callus lines. On the other hand, the Southern blots
of cpDNA in the hybrid calli of 12 combinations of 4 restriction
enzymes and 3 probes, the banding patterns of hybrid callus lines
were the same as those of birdsfoot trefoil. This result shows
unidirectionally nonrandom segregation of cpDNA, despite the fact
that many researchers have reported that the cpDNA of somatic
hybrids are sorted out randomly (Chen et al. 1977, Sidrov e al.
1981, Bonnett and Glimelius 1983, Ashahi et al. 198877 From the
study of somatic hybrid between carrot and tobacco, Smith et al.
(1989) suggested that the irradiationinduced chromosome
instability in carrot nuclei resulted in a tobacco nuclear background
which may provide a selective advantage to tobacco cpDNA. In the
case of somatic hybridization in this study, the chromosomes of
birdsfoot trefoil were predominant in the nuclei of hybrid callus
cells. In addition, original fusion products were cultured in
a modified KM8P medium, which did not allow the division of rice
protoplasts. Therefore, such a medium may be a selective disadvantage
for rice chloroplasts.
Some regenerated plants from the hybrid calli of rice and birdsfoot
trefoil had deep green leaves and vigorous growth under low sunlight
intensity and low temperatures (20°C) in a green house in
the winter season in Hirosaki, Japan (Fig. 1).
It has been shown that the organization of mtDNA is correlative
to the morphological and physiological functions of plants (Bonnett
and Glimelius 1990, Newton et al. 1990, Honda et al. 1991). In
the study of hybrid calli of rice and soybean, we (1985, 1986)
suggested that nucleocytoplasmic interactions determine the colour
of the callus and its susceptibility to streptomycin. Therefore,
it is possible to assume that some kinds of alteration in mtDNA
organization and/or nucleocytoplasmic interaction have an effect
on the traits of callus cells such as the colours of the callus
and its tolerance of low temperatures and low sunlight intensity.
Table 1. Southern blot of mtDNAs in 6 hybrid callus lines (AF) of rice and birdsfoot trefoil
| Restriction | Probe | ||
| enzyme | atpA | rrn26 | coxI |
| EcoRI | A | ND | ND |
| BamHI | ND | ND | ND |
| HindIII | A | A | ND |
| PstI | ND | ND | ND |
| SmaI | A | ND | ND |
| SalI | A | ND | ND |
A: Callus line A shows novel fragments which are different from those of parents. ND: No difference from the fragment pattern of birdsfoot trefoil in all callus lines.
Fig. 1. Shoots under the low temperatures (20°C) and low
sunlight intensity in a green house in the winter season. A: Shoots
of plants derived from the parental calli of birdsfoot trefoil,
leaves of which are etiolated. B: Shoots of hybrid plants, leaves
of which show deep green.
References
1.Asahi, T., T. Kumashiro and T. Kubo 1988. Constitution of mitochondrial
and chloroplast genomes in male sterile tobacco obtained by protoplast
fusion of Nicotiana tabacum and Nicotiana deDneyi.
Plant Cell. Physiol. 29:4349.
2.Bonnett, H.T. and K. Glimelius 1983. Somatic hybridization in
Nicotiana: Behavior of organelles after fusion of protoplast
from malefertile and malesterile cultivars. Theor.
Appl. Genet. 65:213217.
3.Bonnett, H.T. and K. Glimelius 1990. Hybrids of Nicotiana
tabacum and petunia hybrida have an intergeneric mixture of
chloroplasts from P. hybrida and mitochondria identical or similar
to N. tabacum. Theor. Appl. Genet. 79:550555.
4.Chen, K., S.G. Wildman and H.H. Smith 1977. Chloroplast DNA
distribution in parasexual hybrids as shown by polypeptide composition
of fraction I protein. Proc. Natl. Acad. Sci. USA 74:51095112.
5.Cornelissen, M.J., M. De Block, M. Van Montagu, J. Leemanns,
P.H. Schreier and J. Schell 1987. Plastid transformation: A progress
report. In Plant DNA Infections Agents. Hohn T. and J. Schell
Eds. SpringerVerlag Berlin Heidelberg New York Tokyo. pp.
311320.
6.De Block, M., J. Schell and M. Van Montagu 1985. Chloroplast
transformation by Agrobacterium tumefaciens. EMBO J. 4:13671372.
7.Gleba, Y.Y. and K.M. Sytnik 1984. Protoplast fusion and hybridization
of distantly related plant species. In Protoplast Fusion. SpringerVerlag
Berlin Heidelberg New York Tokyo. pp. 115161.
8.Haring, M.A. and M. De Block 1990. New roads towards chloroplast
transformation of higher plants. Physiol. Plant. 79:218220.
9.Honda, H., K. Itoh and A. Hirai 1991. The heterogenous composition
of mitochondrial DNA in somatic hybrid calli and the relatively
simple composition of such DNA in regenerated leaves. Jpn. J.
Genet. 66:279289.
lO.Kao, K.N. 1977. Chromosomal behaviour in somatic hybrids of
soybean-Nicotiana glauca. Mol. Gen. Genet. 150:225230.
ll.Kao, K.N. 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 126:105110.
12.Melchers, G., M.D. Sacristan and A.A. Holder 1978. Somatic
hybrid plants of potato and tomato regenerated from fused protoplasts.
Carlsberg Res. Commun. 43:203218.
13.Milligan, B.G. 1989. Purification of chloroplast DNA using
hexadecyltrimethylammonium bromide. Plant Mol. Biol. Report 7:144149.
14.Murashige, T. and F. Skoog 1962. A revised medium for rapid
growth and bioassays with tobacco tissue culture. Physiol. Planta.
15:473497.
