ISSN 1510-7809
LOTUS NEWSLETTER 2004 Volume 34
Editor: M. Rebuffo INSTITUTO NACIONAL DE INVESTIGACION AGROPECUARIA Editor: M. Rebuffo INSTITUTO NACIONAL DE INVESTIGACION AGROPECUARIA
Editorial Office INIA La Estanzuela Colonia, Uruguay Phone: +598-574-8000 This Newsletter consists of informal Email: [email protected] reports which are presented to further Fax No.: +598-574-8012 the exchange of ideas and information Web: http://www.inia.org.uy/sitios/lnl/ between research workers. Consequently the data presented here are not to be used in publications Front cover: The photographs on the without the consent of the authors. front cover shows a root culture system, Images are copyright of the authors, super growing roots (SR), which was and their reproduction is strictly discovered by Akashi et al. (1998) from prohibited without their consent. Lotus corniculatus L. SR is now available for offer by Legume Base (pp. 26-29). Left: Growth of SR in a flask; middle-up: SR after 7 days of subculture; middle- down: Plant regeneration of SR; right: Plant regeneration of SR in a flask.
The opinions in this publication are those of the authors and not necessarily those of the Lotus Newsletter. The designations employed and the presentation of the material in this publication do not imply the expression of any opinion whatsoever on the part of the Newsletter concerning the legal status of any country, territory, city, or area, or of its autorities, or concerning the delimitation of its frontiers or boundaries. Where trade names are used this does not constitute endorsement of or discrimination against any product by the Newsletter. Lotus Newsletter. (2004). Volume 34.
Contents
Newsletter Announcements and Instructions ii
Personalia iv
Contributions: reports and short communications
P.R. BEUSELINCK. The Lotus Newsletter 1986 (No. 16) to 2001 (No. 32) 1
S. SAREEN. Seed production potential in birdsfoot trefoil (Lotus corniculatus L.) 5
W.F. GRANT. List of Lotus corniculatus (Birdsfoot trefoil), L. uliginosus/ L. pedunculatus (Big trefoil), L. glaber (Narrowleaf trefoil) and L. subbiflorus cultivars. Part 1. Cultivars with known or tentative country origin. 12
S. ISOBE and R. AKASHI. Bean’s Base: A new resource center of Lotus japonicus and Glycine max. 27
T.L. WANG. Lotus research at the John Innes Centre (JIC) and Sainsbury Laboratory (SL). 31
P. INSAUSTI. Substitution of Lotus glaber for the dicots of a natural grassland in the flooding Pampa of Argentina. 34
M. CAUHÉPÉ. Does Lotus glaber improve beef production at the Flooding Pampas? 38
R. LEEP. Birdsfoot trefoil evaluation at Michigan. 44
M.C. DE PABLO, M.S. BARUFALDI, P. SASTRE, H.N. CROSTA, M. ESEIZA and L. DAULERIO. Effect of the induced polyploidy on some characteristics of seed production and quality in Lotus glaber Mill. 45
G. MARTINOIA, A.M. DE HARO and M. BARUFALDI. Preliminary studies on the tolerance of an induced autotetraploid population of Lotus glaber Mill. (= Lotus tenuis) to Tetranychus sp: Tetranychidae (red mite). 51
K. MIYAKE, T. ITO, M. SENDA, R. ISHIKAWA, T. HARADA, M. NIIZEKI and S. AKADA. R2R3-MYB transcription factors from Lotus corniculatus var. japonicus differentially regulated in roots and shoots in response to nitrogen starvation. 54
O. RUIZ (Chairperson). Interdisciplinary workshop on genetic, molecular and ecophysiological aspects of Lotus spp. and their symbionts. [Taller interdisciplinario sobre aspectos genéticos, moleculares y fisioecológicos del Lotus spp. y sus simbiontes]. 60
SEBASTIÁN ARRIVILLAGA, SEBASTIÁN HERNÁNDEZ, JAVIER CILIUTI, MÓNICA REBUFFO, SILVIA GERMÁN and SILVINA STEWART. Evaluation of tetraploid big trefoil (Lotus uliginosus Schkuhr.) for rust resistance. 66
Lotus activities: Background and present research 73
Current list of Lotus researchers. Database last updated September 30 2004 79
Lotus literature. 108
Lotus Newsletter. 2004. Volume 34.
Newsletter Announcements and Instructions
The Lotus Newsletter has been published annually since 1971 by Dr. W.Grant, Dr. R.McGraw and Dr. P.Beuselinck, and it will be published by INIA, Uruguay in the future. It is intended as a worldwide communication link for all those who are interested in the research and development of Lotus species. Persons interested in lotus improvement, genetics, molecular biology, microbiology, production, marketing, or utilization are invited to contribute to the Lotus Newsletter. Previous issues may be used as a guide. It is expected that the work reported will be developed further and formally published later in refereed journals. It is assumed that contributions in LN will not be cited unless no alternative reference is available.
The web site is under transformation, since it will host two databases. The first database will be implemented for year 2004 and it will consist of all researchers’ information. The second database for the Literature will be built throughout year 2005. I would like to encourage everybody to make contributions to the list of bibliography for next year.
Acknowledgement
I would like to express my gratitude to Dr. Paul Beuselinck and Dr. William F. Grant for their permanent support and advice. We can provide electronic copies of all articles that are not available on the web, since Dr. Paul Beuselinck donated all his collection of LN issues. Thanks to Dr. Trevor L. Wang for his suggestions to improve the web site and to develop the “Lotus Network” (collective email list).
The priority of the Newsletter has been the update of the recipient list. I am pleased to inform that the registration increased from 119 researchers by the end of 2003 to 168 researchers in September 2004. Thanks to all for helping me with this task.
I would like to indicate that the session about Lotus literature is a merit of contributors. It is necessary to highlight the contribution of Dr. Jens Stougaard, who supplied an extensive list of bibliography since 1998, including manuals and reviews; Andrea Pedrosa-Harand, who sent the electronic publications and their web links; Søren Bak, Richard Leep, Yousef Papadopoulos, Trevor L. Wang, for providing the latest articles published; P.S. Nagar, who sent the latest publications as well as future book chapters; Mariana Kade, who works at the present time with wheat and sent her last publications about Lotus; William F. Grant, who is permanently contributing with information.
The life of the Newsletter depends on the contributions. Therefore, many thanks to all contributors and those that wrote the summary about their activities.
What to contribute?
Send us the kind of information you would like to see in LN. - Contributions should be current, scholarly, and their inclusion well-justified on the grounds of new information. - Results of recently concluded experiments, recent additions to germplasm collections,
iii Lotus Newsletter (2004). Volume 34.
information on new or tentative cultivars with descriptions. We want to include an adequate cultivar description, including disease reactions and origin if possible. - Notes on acreage, production, varieties, diseases, etc., especially if they represent changing or unusual situations. - Genome maps and information on probe-availability and sequences, and populations synthesized for specific traits being mapped. Glossy black and white prints of maps should be included, if possible. Partial maps can also be submitted. - Short reports of workshops, conferences, symposia, field days, meetings, tours, surveys, network activities, and recently launched or concluded project. - Details of recent publications, with full bibliographic information and short reviews. - Personal news (new appointments, awards, promotions, change of address, etc.)
How to format contributions
- Include the full address with telephone, fax, and e-mail numbers of all authors. - Keep the items brief – remember, LN is a newsletter and not a primary journal. - Give the correct Latin name of every crop, pest, or pathogen at the first mention. - If possible, table should fit within the normal typewritten area of a standard upright page (not a ‘landscape’ page). You may include figures and photographs (black and white or color). Please send disk-files (with all the data) whenever you submit line figures and maps. - Supply the essential information: round off the data-values to just one place of decimal whenever appropriate, choose suitable units to keep the values small (e.g. use tons instead of kg). - All lists of references should have been seen in the original by the author and year. Provide all the details such as author/s, year, title of the article, full title of the journal, volume, issue, and page numbers (for journal articles), and place of publication and publishers (for books and conference proceedings) for every reference. Incomplete references will not be accepted. - The language of the Newsletter is English, but we will do our best to translate articles submitted in other languages. Authors should closely follow the style of the reports in this issue. Contributions that deviate markedly from this style will be returned for revision, and could miss the publication date. If necessary, we will edit communications so as to preserve a uniform style throughout the Newsletter. This may shorten some contributions, but particular care will be taken to ensure that the editing will not change the meaning and scientific content of the article. Wherever we consider that substantial editing is required, we will send a draft copy of the edited version to the contributor for approval before printing. - Contact the Editor for detailed guidelines on how to format text.
Material may be submitted at any time during the year. Deadline for Volume 35 will be 30th June 2005. Please send all contributions and request for inclusion in the recipients list to:
Monica Rebuffo, Lotus Newsletter Editor, c/o INIA La Estancuela, 70000 Colonia, Uruguay Email [email protected] Fax +598-574-8012
iv Lotus Newsletter. 2004. Volume 34.
Personalia
Welcome to the new subscribers of the Lotus Newsletter: P.S. Nagar – India; Graeme Sandral and Mike Ewing – Australia; Nazhar-Ezzamane Noureddine – Algerie; Sangho Jeong, Jeffrey Dukes and Ann Hirsch – USA; Diego Risso, Walter Ayala, Georgget Banchero, Federico Condon, Martin Jaurena and Alejandro La Manna – Uruguay; José Manuel González, Ana María de Haro, Gabriela Martinoia, Ana María Castro, Mariana Melchiorre, Rodolfo Mendoza and Pedro Insausti – Argentina; Jeremy Murray, Martina Stromvik and Ken Richards – Canada; Juan Ramón Acebes Ginovés and Felicia Oliva Tejera – Spain; Francesca Cardinale – Italy; Katharina Pawlowski – Germany; Makoto Hayashi and Shinji Akada – Japan; Adem Kamalak – Turkey; Peter Kemp – New Zealand; Nick Brewin – United Kingdom.
Robert B. McDougall died in 2003. He was a graduate of the University of Saskatchewan, Saskatoon, Saskatchewan. He had a farm south of Edmonton, Alberta where he raised cattle and grew Lotus corniculatus as well as other experimental plants that had economic potential. He collaborated widely with many scientists in this endeavor on a number of crops. One collaboration was with W. F. Grant in which sulfonylurea resistant birdsfoot trefoil germplasm was developed. [Grant, W. F. and McDougall, R. B. 1995. Registration of a sulfonylurea resistant birdsfoot trefoil germplasm. Crop Science 35: 286-287.]
Several Lotus researchers have been recently reassigned or no longer work on the subject. Dr. Susan Wijting – The Netherlands, is presently working with fish; Dr. Carroll Vance – USA, do not have much research on Lotus at the present time, and most of his effort is directed towards Medicago, Lupinus and Phaseolus; the current research interest of Prof. Krystyna M. Urbanska – Switzerland, are focused on restoration ecology in high elevation ecosystems; Dr. Søren Borg, Leif Schauser and Gerhard Saalbach – Denmark, Olga Correa – Argentina, Dr. Helmi Schlaman – The Netherlands, are no longer working with Lotus.
The following personal information has been updated during the current year: Said Amrani – Algerie; Richard Leep, Joe Kirkbride, Joe Brummer, Gary Stacey, C.Jerry Nelson, James English, Gustavo Caetano, David Belesky and Nathan Hartwig – USA; David John Bertioli – Brazil; Ana Arambarri – Argentina; Daniel Real and Ariel Asuaga – Uruguay; Bjarne Jochimsen – Denmark; Andrea Pedrosa-Harand – Austria; Bernie Carroll - Australia; Niels Sandal, Kirsten Gausing – Denmark; Mark Robbins – United Kingdom; William F. Grant – Canada; Martin Parniske – Germany; Emanolis Flementakis – Greece.
v Lotus Newsletter (2004) Volume 34, 1-4.
The Lotus Newsletter 1986 (No. 16) to 2001 (No. 32)
PAUL R. BEUSELINCK* Research Geneticist, USDA-ARS, Plant Genetics Research Unit, 207 Waters Hall, University of Missouri, Columbia, MO 65211, U.S.A. http://www.psu.missouri.edu/lnl/ * Corresponding author.
When I started my Lotus career in 1980, I was very fortunate to have access to the Lotus Newsletter volumes that Dr. Bill Grant had published since 1970. I used those volumes 1-10 to get a perspective of the breadth of current Lotus research conducted internationally. After I had been in Lotus research for a few years, Dr. Grant indicated that he was looking for someone to take the editorial reins for a while. In 1985, Dr. Robert McGraw and I worked very closely on Lotus, even though he was located in Minnesota and I was located in Missouri. So we mutually agreed to share the editorship. Our first volume was the 1986 volume 16. We continued to co-edit the three volumes (vols. 16 – 18), before I took the sole responsibility for the next 13 volumes (vols. 19 – 32).
Our first attempts at editorship resulted in no real changes to the format as conceived and delivered by Dr. Grant. But sometime in the mid-1980s, the electronic revolution started with word processors, personal computers, laser printers, and magnetic disks. These new innovations gave us the chance to take contributions and bibliographic searches and restyle them into a common format. The changes that technology allowed were incorporated in the published volumes until 1995. In 1996 the costs of publishing and mailing concurrent with the widespread acceptance of the World Wide Web, instigated the web-based publication maintained at the University of Missouri, Columbia, Missouri (http://www.psu.missouri.edu/lnl/). From 1996 until 2001 (Vols. 27-32), the Lotus Newsletter was published on the web although I also managed to add previous volumes (Vols. 23-26). I believe the move for the Lotus Newsletter in 1996 from printed to electronic form was a difficult transition for many Lotus researchers and libraries. As humans, we like having hard evidence of our work or communication. Receiving the most recent printed copy of the Lotus Newsletter annually reaffirmed a bond among the Lotus researchers that I believe was lost with the electronic version. That bond was never stronger than when the 1st International Lotus Symposium was organized.
As editor of the Lotus Newsletter, I was privileged to observe the wide breadth of the research that was conducted around the world. Using that perspective, I chose to try to get the international body of Lotus spp. researchers together in order to share our research and insights. The outgrowth of our combined energies was the 1st International Lotus Symposium that was held in St. Louis, Missouri, 22-24 March 1994. A natural outgrowth of the
1 2 Paul R. Beuselinck symposium was a compilation of the research into a special publication “Trefoil: The science and technology of Lotus” published in 1999 by the Crop Science Society of America.
The cover illustration was changed starting with L. edulis in Volume 23. The illustrations used through Volume 32 were created by María Alejandra Migoya who works with Ana Arambarri in Argentina. They were most gracious in letting me use the illustrations. Maria’s illustration of L. corniculatus also graces the cover of “Trefoil: The science and technology of Lotus.”
The advances in Lotus science are documented in the pages (printed or web) of the Lotus Newsletter. The most common Lotus spp. reported were L. corniculatus, L. glaber, and L. uliginosus, usually involving agronomy, pathology, and physiology. Other, lesser known, Lotus spp. received ample attention from reports on evolution, taxonomy, and germplasm collections of the Lotus spp. Within the volumes of the Lotus Newsletter, you will find the first indications of a biotechnology revolution that would eventually identify L. japonicus as a model genetic system syntenic to other organisms.
One of the strengths of the Lotus Newsletter is the extensive bibliographic listing published every year. Many Lotus researchers work at locations or institutes where access to recent literature is difficult. The bibliographic listing provides a citation and abstract, if available.
I sensed a tangible loss when I was reassigned from Lotus research into soybeans in 2001. I miss the interaction and camaraderie with the many Lotus researchers I met and communicated with as editor for the Lotus Newsletter. The Lotus spp. comprise a fascinating genus, but no more fascinating than the cadre of scientists that has researched and compiled their knowledge about Lotus.
Highlights of volumes 16 – 32.
Volume 16. Several authors reported on botanical or chemical characteristics with interests in the origin, evolution, and adaptation of Lotus spp. Other papers reported physiological and reproductive observations on L. glaber, L. corniculatus, and L. uliginosus.
Volume 17. In 1986, rust was reported as being a new disease on L. corniculatus and L. glaber in Argentina. Several reports covered germplasm evaluations, genetic studies, and gene nomenclature.
Volume 18. Several reports reviewed the agronomic importance of L. glaber and L. corniculatus in Argentina. Cytogenetic and agronomic reports were received from Canada and Italy. Plant bug pests affecting L. corniculatus seed production were recognized in another report.
Volume 19. A considerable number of reports demonstrated the breadth of Lotus research across North and South America. Reports covered chemical defenses, reproduction, pollination, physiology, and agronomy topics. Lotus Newsletter vol.16 to 32 3
Volume 20. Reports on evolution of Lotus spp. in the French Alps and a plant exploration in Morocco offered new insights into Lotus speciation. Other reports covered tannins, taxonomy, germplasm, physiology, and agronomic topics.
Volume 21. Two reports covered tissue culture successes making somatic hybrids between Lotus and soybean (Japan) and herbicide tolerance selection (Canada). These were the first Lotus contributions in the new area called “biotechnology.” Agronomy, evolution, and flavonoid research rounded out the remaining reports.
Volume 22. More biotechnology reports were made in this volume and one was suggestive of L. corniculatus becoming a model plant system for genetic studies. Research in Australia was highlighted, but reports from Argentina, United Kingdom, Japan, Canada, and the United States demonstrated the international efforts on Lotus spp.
Volume 23. Additional reports from Australia were presented in this volume. Biotechnology reports on transformed L. corniculatus were received from China and the United Kingdom, while Japan reported on the somatic hybridization of Lotus and rice. The distribution of L. japonicus in Korea was reported. Agronomy, pathology, and flavonoid research rounded out the remaining reports.
Volume 24. Two reports outlined the use of PCR and RAPDs for Lotus research. A number of reports came from Uruguay and Argentina. Three reports described flavonoid or tannin research. Pathology, agronomy, and germplasm research rounded out the remaining reports.
Volume 25. This volume was published after the 1st International Lotus Symposium was held in St. Louis, Missouri, 22-24 March 1994. The symposium gathered the attention of most Lotus researchers and that attention carried over to this volume. A number of reports on L. glaber can be found in this volume. Research topics on Lotus spp. pathology, agronomy, biotechnology, genetics, and physiology comprise the reports.
Volume 26. This was the last volume published in a hard-copy format. Research topics on Lotus spp. germplasm, and L. corniculatus and L. glaber pathology, agronomy, biotechnology, genetics, and physiology comprised the reports.
Volume 27. Moving the Lotus Newsletter in 1996 from printed to electronic form was a difficult transition for many Lotus researchers and libraries. As humans, we like having hard evidence of our work or communication. The number of contributions declined when the printed version ended. Still, reports on genetics, agronomy, taxonomy, and biotechnology were received.
Volume 28. The majority of the reports came from Argentina and pertained to L. glaber, but some included L. corniculatus or L. uliginosus. A report from the International Association for Plant Taxonomy report the nomenclature changes for L. glaber (previously L. tenuis) and L. uliginosus (L. pedunculatus).
4 Paul R. Beuselinck
Volume 29. Lotus disease and insect feeding reports were contributed from Uruguay and Argentina. Plant physiology and chemistry rounded out the remaining reports.
Volume 30. Few reports were received, but all were from Argentina and generally pertained to L. glaber. A report on Lotus spp. taxonomy based on seedling comparisons was included.
Volumes 31 and 32. Only two reports were received for these volumes. One pertained to cultivar differentiation in L. glaber and the other to the dispersal of L. japonicus in Korea.
Lotus Newsletter (2004) Volume 34, 5-11.
Seed production potential in birdsfoot trefoil (Lotus corniculatus L.)
SINDHU SAREEN Indian Grassland and Fodder Research Institute, Regional Research Centre, CSKHPKV Campus, Palampur 176062 (HP) India
Abstract
Lotus corniculatus (birdsfoot trefoil) is a perennial forage legume for pastures and has a high nutritive value. It tolerates infertile, acidic, drought or wet soil conditions. It does not cause bloat even if grazed directly, because of the presence of condensed tannins. It is palatable nutritious, high in protein and very digestible to cattle, sheep and horses. Compared to other pasture species, the forage quality and protein content remains high for longer periods even after the plants start going to seed. However, it suffers from low seed yields and pod shattering has been found to be a major reason for it. During the present study, eighteen populations of Lotus corniculatus were collected from mid hill Himalayas and raised at IGFRI Regional Research Centre, Palampur. The populations were compared for their potential to set seed. The study revealed that in all populations observed seed set was far less than the expected seed yield. Further, it was found that pod shattering is not the sole reason for low seed set in birdsfoot trefoil. Other factors, such as the indeterminate period of flowering, flower bud abortion, lack of pollination and genetic factors may also contribute towards low seed yield.
Introduction
Lotus corniculatus is a perennial plant though not long lived, that has an extensive root system with strong tap root. Under favorable conditions, it may produce up to 100 stems and reach a height of 2 – 3 ft. The growth form can range from prostrate to erect with numerous stems arising from a well developed crown and branches arising from axils. Leaves are pentafoliate, alternately on short stalks with two leaflets at the petiole base resembling stipules.
It is a tetraploid (2n = 24) showing mostly bivalent pairing and tetrasomic inheritance. Flowering is indeterminate continuing for long time. Flowers are borne in an umbel at the end of a short stem. Each umbel consists of five bright yellow flowers. Flowers are bisexual and are largely cross pollinated by bees. Pressure is exerted by bee on the keel causing the staminal filaments to push a quantity of pollen out of tip. The pollen is exerted onto the ventral surface of the bee’s thorax and thereby it is carried from one flower to another resulting in pollination.
Lotus corniculatus is a forage legume for pastures having high nutritive value. It does not cause bloat when direct grazed, because of the presence of condensed tannins. It tolerates
5 6 Sindhu Sareen infertile, acidic, drought or wet soil conditions. It is palatable, nutritious, high in protein and very digestible to cattle, sheep and horses. Even after the plants start going to seed, forage quality and protein content remain high for much longer than other pasture species. However, it suffers from low seed yields and pod shattering has been identified as a major contributing factor. Lack of sufficient yield has prevented its spread (Anderson, 1955; McGraw and Beuselinck, 1983; Metcalfe et al., 1957). During this present study, 18 populations of Lotus were studied for their seed setting potential and to find out the factors responsible for the gap in expected and observed seed yields.