15.Newton, K.J., C. Knudsen, S. GabayLaughnan and J.R. Laughnan
1990. An abnormal growth mutant in maize has a defective mitochondrial
cytochrome oxidase gene. The Plant Cell 2:107113.
16.Niizeki, M., M. Tanaka and K. Saito 1986. Response of somatic
hybrid callus between rice and soybean to streptomycin. Jpn. J.
Breed. 36:7579.
17.Niizeki, M., M. Kihara, K. Cai, R. Ishikawa and K. Saito 1989.
Somatic cell hybridization among gramineous and leguminous species.
Proc. of the 6th Internatl. Congr. of SABRAO. pp.501504.
18.Niizeki, M., M. Tanaka, S. Akada, A. Hirai and K. Saito 1985.
Callus formation of somatic hybridization of rice and soybean
and characteristics of the hybrid callus. Jpn. J. Genet. 60:8192.
l9.Sala, C., M.G. Biasini, C. Morandi, B. Nielsen, B. Parisi and
F. Sala 1985. Selection and nuclear DNA analysis of cell hybrids
between Daucus carota and oryza saliva. J. Plant Physiol.
118:409419.
20.Sidrov, V.A., L. Menczel, F. Nagy and P. Maliga 1981. Chloroplast
transfer in Nicotiana based on metabolic complementation
between irradiated and iodoacetatetreated protoplasts. Planta
152:341345.
21.Smith, M.A., A. Pay and D. Dupid 1989. Analysis of chloroplast
and mitochondrial DNA in asymmetric somatic hybrids between tobacco
and carrot. Theor. Appl. Genet. 77:641644.
22.Staub, J.M. and P. Maliga 1992. Long regions of homologous
DNA are incorporated into the tobacco plastic genome by transformation.
Plant cell 4:3945.
23.Svab, Z., P. Hajdukiewicz and P. Maliga 1990. Stable transformation
of plastics in higher plants. Proc. Natl. Acad. Sci. USA 87:85268530.
AGS4 autogamous broadleaf birdsfoot trefoil (Lotus
corniculatus L.) germplasm was developed and released by the
United States Department of Agriculture, Agricultural Research
Service, in cooperation with the Oregon, Idaho, and Washington
Agricultural Experiment Stations.
AGS4 originated from a single autogamous clone identified
in the broadbased MU81 germplasm, a genetically
diverse, selfincompatible population that was produced by
three cycles of intercrossing plants from 56 foreign introductions
and 35 experimental synthetics, or cultivars. S1 seed resulting
from both unassisted selfing and hand manipulations were collected
from the autogamous clone and grown to flowering in a greenhouse
under longday conditions. S2 and S3 generation were produced
from 11 S1 and 16 S2 plants, respectively, under similar conditions
with only unassisted selfing (no hand manipulations). AGS4
is a composite of equal numbers of S4 seeds from the 12 S3 plants.
Plants from AGS4 are more ovate and lighter green than the
original autogamous clone. AGS4 exhibits no breeding vigor
depression when compared to MU81 and is a prolific seed
producer. Examination of polyacrylamide electrophoresis gels for
highsalt soluble globulin polypeptide patterns from singleseed
extracts indicate that individuals of AGS4 are highly homogeneous.
No seeds are produced from flowers which are emasculated. Somatic
chromosome number, as determined from root tip squashes, was 2n=4x=24.
No other reported sources of autogamous broadleaf birdsfoot
trefoil are available for breeding and experimental purposes for
this predominantly selfincompatible species.
Limited amounts of seed of AGS4 will be provided upon written
request as supplies permit. Recipients are asked to recognize
the source as a matter of open record when this germplasm contributes
to the development of a new germplasm or cultivar or when used
for experimental purposes. Address seed requests to Dr. Jeffrey
J. Steiner, National Forage Seed Production Research Center, USDAARS,
3450 SW Campus Way, Corvallis, OR 97331.
Moon, T.Y.1, J.H. Kim2
1 Korean Entomological Institute,
2 Department of Biology,
Korea University. Seoul 136701, Korea
Lotus corniculatus var. japonicus Regel is the only
species member of genus Lotus in Korea. One of us (TYM)
recently initiated the national survey for distribution of the
species in Korea. As a beginning part, the records from literature
were compiled and databased in cooperation with a plant taxonomist
(JHK) who collated the records between literature. The all records
compiled here are collected from the literature published in Korea
and Japan, while the records from foreign literature mentioned
Korean distribution of the species or the other Lotus species
are yet to be compiling.
Most botanists believe that L. corniculatus var. japonicus
is not native to Korea and is introduced as Trifolium repens
and T. pra sense. It distributes mostly along the coast,
particularly the west and south coastal sands and cliffs. Otherwise,
small riparian colonies scatter along the major rivers.
It is certain that the plants distribute more widely in Korea
than the map suggests, and also include more diverse varieties
than var. japonicus. In Cheju Island (BM,BN,CM,CN) that
is at the southern end of the country and subtropical, the plants
may be composed of heterogeneous populations with various origins
because many parts of the islands were sowed to construct openpastures
with the seedmixtures that were introduced from at least
two different countries(German and Japan), and that, therefore,
are likely to include the seeds of various varieties of L.
corn *u 7atus as usual in commercially supplied seeds.
However, the plants are not quite successful in the main penninsular
Korea. The plants are in the beginning stages remaining small
circluar colonies at the most recorded sites. and also our collections.
The distribution map is based
on the records from the literature listed at the end (also see
Table 1). Some uncertain records were marked
with O, otherwise the rest confirmed by us were marked with .
However, the amount of records are poor due to the resesons that,
firstly Lotus spp. were neglected since most flora surveys
have been designed to investigate forests and woodlands, and secondly
only a few taxonomists worked on the leguminous plants in the
country.