Material and methods
About 25 – 30 plants of eighteen populations of Lotus corniculatus were collected from the mid hills of Himalayas and were raised at IGFRI Regional research Centre, Palampur. In each population of twenty plants in the field, five plants were marked and data on umbel count, pod count and seed count per plant was collected. Data on umbel count and pod count was obtained in the field itself. When the plants were in full bloom, umbels on five plants in each population were counted and an average obtained. Similarly when plants attained maturity, the total number of pods (shattered as well as non shattered) on five plants in each population was counted and an average obtained. Ten non shattered pods from each population were removed and measured with scale to find out pod size. These pods were opened separately and seed/pod counted.
On each marked plant, ten umbels were marked to find ovule count/ovary and ovule fertility. For ovule fertility, the young buds from five umbels in each population were fixed in fixative (ethyl alcohol and acetic acid in the ratio 3:1) for 24 hrs. Then ovaries were separated and mounted in Lewis stain. The cover-slips were pressed gently and unfertilized ovules were counted. The buds from rest of five umbels fixed after pollination and fertilized ovules were counted.
Pollen fertility and chromosome count of these populations were also worked out using aceto- carmine squash technique.
Results and discussion
During the present study of 18 populations, it was found that the single ovary in each floret contains 20 to 46 ovules and within each population ovule number varied. Ovule fertility varied from 60 to 90% in these populations (Table 1). The pods were formed at right angles at the end of peduncle in the shape of a birdsfoot. Three to five pods were formed at each umbel. The pod size varied from 2.0 to 3.5 cm and each pod contained 14 to 25 seeds (Table 2). The total number of pods harvested from each plant varied from 7 to 307. Of these, 5 to 162 were shattered (Table 3) and rest were intact i.e. 0.84% to 79.5% were shattered and rest were intact (Figure1). The total seed harvested from a single plant varied from 112 to 4935.
Lotus corniculatus seed production 7
Table 1. Ovule count and fertility (%) in various Lotus collections.
Collection no. No. of ovules/ovary* Ovule fertility (%) L 125 35.4±2.05 86.40 L 126 37.1±2.32 89.77 L 127 38.9±1.02 67.57 L 128 34.0±1.53 79.11 L 129 37.7±2.00 81.85 L 132 39.4±1.57 75.29 L 133 29.4±2.01 66.45 L 134 36.6±2.13 60.30 L 135 38.3±0.94 85.29 L 136 37.1±2.02 86.39 L 137 37.4±1.91 73.35 L 139 34.9±1.90 64.75 L 141 30.9±1.83 84.06 L 142 35.1±1.93 79.60 L 391 34.3±1.42 86.32 W 1 38.9±0.80 73.69 W 2 39.1±1.08 87.63 W 3 40.0±0.99 90.50 *Mean ± S.E.
Table 2. Average data on pod parameters in various Lotus collections.
Collection No. of No. of No. of Pod Seeds/ Seeds/plant* no. umbels/ pods/ pods/ size(cm)* pod* Plant* Umbel* Plant* L 125 41±2.91 4±0.2 94±4.1 2.5±0.2 20.0±1.7 1257±91.8 L 126 52±2.98 4±0.3 119±7.1 2.6±0.1 22.2±2.8 2129±368.3 L 127 107±4.03 5±0 400±15.0 2.7±0.1 14.6±1.1 4935±409.0 L 128 45±2.81 4±0.2 102±6.1 2.4±0.2 15.0±1.8 1691±55.4 L 129 85±6.45 5±0 307±30.0 2.6±0.1 18.8±2.2 3843±293.4 L 132 77±6.19 5±0 295±15.2 2.8±0.2 21.8±3.0 2835±278.1 L 133 23±3.17 1.5±0.3 7±1.0 2.1±0.2 13.7±2.5 112±26.0 L 134 38±3.28 3.5±0.3 71±10.2 2.6±0.1 18.4±1.9 802±55.1 L 135 58±5.82 4±0.3 150±18.7 2.7±0.1 25.0±3.0 2485±497.5 L 136 44±3.34 3±0.4 80±9.5 2.6±0.2 22.8±1.5 949±106.6 L 137 37±2.86 3±0.4 56±7.1 2.6±0.2 16.6±1.9 763±75.3 L 139 42±6.27 3.5±0.3 76±7.3 2.4±0.2 15.6±1.5 910±316.0 L 141 39±1.92 3±0.4 66±8.9 2.2±0.1 13.6±1.3 480±45.4 L 142 53±4.69 4.5±0.2 149±13.2 2.5±0.1 19.2±2.7 1540±299.1 L 391 45±3.38 3±0.5 80±4.7 2.4±0.1 15.2±2.4 803±72.6 W 1 29±2.16 2.5±0.5 32±6.4 2.7±0.1 16.2±1.6 133±32.4 W 2 33±3.81 2.5±0.6 39±5.2 2.7±0.1 20.6±4.0 868±36.0 W 3 46±4.78 3.5±0.5 92±11.2 2.8±0.1 20.6±1.3 2146±270.3 *Mean ± S.E. 8 Sindhu Sareen
Table 3. Pod shattering (%) in various Lotus collections.
Collection Total no. of Shattered pods % age no. pods/plant* /plant* shattering L 125 94±4.1 52±3.1 55.32 L 126 119±7.1 17±3.7 0.84 L 127 400±15.0 149±8.8 37.25 L 128 102±6.1 39±6.3 38.24 L 129 307±30.0 162±27.1 52.77 L 132 295±15.2 162±10.4 54.92 L 133 7±1.0 5±1.4 71.43 L 134 71±10.2 45±6.3 63.38 L 135 150±18.7 83±8.0 55.33 L 136 80±9.5 26±4.7 32.5 L 137 56±7.1 14±2.1 25.0 L 139 76±7.3 37±3.7 48.68 L 141 66±8.9 43±3.5 65.15 L 142 149±13.2 90±8.0 60.40 L 391 80±4.7 27±1.8 33.75 W 1 32±6.4 22±4.2 68.75 W 2 39±5.2 31±3.8 79.49 W 3 92±11.2 69±6.2 75.00 *Mean ± S.E.
Figure 1. Branches of Lotus corniculatus bearing shattered and intact pods.
Lotus corniculatus seed production 9
During the present study it was found that the ovule count/ovary ranged from 20 to 46 and of these 14 to 25 developed into seeds. Earlier, Bader and Anderson (1962) reported that of 20 to 70 ovules/ovary of Lotus corniculatus, 2 to 35 develop into seeds whilst Hansen (1953) concluded that the average number of seeds per pod is 19.
Theoretically each plant has the potential to produce seed equivalent to fertile ovules/ovary x no. of pods/umbel x umbels/plant. However, during the present study, it was found that none of the populations produced as much seed as expected. Seaney and Henson (1970) reported that the gap between theoretical and actual seed yield is enormous. Seed or pod shattering is a serious problem in Lotus. Winch et al. (1985) stated that even though seed yields can potentially be as much as 750 kg/ha, the average yields obtained in Ontario are around 110 kg/ha i.e. 85% seed is lost due to shattering. Earlier, Winch and MacDonald (1961) have correlated three physiological stages of pod with seed development and shattering. Anderson (1955) and Metcalfe et al. (1957) considered relative humidity as the single most important factor affecting seed pod shattering. The time of pod shattering is related to the maturity of pods (Anderson, 1955; MacDonald and Winch, 1957), the environmental conditions and the individual genetic differences of the plants (Metcalfe et al. 1957).
The eighteen populations, which were studied, differed in the extent of pod shattering. This concurs with the work of Grant (1996), who reported considerable variation in degree of indehiscence among Lotus populations grown under greenhouse conditions. The pod shattering in 18 populations ranged from 0.84% to 79.5% but the observed seed yields are 6 to 37% of expected yield. Even if, the seed loss due to shattering (no. of shattered pods x average no. of seeds/pod) is taken into consideration, the observed seed yields are still 8 to 49% of the expected yield and on average only 35% seed yield is obtained (Table 4). This suggests there is still a gap of 51% to 92% in seed yield attributable to other factors. This can be attributed to indeterminate period of flowering and at the time of harvesting all the pods are not at same stage of maturity. During the present study, all the populations except L 391 started blooming in the month of March and were harvested in May. Since population L 391 came into bloom after harvesting of other populations, it was harvested in July. The other factors, which could be responsible for low seed yields are flower bud abortion, lack of pollination and genetic factors. Another interesting feature that emerges from the present study is the enormous variability among populations. Significant variability for pod shattering, number of umbels/plant was recorded. In the future this can be exploited for isolation of lines with lesser pod shattering and higher seed yield.
10 Sindhu Sareen
Table 4. Expected and observed seed yield/plant in various Lotus collections.
Collection Seed yield/plant Seed loss Expected Gap (%) in No. Expected 1 Observed due to seed yield 2 seed yield shattering3 L 125 5016 1257 1040 3976 68.4 L 126 6928 2129 377 6551 67.5 L 127 14063 4935 2175 11888 58.5 L 128 4842 1691 585 4257 60.3 L 129 13115 3843 3046 10069 61.8 L 132 11421 2835 3532 7889 64.1 L 133 674 112 68 606 81.5 L 134 2935 802 828 2107 61.9 L 135 7577 2485 2075 5502 54.8 L 136 4231 949 593 3638 73.9 L 137 3045 763 232 2813 72.9 L 139 3318 910 577 2741 66.8 L 141 3035 480 585 2450 80.4 L 142 6671 1540 1728 4943 68.8 L 391 3996 803 410 3586 77.6 W 1 2080 133 356 1724 92.3 W 2 2829 868 639 2190 60.4 W 3 5828 2146 1421 4407 51.3 Average 5644.7 1593.4 1126 4518.7 64.7 1 Expected seed yield (1) = Fertile ovule x average no. of umbels/plant x average no. of pods/umbel 2 Expected seed yield (2) = Expected seed yield (1) – seed loss due to shattering 3 Seed loss due to shattering = average no. of seed /pod x no. of shattered pods/plant
References.
ANDERSON S.R. 1955. Development of pods and seeds of birdsfoot trefoil (Lotus corniculatus L.) as related to maturity and to seed yields. Agronomy Journal, 47, 483 – 487.
BADER K.L. and ANDERSON S.R. 1962. Seed yields of birdsfoot trefoil, Lotus corniculatus L. as affected by pre-harvest clipping combined with control of injurious insects. Agronomy Journal, 54, 306 – 309.
GRANT W.F. 1996. Seed pod shattering in the genus Lotus (Fabaceae): a synthesis of diverse evidence. Canadian Journal of Plant Science, 76, 447 – 456.
HANSEN H.W. 1953. Developmental morphology of Lotus corniculatus L. Iowa State College Journal of Science, 27, 563 – 600.
MACDONALD H.A. and WINCH J.E. 1957. Birdsfoot trefoil seed production. Department of Agronomy, New York State Coll. Agric., Cornell Univ., Ithaca, NY no. 57-18, Mimeographed. Lotus corniculatus seed production 11
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 relation between pod dehiscence, relative humidity and moisture equilibrium in birdsfoot trefoil, Lotus corniculatus. Agronomy Journal, 49, 130 – 134.
SEANEY R.R. and HENSON P.R. 1970. Birdsfoot trefoil. Advances in Agronomy, 22, 119 – 157.
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.
WINCH J.E., ROBINSON S.E. and ELLIS C.R. 1985. Birdsfoot trefoil seed production. Factsheet no. 85-111, Ontario Ministry of Agriculture and Food, Toronto, ON pp: 1 – 4.
Lotus Newsletter (2004) Volume 34, 12-26.
List of Lotus corniculatus (Birdsfoot trefoil), L. uliginosus/ L. pedunculatus (Big trefoil), L. glaber (Narrowleaf trefoil) and L. subbiflorus cultivars. Part 1. Cultivars with known or tentative country of origin.
WILLIAM F. GRANT Department of Plant Science, P.O. Box 4000, Macdonald Campus of McGill University, 21,111 Lakeshore Blvd., Ste. Anne de Bellevue, Quebec H9X 3V9, Canada
Table 1. Birdsfoot trefoil (Lotus corniculatus) cultivars and germplasm.
Cultivar Characteristics AC Langille Canada. Superior to Leo in forage yield and seedling vigor in Atlantic Canada. AG-S4 USA. Autogamous plant, no breeding vigor depression compared to MU- 81; prolific seed producer; PI 566818 (Steiner, 1993) Agrosan Trueno Uruguay, registered in Italy Albena Italy Alina Romania Ames 654 USA. PI 340799 Anneforsdammena Anta ARS-2620 USA. First birdsfoot trefoil cultivar with rhizomes, excellent plant vigor, improves long-term stand persistence (Beuselinck and Steiner, 1996). Named Steadfast; PI 542503 ARS-2622 USA. Rhizome-producing population with a broad genetic base; USDA- ARS and Missouri Agric Expt. Station; PI 631492 Au Dewey USA. Prostrate growth habit, good vigor and adaptation; excellent stand persistence; tolerates low pH; high yield potential in southeast US; Alabama selection; developed at Auburn University; PI547080 Ayaksa Baco Italy Balkányia Bästhulta Bérbaltavária aCultivars given in Eurisco (http://eurisco.ecpgr.org). (Continued)
12 List of known Lotus cultivars 13
Table 1. (Continued)
Cultivar Characteristics Biria Bokor Yugoslavia (Nikolic et al., 1997) Bonnie France Borbányaia Boronkaia Bosnalotus Yugoslavia Bull Canada. Leo characteristics; higher yielding, high survival under soil compaction; eastern Canadian selection developed from synthetics by University of Guelph Bursztyn Poland. PI 255176 CAD USA. Missouri, germplasm selected for drought resistance from MU-81 germplasm PI 570669 (Beuselinck and Steiner, 1994) Carroll USA. Iowa, Winter-hardy pasture type; more upright and maturing slightly earlier than Empire; good seedling vigor, adapted to imperfectly drained soils; Iowa-developed; PI 600727 Cascade USA. Early maturing, semi erect to erect; not as winterhardy as Viking; developed from French selections; principal variety for Washington and Oregon; PI 421026 Cateascaa Cornelia Germany Corona de Rey Spain. PI 518447 Cotton ‘alien’ UK. Naturalized (Bullard and Crawford, 1996) Cree Canada. Seed yield superior to Leo; winter-hardiness comparable to Leo and Empire; western Canadian, selected at Saskatoon; PI 592424 Csárdaszállásia Csengeria Cserebökényi ökotipusa CZEBES99a - CZEJES00a - CZEPOD00a - aCultivars given in Eurisco (http://eurisco.ecpgr.org). (Continued) 14 William F. Grant
Table 1. (Continued)
Cultivar Characteristics Dawn USA. Semi-erect Empire type; good forage yields and fall growth, selected for increased resistance to root rots and leaf and stem diseases; wide adaptation within the North Central US; developed by USDA at Missouri; PI 570670 Debrecenia Decubens Greece Dedinovskij Russia Delpon France Dikorastuscija Dikorastushchiia Dikorostuchiia Dinamoa Douglas USA. Vigorous, fast growing; an Oregon-European type selection. Eddei Russia (Nikolaichuk and Yalovs’ka, 1996) Egervölgyia El Boyero MAG Argentina Emlyn United Kingdom Empire USA. New York State. First birdsfoot trefoil cultivar in North America; semi-erect, flowers 10-14 d later than European types; moderately winter hardy; selected for persistence Erdôbényeia Érpatakia Estanzuela Ganador Uruguay. Grown in Argentina, Brazil. Exact Canada Fabio Italy Farmosia Fedikovskiia Fehérgyarmatia Fergus USA. Naturalized semi- prostrate ecotype from Kentucky, good seedling vigor, disease tolerant; adapted to southeast US; PI 592428 (Taylor, 1985). Franco Italy (Mariotti et al., 1984) aCultivars given in Eurisco (http://eurisco.ecpgr.org). (Continued) List of known Lotus cultivars 15
Table 1. (Continued)
Cultivar Characteristics Frilo Italy Fülesdia Gacsályia Gazal Boyn Uzu Turkey. PI 383686 Gelsvis Russia (Mukhina et al., 1972) Gelyovila Georgia 1 USA. Erect-type Mediterranean germplasm that had persisted for many years at Griffin, Georgia; NSL 195509 Geszterédia Ghizdeia Giada Italy Ginestrino Italy (Verna, 1986) Gkeskenylevelu Hungary Granger USA. Oregon, Upright growth habit, good seedling vigor; similar forage production and winterhardiness to Cascade; same source as Cascade; NSL26530 Gran San Gabriele Italy. Derived from San Gabriele Grasslands Goldie New Zealand. Australia Gülzower Germany Gülzower Hornklee Germany Gyömrôia H401-4-4-2 Canada. Macdonald Campus, McGill University, Montreal, source of resistance to the sulfonylurea herbicide thifensulfuron-methyl (Du Pont, DPX-M6316; Harmony; PI 578075) Hoki Belgium, France Horuschottenuleea Ilisestia INIA Draco Uruguay. Erect type, selected for persistency (Rebuffo and Altier, 1997) Jaki Hungary Jaquari Brazil. PI 260011 Jatcinskij Russia aCultivars given in Eurisco (http://eurisco.ecpgr.org). (Continued) 16 William F. Grant
Table 1. (Continued)
Cultivar Characteristics Jiberta Jygevaa K-30, K-37 Serbia and Montenegro KBG Morocco. PI 631539 Kalo USA. Oregon-selected from Dwarf English variety (L. corniculatus var. arvensis ) for grazing, ground cover, and soil erosion; PI 234670 Kállósemjéniva Kemenesszentmárton ia Királyegyházia Kiskôrösia Kisszekeresia Kölcseia Kömörôia Komorvoia Kubanskija Late Roskilde Denmark. PI 237278 Leo Canada. Quebec, Early spring vigor, good winter-hardiness, and seed yield; developed from USSR selection (Morshansk) at Macdonald Campus, McGill University, PI 306182 Lika Croatia Listan Russia (Nikolaichuk and Yalovskaya, 1997) Livada Romania Lotanova Italy Lot Poland Lota Ötoftea Lotar Czechoslovakia Lövôia Mackinaw USA. More vigorous in seedling stage, higher yielding in mature plant stage than Empire; Iowa selected; NSL 85662 Madison France aCultivars given in Eurisco (http://eurisco.ecpgr.org). (Continued) List of known Lotus cultivars 17
Table 1. (Continued)
Cultivar Characteristics Magyaregregyia Magyia Maikopskii Russia (Nikolaichuk and Yalovskaya, 1997) Maitland Canada. Seedling vigor and forage yield greater than Viking, adapted to fair and well-drained soils; University of Guelph selection Malejovsky Czechoslovakia. PI 259512 Mándia Mansfield USA. Early cultivar, NSL 26579 Marcalia Mazelovsky Kraja Melejovskya Mestecanisa Mestnyia Mezôkeresztesia Mezônagymihályia Mirabel Canada. Growth habit intermediate to Leo and Viking, developed from USSR selection (Moscow) at Macdonald Campus, McGill University Mishkovcia Mistseviia MO-20 USA. Germplasm, forage yields higher or equal in nine states, Missouri selection, NSL 13787 Morshanskii Russia. PI 258467 (Chapurin, 1986) Moskovskii Russia. (Mukhina et al., 1972) MU-81 USA. Highly heterogeneous germplasm composed of 3 germplasms and 13 cultivars providing a single genetic source with a broad base, PI 592425 Nagycenki Hungary Nagyecsedi Hungary Nagykállói Nagyszállási Hungary (Nyíregyháza) aCultivars given in Eurisco (http://eurisco.ecpgr.org). (Continued) 18 William F. Grant
Table 1. (Continued)
Cultivar Characteristics Nagyszállási Hungary (Nyíregyháza) ökotipus Nagyszekeresi Hungary Napkoria Napkori ökotipusa Nässjöa Nasu, No. 2 Japan. PI 286468 Nemesborzovaia Nico Romania Nigardelen Norway ecotype Norcen (NC-83) USA. Missouri, Broad-leaved, intermediate type; good forage yield and winter-hardiness; diverse genetic background provides resiliency to different environments within North Central US, PI 592427 Norfolk UK ecotype Nottingham UK ecotype Nueltin USA. Minnesota. Broad-leaved, good spring vigor and excellent winter hardiness. Developed from Leo for glyphosate tolerance for control of Canada thistle; greater seed production, PI 630976 (Ehlke et al., 2003b) Nydalaa Nyíradonyia Nyírbátoria Nyíregyházia OAC Bright Canada. 2% higher seedling vigor and 0.8% higher herbage yield than Leo. Similar maturity and persistence to Leo. Developed at Guelph Oberhaunstadter Germany Odenwale Russia (Andriyanova et al., 1991) Odenwälder Germany. Belgium, France Olaszfaia Old Town USA. NY State, PI 472006 Orosia aCultivars given in Eurisco (http://eurisco.ecpgr.org). (Continued) List of known Lotus cultivars 19
Table 1. (Continued)
Cultivar Characteristics Órségi Hungary. Plants develop rapidly, erect type, good yield, winter hardy but liable to lodge Otocaca Otoftea Óvária Pardee USA. Cornell Univ. developed against fungi, Fusarium wilt; blooms about 10 days earlier than Norcen, PI 634338 (Viands et al. 2004) Pecölia Penyigeia Periceia Perugia 2 Italy. PI 260693 Pietramalo Firenzi Italy. PI 251528 Pishik Yonjasia- Podgorje K'Mjaka Poiana stampeia Polbes99a Polom Slovakia Poltata Populatsiaa Primorskii Russia (Andriyanova et al., 1991) Pskovskij mestnij Russia Pulawska Poland Puspokbolyia Puszta Hungary Quimey Chile RG-BFT USA. Photoperiod insensitive and rapid-flowering autogamous stock (Steiner and Beuselinck, 2001) PI 613539 Reus Kedelmia Rimnicu Vilceaa Rocco Germany aCultivars given in Eurisco (http://eurisco.ecpgr.org). (Continued) 20 William F. Grant
Table 1. (Continued)
Cultivar Characteristics Rodeo France Roseau USA. Minnesota. Intermediate growth habit between Viking and Empire; less winter hardy than Leo, Nueltin, and Carroll, but more hardy than Empire, Fergus and Steadfast. Developed from Norcen for glyphosate tolerance for control of Canada thistle, PI 630977 (Ehlke et al., 2003a) Roskilde Denmark Rosnovsky Kraj Czechoslovakia Rucheroberger Austria. PI 231123 Schotenkler Rumia Sacela Sacel-Ticeroa San Gabriel Brazil. Grown in Uruguay, Argentina, naturalized in Alabama, PI 433923 San Gabriele Italy Simleul Silvanieia Skrzeszowicka Poland Smolens’kii 1 Russia (Nikolaichuk and Yalovskaya, 1997) Somogysimonyia Sorkifaludia Stadil Kirkebya Stanici Russia (Chapurin, 1986) Steadfast See ARS-2620. Stripe 13 Italy. PI 258446 Sturovnik Viglasskya Susan Italy SVKBES99a SVKBUR00a SVKCER99a SVKGEM 98a SVKKAR96a SVKKRE99a aCultivars given in Eurisco (http://eurisco.ecpgr.org). (Continued) List of known Lotus cultivars 21
Table 1. (Continued)
Cultivar Characteristics SVKPOL 96a SVKROH00a SVKSIT 97a Szabadszentkirályia Szabolcs-1 Hungary Szatmari Hungary Szentesia Szentlorincea Szentlôrincia Szirondia Szkrzeszowickaa Szombathelyi Hungary Szomogytúria T-68 USA. Germplasm, 2 to 5 times more tolerant to 2,4-D than Viking, released by USDA and Cornell Táborski Czechoslovakia. Plants resemble Órségi but have firmer stalk, PI 259513 Tana G 19670. See Tretana Targovistel Bulgaria Tatry 9a Terminillo Italy. PI 251827 Timisoaraa Tótújfaluia Trebiczki Czechoslovakia. Flowers 2 to 3 days earlier than Táborski., PI 259514 Tretana USA. Winter-hardy in pasture and hay trails in Montana, replacing Tana; naturalized selection released by Montana Agr. Exp. Sta. and SCS, USDA TREVIG USA. Wisconsin. High yielding, excellent seedling vigor Troubskaa Tygevoky Estonia. PI 285278 Tunyogmatolcsia Turgovishtea aCultivars given in Eurisco (http://eurisco.ecpgr.org). (Continued) 22 William F. Grant
Table 1. (Continued) Cultivar Characteristics Turistvándia Újfehértóia Újszôlôskerti (Kállósemjén) ökotipusa Ukrkara Undia Upstart Canada. Leo characteristics; higher seedling vigor; developed from synthetics by University of Guelph V.Podolyanskiia Varzarii de Josa Vega USA. California: Erect type selected from San Gabriel by Northrup King Co. Vega II USA. California: increased seedling vigor, winter growth, and yield Vicaria Brazil. PI 260013 Vigrassky Czechoslovakia (Uhrikova, 1978) Vígl'assky Kraja Viking USA. Erect, early maturing, 15%-20% greater forage yield persists where Empire types may be lost due to competition; adapted to fair and well- drained soils. Derived from Danish and New York ecotypes at Cornell University; PI 310483 Vilmorina France Virginia Syn No 10 USA, NSL 95724 Wellington Canada; good regrowth Viseul de susa Vladaiaa Vojnica Winnar USA. G22514 WITT USA. Wisconsin. High yielding, large seeded, excellent seedling vigor X 93007-32 China. PI 631598 Yaki szarvaskerea Reported in Ross and Jones (1985) Zamosteava Zora Yugoslavia (Nikolic et al., 1997) aCultivars given in Eurisco (http://eurisco.ecpgr.org). List of known Lotus cultivars 23
Table 2. Big trefoil (Lotus uliginosus/ L. pedunculatus) cultivars and germplasm.
Cultivar Characteristics Barsilvi New Zealand. Tetraploid similar to Maku. Beaver USA. Diploid, smooth or glabrous-leafed, adapted to wet winter and cool summers of coastal Oregon and Washington; developed by Oregon Agric. Exp. Stn. Columbia USA. Diploid, pubescent leaves and stems, adapted to wet winter and cool summers of coastal Oregon and Washington; developed by Oregon Agric. Exp. Stn. Grasslands Maku New Zealand. Tetraploid (2n = 4x = 24); perennial, rhizomatous persisting vegetatively; sown on acid infertile soils in coastal regions of eastern Australia. Grasslands Sunrise New Zealand. Diploid with improved autumn growth, better adapted for semi- intensive to intensive grazing. Kaiser USA. Diploid, suited as a pasture legume in mixed pasture on the fragipan soils of the lower Ohio River valley; developed at Purdue. Marshfield USA. Diploid, survives frequent flooding in winter and is tolerant of brackish overflows. Sharnae Australia. Diploid; morphologically similar to Grasslands Maku, except plant is less hairy. More robust and bulky than Grasslands Maku (Bowman, 1993)
Table 3. Narrow leaf trefoil (Lotus glaber Mill. = L. tenuis Waldst. Et Kit. Ex Wild.) cultivars.
Cultivar Characteristics Aguape Argentina. Angostura Uruguay. Chajá Plus Argentina. Larrañaga Uruguay. Los Banos California. Matrero Uruguay. Pampa INTA Argentina. Toba Argentina. Tresur Chajá Argentina. Soils within a wide range of pH and salt concentrations (Mujica and Rumi, 1999)
24 William F. Grant
Table 4. Lotus subbiflorus cultivar.
Cultivar Characteristics El Rincón Winter annual in Uruguay. Adapted to wide range of soils; low fertilizer requirement; slow establishment and initial growth but adapted to continuous pasture with excellent persistence.
Acknowledgments
I thank the many individuals who gave me information on the Lotus cultivars, including Ian D. Thomas, Genetic Resources Unit, IGER, Aberystwyth, Ceredigion SY23 3EB, Wales; Graeme Sandral, University of Western Australia, 1 Underwood Drive, Shenton Park, Perth, WA 6009, Australia; Richard Leep, Department of Crop and Soil Sciences, Michigan State University, Kellogg Biological Station, 3700 E. Gull Lake Drive, Hickory Corners, MI 49060 and Tatiana E. Kramina, Moscow State University, Biological Faculty, Higher Plants Department, Vorobyevy Gory, 1, Building 12, 199992 Moscow, Russia. I give special thanks for the tremendous help from Mónica Rebuffo, INIA La Estanzuela, Forage Department, Ruta 50 Km 11, 70000 Colonia, Uruguay.
References.
ANDRIYANOVA N.A., PLOTNIKOVA N.V. and TSELOVAL'NIKOV I.K. 1991. Birdsfoot trefoil - a new leguminous crop for pasture in central Ciscaucasia, Novye idei v rastenievodstve i puti ikh realizatsii, Voronezh, 9-13 iyulya, Moscow, Russia, 20-21. [Russian]
BEUSELINCK P.R. and STEINER J.J. 1994. Registration of CAD birdsfoot trefoil germplasm selected for drought resistance. Crop Science, 34, 543.
BEUSELINCK P.R. and STEINER J.J. 1996. Registration of ARS-2620 birdsfoot trefoil. Crop Science, 36, 1414.
BOWMAN A.M. 1993. Sharnae - a new Lotus pedunculatus for Australia. Lotus Newsletter, 24, 13-16.
BULLARD M.J. and CRAWFORD T.J. 1996. Seed yield, size and indeterminacy in diverse accessions of Lotus corniculatus L. grown in the UK. Plant Varieties and Seeds, 9, 21-28.
CHAPURIN V.F. 1986. Initial material for breeding birdsfoot trefoil. Nauchno-tekhnicheskii Byulleten' Vsesoyuznogo Ordena Lenina i Ordena Druzhby Narodov Nauchno- issledovatel'skogo Instituta Rastenievodstva Imeni N. I. Vavilova, 160, 17-20. [Russian].
EHLKE N.J., WYSE D.L. and VELLEKSON D.J. 2003a. Registration of ‘Roseau’ Birdsfoot Trefoil. Crop Science, 43, 732-733. List of known Lotus cultivars 25
EHLKE N.J., WYSE D.L. and VELLEKSON D.J. 2003b. ‘Nuetlin’ birdsfoot trefoil. Crop Science, 43, 733-734.
MARIOTTI D., PEZZOTTI M., FALISTOCCO E. and ARCIONI S. 1984. Plant regeneration from leaf - derived callus of Lotus corniculatus L. cv. Franco. Genetica Agraria, 38, 219-223.
MUJICA M.M. and RUMI C.P. 1993. Effect of chemical and mechanical scarification of Lotus tenuis seeds on germination. Lotus Newsletter, 24, 32-34.
MUKHINA N.A., GORBACHEVA R.G. and KORZHENEVSKAYA N.I. 1972. Effects of early cutting on yields and nutritive value of red clover, lucerne and birdsfoot trefoil. Trudy Po Prikladnoi Botanike Genetike i Selektsii, 48, 131-137. [Russian].
NIKOLAICHUK V.I. and YALOVS'KA G.I. 1996. A character of inheritance of the synthesis of cyanoglucosides in birdsfoot trefoil. Dopovidi Natsional'Noyi Akademiyi Nauk Ukrayiny, 4, 140-142. [Russian].
NIKOLAICHUK V.I. and YALOVSKAYA A.I. 1997. Biochemical composition of the green matter and yield in different varieties of birdsfoot trefoil. Fiziologiya i Biokhimiya Kul'Turnykh Rastenii, 29, 209-214. [Russian].
NIKOLIC R., MITIC N. and NESKOVIC M. 1997. Evaluation of agronomic traits in tissue culture - derived progeny of birds-foot trefoil. Plant Cell Tissue Organ Culture, 48, 67- 69.
REBUFFO M. and ALTIER N. 1997. Breeding for persistence in Lotus corniculatus L. In Proceedings of The XVIII International Grassland Congress. Winnipeg, Manitoba, Saskatoon, Saskatchewan, Canada. 8–19 June 1997. ID 1881.
ROSS M.D. and JONES W.T. 1985. The origin of Lotus corniculatus. Theoretical and Applied Genetics, 71, 284-288.
STEINER J.J. 1993. Registration of AG-S4 autogamous broad-leaf birdsfoot trefoil germplasm. Crop Science, 33, 1424-1425.
STEINER J.J. and BEUSELINCK P.R. 2001. Registration of RG-BFT photoperiod insensitive and rapid-flowering autogamous birdsfoot trefoil genetic stock. Crop Science, 41, 607- 608.
TAYLOR T.H. and TEMPLETON Jr. W.C. 1985. Registration of 'Fergus' birdsfoot trefoil. Crop Science, 25, 712.
UHRIKOVA A. 1978. Index of chromosome numbers of Slovakian flora (Part 6). Acta Fac. Rerum Nat. Univ. Commenianae, Bot., 26, 1-42.
26 William F. Grant
VERNA M. 1986. Composizione chimica e valore nutritivo del fieno di ginestrino (Lotus corniculatus L.). Annali Dell’Istituto Sperimentale Per la Zootecnia, 19, 29-35.
VIANDS D. R., MILLER-GARVIN J.E., HANSEN J.L., BERGSTROM G.C., TILLAPAUGH B.P., LOWE C.C., THOMAS E.M. and NEALLY J.L. 2004. Registration of ‘Pardee’ Birdsfoot Trefoil. Crop Science, 44, 1488-1489.
Appendix.
The author would be pleased to have additions or errors brought to his attention. It would be useful to have the origin of the cultivars for which no data have been given. Further information will be published in the Lotus Newsletter as Part 2. Lotus Newsletter (2004) Volume 34, 27-30.
Legume Base A new resource center of Lotus japonicus and Glycine max –
SACHIKO ISOBE1* and RYO AKASHI2 1 National Agricultural Research Center for Hokkaido Region, Hitsujigaoka 1, Toyohira, Sapporo, 062-8555, Japan 2 Faculty of Agriculture, University of Miyazaki, Miyazaki 889-2192, Japan * Corresponding author.
1. What’s Legume Base
Legume Base (http://www.legumebase.agr.miyazaki-u.ac.jp) was set up in April 2004 as a resource center for Lotus japonicus and Glycine max. The aims of Legume Base are development, collection, conservation and offer of the genetic resource of L. japonicus and G. max, to facilitate the material utilization by the research community.
Legume Base is organized by a core facility, Miyazaki University, and a sub facility, Hokkaido University. Miyazaki University is aimed at the collection, conservation and offer of the L. japonicus genetic resource and Hokkaido University is aiming to handle the G. max genetic resource. Some parts of the reproduction and investigation of data on characteristic features are carried out by following facilities on commission: National Agricultural Research Center for Hokkaido Region, Nihon University College of Bioresource Sciences and RIKEN Plant Science Center. The foundation of Bean’s Base is supported by the National Bioresource Project in Japan (http://www.shigen.nig.ac.jp/shigen/nbrp/nbrp.jsp).
2. Resource type
Lotus japonicus
We distribute two experimental lines, accession lines collected throughout Japan, recombinant inbred lines and a root culture system. The activation tag lines and the EMS mutants are being developed and reproduced for distribution.
One of the experimental lines, Gifu B-129, was established by Stougaard et al. (1996), and another experimental line, Miyakojima MG-20, was established by Kawaguchi (2000). These lines are widely used throughout the world. The accession lines were collected throughout Japan and donated by Japanese researchers (Figure 1). We have reproduced 90 accessions, of
27 28 Sachiko Isobe and Ryo Akachi which 53 lines are available for distribution at present (July 2004).
Figure 1. Phenotypes of the Accessions lines.
The recombinant inbred lines, LjMG RILines, were made by crossing Gifu B-129 and Miyakojima MG-20 (Figure 2). These lines were self pollinated to the F8 generation at Kazusa DNA Research Institute, starting from the F2 seeds offered by Dr. M. Kawaguchi. The number of lines in the final set is going to be 205, of which 172 lines are available for distribution at present (July 2004). The typing data for a total of 48 SSLP markers distributed along the six chromosomes are available from the Kazusa DNA Research Institute (http://www.kazusa.or.jp/lotus/RIline/).
Figure 2. Phenotype segregation of the LjMG RI Lines. Legume base 29
The root culture system, super growing roots (SR), was discovered by Akashi et al. (1998a) from Lotus corniculatus L., which grows efficiently after removing the above-ground organs and cultured in a medium containing no plant hormone (Figure 3). This root possesses high regeneration competence. Also, protoplasts can be extracted easily and these protoplasts show a vital proliferation. These characteristics are maintained until now after 5 years have passed since the discovery (Akashi et al. 1998b, 2000). The SR can be used in physiological researches of the root, and also in functional analysis of genes by transformation.
Figure 3. A root culture system: Super growing root.
Glycine max
Cultivated accession lines, wild accession lines which were collected throughout Japan, RILines between Misuzudaizu and Moshidou Gong 503 and mutants of fatty acid composition are being developed and reproduced for distribution.
3. Conditions for providing the resources
The National Bioresource Project provides resources to domestic and foreign researchers on the basis of the conclusion of the material transfer agreement (MTA). The MTA form will be sent to the researcher by e-mail after the receipt of the request order form. The researcher is then asked to return the MTA after filling the necessary items. The requested resource will be sent to the researcher after the receipt of the MTA. The user of the resource should clarify that the research material was provided by the National Bioresource Project (Lotus japonicus, Glycine max) in his/her presentations and publications in the "materials and methods" or the "acknowledgment" section. Also, the user of the ‘LjMG RILines’ should mention that the lines were established at Kazusa DNA Research Institute. For a resource of which the depositor submits a special condition on its use, additional documents will be sent. 30 Sachiko Isobe and Ryo Akachi
4. To make an order
The order is accepted through the web site. To make an order and for more detail information, please check the web site,
http://www.legumebase.agr.miyazaki-u.ac.jp
References
AKASHI R., HOFFMAN-TSAY S.S. and HOFFMAN F. 1998a. Selection of a super-growing legume root culture that permits controlled switching between root cloning and direct embryogenesis. Theoretical and Applied Genetics, 96, 758-764.
AKASHI R., UCHIYAMA T. and HOFFMAN F. 1998b. High-frequency embryogenesis from cotyledons of bird's-foot trefoil (Lotus corniculatus) and its effective utilization in Agrobacterium tumefaciens-mediated transformation. Journal of Plant Physiology, 152, (1) 84-91.
AKASHI R., HARRIS S., HOFFMAN-TSAY S.S. and HOFFMAN F. 2000. Plants from protoplasts isolated from a long-term root culture (Super Root) of Lotus corniculatus. Journal of Plant Physiology, 157, 215-221.
AKASHI R., KAWANO T., HASHIGUCHI M., KUTSUNA Y., HOFFMAN-TSAY S.S. and HOFFMAN F. 2003. Super roots in Lotus corniculatus: a unique tissue culture and regeneration system in a legume species. Plant and Soil, 255, 27-33.
KAWAGUCHI M. 2000. Nodule Organogenesis in Lotus japonicus. Journal of Plant Research, 113, 507-509.
STOUGAARD J. and BEUSELINCK P.R. 1996. Registration of GIFU B-129-S9 Lotus japonicus germplasm. Crop Science, 36, 476.
Lotus Newsletter (2004) Volume 34, 31-33.
Lotus research at the John Innes Centre (JIC) and Sainsbury Laboratory (SL)
TREVOR L. WANG Metabolic Biology Department, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, United Kingdom
The John Innes Centre has a long and pre-eminent history of research on legumes based around the science of nitrogen fixation and the genetics and molecular biology of peas (Casey and Davies, 1993). More lately, research on legumes worldwide has focussed on the two models amenable to modern genomics analysis, Medicago truncatula (www.medicago.org) and Lotus japonicus (www.lotusjaponicus.org). Both the John Innes and Sainsbury Laboratory have turned their attention towards these models as well.
At our site on the Norwich Research Park, research on Lotus has concentrated on symbioses, both with rhizobium to form nodules and with fungi to make arbuscular mycorrhizae. Much effort, however, has also been made to develop platforms to accelerate our own research and to help the community. A common component to both the symbioses is signalling and extensive work is carried out studying genes involved in the plant signalling pathways that are involved, especially in the overlap between the two (Kistner and Parniske, 2002). Three groups are involved directly with Lotus: Martin Parniske’s (SL), Trevor Wang’s (Metabolic Biology, JIC) and Allan Downie’s (Molecular Microbiology, JIC) and collaborations exist between all three groups (e.g. Stracke et al., 2002). In the Parniske group, the early signalling events in arbuscular mycorrhizal interactions are the most important, particularly those that are common to both symbioses (Downie and Parniske, 2002). In the Downie group, the centre of attention is calcium spiking (and its role in nodulation signalling; Oldroyd and Downie, 2004), and mutants affected in early stages of infection thread growth. The Wang group is focused on later metabolic events, especially carbon partitioning enzymes such as sucrose synthase (Craig et al., 1999).
Trevor Wang and Martin Parniske have collaborated on an exciting new development for legumes – TILLING (Targeting Induced Local Lesions IN Genomes; Colbert et al., 2001). This technology was pioneered in plants by the Henikoff group in Seattle. We have developed a similar platform for Lotus japonicus over the last three years (Perry et al., 2003). We set up an ethylmethane sulphonate (EMS) mutation programme to generate a population of about 5000 M2 families that would be used in both forward screens and to collect DNA samples. The initial ca. 40000 plants were screened for a large number of different phenotypes. A further group of about 1400 M2 families was screened for starch mutants to use in research on sugar metabolism (http://www.jic.bbsrc.ac.uk/staff/trevor-wang/starchscreen.ppt). This collection has been entered onto a web-accessible database for people to browse and order mutants (http://data.jic.bbsrc.ac.uk/cgi-bin/lotusjaponicus/). In total more than 60000 M2 individuals have been screened for nodulation-defective mutants. Each M2 family had one
31 32 Trevor L. Wang representative that was used to collect DNA and this DNA has been employed subsequently for developing the reverse genetic, TILLING, tool.
TILLING allows you to identify plants bearing a mutation in your gene of choice. It relies on the ability of the CEL1 endonuclease to detect mismatches between normal and mutant DNA strands when they are annealed. CEL1, which can be isolated readily from celery, permits the detection of single point mutations of the type induced by chemicals, such as EMS, that are conventionally used in mutation breeding programmes.
Using ‘Coddle’ a bioinformatics tool developed by the Seattle groups, regions of your favourite gene can be selected where it is more likely that a mutation will cause a change in the corresponding protein’s activity. DNA primers can then be engineered to amplify only that portion of the gene. The DNA from the M2 individuals represents the whole spectrum of variation in the population and is mixed with DNA from normal plants. Special fluorescently- labelled DNA ‘primers’ are used to amplify specific regions of the known gene in which a mutant is desired. The amplified DNA is then heated and cooled to separate and re-anneal the strands. The DNA is then cleaved with CEL1 at the mismatch between the normal strand and the mutant strand. The strands are then separated on a DNA sequencing gel. Since the enzyme can only cleave in a single direction, the two cleaved strands have reciprocal sizes that can easily be recognised on the gel from their fluorescence and position thus identifying the DNA sample and the corresponding plant with a mutation in the gene of interest. This plant is then used to study the function of the gene or it can be used directly and incorporated into a breeding programme. The whole process can be viewed in cartoon format at http://www.lotusjaponicus.org/Illustration.html (remember to have sound on!).
Collaborations like this one underpin much of the work at the Centre and have led to a successful bid for an EU Marie Curie Research and Training Network, INTEGRAL (www.lotusjaponicus.org/integral), coordinated by Martin Parniske. Its predecessor, LOTUS, finished this year and will culminate in the production of ‘The Lotus japonicus Handbook’ that details background information and techniques to aid research on this model plant. It is hoped that the volume will be published next year.
References.
CASEY R. and DAVIES D.R. 1993. Peas: Genetics, Molecular Biology and Biotechnology. CAB International, Wallingford. 323 p.