The better understanding the distribution of L. corniculatus
var. japonicus would provide various merits to work
with the plants. Together with the distribution survey, the other
aspects of ecology, in particular on the insect herbivores, are
also under investigation.
Table 1. The distribution records of Lotus cornicuulatus var. japonicus in Korea,
SC: Seoul; KG: Kyonggido; KW: Kangwondo; CB: Ch'ungchongbukdo; CN: Ch'ungchongnamdo; Cb: Chollabukdo; Cn: Chollanamdo; KB: Kyongsangbukdo; KN: Kyongsangnamdo;
CJ: CheJudo
| Locality | |||||||||||
| ref.no | |||||||||||
| 1 | |||||||||||
| 2 | |||||||||||
| 3 | |||||||||||
| 4 | |||||||||||
| 5 | |||||||||||
| 6 | |||||||||||
| 7 | |||||||||||
| 8 | |||||||||||
| 9 | |||||||||||
| 10 | |||||||||||
| 11 | |||||||||||
| 12 | |||||||||||
| 13 | |||||||||||
| 14 | |||||||||||
| 15 | |||||||||||
| 16 | |||||||||||
| 17 | |||||||||||
| 18 | |||||||||||
| 19 | |||||||||||
| 20 | |||||||||||
| 21 | |||||||||||
| 22 | |||||||||||
| 23 | |||||||||||
| 24 | |||||||||||
| 25 | |||||||||||
| 26 | |||||||||||
| 27 | |||||||||||
| 28 | |||||||||||
| 29 | |||||||||||
| 30 | |||||||||||
| 31 | |||||||||||
| 32 | |||||||||||
| 33 | |||||||||||
| 34 | |||||||||||
| 35 | |||||||||||
| 36 | |||||||||||
| 37 | |||||||||||
| 38 | |||||||||||
| 39 | |||||||||||
| 40 | |||||||||||
| 41 | |||||||||||
| 42 | |||||||||||
| 43 | |||||||||||
| 44 | |||||||||||
| Total | |||||||||||
Literature from Lotus Database
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Chosen, Plant Soc. Kyungsung Pharm.School (in Japanese)
2. Anonymus, 1984, A Footmark of Prof, Changbok Lee, VII,
An Account of Herbaria (SNU), pil3314 (in Korean)
3. Anonymus, 1991, An Account of Herbaria of Natural History Museum,
Hannam Univ, vol 1, Nat.His.Mus.Hannam Univ. 236pp (in Korean)
4. Chung,M.H, 1976, A Study on the Pharmacy Plant Resources of
Mt. Chiri, J. Chosun Univ. 189 (in Korean)
5. Chung,Y.H, 1989, An Account of Herbaria of Department of Botany
in Seoul National University (IV), Bull.Dept.Bot.Seoul Nat l.Univ.
1189
6. Chung,Y.H, C.W.Park & B.Y.Sun, 1980, An Account of Herbaria
of Department of Botany in Seoul National University (II), Kor.J.Bot.
22: 1163
7. Chung,Y.H. g Y.M. Kang, 1971, A Taxonomic Study in Kangwha
Island, J.Nat. Acad. Sci. Korea, 10:91299 (in Korean)
8. Hong,W.S, 1958. A Study on the Flora of Youngjong Isl,
Bull.Sci.Tech. Yonsei Univ. 2:5285 (in Korean)
9. Joo,S,W, 1963, Report on the Vegetation of Pusan Area. Korean
Ministry of Education
10. Kil,B.S, 1989, The Flora of Pyonsan Penninsula National Park,
Buan, J.Sci. WonKang Univ. 9(3):5794
11. Kim,T.W. & S.S.Kim, 1979, A Study on the Distribution
of Vascular Plants at Six Islands near Geoje Island, The Report
of the KACN, 14:3558
12. Kim,Y.S, J.K.Sim & B.U.Oh, 1985, Studies on the Flora
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13. Kim,Y.S. & S.N.Son, 1984, A Study on the Flora of Chungmu
Area, Sci.Tech. Korea Univ. 25:83110
14. Kim,Y.S. & S.W. Park, 1984, A Study of the Flora of Kayasan
(Ch'ungnam) , J. Educ.Korea Univ. 11:95116
15. Kim,Y.S. & Y.P.Hong, 1986, Report on the Vegetation of
Mt. Sanbang, Chejudo, J. Nat. Sci.Korea Univ. 27:4363
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Sci.Tech. Korea Univ. 23:89104
17. Kim, Y.S, K.K. Kim, W.B. Lee & K.S. Koh, 1988, A Study
on the Flora of Kadok Isl., J.Nat.Sci.Korea Univ. 29:93120
18. Kim,Y.S, W.K.Kim 8 H.J.Lee, 1980, Report on the Vegetation
of Jodo Isl., Sci. Tech, Korea Univ. 21:99114
19. Kim,Y.S, Y.H.Chun & K.O.Ro, 1981, A Study of the Flora
of Mt. Wangbang, J.Educ.Korea Univ. 11:127148
20. Kim,Y.S, Y.K.Kim, W.B.Lee & J.K.Sim, 1988, An Investigation
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Korean)