COLBERT T., TILL B.J., TOMPA R., REYNOLDS S., STEINE M.N., YEUNG A.T., MCCALLUM C.M., COMAI,L. and HENIKOFF S. 2001. High-throughput screening for induced point mutations. Plant Physiology, 126, 480-484.
CRAIG J., BARRATT P., TATGE H., DÉJARDIN A., HANDLEY L., GARDNER C.D., BARBER L., WANG T.L., HEDLEY C.L., MARTIN C. and SMITH A.M. 1999. Mutations at the rug4 locus alter the carbon and nitrogen metabolism of pea plants through an effect on sucrose synthase. Plant Journal, 17, 353-362.
Lotus research at JIC and SL 33
DOWNIE J.A. and PARNISKE M. 2002. Fixation with regulation. Nature, 420, 369-370.
KISTNER C. and PARNISKE M. 2002. Evolution of signal transduction in intracellular symbiosis. Trends in Plant Science, 7, 511-518.
OLDROYD G.E. and DOWNIE J.A. 2004. Calcium, kinases and nodulation signalling in legumes. Nature Reviews. Molecular Cell Biology, 5, 566-576.
PERRY J.A., WANG T.L., WELHAM T.J., GARDNER S., PIKE J.M., YOSHIDA S. and PARNISKE M. 2003. A TILLING reverse genetics tool and a web accessible collection of mutants of the legume Lotus japonicus. Plant Physiology, 131, 866-871.
STRACKE S., KISTNER C., YOSHIDA S., MULDER L., SATO S., KANETO T., TABATA S., SANDAL N., STOUGAARD J., SZCZYGLOWSKI K. and PARNISKE M. 2002. A plant receptor-like kinase required for both bacterial and fungal symbiosis. Nature, 417, 959- 962.
Lotus Newsletter (2004) Volume 34, 34-37.
Substitution of Lotus glaber for the dicots of a natural grassland in the flooding Pampa of Argentina.
PEDRO INSAUSTI IFEVA – CONICET, Facultad de Agronomía, Universidad de Buenos Aires, Av. San Martín 4453, CP1417DSQ Buenos Aires, Argentina.
Introduction
Gap opening and colonization are considered substantial mechanisms of community dynamics (Grubb, 1977). They represent opportunities for new trade-off between the species already present or for newcomers to be established. Every new gap represents an available volume of resources (light, water and nutrients) for the encircling plants and for new individuals naturally dispersed or purposely introduced in it (Tilman, 1982).
The objective of this work was to assess the ability of Lotus glaber to colonize gaps opened as a result of dicots death and to appraise the degree to which an individual species is able to fill the niche of a whole guild (dicots) and even to enlarge another guild (graminoids). Under the effect of livestock grazing, the grasslands of the Pampa lowlands, currently dominated by a rich array of grass species, modify their structure leaving way to an increase in cover of dicots (Sala et al., 1986), most of them introduced species of low forage value. Flooding, a frequent but aleatoric disturbance in this area, has the opposite effect, namely, the removal of most of the introduced dicots and also of some native herbs (Insausti et al., 1999), resulting in the opening of a high number of gaps.
The hypothesis was that L. glaber introduced immediately after gap opening, would outcompete the dicots (Tilman, 1982) associated to its ability to fix atmospheric N. Also, this ability would enlarge the graminoids niche, more than compensating for the elimination of dicots competition. Thus, the hypothesis embodies the assumption that L. glaber would fill the role of a keystone or an engineer species in the modified system (Lawton and Jones, 1995; Power et al., 1996).
Materials and Methods
The experiment was carried out in a natural grassland located in flooding Pampa (Argentina). Two kinds of manipulations were generated inside an exclosure: 1) removal of dicots with herbicide application and 2) introduction of L. glaber, resulting in a combination of four treatments: (a) grassland without dicots, (b) grassland without dicots in which L. glaber was introduced, (c) L. glaber intersown controls (with dicots) and (d) undisturbed controls, arranged in a 2 x 2 factorial randomized blocks design. Six 3 x 6 m experimental plots were assigned at random to each treatment. Seeds of L. glaber were intersown one month after herbicide application.
34 Lotus glaber establishment 35
Seedling density of L. glaber was recorded 10 days after sowing; plant density and the number of shoots per plant was recorded three times: 2, 6 and 28 months after sowing. Aboveground living biomass was sampled in each plot 12 months after sowing (autumn); the clipped material was sorted to dicots, graminoids and L. glaber and was dried at 70 ºC for 72 hours and weighed. Dried aboveground biomass from each guild was analyzed for total tissue N. The measure of total nitrogen content of plant material was determined with the Kjeldahl-method. Data were analyzed by means of the single classification analysis of variance. The density data were transformed by the factor √(x+0.5), to attain the ANOVA assumptions.
Results and Discussion
Seedling density of L. glaber 10 days after sowing was similar in plots with dicots or without them, but differences in the number of established plants between those two treatments progressively increased up to 28 months (Table 1). The number of shoots per plant was larger (P<0.01) six months after sowing in plots in which dicots had been killed, but this difference was not apparent 28 months after.
Table 1. Plant density and number of shoots per plant of Lotus glaber, 2, 6 and 28 months after sowing in plots with dicots or without them. Values followed by different letters are significantly different (P<0.05).
Time Plants/m2 Shoots/Plant With Dicots Without Dicots With Dicots Without Dicots 2 months 210a 306b 1h 1h 6 months 173c 225d 1.1h 1.7i 28 months 4f 48g 30j 29j
Twelve months after sowing (autumn), aboveground biomass of L. glaber in plots previously treated with herbicides was 25 times higher than in plots with dicots (Table 2). At this time, dicots biomass was significantly lower in intersown plots, with herbicides applied treatment, than all the others (P<0.01; Table 2). The biomass of grasses was larger in plots in which dicots had been killed than in those in which they were not (Table 2). However, grasses biomass in plots in which herbicides had been applied as the only treatment (no L. glaber sowing), differ from those with L. glaber sown after herbicides application (P<0.05). Total aboveground biomass in this last treatment was also larger than all the others (P<0.01; Table 2). Content of N expressed as the amount of the element per unit area was higher in graminoids when dicots were killed by herbicides application (Table 3). Moreover, N content in total biomass and in graminoids biomass was higher in plots with L. glaber, previously treated with herbicides than in the other three treatments (Table 3).
36 Pedro Insausti
Table 2. Aboveground biomass (g.m-2) twelve months after sowing (autumn). Values are means of six replicates with standard errors in brackets.
Herbicide Control Herbicide Control +L.glaber +L.glaber Graminoids 71 (2.2) 39.1 (2.1) 60.7 (1.9) 38 (1.3) Lotus glaber 32.4 (1.8) 1.3 (0.2) ------Dicots 1.6 (0.3) 24.5 (2.5) 5.6 (0.6) 26.3 (0.9) Total 105 (5.3) 64.9 (3.3) 66.3 (3.8) 64.3 (2.7)
Table 3. Total nitrogen expressed as the amount of the element content per unit area (mg.m-2). Values followed by different letters are significantly different (P<0.05).
Herbicide Control Herbicide Control +L.glaber +L.glaber Graminoids 1.95a 0.86b 1.54c 0.84b Lotus glaber 0.81b 0.03d ------Dicots 0.12e 1.02b 0.36f 1.12b Total 2.88 1.91 1.90 1.96
In these grasslands, L. glaber introduced after gap opening outcompeted the dicots guild. Moreover, biomass enhancement of graminoids is interpreted as the result of interaction of the facilitation mediated by L. glaber (Quinos et al., 1998) and the release of competition exerted by the dicots guild. L. glaber reduces the shortages in N supply and enhances vegetative growth of graminoids, resulting in an increase in the competitive ability of this guild. It is on these grounds that L. glaber might be considered as a keystone species in the managed grassland.
References.
GRUBB P.J. 1977. The maintenance of species-richness in plant communities: The importance of the regeneration niche. Biological Reviews, 52, 107-145.
INSAUSTI P., CHANETON E.J. and SORIANO A. 1999. Flooding reverted grazing effects on plant community structure in mesocosms of lowland grassland. Oikos, 84, 266-278.
LAWTON J.H and JONES C.G. 1995. Linking species and ecosystems: organisms as ecosystem engineers. In JONES C.G. and LAWTON J.H. (Eds.) Linking species and ecosystems. Chapman and Hall, New York. pp. 141-150.
POWER M.E., TILMAN D., ESTES J.A., MENGE B.A., BOND W.J., SCOTT MILLS L., DAILY G., CASTILLA J.C., LUBCHENCO J. and PAINE R.T. 1996. Challenges in the quest for keystones. BioScience, 52, 609-620.
Lotus glaber establishment 37
QUINOS P., INSAUSTI P. and SORIANO A. 1998. Facilitative effect of Lotus tenuis on Paspalum dilatatum in a lowland grassland of Argentine. Oecologia, 114, 427-431.
SALA O.E., OESTERHELD M., LEÓN R.J.C. and SORIANO A. 1986. Grazing effects upon plant community structure in subhumid grasslands of Argentina. Vegetatio, 67, 27-32.
TILMAN D. 1982. Resource competition and community structure. Princeton University Press, Princeton, New Jersey, USA. 296 pp.
Lotus Newsletter (2004) Volume 34, 38-43.
Does Lotus glaber improve beef production at the Flooding Pampas?
MIGUEL CAUHÉPÉ* Private Researcher and Consultant on Range and Pasture Management, Cereijo 979, 7620 Balcarce, Prov. de Buenos Aires, Argentina * Corresponding author.
Introduction
The Flooding Pampas is an extensive and flat region whose natural grasslands offer an appropriate set of nutrients for the beef cow-calf operations. More than 50% of the beef calves of Argentina are produced in almost 6 million ha of this area. Although most of the ranches of the Flooding Pampas have introduced some improved technologies regarding cattle and grasslands management, the low forage quality in winter, and the frequent and extensive floodings it suffers, keep extensive and conservative management as a dominant feature.
The natural grasslands of the Flooding Pampa are dominated by perennial grasses that could be classed in two groups according with their seasonal growth: winter-spring grasses and summer grasses. Winter growth is limited by low solar radiation and temperature. Growth rates of about 4-5 kg MS/ha/day are common. During spring and summer, growth reaches 20- 30 kg DM/ha/day and it extends through the autumn until air temperature decreases (Figure 1). Primary productivity of the main forage communities ranges among 6000 and 2500 kg DM/ha/year along the soil gradient they grow (Hidalgo and Cauhépé, 1991a). Forage quality varies markedly with its minimum values during winter and top quality during spring and summer.
25
20
day 15
10 kg DM/ha/day kg/DM/
5
0 Winter Spring Summer Fall
Figure 1. Forage production of a flooding Pampa grassland (Source: Hidalgo and Cauhépé, 1991a)
38 Lotus glaber and beef production 39
Lotus (Lotus glaber) was introduced long ago in the Flooding Pampa and naturally expanded favored by floodings. At present, covers an extense area. Ranchers see Lotus as a valuable species and a significant number of researches give some support to this rancher’s opinion (Mazzanti et al., 1988; Miñón et al., 1990). Thus, the objective of this review is to evaluate the possible impact of Lotus glaber on the beef cattle production of the Flooding Pampas through the research results produced on the region.
Could Lotus improve the forage production?
The Nitrogen fixation of Lotus growing in a fescue (Festuca arundinacea) pasture varied between 39-42 kg N/ha/year (Refi et al., 1989). Lotus coverage ranged in this experiment from 7-24%. As in temperate grasslands nitrogen is related directly to forage production, the presence of Lotus is expected to increase the forage production of grasslands. The winter forage production of a fescue-lotus mixture (Miñón et al., 1990) almost tripled the daily growth rates: of native grasslands: 15 kg DM/ha/day as compared with 4-5 kg DM/ha/day (Hidalgo and Cauhépé, 1991a). In other experiment conducted in humid native grassland, Hidalgo and Rimoldi (1992) found higher forage production in a pasture with Lotus than others without Lotus and attributed this result to the N fixation of Lotus.
Could Lotus improve the forage quality?
Native grassland at the Flooding Pampa has good nutritional quality for breeding cattle all year around except during winter. Pigurina et al. (1998) showed that pregnant heifers grazing native grasslands of Uruguay lost between 15-20 kg live weight from June through August.
Figures 2 and 3 shows the diet quality of steers grazing a native grassland of the Flooding Pampa and the impact of Lotus on the forage quality of a native grassland. It is apparent that the main effect of Lotus on the nutritive value of grasslands is during the winter months when the diet reaches their poorest nutritional values. Moreover the impact on the pregnant cows and especially on pregnant heifers is stressed as a consequence of the high nutrient requirements of the final pregnancy and early lactations phases.
Rosso and Chifflet (1991) found an increase of live weight gains of steers grazing a tall wheatgrass without and with lotus from 400 g/head/day to 700 g/head/day, respectively. Freddi et al. (1991) and Rosso and Gómez (1995) also found improvement of nutritional quality related with the presence of Lotus in pastures of the Flooding Pampa.
40 Miguel Cauhépé
80 3
70 2.5 60 2 50 Digest. In vitro 40 1.5 Nitrogen % 30 1 Nitrogen % 20
Dry matter digestility % 0.5 10 0 0 Winter Spring Summer Fall
Figure 2. Digestibility and Nitrogen content (%) in diets of steers grazing a native grassland of the flooding Pampa (Source: Hidalgo and Cauhépé, 1991b)
Further comments on the possible impact of Lotus on beef production
So far, the available data allows us to tentatively support the hypothesis that the increase of Lotus in the Flooding Pampa grasslands will have a positive effect on beef production as a result of the expected increase in forage production and in forage quality. Moreover it seems to be other processes related to internal parasites on which Lotus genera has anti-helmintic effect: the condensed tannins, common in the Lotus genera, would compensates the loss of protein absorption as a consequence of the action of internal parasites (Otero 1).
The resistance of Lotus glaber to flooding has been proved by Vignolio and Fernández (1994) and Vignolio and Fernández (1997). Lotus plants formed aerenchyma which provided them the oxygen necessary to survival and growth under flooding. Also the lower phosphorus requirements of Lotus is an economic advantage o this specie. The response to phospahte fertilizers has been erratic (Quadrelli de Escuder et al., 1997). Resistance of Lotus to soil salinity has been shown by Bañuelos and Beuselinck (2003).
Lotus promotion through phosphate fertilization and herbicides, grazing management or aerial post-burning seeding (Juan et al., 2000) and sod seeding (Miñón and Colabelli, 1993; Laterra, 1997) could be a ways to increase its biomass and the beef production in Flooding Pampa grasslands.
1 OTERO M.J. Efecto de distintas especies forrajeras que contienen taninos condensados sobre la productividad de ovinos parasitados. Monografía. Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires, Argentina. 22 p. [Spanish] Lotus glaber and beef production 41
90
80
70
60
50
40
30
20
10
0 Fall Winter Spring Summer With Lotus Without Lotus
Figure 3. Effect of Lotus glaber on the forage quality (dry matter digestibility, %) of a flooding Pampa grassland (Source: Hidalgo and Rimoldi, 1992)
Considering beef cattle management, the nutrition of pregnant premature heifers, especially during their last third of pregnancy, the nutritive value of native Flooding Pampa grasslands is inadequate in terms of metabolizable energy concentration and protein content (Hidalgo and Cauhépé, 1991b; Pigurina et al., 1998). The use of native grasslands with Lotus has been successful to manage this kind of heifers, although no scientific demonstration is available. However, it will be necessary to conduct more intensive research to prove the overall advantages in beef production of the inclusion of Lotus on native grasslands under the Flooding Pampa conditions.
References
BAÑUELOS G.S. and BEUSELINCK P.R. 2003. Growth of three forage species in saline conditions. Arid Land Research and Management, 17, 13-22.
FREDDI A.J., ESCUDER C.J., CHIFFLET DE VERDE S., ROSSO O.R. and. CANGIANO C.A. 1991. Evaluación de la ganancia de peso, consumo, selectividad de la dieta y ambiente ruminal en novillos sobre una pastura de Lotus tenuis y Thynopyrum ponticum. Simposio Argentino del Género Lotus, Chascomús, Intech, pp 22-23. [Spanish] 42 Miguel Cauhépé
HIDALGO L. and CAUHÉPÉ M.A. 1991a. Producción de forraje de las comunidades forrajeras de la Pampa Deprimida. Revista de los CREA, 145, 58-64. [Spanish]
HIDALGO L. and CAUHÉPÉ M.A. 1991b. Effects of seasonal rest in a above-ground biomass for a native grassland of the Flooding Pampa, Argentina. Journal of Range Management, 44, 471-475.
HIDALGO L.G. and RIMOLDI P.O. 1992. Lotus tenuis en pastizales templado sub húmedos: su efecto en el valor nutritivo de la vegetación. Congreso Latinoamericano de Ecología. 2. (Resúmenes), Carambú, Mina Gerais, Brasil, disc 6-11. SEB. pp. 540-542. [Spanish]
JUAN V., MONTERROSO L., SACIDO M. and CAUHÉPÉ M. 2000. Postburning legume seeding on a Paspalum quadrifarium community in the Flooding Pampas, Argentina. Journal of Range Management, 53, 300-304.
LATERRA P. 1997. Post burn recovery in the Flooding Pampa: impact of an invasive legume. Journal of Range Management, 50, 274-277.
MAZZANTI A.E., MONTES L. and MIÑÓN D.P. 1988. Utilización de Lotus tenuis en establecimientos ganaderos de la Pampa Deprimida: resultados de una encuesta. Revista Argentina de Producción Animal, 8, 301-305. [Spanish]
MIÑÓN D.P. and COLABELLI M.R. 1993. Intersiembra de Lotus tenuis en tres comunidades nativas de la Pampa Deprimida. Revista Argentina de Producción Animal, 13, 133-140. [Spanish]
MIÑÓN D.P.G., SEVILLA H., MONTES L. and FERNÁNDEZ O.N. 1990. Lotus tenuis: leguminosa forrajera para la Pampa Deprimida. EEA Balcarce, INTA, Argentina. Boletín Técnico, N° 98, 15 pp. [Spanish]
PIGURINA G., BEMHAJA M. and BRITO G. 1998. Utilización de heno de campo natural diferido en areniscas de Tacuarembó. II Degradación ruminal y relación con comportamiento animal. INIA, Uruguay, Serie Técnica, N° 94, 127-130. [Spanish]
QUADRELLI DE ESCUDER A.M., LAICH F.S., ANDREOLI Y.E. and ECHEVERRÍA H.E. 1997. Respuesta de Lotus tenuis Waldst a la inoculación con Rhizobium loti y a la fertilización fosfatada. Ciencia del Suelo, 15, 22-27. [Spanish]
REFI R.O., NAVARRO C.A. and ESCUDER C.J. 1989. Estimaciones de la fijación simbiótica de nitrógeno en pasturas bajo pastoreo de trébol blanco, Lotus tenuis y festuca alta. Revista Argentina de Producción Animal, 9, Sup 1, 31-36. [Spanish]
ROSSO O.R. and CHIFLET S. 1991. Ganancia de peso en pastura mezcla de Agropiro y Lotus tenuis y agropiro solo (Resumen). Simposio Argentino del Género Lotus, Chascomús, Intech. p.21. [Spanish] Lotus glaber and beef production 43
ROSSO O.R. and GÓMEZ P.O. 1995. Selectividad de novillos en una pastura de agropiro y Lotus tenuis. Revista Argentina de Producción Animal, 15, 364-366. [Spanish]
VIGNOLIO O.R. and FERNÁNDEZ O.N. 1994. Efecto del anegamiento en invierno y verano sobre el crecimiento y supervivencia de Lotus tenuis y L. corniculatus. Ecología Austral, 4, 19-28. [Spanish]
VIGNOLIO O.R. and FERNÁNDEZ O.N. 1997. Cambios anatómicos y morfológicos en tallos de Lotus tenuis y L. corniculatus (Leguminosae) generados por el anegamiento. Boletín de la Sociedad Argentina de Botánica, 32, 147-151. [Spanish]
Lotus Newsletter (2004) Volume 34, 44.
Birdsfoot trefoil evaluation at Michigan.
RICHARD LEEP Crop and Soil Sciences, Michigan State University, W. K. Kellogg Biological Station, 3700 E. Gull Lake Drive, Hickory Corners, MI 49060, U.S.A.
Table 1. Experiment description.
Location: Lake City Exp. Station, Lake City Design: RCB, plot size: 3 x 23(3 x20 harvested) Seeded: May 2001, 12 lbs PLS/A. Soil Type: Kent Silt loam Cuttings: One in 2001, two in 2002
Table 2. 2001 Lake City Birdsfoot trefoil Variety Trial.
DM tons/acre 2003 2002 2001 3-yr. Variety Marketer/Breeder 17-Jun 24-Jul Total Total Total Total
AU Dewey public 0.96 0.58 1.54 1.70 0.71 3.95 Viking public 1.03 0.45 1.48 1.78 0.62 3.88 Leo Werner Farm Seeds 0.84 0.46 1.30 1.85 0.69 3.84 OAC Bright Univ. of Guelph 0.97 0.46 1.43 1.77 0.64 3.84 Norcen public 0.88 0.53 1.41 1.80 0.62 3.83 Dawn Deer Creek Seed 0.85 0.48 1.33 1.86 0.63 3.82 Mirabel public 0.85 0.48 1.33 1.74 0.72 3.79 Empire public 0.85 0.51 1.36 1.68 0.61 3.65 Georgia One Deer Creek Seed 0.84 0.42 1.26 1.68 0.62 3.56 Pardee Cornell Univ. 0.78 0.39 1.17 1.59 0.70 3.46
Mean 1.36 1.74 0.65 3.76 CV 30 9 15 12 LSD NS 0.23 NS 0.42
Further evaluation trials: http://www.msue.msu.edu/fis/extension_documents/extpub01.pdf
44 Lotus Newsletter (2004) Volume 34, 45-50.
Effect of induced polyploidy on some characteristics of seed production and quality in Lotus glaber Mill.