23. Lee,D.B, 1957, Flora of Cheju Island, J.Cult.Sci.Korea Univ.
2 (in Korean)
24. Lee,T.B, Y.B.Lee & C.H.Lee, 1980, Vegetation of the Islands in the Sinan county, near Mokpo. The RePort of the KACN, 16:3154
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of Songji Ho, Jour. Kor.Pl.Tax. 10(1,2):105111
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Nat'l.Univ. (in Korean)
27. Lee,Y.N, 1968, Report of Academic Survey of Mt. Hanlasan
& Isl.Hong do, 6782, Ministry of Culture, Korea
(in Korean)
28. Lee,Y.N, 1979, A Study of Conservative Condition on Hallamountain
Top Flora, Bull. KACN, 1:6377
29. Lee,Y.N. 8 Y.C.Oh, 1970, Limestone Flora of Todam, Province
Chungbuk in South Korea, J.Kor.Rehab.Indust. 5:256
30. Lee,Y.N. & Y.C.Oh, 1985, An Investigation on Flora Chuwangsan
National Park, The Report of the KACN, 23:3761
31. Moon,T.Y,1992, Cyanogenic Polymorphism in the Leaves of Lotus
corniculatus var. japonicus Regel (Leguminosse) in
South Korea, Kor.J.Ecol. 15(1):7580
32. Na.Y.J, 1991, A Study on the Flora of Mt. Sodae, Thesis
of MS, Hannam Univ. 156pp
33. Nakai,T, 1915, Report on the Flora of Mt. Chiri. pp52
(in Japanese)
34. Nakai,T. 1919, Report on the Vegetation of the Island Ooryongto
or Daglet Island, Corea, Govern.Chos.Corea, pp86
35. Oh,S.Y, 1973, Report on the Vegetation of Mt. Gaya,
J.Andong Nat' l.Teach. Coll. 6:153
36. Oh.S.Y, 1974, Report on the Vegetation of Mt. Gaji,
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37. Oh,S.Y, 1982, Floristic and Phytogeographical Studies of Mt.
Naeyeonsan and Its Neighbourhood, Res.Rev.Kyungpuk Nat'l.Univ.
33:367413
38. Oh,S.Y. & W.Kim, 1978, Taxonomic and Ecological Studies
on the Flora of Taegu Area. Nature 8 Life, 8:153
39. Park,K.H, 1985, A Study on the Flora of Oyoujong
Isl. in Kyunggi Bay, Nature Conservation, 52:3548
40. Park,K.H, 1987, The Islands Adjacent to Paengnyongdo, Report,Survey
Nat. Environ. Korea, 7:137169
41. Hatusima,S, 1934, Preliminary Report on the Flowering Plants
and Ferns Collected in the Nansen Experimental Forest of Kyushu
Imperial University, Bull.Kyushu Imp.Univ. 5:1281
42. Toh,S.H. & S.H.Park, 1971, Plant Resources of Mt. Duryun,
Kor.J.Fharmacy, 2(2):99118
43. Yang,I.S, 1958, An Investigation Report of the Plant in Jindo
Isl. J. Kyungpuk Nat 1.Univ. 2:323349
44. Yang,I.S, 1963, An Investigation of Kyungpuk Flora,
Kyungpuk Nat'l.Univ. Press, 141pp
Acknowledgasnt
We are grateful to Dr. JungKi Sim (MoleWon University)
who kindly allowed us using
his personal library during the literature survey.
Regeneration of GUS , Bt-toxin and HNP transgenic
plants of Lotus corniculatus.
Lu DY. , Shao JH., Zhang L, Wang Q, Yu MM, Xu SM, Fan YL.
1 Instutute of Genetics, Academia Sinica, Beijing 100101, China.
2 Biotechnology Research Center, Agricultural Academy of Sciences
in China, Beijing 100101, China.
Lotus corniculatus L (birds-foot trefoil) is an important
forage legume with rich nutrients and "blot safety".
However, some characteristics of L. corniculatus, such as deficiency
of sulfur amino acid need to be improved. Content of sulfur amino
acids in forage plants is positively related to wool growthin
sheep.
Previous reports of transformation of L corniculatus described
the incorporation of NPT11 (Yu and Shao 1990), cat (Hansen et
al.1987,Petit et al 1987, Jorgensen et al. 1988), luc (Jensen
et al. 1986, Hansen et al.1989), Gus (Hansen et al. 1989). Here
we report regeneration of GUS ., BT-toxin and HNP(sulfursrich
amino acid )transgenic plants from seedling cotyledon explants
of L corniculatus via an Agrobacterium vector.
Materials and methods
Axenic seedling cotyledons(7-10 days) of L corniculatus were cut
transversely into two pieces, and placed into a suspension of
A. tumefaciens strain A3 with plasmid pBI121 containing
NPT11 gene ( nos promoter) and GUS gene (CaMC35 Promoter), LBA4404
with plasmid pB48.215 containing NPT11 gene and B toxin gene (CaMv35s
promoter) or A2881 with plasmid pBF649 containing NPT11gene (nos
promoter)and HNP(sulfursrich aminoacid) gene (Ca MB35s promoter)
for 0.5 h, and co-cultured for four days on MS1-2(Lu et al. 1986).
Then the segments were transferred to MS1-2with 25 Mg/lKnamycin
and 300 mg/l cefotaxime. The control were not infected by A281
with other conditions same as the treatment samples. Shoot and
plants from cotyledon explants were removed and cultured on fresh
MS1-2 with 100 mg/l kanamycin and 300mg/l cefotaxime at one month
interval, for 5-6 times. Roots or stems from kanamycin-resistant
plants were cut into0.5-1 cm long and cultured on MS1-2 with 25-50
mg/l kanamycin. Samples of cotyledon, leaf and stem segments were
made for observation of scanning electron microscopy according
to report of Ho and Vasil (1983). The kanamycin-resistant plants
ere cut into small pieces and inoculated to YEB medium for axenic
examination of the plants. The plants were transferred to pots
and greenhouse.
Determination of NPT11 enzyme activity was carried out according
to the method of Angenon et al (1987), Schreier et al, (1985),
and Roy and Nirmala (1990).
DNA molecular hybridization (dot blot and southern blot) was carried
out referring to the method of Maniatis et al (1982).
Results and Discussions
Plants were regenerated from cotyledon explants on selection medium
with about 10 per cent regeneration frequencies, and form root
or stem segments of the regenerated plants on MS1-2 with 25-50
mg/l kanamycin. Untreatment cotyledon explants all died on selection
medium. It will shorten period of selection the transformation
plants to obtain plants from root or stem segment of the regeneration
plants.
Scanning electron microscopic observation showed that Agrobacterium
number attached on cut face of cotyledon segments were much more
than it on surface of them.
There was no Agrobacterium in and out kanamycin-resistant plants
after 5-6 times selection .
Enzyme assay showed NPT11 or Gus enzyme activity in the plants
(fig. 1-1, 3) Results of dot and southern blotting of the plant
NA demonstrated that there were homogenic fragments of GUS, Bt-toxin
and HNP genes in the plants, but there were not them in the controls
(fig. 1-2, 4). These confirmed that GUS, Bt-toxin and HNP genes
had been integrated into genomes of L. corniculatus cells. The
transformed plants have survived , continued grow since being
transferred to pots and greenhouse, and showed no phenotypic abnormalities.
References
1 Angenon G et al.(1987) EMBO) Course on Plant Molecular Biology Genet.
2 Hansen J et al.(1988) . Plant Cell Rep. 8:12-15.
3 Ho WJ and Vasil IK. 91983). Ann Bot. 51:719-726.
4 Lu Dy et al.(1986) Kexue Tongbao (Chinese Sci Bul) . 31:1647-1650.
5 Jensen JS et al. (1986). Nature. 321:669-674.
6. Jorgensin Jan-Elo et al.(1988). Nucl Aci Res. 16:40-50
7 Petit et al.(1987) Mol Gen Genet. 207:245-250.
8 Roy Pand Nirmala S .(1990) . Plant Mol Biol. 14:873-876
9 Schreier P et al.(1985) EMBO J. 4:25-32
10 YU JP and Shao QQ. (1990) . Science in China (Series B). 34:932-937.
Acknowledgment
We thank Dr. Tian YC and Dr. Hen SY for Agrobacterium and work
support.
1 USDAARS, Water Management Research Laboratoroy, Fresno, CA, USA
2 USDAARS, Department of Biometrics, Albany, CA, USA
3 BenGurion University of the Negev, Sedi Boqer Campus, Israel
4 UC Davis, Department of Environmental Science, Davis, CA, USA
5 USDAARS, Plant Genetics Research Unit, Columbia, M0, USA
INTRODUCTION
High concentrations of boron (B) and selenium (Se) found in the
soils are detrimental to sustainable agriculture in the western
USA. Vegetation management may be a remediation strategy designed
to reduce soil B and Se concentrations to nontoxic levels,
since extensive leaching with water is a practice which consumes
excessive water in drought stricken areas and causes drainage
water and runoff problems in perched water table farming areas.
In this regard, Parker et al. (1991) suggested using perennial
grasses and/or legumes as candidate crops for removal of Se from
soils high in B. Plant removal of B with B and Se tolerant plants
may economically be more feasible and more environmentally sound
than physically removing B contaminated soil or taking Bladen
regions out of crop production. The objective of this field study
was to determine if birdsfoot trefoil (Lotus corniculatus L.
) tolerates high B soils and simultaneously lowers extractable
B and Se in soil by plant uptake of each respective ion.
METHODS AND MATERIALS
Field experiments were established between May and November of
1990 and 1991 in the west side of San Joaquin Valley, California.
The site was chosen because of the high levels of B in the soil.
The soil belongs to the Los Banos Clayloam, fine mixed thermic
Typic Haploxeralfs. The treatment design for both years on two
similar sites was a completely randomized design with two treatments;
a) planted to Lotus corniculatus L. in 1991 and b) bare
plot where no plants were planted. In 1990, each 5X5 m plot was
replicated nine times and in 1991, each 10X10 m plot was replicated
three times. Triplicate soil cores were collected within each
plot from depths of 030 and 3060 cm, respectively,
prior to planting and at harvest. Selected ions and other parameters
were extracted from saturated soil extract (mean values are shown
inTable 1 ) . Water extractable B (any mention
of soil B references water extractable B) was determined spectrophotometrically
by the azomethine H method (Bingham, 1982). Total Se was
determined by atomic absorption with continuous hydride generation
after soil samples were wet digested with HNO3/H2O2.
| Saturated water | |||||||||
| Soil depth
(cm) | Ca | Mg | Na
(mg/L) | PO4 | SO4 | Cl
(mMol) | Ec
(d/Sm) | pH | Percentage
% |
| Experiment 1990 * | |||||||||
| 030 | 240(95) | 85(31) | 267(34) | 22(34) | 800(245) | 6(2) | 2.8(.7) | 7.9(.2) | 47 |
| 3060 | 300(125) | 70(25) | 305(55) | 16(5) | 667(104) | 4(1) | 2.2(.6) | 7.8(.2) | 48 |
| Experiment 1991 | |||||||||
| 030 | 225(80) | 79(25) | 258(Z9) | 19(4) | 756(158) | 6(3) | 3.1(.5) | 7.8(.2) | 48 |
| 3060 | 273(69) | 71(26) | 295(39) | 16(6) | 692(111) | 5(2) | 2.5(.6) | 7.7(.3) | 49 |
*Men values presented with standard deviations in parenthesis.