M.CRISTINA DE PABLO*, MÓNICA S. BARUFALDI, PATRICIA SASTRE, HILDA N. CROSTA, MARCELO ESEIZA and LORENA DAULERIO Departamento de Ciencias Básicas Agronómicas y Biológicas, Laboratorio Regional de Análisis de Semillas y Granos, Facultad de Agronomía de Azul, UNCPBA. * Corresponding author.
Introduction
In 1998 in the Faculty of Agronomy in Azul, UNCPBA, a research project was initiated aimed at the production of artificially induced tetraploids in L. glaber, using colchicine as the polyploidising antimitotic agent. In this way, tetraploid germplasm was obtained that derived from the chromosomal duplication of seedlings belonging to four naturalised populations found in the Province of Buenos Aires (Barufaldi et al., 2000). Chromosomal counts have been carried out in this tetraploid population, and they continue to be made in order to verify the stability of the ploidy level of different random-crossing cycles derived from it.
The induced polyploids have been used in some forage species such as Dactylis glomerata ssp. lusitanica, Lolium perenne, Lolium multiflorum, Trifolium pratense, Trifolium hybridum, Lotus pedunculatus and Secale cereale, with the purpose of obtaining improved autotetraploid cultivars or to generate interspecific hybrids. In general, the induced autotetraploids in ryegrass, rye, clovers and Lotus pedunculatus show better establishment, higher in vitro digestibility and forage production, and better performance in response to such adverse factors as disease, frost and drought, than the corresponding diploids (Evans, 1955; Ahloowalia, 1971; Armstrong, 1974; Simonsen, 1976; Luchini, 1979; Dennis, 1980; Pérez, 1980; Herrera et al., 1988; Carámbula et al., 1994).
However, gamete fertility of induced autotetraploid plants is reduced compared with that of the diploid forms. The cause of this decline and, therefore, of the reduction in seed production, is mainly due to meiotic irregularities that lead to the formation of chromosomally unbalanced gametes. These gametes are commonly sterile or, on those occasions where their fertility permits zygote formation, produce viable abnormal zygotes that give origin to aneuploids of reduced fertility. In the induced tetraploid forms of species like red clover and rye, seed yields increased after the first generations, due to a both natural and artificial selection. Nevertheless, seed production was considerably lower than in similar diploid forms (Dennis, 1980; Funes et al., 2001).
The present work analyses the effect that chromosomal duplication in Lotus glaber produces on the following characteristics related to seed production and quality: - Number of flowers per umbel (size of the umbel).
45 46 M.Cristina De Pablo, Mónica S. Barufaldi, Patricia Sastre, Hilda N. Crosta, Marcelo Eseiza and Lorena Daulerio - Number of pods per umbel. - Number of seeds per pod. - 1000 seed weight. - Percentage of normal seedlings (germinative power) - Percentage of hard seeds. - Percentage of ungerminated soft seeds. - Percentage of abnormal seedlings. - Percentage of dead seeds.
Material and methods
The research was carried out in the experimental field of the Faculty of Agronomy of Azul, using spaced planted. The evaluations were carried out on diploid plants of a naturalised population denominated "Azul" and plants of the tetraploid germplasm under study.
Determination of the number of flowers per umbel and the number of pods per umbel. Starting from 20th January 2002, samples were taken at random: - 284 and 254 umbels in the tetraploid and the diploid populations, respectively, to determine the number of flowers per umbel. - The same number of umbels with mature pods were harvested at random the following week for the pod count.
Determination of the number of seeds per pod, 1000 seed weight and seed quality. The parameters of seed quality were evaluated in both cytotypes on the samples harvested previously, in the following way: - 20 pods were taken at random from each population and the number of seeds per pod counted. - 1000 seed weight was determined taking the average of eight repetitions of 100 seeds each, according to the rules of the International Seed Testing Association (ISTA, 2002).
Samples were taken at random for the determination of seed quality for each cytotype. After mechanical scarification, 100 seeds per tray were laid out with 4 repetitions (400 seeds) and were pre-chilled during 7 days at 8º C. Later on, they were located in a germination camera during 12 days at 20º C (according to ISTA norms). The percentage of normal and abnormal seedlings, as well as the percentage of hard, fresh and dead seeds, were recorded at the end of this period.
Statistical Analysis Pearson chi-square analysis was performed for testing the independence between the number of flowers per umbel and cytotype, as well as between the number of pods per umbel and cytotype. A t-test was carried out to compare the means of the variables: number of seeds per pod and the characteristics of seed quality in both cytotypes.
Seed production of tetraploid Lotus glaber 47
Results and discussion
The existence of a dependence relationship between the number of flowers per umbel and the diploid and tetraploid cytotypes was detected by means of the chi-square test.
The highest frequency for both cytotypes corresponded to 5 flowers per umbel (Table 1). However, the diploid cytotype showed a distribution closer to normal than did the tetraploid cytotype, with the latter presenting a higher concentration of umbels with low numbers of flowers. Also, 2.4% of the umbels of the diploid had 8 -10 flowers, while the tetraploid did not produce umbels that had more than 7 flowers.
Table 1. Number of flowers per umbel in the tetraploid and diploid cytotypes of L. glaber Mill.
Cytotype Number of flowers per umbel
1 2 3 4 5 6 7 8 9 10 Total
Tetraploid
Absolute frequency 6 23 45 86 90 31 3 0 0 0 284 Relative frequency (%) 2.1 8.1 15.8 30.3 31.7 10.9 1.1 - - - 100 Accumulated relative frequency (%) 2.1 10.2 26.0 56.3 88.0 98.9 100
Diploid
Absolute frequency 3 12 27 54 78 59 15 3 2 1 254 Relative frequency (%) 1.2 4.7 10.6 21.3 30.7 23.2 5.9 1.2 0.8 0.4 100 Accumulated relative frequency (%) 1.2 5.9 16.5 37.8 68.5 91.7 97.6 98.8 99.6 100
The chi-square test confirmed the existence of a dependence relationship between the number of pods per umbel and the diploide and tetraploide cytotypes. The highest frequency in the number of pods corresponded to 1 pod per umbel in the tetraploid, while it corresponded to 4 pods in the diploid (Table 2). The diploid cytotype concentrated 50% of the umbels with 4 and 5 pods, whereas 80% of the umbels had between 1 and 2 pods in the tetraploid. From the analysis of these results it could be concluded that, although a reduction existed in the number of pods compared to the number of flowers per umbel in both cytotypes, this reduction was significantly higher in the tetraploid than in the diploid.
48 M.Cristina De Pablo, Mónica S. Barufaldi, Patricia Sastre, Hilda N. Crosta, Marcelo Eseiza and Lorena Daulerio Table 2. Number of pods per umbel in the tetraploid and diploid cytotypes of L. glaber Mill.
Cytotype Number of pods per umbel
1 2 3 4 5 6 7 Total
Tetraploid
Absolute frequency 132 95 40 9 7 1 0 284 Relative frequency (%) 46.5 33.5 14 3.1 2.5 0.4 - 100 Accumulated relative 46.5 80 94 97.1 99.6 100 frequency (%)
Diploid
Absolute frequency 16 37 46 68 58 28 1 254 Relative frequency (%) 6.3 14.6 18.1 26.8 22.8 11 0.4 100 Acumulated relative 6.3 20.9 39 65.8 88.6 99.6 100 frequency (%)
With regard to the number of seeds per pod and 1000 seed weight, significant differences were detected (P <0.001) between the cytotypes. The tetraploid produced half the seeds per pod with approximately double the seed weight, compared with the diploid cytotype (Table 3).
Table 3. Comparison of characters between the diploid and tetraploid cytotype of L. glaber Mill.
Character Diploid Tetraploid Differences Cytotype§ Cytotype§ between means Number of seeds per pod 17 ± 7.25 8 ± 4.6 *** 1000 seed weight (g) 0.94 ± 0.06 1.60 ± 0.02 *** Normal seedlings (%) 76 ± 2.29 74 ± 1.70 ns Hard seeds (%) 13 ± 2.54 11 ± 0.96 ns Soft seeds (%) 5 ± 1.03 3 ± 2.22 ns Abnormal seedlings (%) 2 ± 0.92 3 ± 1.15 ns Dead seedlings (%) 4 ± 0.75 9 ± 0.96 *** § Mean values ± standard deviation ns: differences non significant P <0.05 *** significant differences P <0.001
With reference to the characters that define the quality of the seeds: for the percentage of Seed production of tetraploid Lotus glaber 49 normal and abnormal seedlings, and for the percentage of hard and soft seeds, significant differences were not observed; however, for the percentage of dead seeds, a significant difference was obtained (Table 3), with the tetraploid population presenting a higher percentage of dead seeds (9%) compared with the diploid population (4%). The high percentage of dead seeds of the germplasm under study was also obtained when it was compared with the diploid cultivar Chajá Tresur (Barufaldi et al., 2003). Therefore, an effect of the induced polyploidy would appear to be an increase in the number of non-viable seeds produced. This suggests the need for future study of the causes that affect seed viability, by means of the topographical test with tetrazolium.
Conclusions
Among the components of seed production analysed in this work, it was observed that polyploidy had a negative effect on the number of flowers per umbel, on the number of pods per umbel and on the number of seeds per pod, which would appear to affect seed production markedly.
Due to the induced polyploidy, the seed weight was superior in the tetraploid cytotype. Detrimental effects were not detected in the majority of characteristics related to seed quality, such as the percentage of normal and abnormal seedlings, and the percentage of hard and soft seeds, although seed viability was affected.
References.
AHLOOWALIA B.S. 1971. Performance of diploid varieties and their tetraploid progenies in perennial ryegrass. Irish Journal of Agricultural Research, 10, 330-340.
ARMSTRONG C.S. 1974. ‘Grasslands Maku’ tetraploid lotus (Lotus pedunculatus Cav.). N.Z. Journal of Experimental Agriculture, 2, 333-6.
BARUFALDI M.S.; ANDRÉS A.; CROSTA H. and ESEIZA M. 2000. Obtención de una población autotetraploide de Lotus glaber Mill. (Lotus tenuis Waldst. & Kit). Revista de Tecnología Agropecuaria INTA Pergamino, Vol. V (15), 45-50. ISSN 0328-7750. [Spanish]
BARUFALDI M.S.; DE PABLO M.C.; RODRÍGUEZ R.H.; MONTAGNA M.V.; PÉREZ R.D.; DAULERIO L. and HIDALGO E. 2003. Differences in pollen fertility and seed quality between diploid and tetraploid cytotypes of Lotus glaber Mill. Lotus Newsletter, 33, 9-10. ISSN 1510- 7809. http://www.inia.org.uy/sitios/lnl/.
CARÁMBULA M.; AYALA W. and CARRIQUIRY E. 1994. Lotus pedunculatus. Adelantos sobre una forrajera que promete. INIA, Uruguay. Serie Técnica Nº 45. 14p. [Spanish]
DENNIS B.A. 1980. Mejoramiento genético de la producción de semilla de Trifolium pratense L. autotetraploide. In: Producción moderna de semillas. Ed. Hemisferio Sur S.R.L.Montevideo, Uruguay. pp.276-290. [Spanish] 50 M.Cristina De Pablo, Mónica S. Barufaldi, Patricia Sastre, Hilda N. Crosta, Marcelo Eseiza and Lorena Daulerio
EVANS A. 1955. The production and identification of polyploids in red clover, white clover and lucerne. New Phitologist, 54, 149 - 162.
FUNES N.; SALABERRY M.T.; ECHEVERRÍA M.M ; LUCHINI A and RODRÍGUEZ R.H. 2001. Efecto de la duplicación cromosómica en un cultivar de centeno (Secale cerale L.). Actas XXX Congreso Argentino de Genética. Mar del Plata, 16-19 setiembre. p. 115. [Spanish]
HERRERA L.M.; SALABERRY M.T.; ECHEVERRÍA M.M. and RODRÍGUEZ R.H. 1988. Estabilidad del nivel de ploidía del cultivar de centeno Tetrabal Inta. Actas IV Congreso Nacional de Trigo - II Simposio Nacional de Cereales de siembra otoño-invernal. Mar del Plata, 11-13 noviembre. I-26. [Spanish]
International Seed Testing Association. Suiza 2002. International Rules for seed testing. Chapter 5: Germination 5 A1- 5 A27.
LUCHINI A.E. 1979. Obtención de tetraploides en L.multiflorum y L. perenne. Trabajo de Graduación. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias. 30 p. [Spanish]
PÉREZ J. 1980. La poliploidía inducida como método de mejora del centeno forrajero. Actas IV Congreso Latinoamericano de Genética. 2: 457-463. [Spanish]
SIMONSEN O. 1976. Genetic variation in diploid and tetraploid populations of Lolium perenne L. Hereditas, 84,133-156.
Lotus Newsletter (2004) Volume 34, 51-53.
Preliminary studies on the tolerance of an induced autotetraploid population of Lotus glaber Mill. (= Lotus tenuis) to Tetranychus sp: Tetranychidae (red mite).
GABRIELA MARTINOIA*, ANA MARÍA DE HARO and MÓNICA BARUFALDI. Universidad Nacional del Centro de la Provincia de Buenos Aires, Facultad de Agronomía Departamento de Producción Vegetal, Av. República Italia 780, C.C. 47, (7300) Azul, Provincia de Buenos Aires, Argentina. * Corresponding author.
Introduction
The genera of red mites Tetranychus is found on a great variety of leguminous species. Its attack is initially visualised as chlorotic points that, upon fusion, become yellowish sectors before finally adopting a tanned appearance. It also produces a web that accumulates dust that adversely affects photosynthesis (Quintanilla, 1978).
The Genetics and Plant Breeding Group and the Agricultural Botany Group of the Faculty of Agronomy of Azul, UNCPBA, Province of Buenos Aires, which are responsible for a breeding programme involving the species Lotus glaber Mill., obtained induced autotetraploid germplasm (2n=4x=24) by means of colchicine treatment of diploid plants (2n=2x=12). The induced tetraploid currently comprises a relatively stable population at ploidy level (4x), from which the selection process for productivity traits has been initiated (Barufaldi et al., 2000; Andrés et al., 2001).
Induced autotetraploid cultivars obtained in other forage species such as Lolium perenne, L. multiflorum, Lotus uliginosus (L. pedunculatus) and Trifolium pratense, in general show good establishment, enhanced resistance to adverse factors, and better persistence and forage production compared to the diploid parents (Ahloowalia, 1971; Carámbula et al., 1994; Evans, 1955; Herrera et al., 1988; Luchini, 1979; Pérez, 1980; Simonsen, 1976).
The presence of the mite was observed in some autotetraploid plants of Lotus glaber in both the greenhouse and the field. For this reason it was considered desirable to perform experiments in order to detect tolerance in tetraploid genotypes; preliminary studies for this purpose began in 2003.
Objective
To select individuals that present better behaviour in response to acarus attack, and to achieve their vegetative multiplication as a first step towards verifying their tolerance.
51 52 Gabriela Martinoia, Ana María de Haro and Mónica Barufaldi
Materials and Methods
Observations began in July of 2003 on 70 different genotypes used in experiments designed to assess tolerance to water deficit and evaluate the response to Phosphorous fertilisation, since acarus presence was detected on these experiments. For this purpose, a low Phosphorous soil was used (- P; 4 ppm Bray and Kutz Number 1) from the Parish Area, District of Azul, in the Salado Depression, as well as the same soil fertilised with Phosphorous (+P; 100ppm calcium triple superphosphate). The plants, which had been previously maintained in 800g pots, were transplanted to 3000g pots and subsequently subjected to permanent irrigation. Weekly evaluations of the attack were made until December of the same year, whereupon plants that had not been attacked hitherto were selected.
Results
It was possible to observe that all plants affected from July until October of 2003 had been fertilised with Phosphorous. The same tendency was detected when they were evaluated by their response to rust Uromyces loti (M. Barufaldi, personal communication).
After a strong drought coinciding with raised temperatures (November 2003), conditions that benefited the development of the mites, plants suffering attack were detected with and without fertilisation. The formation of abundant web and infection spread among the different genotypes could also be observed.
Of the 22 plants selected in December of 2003, three genotypes were detected in February 2004 that displayed superior behaviour. They will be cloned and later transplanted to the field in order to follow their behaviour during the next cycle.
References.
AHLOOWALIA B.S. 1971. Performance of diploid varieties and their tetraploid progenies in perennial ryegrass. Irish Journal of Agricultural Research, 10, 330-340.
ANDRÉS A., BARUFALDI M., CROSTA H., ESEIZA M. and CARRETE J. 2001. Variaciones en algunos aspectos productivos y de fertilidad de genotipos tetraploides de Lotus glaber Mill. (Lotus tenuis Waldst. et Kit) provenientes de dos ciclos de multiplicación. Revista de Tecnología Agropecuaria INTA Pergamino, vol. VI (17), 40-46. [Spanish]
BARUFALDI M.S., ANDRÉS A., CROSTA H.N. and ESEIZA M.F. 2000. Obtención de una población autotetraploide de Lotus glaber Mill. (Lotus tenuis Waldst. et Kit). Revista de Tecnología Agropecuaria INTA Pergamino, vol. V (15), 45-50. [Spanish]
CARÁMBULA M., AYALA W. and CARRIQUIRY E. 1994. Lotus pedunculatus. Adelantos sobre una forrajera que promete. INIA, Uruguay. Serie Técnica, Nº 45, 14p. [Spanish]
Tetranychus in Lotus glaber 53
EVANS A. 1955. The production and identification of polyploids in red clover, white clover and lucerne. New Phytologist, 54, 149 - 162.
HERRERA L.M., SALABERRY M.T., ECHEVERRÍA M.M. and RODRÍGUEZ R.H. 1988. Estabilidad del nivel de ploidía del cultivar de centeno Tetrabal INTA. Actas IV Congreso Nacional de Trigo - II Simposio Nacional de Cereales de Siembra Otoño-invernal (Mar del Plata, Argentina. 11-13 de noviembre de 1988). I-26. [Spanish]
LUCHINI A.E. 1979. Obtención de tetraploides en L. multiflorum y L. perenne. Trabajo de Graduación. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias. 30p. [Spanish]
PÉREZ J. 1980. La poliploidía inducida como método de mejora del centeno forrajero. Actas IV Congreso Latinoamericano de Genética (Mendoza, Argentina. 21-27 de septiembre1979) 2, 457-463. [Spanish]
QUINTANILLA R.H.1978. Ácaros Fitófagos. Editorial Hemisferio Sur. Buenos aires. 73 p. [Spanish] Lotus Newsletter (2004) Volume 34, 54-59.
R2R3-MYB transcription factors from Lotus corniculatus var. japonicus differentially regulated in roots and shoots in response to nitrogen starvation.
KUNIHIKO MIYAKE1, TAKURO ITO1, MINEO SENDA2, RYUJI ISHIKAWA1, TAKEO HARADA1, MINORU NIIZEKI1 and SHINJI AKADA2* 1 Plant Breeding Laboratory, Faculty of Agriculture, Hirosaki University, Hirosaki, Aomori 036-8561, Japan 2 Gene Research Center, Hirosaki University, Hirosaki, Aomori 036-8561, Japan * Corresponding author
In leguminous plants, a species-specific type of (iso)flavonoid secreted out from roots functions as a trigger for the nitrogen-fixing symbiosis of rhizobacteria. The pathway of (iso)flavonoid biosynthesis is significantly down-regulated by exogenously supplied nitrate in alfalfa (Coronado et al., 1995). This suggested a possibility that inhibitory effects of nitrate on nodulation, in part, operate via regulation of the pathway of flavonoid biosynthesis. On the other hand, it has been reported that pathways of flavonoid biosynthesis are regulated by various members of R2R3-MYB superfamily in different organs and in different conditions (Paz-Ares et al., 1987; Grotewold et al., 1994; Quattrocchio et al., 1999; Jin et al., 2000; Nesi et al., 2001). In this study, we attempted to isolate members of the R2R3-MYB superfamily, which may be involved in regulation of nitrogen fixation or of nitrogen metabolism in legumes.
A library of 300 recombinant plasmid clones containing the PCR fragments amplified by using degenerate primers for the R2R3-MYB superfamily was screened by differential hybridization to isolate R2R3-MYB genes whose expression was up-regulated under nitrogen nutrient limited conditions. Two groups of clones were identified, each of which seemed to represent a gene responsive to the nitrogen starvation. The entire coding regions for the genes were further isolated by PCR and were designated as LjMYB101 and LjMYB102. By screening a genomic library of Lotus japonicus with a probe derived from LjMYB101, the third gene, LjMYB103, was isolated. In addition, a candidate for the soybean orthologue of LjMYB101 was isolated and designated as GmMYB101 (Miyake et al., 2003). Sequence alignment of the genes with members of the plant R2R3-MYB superfamily showed that they all belonged to the subgroup 10 (Stracke et al., 2001) of the R2R3-MYB superfamily (Figure 1). Among these genes a contrasting expression pattern has been observed between LjMYB101 and LjMYB103. That is, the expression level of LjMYB101 was higher in roots and it was up-regulated under a nitrate limited condition (N-) in the roots, but not in the shoots. On the other hand, expression of LjMYB103 was higher in shoots and was also up-regulated only in the shoot (Figure 2). Considering their structural similarity in the coding region, LjMYB101 and LjMYB103 seem to have diverged through acquisition of their
54 R2R3-MYB transcription factors 55 organ specific responsiveness, which might have differentiated their physiological roles. Therefore, the primary role of LjMYB103 might have been assigned to the shoot while the same kind of role fulfilled by LjMYB101 in the roots.