Plants were first established under controlled greenhouse conditions,
hardened for two weeks under field conditions and then transplanted
as clumps every 20 cm with 125150 plants/m2. L. corniculatus
plots were each hand clipped at 60, 85, and 115 days after
the plants were at least 5.0 cm in height. Subsamples were taken
from four one square meter sites within each plot, washed three
times, ovendried at 45°C for 7 d, weighed, and ground
in a stainless steel Wiley mill. Plant B was determined spectrophotometrically
after wet acid digestion and plant Se was determined by atomic
absorption with continuous hydride generation. Irrigation scheduling
was based on the local California Irrigation Management Information
System (CIMIS) weather station. Irrigation was performed with
a sprinkler system. Spacing of 9 m x 8 m resulted in an irrigation
rate of about 10 mm/ hr. Mean irrigation depth for the two years
was 825 mm. The total amount of evaporation throughout the growing
season averaged 1000 mm, which resulted in an irrigation coefficient
of 0.82. Irrigation water contained negligible concentrations
of both B and Se.
RESULTS
The mean dry matter yield of L. corniculatus was O.9kg/m2
in 1990 and 1 kg/m2 in 1991.
Yields might have been higher if the plants had been grown longer
than one year (only three clippings made). The mean tissue B and
Se concentrations are shown in Table 2 for
each clipping. Each clipping removes additional B and Se from
the soil. Table 3 shows the reductions of
soil B and Se after the final harvest of L. corniculatus. Reductions
of soil B and Se in bare plots were probably due to some leaching
from the sprinkler irrigation and/or the partial transformation
of soil B to other forms of B which were not extractable by water.
| Boron | Selenium Concentration | ||||
| Clipping | Shoot | Root (mg kg/DM) | Shoot | Root | |
| 1990 Experiment*! | I | 84(6)a | -- | 0.44(.03)b | -- |
| II | 131(12)b | -- | O.B7(.09)a | -- | |
| III | 135(14)b | 95(10) | O.90(.10)b | 0.10(.08) | |
| 1991 Experiment | |||||
| I | 116(11)b | -- | 0.36(.06)bc | -- | |
| II | 116(6)b | -- | O.Z9(.04)bc | -- | |
| III | 118(8)b | 110(9) | 0.22(.03)bc | 0.14(.01) | |
* Values presented represent means followed by standard error of mean in parenthesis from a minimum of 20 separate samplings in 1990 and 12 samplings in 1991. Means are separation within columns and within each experiment by a Tukey's range teat. The same literature represents no significant difference between means at the P0.05 level.
! Root sample were not taken, for first2 clippings in each experiment.
| 1990 Experiment * | 1991 Experiment ! | |||||||
| Species | B | Se | B | Se | ||||
| Preplant | Postharvest | Preplant | Postharvest | Preplant | Postharvest | Preplant | Postharvest | |
| (mg / L) | (mg / L) | (mg / L) | (mg / L) | |||||
| Control
(Bare points) | 4.47(0.52) | 3.99(0.66)a $ | 0.49(0.11) | 0.43(0.01)a | 3.57(0.60) | 3.43(0.11)a | 0.88(0.046) | 0.86(0.02)a |
| Birdsfoot
trefoil plots | 5.14(0.57) | 2.26(0.38)b | 0.39(0.10) | 0.12(0.07)b | 4.16(0.71) | 2.98(0.22)b | 0.82(0.075) | 0.71(0.03)b |
* Values represent the mean from 36 soil samples followed by the standard error of mean in parenthesis.
! Values represent the mean from 16 soil samples followed by the standard error of mean in parenthesis.
$ Mean separation in columns obtained by Tukey's range test. The
same letters represent no significant difference between species
at the P= .05 level.
We suggest that utilizing harvested L. corniculatus as
animal forage is worthy of economic consideration, especially
because of its high quality as animal forage. More importantly,
tissue levels of B and Se were under the maximum tolerable limits
established by the National Research Council (1980) for plant
material used as animal forage. Using vegetation management in
conjunction with efficient irrigation management may be a critical
component of remeditation strategy to reduce soil and drainage
water concentrations of B and Se to safe levels.
LITERATURE
National Research Council. 1980. Mineral Tolerances of domestic
animals. National Academy of Sciences, Washington, D. C.
gingham, J.T. 1987. Boron. p. 431447. In A.L. Page
et al. (eds.) Methods of Soil Analysis . Part 2: Chemcial Microbiological
properties . Am. Soc. Agron ., Madison, WI .
Parker, D . R ., A. L. Page, and D . N. Thomas . 1991. Salinity
and boron tolerances of candidate plants for the removal of selenium
from soils. J. Envir. Qual. 20:157164.
The Desert Legume Program (DELEP) was established in 1988 as a
joint project of the Boyce Thompson Southwestern Arboretum and
the University of Arizona College of Agriculture. DELEP is developing
a comprehensive germplasm collection of species of Leguminosae
which are native to, or adapted to arid and semiarid climates.
The collection presently contains 938 species. The program seeks
to evaluate these plants for any potential uses including food,
fuelwood, forage, fodder, cover crops/green manure, erosion control,
industrial compounds, pharmaceutical properties, and landscape
plants. Much of this work will be accomplished through collaborative
arrangements with academia, movement agencies, and private industry.
Samples of seeds are available free of charge to individuals and
organizations working with these plants. A quarterly newsletter,
ARIDUS, and periodic special publications including an Index Seminum
are available at no cost.
DELEP has recently initiated a greenhouse propagation program
for seed increase of various herbaceous species for which a limited
stock of seeds exist in the seed bank. Included in this project
are species of Lotus. Twentytwo taxa of Lotus
have been successfully propagated to date. The plants are
grown in standard, plastic onegallon nursery containers
in a mixture of peat moss, vermiculite, perlite, and sand. The
greenhouse thermostat is set to maintain a temperature of 33°
C in the daytime and 16° C at night. Many of these plants
are beginning to produce flowers as of November 1992.
Samples of seeds of the following taxa of Lotus, with field
collection data, are available free of charge: L. haydonii,
L. humistratus, L. oroboides, L. rigidus, L. salsuginosus var.
brevivexillus, L. strigosus var. tomentellus, and
L. wrightii. Individuals interested in receiving seed samples
and those who may have seeds to contribute, are invited to contact
the Desert Legume Program.