Figure 1. Phylogeny of the members in the subgroup 10 as well as those in its related subgroups (24, 4, and 11) of the plant MYB superfamily. The amino acid sequences of LjMYB101, LjMYB102, LjMYB103, and GmMYB101 (indicated with arrowheads) along with those of the genes of which sequences are currently in GenBank/EMBL/DDBJ (accession numbers are indicated in parenthesis) were aligned with the ClustalW (Thompson et al., 1994) and, at the same time, reconstruction of a phylogenic tree with neighbor joining method and calculation the bootstrap values were carried out with the program package of DDBJ. Members of each subgroup are indicated with side brackets. 56 Kunihiko Miyake, Takuro Ito, Mineo Senda, Ryuji Ishikawa, Takeo Harada, Minoru Niizeki and Shinji Akada
Figure 2. Differential expression of LjMYB101 and LjMYB103 in the roots and shoots of L. japonicus grown under the N- or N+ conditions. RT-PCR/Southern analysis was carried out using the specific primers for each gene. The total RNA samples as templates were extracted from seedlings grown with the agar medium containing the basic MS salts (Murashige and Skoog, 1962) with its nitrogenous components (NH4NO3 plus KNO3) substituted for by the equal molars (39.4 mM) of KCl (N-) or KNO3 (N+). The expressed levels of LjACTIN transcripts were analyzed as the reaction controls.
At this point, we don’t have any evidence defining the cis-elements that conferred their organ specificity, but the sequence information of the promoters seems informative. The 5' upstream fragment (524 bp) of LjMYB101 was isolated by inversed PCR and that (1569 bp) of LjMYB103 was obtained from the lambda genomic clone, λLj72302-2. On the analysis of the sequence information, a striking similarity was found between the promoter segments of LjMYB101 and LjGln1, a member of the Gln synthetase gene family in Lotus corniculatus var. japonicus (Thykjær et al., 1997; Figure 3). The homologous region spans over a stretch of 138 bp with similarity of 89 %. It is located from 374 to 511 bp upstream of ATG in LjMYB101, or from 1517 to 1654 bp upstream in LjGln1, respectively. It would be interesting to speculate that insertion or deletion of this element in the promoter of the common ancestor of LjMYB101 and LjMYB103 might have caused diversion of the two genes. Because nitrate regulated expression of LjGln1 was also restricted to the root tissues, the homologous region of LjMYB101 and LjGln1 may be related to their organ specific regulation. When a blast search was performed with the element of 138 bp, a list of 27 sequences containing this element was found in the genome of Lotus corniculatus var. japonicus. We found ORFs of various sizes in many of their proximal regions, but we did not find any genes closely relevant to nitrogen metabolism. Further work will be required to verify the significance of this element.
R2R3-MYB transcription factors 57
Figure 3. Sequence alignment of the promoters of LjMYB101 (LjMYB101 PRO) and LjMYB103 (LjMYB103 PRO), as well as the homologous sequence element found in the promoter of LjGln1 (GLN1 ELEMENT). The sequences were aligned using the Higgins’ multiple alignment program of DNASIS V3.6 (HITACH SOFTWARE). The TATA-boxes are underlined and the homologous elements of 138 bp are lined above the sequences.
58 Kunihiko Miyake, Takuro Ito, Mineo Senda, Ryuji Ishikawa, Takeo Harada, Minoru Niizeki and Shinji Akada Downstream of this promoter element is a promoter core region including 332 bp upstream of the ATG initiation codon in LjMYB101 or 341 bp upstream of LjMYB103, respectively. The putative TATA-box was found at 190 bp or 166 bp upstream of ATG for each of the genes. Sequence alignment of the two promoters revealed that LjMYB101 and LjMYB103 shared continually homologous sequences around the regions surrounding the TATA-box (Figure 3). Thus, the two sequences seem to have retained substantial homology, while little homology was detected between the two in their further upstream regions, because of presence of the element of 138 bp in the promoter of LjMYB101. Albeit their difference in tissue specificity, the two genes are both up-regulated under nitrogen-limited conditions. Thus, it is possible that the sequence element conserved between the promoters of LjMYB101 and LjMYB103might act as cis-elements for the nitrogen regulation.
We speculated that LjMYB101 might function as a transcriptional activator of genes such as members of chalcone synthase gene family (LjCHS) and/or LjGln1, because both these genes are also up-regulated specifically in roots under nitrogen-limited conditions (Miyake et al., 2003). In order to verify if the MYB transcription factor targets to the promoters of these genes, the DNA binding domain of LjMYB101 was synthesized using an E.coli protein expression system. The protein was used for the electrophoretic mobility-shift assay (EMSA) and it was found that the protein bound to the probe DNA fragments containing the sequence of CACCTACC or CACCAACC, which coincided very well with the type IIG (H-box like) consensus of YACCWACC as a recognition sequence for a class of MYB transcription factors. The sequence perfectly matching YACCWACC is found in three members (accession numbers; AP006705, AP006706, and AP006709) of LjCHS, among eight members of the gene family available from the database. This motif locates upstream of the TATA-box and the distance ranges from 22 to 34 bp, which is very similar to the distance of the same element to the TATA-box on the promoter regions of the soybean CHS gene family, ranging from 22 to 38 bp (Shimizu et al., 1999). These suggest that LjMYB101 may target to the promoters of members of CHS gene family in Lotus while GmMYB101 may do the same in soybean. On the other hand, the sequence of CACCTACC also exists at 43 bp upstream of TATA-box on the promoter of LjGln1, inspiring another possibility that LjMYB101 may target to LjGln1. In vivo analysis of the gene function will be required to further investigate the physiological roles for the newly isolated R2R3-MYB genes, in relation to flavonoid biosynthesis or to nitrogen metabolism.
References.
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R2R3-MYB transcription factors 59
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SHIMIZU T., AKADA S., SENDA M., ISHIKAWA R., HARADA T., NIIZEKI M. and DUBE S.K. 1999. Enhanced expression and differential inducibility of soybean chalcone synthase genes by supplemental UV-B in dark-grown seedlings. Plant Molecular Biology, 39, 785-795.
STRACKE R., WERBER M. and WEISSHAAR B. 2001. The R2R3-MYB gene family in Arabidopsis thaliana. Current Opinion in Plant Biology, 4, 447-56.
THOMPSON L.D., HIGGINS D.G. and GIBSON T.J. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice. Nucleic Acids Research, 22, 4673-4680.
THYKJÆR T., DANIELSEN D., SHE Q. and STOUGAAD J. 1997. Organization and expression of genes in the genomic region surrounding the glutamine synthetase gene Gln1 from Lotus japonicus. Molecular Genetics and Genomics, 255, 628-636.
Lotus Newsletter (2004) Volume 34, 60-65.
Interdisciplinary workshop on genetic, molecular and ecophysiological aspects of Lotus spp. and their symbionts. [Taller interdisciplinario sobre aspectos genéticos, moleculares y fisioecológicos del Lotus spp. y sus simbiontes].
Chairperson: OSCAR A. RUIZ*
Held at: Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús. IIB-INTECh. (UNSAM-CONICET). Chascomús. Provincia de Buenos Aires. Argentina. 11 –13 September 2002
* Corresponding author
Proposal and program
The introduction and sustainable management of highly valuable forage legumes in the “Pampa Deprimida” is a promissory alternative to increase significantly the cattle production output. In particular, Lotus glaber (former L. tenuis), species naturalized species from the Mediterranean Basin, has evidenced high adaptability to the environmental restrictions to the plant growth vegetable in these grasslands (flooding, salinity, alkalinity, low concentrations of assimilable Phosphorus, droughts, grazing by cattle, fire, parasites and predators), prospering and spreading in spontaneous form.
The proposed Workshop has an antecedent in the “1er Simposio Argentino del Género Lotus” [1st Argentinean Symposium of the Genus Lotus] that was developed in the INTECh (at the present time IIB-INTECh) in 1992. From then on, and especially during the last years, the advances in studies of molecular biology and bio-engineering have enlarged and enhanced the opportunities to advance in the efficient use of these strategic resources for our beef production. On the other side, they outline an enormous challenge to integrate these advances in effective form, not only in terms of increasing the cattle production but of making it in the mark of a commitment irreducible with the conservation of the “Pampas” ecosystem.
In this context, the proposed objectives of this Workshop are: 1) to update, exchange and integrate knowledge from different disciplines on biological processes - from molecular to population level-, committed with the adaptability, the production and the persistence of legume populations in grasslands of the Pampa Deprimida (in particular on the Genus Lotus); 2) to propitiate the generation of links between disciplines and institutions in order to maximise the investigation efforts that are carried out in the country and overseas; and 3) to evaluate critically and strategically, the new opportunities that the biotechnology advances can offer for the development of a sustainable cattle production.
60 Lotus Workshop, Argentina 61
Program
Day 11 September:
Workshop of Microbiology
Agronomic management of the symbiosis in species of the Genus Lotus. [Manejo agronómico de la simbiosis en especies del género Lotus.] CARLOS LABANDERA, Laboratorio de Microbiología de Suelos, Ministerio de Agricultura y Pesca, Uruguay.
Characterization of strains of Rhizobium loti of the Pampa Deprimida del Salado. [Caracterización de cepas de Rhizobium loti de la Pampa Deprimida del Salado.] JULIA ESTRELLA. Instituto de Investigaciones Biotecnológicas, Instituto Tecnológico de Chascomús IIB-INTECh, UNSAM-CONICET, Argentina.
Arbuscular Mycorrhiza in Lotus glaber of the “Depresión del Salado” and their possible influence on the mechanism of resistance to plant saline stress. [Micorrizas arbusculares en Lotus glaber de la Depresión del Salado y su posible influencia sobre el mecanismo de resistencia a estres salino de la planta.] ANA MENENDEZ, Departamento de Microbiología, Facultad de Biología, Universidad de Buenos Aires (UBA), Argentina.
Molecular aspects of the symbiosis Rhizobium loti - Lotus glaber. [Aspectos moleculares de la simbiosis Rhizobium loti- Lotus glaber.] VIVIANA LEPEK, Instituto de Investigaciones Biotecnológicas, Instituto Tecnológico de Chascomús IIB-INTECh, UNSAM-CONICET, Argentina
Physiological conditions of Rhizobium in the nodules of legumes. [Condiciones fisiológicas de Rhizobium en el interior de nódulos de leguminosas.] JUAN SANJUAN, Departamento de Microbiología del Suelo y Sistemas Simbióticos, Estación Experimental del Zaidin CSIC, España.
Day 12 September:
Workshop of Biology, Biochemistry, Physiology and Biotechnology.
Assimilation of Nitrogen in nodules of legumes: molecular aspects of the enzymes involved. [Asimilación de nitrógeno en nódulos de leguminosas: aspectos moleculares de las enzimas implicadas.] CARMEN LLUCH, Departamento de Fisiología Vegetal, Facultad de Ciencias, Universidad de Granada, España.
Prosopis strambulifera, a halophyte legume with potential use in biotechnology. [Prosopis strambulifera, una leguminosa halófita con potencial uso en biotecnología.] VIRGINIA LUNA, Departamento de Fisiología Vegetal, Universidad Nacional de Río Cuarto (UNRC), Córdoba, Argentina.
62 O. Ruiz
Populational biology de Lotus glaber in the “Pampa Deprimida”. [Biología poblacional de Lotus glaber en la Pampa Deprimida.] OSVALDO VIGNOLIO, Cátedra de Ecología, Universidad Nacional de Mar del Plata (UNMdP), Argentina.
The inundation in the “Pampas Deprimidas” as a benefit of the nature. Its possible utility for the introduction of Lotus glaber. [Las inundaciones en la Pampa Deprimida como servicio de la naturaleza. Su posible utilidad para la introducción de Lotus glaber.] PEDRO INSAUSTI, Cátedra de Fisiología Vegetal, Facultad de Agronomía, Universidad de Buenos Aires (UBA), Argentina.
An evaluation on the list of the polyamines in Lotus spp. under saline stress, as study tool of their mechanisms of tolerance. [Una evaluación sobre el rol de las poliaminas en Lotus spp. bajo estres salino, como herramienta de estudio de sus mecanismos de tolerancia.] OSCAR A. RUIZ, Instituto de Investigaciones Biotecnológicas, Instituto Tecnológico de Chascomús IIB-INTECh, UNSAM-CONICET, Argentina.
Workshop of Genetic Improvement and Cultivar Management
Genetic resources and breeding de Lotus glaber, ideas for the discussion of a regional strategy. [Recursos genéticos y mejoramiento de Lotus glaber, pautas para la discusión de una estrategia regional.] MARÍA DE LA MERCED MUJICA, Mejoramiento Genético, Facultad de Ciencias Agrarias y Forestales, Universidad Nacional de La Plata (UNLP), Argentina.
Pampa INTA, a new cultivar of Lotus glaber obtained by phenotypic recurrent selection. [Pampa INTA, un nuevo cultivar de Lotus glaber obtenido por selección recurrente fenotípica.] MANUEL GÓNZALEZ GARCÍA, Estación Experimental Agropecuaria de Balcarce, Instituto de Tecnología Agropecuaria (EEA-INTA), Pcia de Buenos Aires, Argentina.
Management and utilization of Lotus spp in the “Cuenca del Salado”. [Manejo y utilización del Lotus spp en la Cuenca del Salado.] MATÍAS BAILLERES, Estación Experimental Manantiales, Ministerio de Asuntos Agrarios de la Provincia de Buenos Aires, Argentina.
The induced poliploidy in Lotus glaber Mill. [La poliploidía inducida en Lotus glaber Mill.] MÓNICA BARUFALDI, Facultad de Ciencias Agrarias, Universidad Nacional del Centro de la Provincia de Buenos Aires (UNCPBA), Azul, Pcia de Buenos Aires, Argentina.
Breeding of the Genus Lotus in Uruguay. [Mejoramiento genético del género Lotus en Uruguay.] MÓNICA REBUFFO, INIA La Estanzuela, Estación Experimental “Alberto Böerger”, Instituto Nacional de Investigación Agropecuaria (INIA), Uruguay.
Lotus Workshop, Argentina 63
Day 13 September:
Workshop of Production and Agronomic Evaluation
Space and temporal heterogeneity of the Pampean vegetation. [Heterogeneidad espacial y temporal de la vegetación pampeana.] ROLANDO LEON, Facultad de Agronomía, Universidad de Buenos Aires (UBA), Argentina.
Perception of the farmer on the intensive use of the L. glaber in the “Pampa Deprimida del Salado”. [Una visión del productor sobre el uso intensivo del L. glaber en la Pampa Deprimida del Salado.] SYLVIO CORTINA, Productor asociado de la Confederación Agropecuaria de Productores de Buenos Aires y La Pampa (CARBAP) y Sociedad Rural Argentina (SRA), Argentina.
The contribution de L. glaber to the bovine rearing in the grasslands of the inundated pampas. [El aporte de L. glaber a la cría vacuna en los pastizales de la pampa inundable.] MIGUEL CAUHÉPÉ, Facultad de Ciencias Agrarias, Unidad Integrada Universidad Nacional de Mar del Plata, Instituto de Tecnología Agropecuaria, UNMdP-INTA-Balcarce, Pcia. de Buenos Aires, Argentina.
Agronomy, cultivar evaluation, and characterization of naturalized germplasm of three species de Lotus in Chile. [Agronomía, evaluación de variedades, y caracterización de germoplasma naturalizado de tres especies de Lotus en Chile.] HERNÁN ACUÑA, Centro Regional de Investigación Quilamapu, Instituto de Investigaciones Agropecuarias (INIA), Chile.
Final Conclusions of the Workshop
JUAN SANJUAN1, OSVALDO VIGNOLIO2, CARLOS LABANDERA3 and MÓNICA REBUFFO4.
1 Consejo Superior de Investigaciones Científicas (CSIC), Estación Experimental del Zaidin, Departamento de Microbiologia del Suelo y Sistemas Simbióticos, Prof. Albareda 1, E-18008 Granada, Spain. 2 EEA INTA Balcarce, Producción Vegetal, Ruta 226, Km 73.5, CC 276 (7620) Balcarce, Argentina. 3 Laboratorio de Microbiología de Suelos, Ministerio de Agricultura y Pesca, Uruguay. 4 INIA La Estanzuela, Estación Experimental “Alberto Böerger”, Instituto Nacional de Investigación Agropecuaria (INIA), Uruguay.
After 3 intense days of conferences and discussions, we can say that the execution of this workshop has been a complete success. The implementation of the workshop was itself a challenge, taking into account the difficulty of gathering together scientists, technologists, agronomists and farmers. Despite the difficulties the meeting was extremely dynamic and co-operative. Furthermore, it allowed to evidence the enormous potential of Lotus for the 64 O. Ruiz production system in the Pampa Deprimida del Salado, as a real alternative to increase cattle production in this region. The species Lotus glaber (previously L. tenuis) appeared with a higher adaptive potential to the regional geo-productive conditions.
Thanks to the multidisciplinary character of the workshop, as well as to the great interest of all the participants for the potentiality of this system, it was possible to identify a series of organisational, biological, agronomical and environmental factors which limit or can be limiting crop productivity. The diversity of these factors reflects the intrinsic complexity of a serious and rigorous study towards the sustainable production of this forage legume.
1) The excessive compartmentalization of the existing knowledge on the geographical, meteorological and productive conditions of the region. There are a number of partial studies on different aspects of the system. Much of this information is not optimally reflected in publications easily accessible to the interested parties. On the other hand, it is necessary to integrate all that knowledge, so to facilitate the development of appropriate technological strategies.
2) The absence of systematic studies of the productive system. It was detected the necessity to generate and/or gather evaluation data on the impact of Lotus spp. on the beef farming system, with a diagnosis of the potentialities and of the limiting factors in each situation. The apparent environmental heterogeneity that exists in the region imposes the need for this type of studies.
3) The urgent need of characterization and evaluation of the existing biological resources. It is deduced from the available information that a wide biological diversity exists in the region, either for the plant (species and genotypes of Lotus), as well as of beneficial micro-organisms important for its nutrition and growth (Nitrogen fixing bacteria, mycorrhiza, etc). Nevertheless, it is necessary an appropriate evaluation of this bio-diversity towards the selection of the best genotypes, particularly in relation to biomass production as well as to those factors limiting for the production, such as: • low levels of Phosphorus and Nitrogen in the soils of the region. • alkalinity / salinity of the soils • the flooding and drought cycles • the extreme temperatures • the legume persistence • agronomic management (the need of optimization of the legume establishment, grazing and the seed production).
4) Necessity of improvement of the resources Particularly in the case of L. glaber, there seem to exist only a few cultivars selected for the conditions of the region. Neither there are bio-fertilizers specifically selected for the regional needs. There is a need to identify those cultivars and/or to improve them, as well as to develop specific bio-fertilizers so to improve the Lotus adaptive capacity and towards its productive optimization in the region. In this sense, the generation of genotypes with Lotus Workshop, Argentina 65 increased ploidy and the generation of molecular markers to accelerate plant breeding programs are of supreme importance.
5) Necessity of appropriate strategies of R+D (Research + Development) The complexity of factors that determine the productive capacity of Lotus spp. in the region makes necessary the establishment of a R+D program that agglutinates all the necessary efforts. The following activities are identified as high-priority: • the co-ordination of the different participants and activities. • design of strategies for the obtaining of the necessary human and material resources. Integration with European scientific programs and at Ibero-American level. • the elaboration of strategies for the implementation and diffusion of results (newsletter, etc.). Lotus Newsletter (2004) Volume 34, 66 – 72.
Evaluation of tetraploid big trefoil (Lotus uliginosus Schkuhr.) for rust resistance
SEBASTIÁN ARRIVILLAGA, SEBASTIÁN HERNÁNDEZ, JAVIER CILIUTI, MÓNICA REBUFFO, SILVIA GERMÁN and SILVINA STEWART.
Instituto Nacional de Investigación Agropecuaria, INIA La Estanzuela, Colonia, Uruguay.
Introduction
Several species of Lotus are cultivated in Uruguay; however, the prevalence of diseases has only been thoroughly studied on birdsfoot trefoil (Lotus corniculatus L.; Altier, 1997). The widespread cultivation of susceptible cultivars almost always results in an increase of disease severity. The area of big trefoil (Lotus uliginosus Schkuhr.) has increased in recent years, mainly due to the good adaptation of the tetraploid cultivar Grasslands Maku. This synthetic cultivar developed in New Zealand is based on 13 elite plants selected after three generations of recurrent selection of an induced tetraploid developed by colchicine treatment (Charlton, 1983). Rust outbreaks caused by Uromyces spp. have been observed on this cultivar since 2000 and its severity ranged from trace to severe, where all the lower leaves were killed by the rust and upper leaves had open pustules (Ciliuti et al, 2003).
Rust on birdsfoot trefoil develops mostly in summer, from February to April (Altier, 1997), whereas the disease has been observed on big trefoil from November onwards (Ciliuti et al, 2003). Mild winters allow overwintering of the pathogen in Uruguay; therefore the disease may appear at any time during warm and wet weather conditions. Host specificity experiments have identified two races of rust on big trefoil in Uruguay (Ciliuti et al., 2003).
The recent appearance of rust on the main big trefoil cultivar utilized in Uruguay and the lack of information of the pathogen reaction within the tetraploid cultivar raised the need to study its genetic variability and to compare the effectiveness of field selection for rust tolerance with selection under artificial inoculation in greenhouse conditions.
Materials and Methods
Big trefoil germplasm utilized in this study came from seed stocks of tetraploid cultivar Grasslands Maku harvested either in New Zealand (NZ 1 to NZ 3) or Uruguay (UR 1, UR 2), and a sample of the diploid accession G4701 (D) from New Zealand that gave origin to the tetraploid cultivar (Charlton, 1983). An experiment was set on the Forage Nursery Experimental Field at La Estanzuela, Colonia, Uruguay. Plots of 25 plants, spaced 1 m apart, were distributed in a complete randomized block design with 10 replicates. Seedlings were established in the greenhouse in June 2000 and transplanted to the field on the second week of August 2000.
66 Rust resistance evaluation in Lotus uliginosus 67
Individual plants were characterized in the field during the second year. Date of flowering was recorded twice a week (coded as initial date 1= November 27 2001 to 14= January 28 2002). Plants were individually harvested on September 19 2001 to evaluate early spring forage yield. Mature pods were harvested twice a week from the first week of January to the end of February and the seed accumulated for total yield. Rust was scored in mature plants during the second week of December, when epidemic was evenly established and the most susceptible plants showed symptoms of the disease in all leaves. Plants were rated on a 1 to 5 visual scale (1 – no foliar symptoms, 2 – few pustules on lower leaves, 3 – many pustules on lower leaves, 4 – pustules in mid leaves, 5 – pustules in all leaves). Seed stocks were compared on a plot- mean basis and the association between rust and other characteristics was estimated.
Selected plants were vegetatively multiplied in March 2002. Chromosomes dyed with acetocarmine were counted from the root tips of these plants to verify the ploidy level after conservation in a 3:1 solution of alcohol and acetic acid in the fridge (Sass, 1958). Tetraploid plants were separated into three groups, according to rust resistance: MR - moderately resistant (classes 1 and 2), MS – moderately susceptible (class 3) and S - susceptible (classes 4 and 5). Diploid plants with rust scores between 1 and 2 were grouped in a forth category (D). Each group was polycrossed by manual pollination and seed harvested from individual plants. Rust resistance was evaluated for each half-sib family, artificially inoculating at least 40 seedlings distributed in 10 pots. Urediniospores from cv Grassland Maku were collected at La Estanzuela, Colonia. Rust collection and inoculation procedures were described by Ciliuti et al (2003). Inoculation was carried out on January 2 2003 at the seedling stage (5 to 8 leaflets). First symptoms of the disease appeared 10 days after inoculation. Rust was evaluated 14 and 17 days after inoculation (13 January and 16 January) in the most severely infected leaf per individual seedlings. The scale described by Skinner and Stuteville (1995) was used: 1 - resistant (no symptoms), 2 - moderately resistant (flecks and closed pustules), 3 - moderately susceptible (closed pustules and small open pustules), 4 - susceptible (small open pustules), 5 - highly susceptible (medium to large open pustules). Plants recorded as class 1 and 2 were inoculated again on February 11 2003. Polycross groups were compared on a half sib family mean basis.
Results and Discussion
Field observations of the pathogen have been reported on several Lotus species in the region (Altier, 1997; Ciliuti et al, 2003; Juan et al, 2000). The disease severely damaged spaced plants of Grassland Maku in 2002 (Figure 1). Uredinia were mostly found on leaflets and stems, which turned yellow and drop or dried off (Figure 2). Maximum damage was observed during dry spells, when vegetative growth was stunted and pustules developed up to the top leaves. Seed stocks were ranked based on the mean rust leaf ratings averaged over 10 replications (Table 1). Rust ratings were lower (P<0.01) in one of the seed stocks of G.Maku harvested in Uruguay (UR 2) and the diploid accession (D). The latter was easily recognized by its small leaves and late flowering, in addition to the lower rust ratings compared with Grassland Maku. Based on the rust ratings, 28% of plants were resistant (class 1) for the diploid, whereas the resistant plants ranged from 6 to 20% within G.Maku seed stocks (Figure 68 Sebastián Arrivillaga, Sebastián Hernández, Javier Ciliuti, Mónica Rebuffo, Silvia Germán and Silvina Stewart.
3). With the exception of UR 2, all seed stocks of G.Maku had 50% or higher proportion of susceptible and highly susceptible plants (classes 4 and 5). Despite the differences in rust rating under field conditions, there were no differences in other evaluated traits for the tetraploid seed stocks (Table 1).
Figure 1. Spaced plants in early December. Figure 2. Foliar damage caused by rust.
Table 1. Field rust ratings, date of flowering, spring forage yield (g DM per plant) and seed yield (g per plant). Average data for seed stocks.
Seed Rust Date of Forage Seed stocks ratings flowering yield yield NZ 1 3.62 a 7.23 bc 19.3 3.6 NZ 2 3.61 a 7.54 b 17.7 3.0 NZ 3 3.46 a 7.06 bc 18.1 2.9 UR 1 3.55 a 6.88 c 15 4.2 UR 2 2.86 b 7.57 b 17.5 3.0 D 2.78 b 9.33 a 9.2 2.0 F-test 0.003 <.001 n.s. n.s. s.e.d. 0.280 0.315 3.97 0.94 For each column, any value with the same letter were not significantly different (P=0.05) with Fisher LSD test.
Rust resistance evaluation in Lotus uliginosus 69
100%
80% 5 4 60% 3 40% 2 20% 1 Proportion of plants of Proportion 0% NZ 1NZ 2NZ 3UR 1UR 2D
Figure 3. Proportion of plants with rust scores within the different seed stocks (Scale 1 – resistant to 5 – highly susceptible).
Severe damage caused by foliar diseases usually affects plant performance. Barbetti and Nichols (1995) studied forage yield and seed losses for several cultivars of Trifolium subterraneum associated with rust scores. However, in this study rust severity was not correlated (P<0.05) with growth or seed production, despite the fact that the disease progress throughout all the flowering period. This result probably reflects the low potential for seed production that the tetraploid cultivar has shown in Uruguay (Formoso, 2001). Seed yield correlated with forage yield (r= 0.65, P<0.05) and date of flowering (r= -0.43), P<0,05), indicating higher seed production in larger plants with early flowering.
The severity of foliar diseases under field conditions could be affected by factors such as the physiological stage and the microenvironment of the plant (density of foliage), climatic conditions, etc. Emery and English (1994) demonstrated that the incidence and severity of alfalfa foliar diseases, including rust, depended on moisture conditions and so, varied significantly along the growing season. Disease on the lower half of the plant was correlated with leaf wetness duration and atmospheric moisture conditions while disease on the upper half of the plants was correlated with cumulative rainfall. Furthermore, climatic factors such as temperature altered the genetic behavior of alleles involved in alfalfa resistance to rust (Skinner and Stuteville, 1989). In this field study, rust rating was negatively correlated with flowering date (r= -0.39, P<0.05), indicating that the disease severity was higher in early materials. Since rust was evaluated only once during the growth season in this study, it is not possible to attribute this relationship to either environment or physiological stage.
Variation for field rust rating suggested that opportunities may exist for improvement of rust resistance in the tetraploid cultivar (Figure 3). However, since rust severity could have been affected by the interaction of plant physiological stage and environmental conditions previous to the field evaluation, progenies of selected plants were further tested with artificial inoculation. The selected groups exhibited different patterns of rust scoring under artificial inoculation. The dipliod had lower values of rust score (P<0.001) than all three tetraploid 70 Sebastián Arrivillaga, Sebastián Hernández, Javier Ciliuti, Mónica Rebuffo, Silvia Germán and Silvina Stewart. groups 14 days after inoculation (Table 2), indicating a similar ranking as in the field evaluation. However, three days later rust score was over 4 for all groups giving a different group ranking. Although there were significant differences (P<0.001) between groups at this stage, all of them were ranked susceptible (class 4) to highly susceptible (class 5) for most of the plants. Few plants classified as moderately resistant (class 2) in the first inoculation, were reclassified as susceptible after the second inoculation. These plants may have not been properly inoculated previously because their foliage was covered by other plants.
Table 2. Average data and range of rust rating at 14 and 17 days after artificial inoculation (Scale 1 to 5) for half sib families within crossing groups.
Day 14 Day 17 Mean Range Mean Range S 3.53 a 3.41-3.62 4.36 b 4.20-4.67 MS 3.54 a 2.46-4.52 4.68 a 4.40-4.96 MR 3.50 a 2.65-4.42 4.20 b 3.37-4.84 D 2.80 b 2.38-3.03 4.84 a 4.48-5.0 F- test <.001 <.001 sed 0.14 0.09 cv 26.4 13.5 For each column, any value with the same letter were not significantly different (P=0.05) with Fisher LSD test.
Genetic resistance is the only practical method to deal with diseases in perennial forage species in the region. The availability of genetic diversity and a proper method of evaluation and genetic diversity are the basic tools to select for acceptable levels of resistance. The results of the present study indicate that phenotypic evaluation of rust severity in the field is not an appropriate method to identify resistant genotypes. This is possibly due to the existence of an important environmental effect that generates microenvironment variability in the field that hides the genotypic performance. The artificial inoculation made under uniform environmental conditions and performed at a uniform physiological stage (seedlings is more appropriate for the characterization of genetic resistance to Uromyces than field evaluation.
Genetic uniformity can result in Lotus cultivars being vulnerable to the increase and spread of Uromyces pathotypes with virulence to deployed resistance genes. The present study determined that G.Maku was susceptible to the rust strain collected in commercial fields, since all half-sib families tested had high values of rust ratings. This uniformity could be explained by the narrow genetic base of this cultivar (Charlton, 1983). The disease has not been reported in New Zealand, where the tetraploid cultivar was bred. Therefore, selection did not take place during the breeding process in the absence of the pathogen.
No reference about genetic resistance to rust in Lotus uliginosus has been found in the literature. However, Medicago and Trifolium rusts are good examples of case studies where resistance has been identified and the genetic basis has been determined (Diachun and Henson, 1974; Engelke et al, 1977; Elgin et al, 1988; Barbetti and Nichols, 1991). Future Rust resistance evaluation in Lotus uliginosus 71 studies should involve accessions from different geographical origins in order to expand the possibilities to identify genotypes with rust resistance, since this research only involved two accessions. Breeding in Uruguay and neighbouring countries is likely to be based on parents adapted to the regional specific environment. However, this study further emphasises the need to have variability in the basis of effective genetic resistance, which reduce the selection for virulence in the pathogen and minimize the possibility of appearance of a pathotype that affect all cultivars at the commercial level.
Acknowledgements Richard García and Dinorah Rey for his support on the greenhouse testing.
References
ALTIER N. 1997. Enfermedades del Lotus en Uruguay. INIA, Montevideo. Serie Técnica N° 93, 16 p.
BARBETTI M.J. and NICHOLS P.G.H. 1991. Susceptibility of subterranean clover varieties to rust under controlled environment and field conditions. Australian Journal of Experimental Agriculture, 31, 77-80.
BARBETTI M.J. and NICHOLS P.G.H. 1995. Resistance and associated yield losses from rust (Uromyces trifolii-repentis Liro) in midseason cultivars of Trifolium subterraneum L. var. subterraneum L. and var. yanninicum Katzn. et Morley. Australian Journal of Experimental Agriculture, 35, 73-77
CILIUTI J., ARRIVILLAGA S., GERMÁN S., STEWART S., REBUFFO M. and HERNÁNDEZ S. 2003. Studies of rust fungi on Lotus subbiflorus and L.uliginosus. Lotus Newsletter, 33, 19-24.
CHARLTON J.F.L. 1983. Lotus and other legumes. In Wratt G.S. and Smith H.C. (eds) Plant Breeding in New Zealand. Butterworths of New Zealand (Ltd), pp 253-262.
DIACHUN S. and HENSON L. 1974. Dominant resistance to rust in red clover. Phytopathology, 64, 758-759.
ELGIN J.H.J., WELTY R.E., GILCHRIST D.B., HANSON A.A., BARNES D.K. and HILL R.R.J. 1988. Breeding for disease and nematode resistance Alfalfa and alfalfa improvement. Agronomy Monograph No. 29, pp 827-858.
EMERY K.M. and ENGLISH J.T. 1994. Development of foliar diseases of alfalfa in relation to microclimate, host growth, and fertility. Phytopathology, 84, 1263-1269.
ENGELKE M.C., SMITH R.R. and MAXWELL D.P. 1977. Monogenic resistance to red clover leaf rust associated with seedling lethality. Crop Science, 17, 465-468.
72 Sebastián Arrivillaga, Sebastián Hernández, Javier Ciliuti, Mónica Rebuffo, Silvia Germán and Silvina Stewart.
FORMOSO A. 2001. Producción de semillas de lotus Maku. [Seed production of Lotus Maku] In Risso D.F. y albicette M.M (eds) Lotus Maku: manejo, utilización y producción de semillas. [Lotus Maku: management, utilization and seed production]INIA, Montevideo. Serie Técnica N° 119, pp. 39-69. [in Spanish]
JUAN V.F., MONTERROSO L., SACIDO M.B. and CAUHÉPÉ M.A. 2000. Postburning legume seeding in the flooding pampas, Argentina. Journal of Range Management, 53, 300- 304.
SASS J.E. 1958. Acetocarmine in Botanical Microtechnique. 3rd Ed. Ames, Iowa State University Press. pp. 106-108.
SKINNER D.Z. and STUTEVILLE D.L. 1989. Influence of temperature on expression of resistance to rust in diploid alfalfa. Crop Science, 29, 675-677.
Lotus Newsletter (2004) Volume 34, 73-78.
Lotus activities: Background and present research
Juan Ramón Acebes Ginovés Shinji Akada Shinji Akada Said Amrani JAPAN David P. Belesky Ana María Castro We are interested in functional Emmanouil Flemetakis characterization of some members of MYB Ana María de Haro superfamily from Lotus and soybean, which Pedro Insausti may be involved in flavonoid biosynthesis Sangho Jeong and some physiological aspects, such as Richard H. Leep nodulation, nitrogen metabolism, and UV-B Gabriela Martinoia response. Rodolfo Mendoza
Jeremy Murray Said Amrani P.S. Nagar ALGERIA Nazhat-Ezzamane Noureddine Felicia Oliva Tejera I am studying “Survey of nodulation and Graeme Sandral Nitrogen fixation among legumes of Dmitry Sokoloff Algeria”. My interest is in the phylogenetic aspects of this symbiosis and coevolution
threads of the two symbionts. Even if I have a soil microbiology background I'm very interested by the taxonomy and breeding of Juan Ramón Acebes Ginovés Lotus. SPAIN
I am involved, together with my colleagues David P. Belesky Felicia Oliva Tejera, J. Caujape-Castell, and U.S.A. D. Bramwell in relation to the genetic variation (iso-enzimes) and the taxonomy of Earned degrees at Pennsylvania State Lotus spartioides Webb & Berthel., Lotus University (B. Sc., 1973; & M. Sc., 1975) in holosericeus Webb & Berthel, Lotus Agronomy, and West Virginia University campylocladus Webb & Berthel and Lotus (Ph. D., 1978) in Plant Sciences with strong hillebrandii Christ, endemic of the Canary multidisciplinary training in grazing Islands. livestock nutrition. Employed as Research Agronomist with USDA-ARS Research Agronomist for 26 years with10 at Southern Piedmont Conservation Research Center,
73 74 Lotus activities.
Watkinsville, Georgia and the past 16 at in the Agricultural University of Athens. Appalachian Farming Systems Research During the last years his research has been Center, Beaver, West Virginia. Research focused in the molecular mechanisms interests and experience address tall fescue- governing the legume nodule organogenesis endophyte relartionships in terms of forage and function. To this purpose the model production and ecology of tall fescue- legume Lotus japonicus has been used as it endophyte associations. Current interests accumulates several advantages including a address management of forages in diploid relatively small genome size, short silvopastoral systems and forage resources life cycle, and it is amenable to forward and to extend the spatial and temporal reverse genetics. Special attention has been availability of herbage. Currently serve as given in the identification of genes involved Research Leader of the USDA-ARS, in carbon and nitrogen metabolism during Appalachian Farming Systems Research nodule development and function. Several Center, Beaver, West Virginia, with a strong cDNA clones coding for nodule-enhanced multidisciplinary research program isoforms of sucrose cleaving enzymes addressing issues in the soil-plant-grazing (Sucrose synthase and Invertase), sugar animal continuum of pasture in the metabolizing enzymes, isoforms of carbonic Appalachian Region. anhydrase (CA), sugar transporters, trehalose biosynthesis and breakdown enzymes, polyamine biosynthesis enzymes, Ana María Castro and ammonia assimilation enzymes have ARGENTINA been isolated and characterized from L. japonicus nodule specific cDNA libraries. Lotus glaber is a naturalized species in the Real-Time quantitative RT-PCR and in situ Salad River (Buenos Aires province). It is RNA-RNA hybridization techniques have the unique legume forage that grows in soils been used to study the temporal and spatial with saline restrictions, becoming of great expression patterns of these genes. This importance for animal production. Selection research has provided us with new insights of genotypes with tolerance to saline stress into the temporal and spatial organization of will enable to spread out the sowing of carbon and nitrogen metabolism within the Lotus glaber in those soils with low fertility, nodule. As the Molecular Biology research improving the quality of cow food. We was entered a post-genomic phase, the found molecular markers associated with current work is aimed towards the genotypes tolerant to saline conditions. elucidation of the biochemical and These findings will enable the marker physiological role of these enzymes during assisted selection of germplasm better the process of the symbiotic nitrogen adapted to these limiting conditions. Our fixation. To this effort, we are employing group also has determined the prevalent novel experimental approaches, including mechanism of tolerance to saline stress in reverse and forward genetics, high this species (Barragán et al., 2004). throughput metabolite analysis and protein function and structure analysis. Emmanouil Flemetakis
GREECE
Dr. Flemetakis is a lecturer in the Department of Agricultural Biotechnology Lotus activities. 75
Ana María de Haro genetic and biochemical tools. Now I am a ARGENTINA postdoc in Dr. Yanofsky’s lab at UCSD. Our lab has been working on flower and The project “Genetic Improvement of an fruit development using primarily induced tetraploid population of Lotus Arabidopsis. I am interested in expanding glaber Mill. (=Lotus tenuis)” is based at the our work into legume fruit development. Faculty of Agronomy, National University Pod characters such as size, shattering, of the Centre of the Province of Buenos shape and fiber content are not only Aires. A colchicine-induced autotetraploid economically important but also interesting population was generated with the objective in developmental point of view. While of carrying out genetic, cytogenetical, genetic diversity for the characters already physiological, microbiological (symbiotic exist in peas and beans, they are difficult to associations), agronomical and biochemical study. Therefore I have decided to use the studies. Furthermore, this germplasm will be Lotus as a model system. As part of our evaluated with the aim of obtaining an studies, our lab has identified some of the improved population that contemplates the major regulators of pod development in possibility of obtaining tetraploid cultivars, Arabidopsis. I have initiated a project to selected for the pastureland conditions of determine the possible roles that these genes the Depressed Pampa of the Province of may play during pod development in Buenos Aires. We will investigate the legumes, using reverse genetic approaches. arthropod species that feed on Lotus glaber. EMS mutagenesis screens using Lotus We will identify the species, study their japonicus are also in progress. Interesting habits and describe the damage they cause. mutations will be mapped and the corresponding genes will be cloned. I
believe that this work will provide valuable Pedro Insausti insights into legume pod development, and ARGENTINA allow us to compare development/patterning We study the impacts of stressors on Lotus of two dry dehiscent fruit types, pods and glaber and Lotus corniculatus and on siliques. grassland functioning. At the physiological level we seek to understand the mechanisms Richard H. Leep whereby Lotus plants respond to one or U.S.A. more stress factors like flooding, grazing, trampling and water stress. At the ecological I have been from 1976 to 1996 Research level the research interests are the study of and Extension Specialist and Professor at Lotus in a natural grassland and its the Upper Peninsula Experiment Station, responses to variations in disturbance Department of Crop and Soil Sciences, regimes on grasslands of the flooding Michigan State University. At the present Pampa, in Argentina. time, I am Professor and Extension Specialist, Professor at the Department of
Sangho Jeong Crop and Soil Sciences, Michigan State U.S.A. University. Research programs include: 1. Managed rotational grazing, 2. Forage During my Ph.D., I studied Arabidopsis management, 3. Alfalfa persistence, and 4. meristem development using molecular, Dairy-forage cropping systems. Extension 76 Lotus activities. programs include: 1. Providing education plant community sites. Its roots were highly and learning experiences to extension staff colonised by AM fungi. Temporal variations and producers, 2. Sustainable forage in spore density, spore type, AM root systems, 3. Co-chair of Michigan State colonisation, floristic composition and soil University Forage Area of Expertise team. chemical characteristics occurred in each Teaching consists of Forage Crops 201 and site and were different among sites. The advising 5 graduate students and advisory period over which the soil site was subjected committee of 6 graduate students. to flooding influenced the dynamic of the fungal community. Eleven different types of spores were recognized and four were Gabriela Martinoia identified. Two species dominated at the ARGENTINA four sites: Glomus fasciculatum and Glomus intraradices. Spore density was highest in The project "Genetic Improvement of an summer (dry season) and lowest in winter induced tetraploid population of Lotus (wet season) with intermediate values in glaber Mill. (=Lotus tenuis)” is based at the autumn and spring. Colonization of Lotus Faculty of Agronomy, National University glaber roots was highest in summer or of the Centre of the Province of Buenos spring and lowest in winter or autumn. The Aires. A colchicine-induced autotetraploid duration of flooding was associated with population was generated with the objective decreases in spore density and in root of carrying out genetic, cytogenetical, colonization of Lotus glaber, but the main physiological, microbiological (symbiotic change was in the relative density of associations), agronomical and biochemical different spore types. The relative density of studies. Furthermore, this germplasm will be Glomus fasciculatum and Glomus evaluated with the aim of obtaining an intraradices versus Glomus sp. and improved population that contemplates the Acaulospora sp. had distinctive seasonal possibility of obtaining tetraploid cultivars, peaks. These seasonal peaks occurred at the selected for the pastureland conditions of four sites, suggesting differences among the Depressed Pampa of the Province of AM fungus species with respect to the Buenos Aires. We will investigate the seasonality of sporulation. Spore density arthropod species that feed on Lotus glaber. and AM root colonization when measured at We will identify the species, study their one time were poorly related to each other. habits and describe the damage they cause. However, spore density was significantly correlated with root colonization three Rodolfo Mendoza month prior suggesting that high ARGENTINA colonization in one season precedes high sporulation in the next season. We studied seasonal variation in population attributes of AM fungi over two years in four sites of temperate grasslands of the Jeremy Murray Argentinean Flooding Pampas. The sites CANADA represent a wide range in soil conditions, I am currently a member of Krzysztof hydrologic gradient, and floristic Szczyglowski's laboratory. Our work composition. Lotus glaber, a perennial involves mapping and characterization of herbaceous legume naturalised in the nodulation mutants in Lotus japonicus. In Flooding Pampas, was dominant at the four Lotus activities. 77 particular we are looking at mutants Nazhat-Ezzamane Noureddine defective in nodulation that were identified ALGERIA in the har1 mutant supernodulating background. We intend to positionally clone I am studying taxonomy of rhizobia the genes responsible for selected mutations associated to Algerian legumes including in order to better understand the molecular the genus Lotus. This study is done by a processes underlying nodule development numerical taxonomy and biomolecular and autoregulation of nodulation. In means. Even if I have a soil microbiology addition, we are interested in exploring background, I am very interested by the Lotus japonicus as a model system for taxonomy and breeding of Lotus. studies in crop species. Towards this end we are studying the inheritance of quantitative Felicia Oliva Tejera trait loci that effect the protein SPAIN content in seeds and leaves using recombinant inbred lines derived from a I am involved in the studies carried out by Lotus japonicus and Lotus filicaulis cross the group formed by Dr. J. Caujape-Castell, which were created by Niels Sandal at the Juan Ramón Acebes Ginovés and D. University of Aarhus in Denmark as a Bramwell in relation to the genetic variation resource for the Lotus community. (iso-enzimes) and the taxonomy of Lotus Ultimately we hope to clone the genes spartioides Webb & Berthel., Lotus underlying the identified QTL which will holosericeus Webb & Berthel, Lotus subsequently allow us to extend these campylocladus Webb & Berthel and Lotus studies to important legume crop species hillebrandii Christ, endemic of the Canary such as soybean and alfalfa. Islands.