Introduction
The initial objectives in assembling a small collection of Lotus
species and varieties at North Wyke Research Station, Devon
and commencing to screen them on an acid (pH 5.4), lowphosphate
status (P<10 ppm) site were described in Lotus Newsletter
1991, pages 3739. When planning lowinput, sustainable
grassland systems, whiteclover based technology may provide
a solution on the better soil types, though it is increasingly
evident that regular inputs of lime and phosphate will
be needed, particularly under high rainfall conditions. In such
circumstances, as well as on all poorer soils, Lotus species
may provide a better option.
The programme of Lotus assessment at North Wyke will be
developed to include investigation of productivity and persistence
under grazing. However, before attempting the grazing experiment,
a smallplot experiment to investigate the influence of four
possible companion grasses on Lotus performance was planned
and sown in 1991. This article reports some quality determinations
on material from the screening trial in 1991, together with the
1992 yield data. In addition, yield data from the first cut of
the companion grass experiment are reported.
As mentioned in the earlier Newsletter report, North Wyke Research
Station is situated in S.W. England and has a mild maritime climate.
Mean air temperature in January is 4.5°C and in July 15.3°C.
Average rainfall is 1035 mm, with approximately 200 rain days.
The soil is a poorly drained, seasonally waterlogged silty
clay loam (pelostagnogley).
Initial Screening Trial 1991 Quality Data
The establishment of this trial and the 1991 yields were reported
in the Lotus Newsletter 1991, 3739. Lotus samples
from Cut 2 (31 August 1991) for the five best yielding cultivars
were analyzed for digestibility (predicted from pepsintcellulase
solubility) and N content (acid digestion followed by colorimetric
assay). The data appear in Table 1, and show
that at this cut (8 weeks regrowth) both of the L. uliginosus
cvs. and the L. tenuis were of lower (P<0.001) digestibility
than the two L. corniculatus cvs. tested. However, L.
corniculatus cv. Cascade had the lowest nitrogen concentration
(P<0.001).
Table 1. Lotus quality data at Cut 2 (31 Au~ust) 1991.
| Species | Cultivar | DOMD* | N (g /kg ) |
| L. uliginosus | Maku | 0.515 | 39.7 |
| Marshfield | 0.540 | 36.3 | |
| L. corniculatus | Cascade | 0.619 | 29.2 |
| Norcen | 0.656 | 36.1 | |
| L. tenuis | Blenheim | 0.536 | 33.4 |
| s.e.d. (8 DF) | 0.0083 | 1.34 | |
| Level of significance | *** | *** |
* digestible organic matter fraction in the dry matter.
Initial Screening Trial 1992 Yield Data
The screening trial was continued in 1992, the third year since
sowing in 1989. Growing
conditions were dry and sunny for the first part of the 1992 season, though dull and moist from
July onwards. Cuts were taken from this experiment on 16 June and 17 September 1992. Fresh
herbage yields were recorded on the plotharvester, and two subsamples collected. The first
subsample was dried in a forceddraught oven at 85°C to determine the dry matter (DM) content
of the herbage, and the second subsequently sorted to determine the proportion of Lotus and
grass in the sample. Table 2 shows the comparative yields of Lotus DM at Cuts 1 and 2, as well
as the annual total yields of combined forage in the same rank order as the 1991 data were
displayed.
Table 2. DM yields of Lotus species and varieties. 1992 (t /ha ).
| Species Species | Cultivar | Lotus DM | Annual DM Yield | ||
| Cut 1 | Cut 2 | Lotus | Total * | ||
| L. uliginosus | Maku | 1.64 | 0.71 | 2.35 | 10.79 |
| Marshfield | 0.70 | 0.63 | 1.33 | 8.17 | |
| L. corniculatus | Cascade | 0.65 | 0.59 | 1.24 | 10.51 |
| Norcen | 0.21 | 0.36 | 0.58 | 9.94 | |
| L. tenuis | Blenheim | 1.23 | 0.43 | 1.66 | 9.97 |
| L. corniculatus | Empire | 0.18 | 0.43 | 0.61 | 9.35 |
| L.c. ssp. arvensis | Kalo | 0.56 | 0.67 | 1.23 | 9.14 |
| L. corniculatus | GA1 | 0.19 | 0.47 | 0.66 | 8.71 |
| AUDewey | 0.10 | 0.36 | 0.45 | 9.03 | |
| Fergus | 0.22 | 0.81 | 1.03 | 9.34 | |
| s.e.d. (18 df) | 0.353 | - | 0.482 | - | |
| Level of significance | ** | NS | * | NS | |
* includes grass and other plant DM.
With a few exceptions, yields of Lotus forage declined
since 1991, though with the good grass growing conditions for
Cut 2, the total annual yields were above those of 1991, so that
the general percentage legume contribution fell to 1020%
of DM. As in 1991, top forage yields came from L. uliginosus
cv. Maku. There was marked improvement in the ranking of L.
tenuis cv Rlenheim and L. corniculatus ssp. arvensis cv.
Kalo.
Companion Grass Experiment 1992
This experiment was sown in July 1991, with the two Lotus and
the four companion grasses listed in Table 3. Perennial
ryegrass (Lolium perenne) in UK is generally too competitive
and densely tillered to form stable associations with birdsfoot
trefoil, so these possible alternative grasses were chosen. The
Lotus was sown at 10 kg ha' and the grasses at either
2 or 4 kg ha', excepting Festuca pratensis which
was sown at 3 or 6 kg ha'. Appropriate quantities of grass
and inoculated Lotus seed were thoroughly mixed, broadcast
on to a harrowed seedbed and rolled in, using a layout of
3 randomized blocks of plots 1.5 m x 5.0 m, with all factorial
combinations of Lotus variety, grass species and grass
seedrate. Data from the first cut, taken on 11 June 1992
are shown in Table 3.
Table 3. Lotusperformance when grown with foiur companion grasses at two seed - rates.
Sown July 1991, cut 11 June 1992.
| Comparisons: | Lotus DM (t/ha) | Sown grass(t/ha) | Legume(%) | Total DM(t/ha) |
| L. corniculatus cv. Leo | 3.35 | 0 90 | 73.0 | 4.61 |
| L. uliginosus cv. Maku | 1.00 | 0.98 | 43.6 | 2.48 |
| s.e.d. (23 df) | 0.127 | 0.128 | 2.51 | 0.216 |
| Phleutn pratense cv. S.48 | 1.64 | 1.55 | 42.8 | 3.53 |
| Agrostis capillaris cv. Muster | 2.34 | 1.44 | 52.9 | 3.99 |
| Festuca pratensis cv. Senu | 2.26 | 0.70 | 57.0 | 3.71 |
| Poa pratensis cv. Asset | 2.48 | 0.06 | 80.5 | 2.96 |
| s.e.d. (11 df) | 0.179 | 0.181 | 3.55 | 0.305 |
| High seedrate (4 or 6 kg/ ha) | 2.02 | 1.08 | 52.7 | 3.53 |
| Low seedrate (2 or 3 kg/ ha ) | 2.33 | 0.80 | 63.9 | 3.57 |
These data show that, as anticipated, L. corniculatus established
more rapidly than L. uliginosus resulting in higher yields
of Lotus DM (P<0.001) and, because there was no compensatory
effect on grass yield, higher overall yields of forage (P<0.001)
at this first cut. Phleum pratense depressed Lotus DM
yield (P<0.001); Poa pratensis gave the lowest sown
grass yield (P<0.001) and overall forage yield (P<0.05)
but consequently the highest (P<0.001) legume contribution
to yield. The higher level of companion grass seeding depressed
Lotus DM yield (P<0.05), raised grass DM yield (P<0.05),
lowered the legume contribution (P<0.001), but had no overall
effect on total forage yield. It remains to be seen whether L.
uliginosus] will yield more highly when fully established
next year, and whether the higher yielding companion grasses and
the higher seedrates will have an adverse effect on the
longer term performance of the Lotus.
Future Research
The plots of the screening trial will not be harvested after 1992,
but it is anticipated that the companion grass experiment will
provide valuable information through 1993 and 1994. It is planned
to sown the swards for the grazing assessment during 1993, for
an experiment to run through 1994 and 1995. Grazing will involve
contrasts between sheep and cattle, probably in rotational systems
with differing criteria for moving the stock on. Inevitably sward
carrying capacity will be low, but properly researched guidelines
for managing grassLotus associations could provide
a valuable alternative to white cloverbased technology for
marginal land situations.
Any reader who would like to offer comment on the design of the
grazing experiment is invited to write (or FAX (+44) 837 82139)
to the second author.
LOTUSDEV. PAP. RDS1
In March, 1992, 164 accessions of Lotus were acquired from
the USDAARS Regional Plant Introduction Station in Geneva, N.Y.
Accessions included the species L. tenuis, L. uliginosus, and
L. corniculatus. These accessions were acquired to be evaluated
for condensed tannins, as approved and funded by the Clover and
Special Purpose Legume CAC.
Methods
Seed were mechanically scarified and planted in a greenhouse in
April, 1992, then allowed to grow for 3 weeks. After 3 weeks,
plants were clipped to a uniform height of 10 cm and allowed to
grow for another month. After the initial clipping, plants were
allowed to grow for another month, then transplanted to the University
of Missouri Agronomy Research Center near Columbia, MO. The plants
were arranged in a randomized complete block design with three
replications, and 3 accessions per replication.
In September, 1992, foliage was harvested and stored at 5
C, freezedried, and ground to 1mm using a cyclone type grinding
mill. Dried material was analyzed using near infrared reflectance
spectroscopy, as reported by Roberts et al. (1992); condensed
tannins were quantified and expressed as % catechin equivalents
(CE). Significance of treatment effects was assessed at the 0.05
alfa level using analysis of variance techniques. Samples were
clustered by the single variable, tannin, according to the ScottKnott
nonoverlapping means separation technique.
Results
Data from this study indicated that tannins ranged from 1.67 to
11.44 9e CE. As expected from previous reports, L. uliginosus
contained higher tannin concentrations than other species.
In fact, L. uliginosus comprised all but one of the 42
accessions in the highest cluster, as well as the top 27 in the
next highest cluster. Only 9 L. uliginosus appeared in
the third cluster, and none appeared in the lowest cluster.
Also as expected, L. corniculatus contained both moderate
low levels of tannin concentrations. However, L. tenuis, a
species reported as tanninnegative, also contained moderate
and low levels.
These data will be entered in the GRIN system, as well as used
in our germplasm development studies at the University of Missouri.
Birdsfoot Trefoil (Lotus corniculatus) cultivars
Forage yields were obtained on four birdsfoot trefoil cultivars
at three Georgia locations: Mountains, Blairsville (2200 feet
elevation); Piedmont, Athens (980 feet); and Central, Eatonton
(490 feet). Plots 4 x 20 feet with 4 replications were planted
in September 1989 on prepared land.
| Pounds/acre oven dry forage | |||||
| Entry | |||||
| AU Dewey | |||||
| Bonnie | |||||
| Georgia 1 | |||||
| Norcen | 2683 c | 2953 a | 3072 b | ||
| Number of harvests | |||||
*Means within a column followed by the same letter are not significantly
different at the 5% level.
| Entry | |||||
| AU Dewey | |||||
| Bonnie | |||||