P.S. Nagar Graeme Sandral INDIA AUSTRALIA
I worked as Principal investigator for the I along with Dr Daniel Real have funding studies of “Floristic Diversity the Barda from Australian Wool Innovation and the Sanctuary, Saurashtra”, 2000-2001, GEER Rural Research and Development Foundation, Gandhinagar, “Medicinal plants Corporation to develop commercial cultivar of Saurashtra”, 2001-2002, GEER of Lotus suitable for environments that Foundation, Gandhinagar (with prior experience up to 11 to 13 hours day lengths permission and instructions of the Vice and summer droughts of 5 to 7 months chancellor, Saurashtra University, Rajkot), where the growing season rainfall is 450 to “Threatened trees of Gujarat”, 2002-2003, 650 mm and winter dominant. The primary GEER Foundation, Gandhinagar (with prior target species for this activity are Lotus permission of the Vice chancellor, corniculatus for slightly acid soils and Lotus Saurashtra University, Rajkot). glaber for slightly saline waterlogged soils.
This program has linkages to another in Glen Innes, New South Wales, Australia, run by Dr John Ayres. Primarily our linkage is to provide early prolific flowering populations of Lotus corniculatus to John’s 78 Lotus activities. program which is focused on summer dominant rainfall environments that receive greater than 650 mm and experience short day lengths. Other important linkages have been established with Dr John Howieson from the Centre for Rhizobium Studies, Murdoch University, Western Australia, Australia. The aim of this activity is to ensure that commercial inoculants released for Lotus have a broad and effective host range across species and life forms. We are also keen to develop linkages with other breeding programs where it can be identified that benefits will exist for both programs. The breeding programs described are apart of the CRC for Plant Based Management of Dryland Salinity. They are aimed at reducing soil water recharge (drainage) which if effective will reduce the movement of soil stored salts to lower parts of the landscape.
Dmitry Sokoloff RUSSIA
I study, together with Dr. Tatiana Kramina, sectional delimitation in the genus Lotus as well as generic limits of Lotus. We suggest to exclude all New World species from Lotus. In the Old World, we accept segregate genera Podolotus, Pseudolotus, and Kebirita. We treat Dorycnium and Tetragonolobus as synonyms of Lotus. I am involved also in study of some groups of white and red-flowered Lotus species.
Lotus Newsletter (2004) Volume 34, 79 – 107.
Current list of Lotus researchers
Database last updated September 30 2004
Juan Ramón Acebes Ginovés http://nature.cc.hirosaki-u.ac.jp/gene/ Professor Phone: +81-172-3892 Universidad De La Laguna Fax No.: +81-172-3894 Biología Vegetal (Botánica) Lotus corniculatus, Lotus japonicus. Genetics, 38071 La Laguna- Tenerife Breeding, Physiology, Tissue Culture, Islas Canarias Molecular Biology Spain entry last revised Aug 26 2004 [email protected]
Phone: +34-922-318606 Kenneth A. Albrecht Fax No.: +34-922-318447 Professor Lotus sect. Pedrosia. Taxonomy, Ecology, Univ. of Wisconsin-Madison Molecular Biology. Department of Agronomy entry last revised Jun 2 2004 1575 Linden Dr. Madison WI 53706 Hernan Acuña U.S.A. Director [email protected] Centro Regional de Investigación Quilamapu, Phone: +1-608-262-2314 INIA Fax No.: +1-608-262-5217 Casilla 426 Lotus corniculatus, Lotus uliginosus. Ecology; Chillan forage production; utilization. Chile entry last revised Oct 23 2003 [email protected] Phone: 56-42-211177 Nora Altier Fax No.: 56-42-217852 Researcher Lotus corniculatus, Lotus glaber, Lotus INIA, Nacional Institute for Agricultural uliginosus. Breeding, utilization, germplasm, Research seed, reclamation, physiology, forage. Department of Plant Pathology Germination, emergence, establishment and INIA Las Brujas vegetative growth of Lotus sp. in clay soils. Ruta 48 km.10 entry last revised Nov 7 2003 Canelones CP 90200 Uruguay Shinji Akada [email protected] Associate Professor http://www.inia.org.uy Hirosaki University Phone: +598-2-3677641 Gene Research Center Fax No.: +598-2-3677609 3 Bunkyo-cho Lotus corniculatus, Lotus subbiflorus, Lotus Hirosaki 036-8561 uliginosus. Pathology. Japan entry last revised Nov 7 2003 [email protected]
79
80 Lotus researchers.
Ana Arambarri Rosario Alzugaray Profesora Researcher Facultad de Ciencias Agrarias y Forestales INIA Uruguay Universidad Nacional de La Plata Plant Protection Calles 60 y 118 - C.C. 31 CC 39173 Colonia C. P. 1900 La Plata Uruguay Prov. Buenos Aires [email protected] Argentina http://www.inia.org.uy [email protected] Phone: +598-574-8000 ext 1464 [email protected] Fax No.: +598-574-8012 [email protected] Lotus corniculatus. Entomology. Phone: +54-221-423-6618 entry last revised Oct 29 2003 Fax No.: +54-221-425-2346 Lotus species, Acmispon, Hosackia and
Syrmatium. Plant taxonomy, seeds. Old and Said Amrani New World Lotus species, epidermal Project Manager characteristics. Taxonomy of the New World Laboratorie de Biologie du Sol species, known as: Acmispon, Hosackia and Faculté des Sciences Biologiques Syrmatium. USTHB - BP 32 El Alia - Bab Ezzouar entry last revised Oct 17 2003 16111 – Alger Algeria [email protected] Alberto Artola Phone: +213-21-24-79-50 to 64 ext 936 Breeder Fax No.: +213-21-24-72-17 IPB Semillas All species present in Algeria (Lotus Crop Department conimbricensis, Lotus creticus, Lotus Enrique Hurtado Nº 11 roudairei, Lotus jolyi, Lotus glinoides, Lotus Colonia del Sacramento. CP 70.000 palustris) Microbiology, symbiosis, biological Uruguay Nitrogen fixation. Taxonomy, seed, utilization. [email protected] entry last revised Mar 5 2004 Phone: +598-52-23742 Lotus corniculatus. Seed Production, Crop
Establishment. Development of seed vigor tests Toshio Aoki and seed enhancement methods. Assistant Professor entry last revised Nov 24 2003 Nihom University College of Bioresource Sciences Department of Applied Biological Sciences Ariel Asuaga Kameino 1866 Ingeniero Agrónomo Fujisawa Lancasteriana 2284 Kanagawa 252-8510 Montevideo Japan Uruguay [email protected] [email protected] Phone: +81-466-843703 Phone: +598-2-6007821 Fax No.: +81-466-843353 Fax No.: +598-5325200 Lotus japonicus. Genetics, Physiology, Tissue Lotus corniculatus, Lotus glaber, Lotus Culture, Molecular Biology uliginosus, Lotus subbiflorus. Seed production entry last revised Oct 13 2003 and trade. entry last revised May 5 2004
Lotus researchers. 81
Walter Ayala Researcher INIA Uruguay Søren Bak Forage Department Associate Professor Casilla de Correo 42 The Royal Veterinary and Agricultural CP 33000 Treinta y Tres University, Department Plant Biology Uruguay DK-1871Thorvaldsensvej 40 [email protected] Frederiksberg C, Copenhagen Phone: +598-452-2023 Denmark Fax No.: +598-452-5701 [email protected] Lotus corniculatus, Lotus uliginosus, Lotus http://www.biobase.dk/P450 glaber, Lotus angustissimus. Ecology, http://plbio.kvl.dk http://www.place.kvl.dk Physiology, Forage Production, Utilization, Phone: +45-35283346 Seed Production. Fax No.: +45-38283333 entry last revised Mar 25 2004 Lotus japonicus. Metabolite pathways. Metabolite profiling, transcriptome. entry last revised Oct 16 2003 John F. Ayres Supervisor of Research & Principal Research Scientist Georgget Banchero Agricultural Research & Advisory Station, Senior researcher ‘Centre for Perennial Grazing Systems’ INIA NSW Agriculture Department of Animal Production PMB Glen Innes INIA La Estanzuela New South Wales 2370 Ruta 50 Km 12 Australia 70000 Colonia [email protected] Uruguay Phone: +61-2-67301930 [email protected] Fax No.: +61-2-67301999 Phone: ++598 574 8000 Lotus uliginosus, Lotus corniculatus. Fax No.: ++ 598 574 8012 Breeding, Ecology, Forage Production, Lotus uliginosus. Forage production and Utilization. utilization for strategic nutrition to improve entry last revised Nov 18 2003 sheep reproduction (increase of ovulation rate/improve on lactogenesis). entry last revised Mar 25 2004 Andreas Bachmair Group leader Max Planck Institute for Plant Breeding Gary S. Bañuelos Research USDA-ARS Plant Developmental Biology Water Management Research Laboratory Carl-von-Linné-Weg 10 9611 S. Riverbend Ave. D-50829 Cologne Parlier CA 93648 Germany U.S.A. [email protected] [email protected] http://www.mpiz-koeln.mpg.de/ Phone: +1-559-596-2880 Phone: +49-221-5062265/266 Fax No.: +1-559-596-2851 Fax No.: +49-221-5062207 Lotus corniculatus, Lotus glaber. Remediation Lotus japonicus (currently no research activity of trace element-laden soils with plants. entry last revised Oct 13 2003 with Lotus). Genome organization, retrotransposons. entry last revised Oct 30 2003
82 Lotus researchers.
Mónica Susana Barufaldi Lotus corniculatus. Forage production, Ayudante graduado ecology. Universidad Nacional del Centro de la entry last revised Aug 23 2004 Provincia de Buenos Aires (UNCPBA)
Facultad de Agronomía María Bemhaja Ciencias Básicas Agronómicas y Biológicas Forage Researcher Av. República Italia 780, C.C. 47 INIA Uruguay (7300) Azul, Provincia de Buenos Aires Forage Department, Animal Production Argentina R. 5 Km 386 [email protected] Tacuarembó 45000 http://www.faa.unicen.edu.ar/ Uruguay Phone: +54-2281-433291/92/93 [email protected] Fax No.: +54-2281-43329/92/93 http://www.inia.org.uy Lotus glaber, Lotus corniculatus. Genetics, Phone: +598-632-4560 breeding, forage production, seed production. Fax No.: +598-632-3969 Generate, verify and evaluate autotetraploid Lotus corniculatus, Lotus uliginosus. (via colchicine) populations of L.glaber. entry last revised Oct 31 2003 Physiology, Forage and seed production, Utilization. entry last revised Nov 5 2003 Manuel Becana Professor Estacion Experimental de Aula Dei, CSIC David John Bertioli Department Plant Nutrition Lecturer/Researcher Avda Montañana 1005, Apdo 202 Universidade Catolica de Brasilia 50080 Zaragoza Departamento de Biotecnologia e Ciencias Spain Genomicas [email protected] EMBRAPA-CENARGEN Phone: +34-976-716055 Parque Estação Biológica-pqEB Fax No.: +34-976-716145 Final Av. W5 Norte Lotus japonicus. Biochemistry, Molecular Brasília-DF Biology. Antioxidants –Abiotic stress (drought, CEP: 70770-900 salinity, heavy metals), Free radicals, Thiol Brazil metabolism, Nodule senescence, Oxidative [email protected] stress. http://www.bioinformatica.ucb.br/arachisne entry last revised Oct 16 2003 t/Arachis_Net.htm Phone: +55-61-4484735 Lotus japonicus. Genome synteny of peanut David P. Belesky with Lotus. Research Agronomist/Research Leader entry last revised Jan 2 2004 USDA-ARS Appalachian Farming Systems Research Center Paul R. Beuselinck 1224 Airport Road USDA-ARS Plant Genetics Research Unit Beaver University of Missouri West Virginia 25813-9423 207 Waters Hall U.S.A. Columbia, MO 65211 [email protected] U.S.A. http://www.arserrc.gov/Beckley/ [email protected] Phone: +1-304-256-2841 Phone: +1-314-268-3114 Fax No.: +1-304-256-2921 Fax No.: +1-314-882-1467
Lotus researchers. 83
Lotus corniculatus, Lotus glaber, Lotus uliginosus, Lotus japonicus. Formerly Lotus spp. genetics; breeding; germplasm. Current Nick Brewin research emphasis is on soybean seed Project Leader composition and seed physiology. Former John Innes Centre research was on Lotus breeding and selection Department of Symbiosis Research for improved persistence and evaluation of Norwich Research Park exotic germplasm. Colney entry last revised Nov 7 2003 Norwich NR4 7UH
United Kingdom
[email protected] Arvid A. Boe Phone: +44-1603-450273 Professor of Plant Science Fax No. : +44-1603-450045 Plant Science Department Lotus japonicus. Symbiosis. South Dakota State University entry last revised Aug 10 2004 NPB 244A, Box 2140C Brookings, SD 57007 U.S.A. E. Charlie Brummer [email protected] Associate Professor Phone: +1-605-688-4759 Iowa State University Fax No.: +1-605-688-4452 1204 Agronomy Hall Lotus corniculatus, Lotus purshianus. Ames Iowa 50011 Genetics; breeding; seed production; U.S.A. entomology. [email protected] entry last revised Nov 6 2003 http://www.public.iastate.edu/~brummer
Phone: +1-515-294-1415
Fax No. : +1-515-294-6505 Omar Borsani Lotus corniculatus. Genetics; breeding; forage; Assistant Professor utilization; ecology. Facultad de Agronomía entry last revised Oct 23 2003 Departamento de Biología Vegetal, Bioquímica Av. Garzón 780, Montevideo Uruguay Joe Brummer [email protected] Colorado State University http://www.fagro.edu.uy/bioquimica Western Colorado Research Center Phone: +598-2-3540229 P.O. Box 598 Fax No.: ++ 598-2-3543004 Gunnison, CO 81230 Lotus corniculatus, Lotus uliginosus, Lotus U.S.A. japonicus. Biology, Physiology, Tissue [email protected] Culture, Molecular Biology. Phone: +1-970-641-2515 entry last revised Nov 18 2003 Fax No.: +1-970-641-0653
Lotus corniculatus, Lotus wrightii. Genetics,
breeding; forage, utilization, ecology. Caroline Bowsher Improvement of forage production and quality Dept. of Cell and Structural Biology in mountain meadows. School of Biological Sciences entry last revised Mar 11 2004 University of Manchester Williamson Bd. Oxford Rd. Manchester M13 9PL UK [email protected]
84 Lotus researchers.
Anton van Brussel Assistant professor Bernie Carroll Leiden University Biochemistry and Molecular Biology Institute of Molecular Plant Sciences The University of Queensland Postbus 9505 Brisbane, 4072 2300 RA Leiden Queensland The Netherlands Australia [email protected] [email protected] Phone: +31-71-5275068 Phone: +61 7 3365 2131 Fax No.: +31-71-5275088 Fax No.: +61 7 3365 4699 Lotus japonicus, Lotus pressli. Physiology, entry last revised Mar 15 2004 Microbiology, Molecular Biology (currently no research activity with Lotus). Autoregulation of nodulation of Vetch (Vicia) and Rhizobium Fabricio Dario Cassán leguminosarum. Posdoctoral, CONICET entry last revised Oct 30 2003 Laboratorio de Fisiología Vegetal-UNRC Laboratorio de Biotecnología 1-IIB-INTECh. Universidad Nacional de Rio Cuarto Gustavo Caetano-Anolles Campus Universitario, Ruta 36, km 601 Associate Professor of Bioinformatics (5800) Rio Cuarto University of Illinois Argentina Department of Crop Sciences [email protected] 332 NSRC, 1101 West Peabody Drive Phone: +54-358-4676103 Urbana, IL 61801 Fax No.: +54-358-4676230 U.S.A. Lotus glaber. Microbiology. [email protected] entry last revised Nov 12 2003 http://www.cropsci.uiuc.edu/faculty/gca/ Phone: +1-217-333-8172 Fax No.: +1-217-333-8046 Ana María Castro entry last revised Mar 23 2004 Genetics Proffesor Faculty of Agricultural Sciences Department of Biological Sciences Francesca Cardinale CC31, 1900-La Plata Staff Scientist Argentina University of Turin Phone: ++54-221-423-6758, ext. 421 Di.Va.P.R.A. – Plant Pathology Fax No: ++54-221-423-3698 via Leonardo da Vinci [email protected] 44 Grugliasco (TO)10095 Lotus glaber. Genetics, Breeding, Physiology, Italy Molecular Biology [email protected] entry last revised Aug 4 2004 http://www.divapra.unito.it/personale/dbas e/visualizza_singolo.php?ing=yes&singolo =Cardinale%20Francesca Miguel Cauhépé Phone: ++39-011-670 8701 or 8875 Private Researcher and Consultant on Range Fax No.: ++39-011-236 8875 and Pasture Management Lotus japonicus. Physiology, Pathology, Cereijo 979 Molecular Biology 7620 Balcarce (Prov. de Buenos Aires) entry last revised June 23 2004 Argentina [email protected]
Phone: +54-266-430668 Fax No.: +54-266-430908
Lotus researchers. 85
Lotus glaber. Ecology, Forage Production, Utilization entry last revised Nov 3 2003 Quentin Cronk Professor of Plant Science and Director University of British Columbia Maurizio Chiurazzi UBC Botanical Garden and Centre for Plant Institute of Genetics and Biophysics “A. Research Buzzati Traverso” 6804 SW Marine Drive Via Marconi 12 Vancouver B.C., V6T 1Z4 80125 Naples Canada Italy [email protected] [email protected] http://www.ubcbotanicalgarden.org http://www.iigb.na.cnr.it/ Fax No.: +1-604-822-2016 Phone: +39-081-7257256 Lotus japonicus, Lotus bertholletii, Lotus Fax No.: +39-081-5936123 uliginosus, etc (including N. American Lotus). Lotus japonicus. Gene phylogenies, evolution of florally entry last revised Oct 13 2003 expressed genes. entry last revised Oct 23 2003
Daniel H. Cogliatti Professor of Plant Physiology Hilda Nélida Crosta Facultad de Agronomía, Universidad Nacional Profesor Adjunto del Centro de la Provincia de Buenos Aires Facultad de Agronomía Departamento de Ciencias Básicas Universidad Nacional del Centro de la Agronómicas y Biológicas Provincia de Buenos Aires (UNCPBA). Av. República Italia 780 C.C. 47, (7300) Azul Ciencias Básicas Agronómicas y Biológicas. Provincia de Buenos Aires. Av. República Italia 780, C.C. 47. Argentina (7300) Azul, Provincia de Buenos Aires. [email protected] Argentina http://www.faa.unicen.edu.ar [email protected] Phone: +54-2281-433292 http://www.faa.unicen.edu.ar Fax No.: +54-2281-433292 Phone: +54-2281-433291/92/93 Lotus glaber. Genetics, Physiology, Mineral Fax No.: +54-2281-433291/92/93 nutrition. Lotus glaber, Lotus corniculatus. Biology, entry last revised Nov 17 2003 Taxonomy, Genetics, Tissue Culture, seed production, Molecular Biology. entry last revised Nov 25 2003 Federico Condon Researcher INIA Uruguay Miguel Dall'Agnol Department of Genetic Resources Faculdade de Agronomia INIA La Estanzuela Univ. Federal of Rio Grande do Sul CC 39173, 70000 Colonia Caixa Postal 776 Uruguay 91501-970 Porto Alegre-RS [email protected] Brazil Phone: +598-574-8000 [email protected] Fax No.: +598-574-8012 Phone: +51-3316-7405 Lotus corniculatus, Lotus glaber, Lotus Fax No.: +51-3316-6045 uliginosus, Lotus japonicus. Genetic Diversity, Lotus corniculatus, Lotus uliginosus. Plant Genetics. Allelic diversity, population structure Breeding and Genetics. and genetic resources utilization. entry last revised Oct 8 2003 entry last revised Apr 12 2004
86 Lotus researchers.
Lotus corniculatus, Lotus glaber, Lotus Francesco Damiani subbiflorus. Breeding, Forage production Researcher and utilization. Consiglio Nazionale delle Ricerche entry last revised Nov 12 2003 Istituto Genetica Vegetale sez Perugia via della Madonna Alta 130 06128 Perugia Guilhem Desbrosses Italy Lecturer [email protected] University of Montpellier II http://www.irmgpf.pg.cnr.it UMR113 / Laboratoire des Symbioses Phone: +39-075-5014862 Tropicales et Méditerranéennes / Réponse des Fax No.: +39-075-5014869 plantes aux micro-organismes Lotus corniculatus, Lotus glaber, Lotus UMR113 / CC002 uliginosus, Lotus japonicus. Genetics, Place Eugène Bataillon Breeding, Molecular Biology, Physiology, F-34095 Tissue Culture. Montpellier Cedex 05 entry last revised Oct 10 2003 France [email protected] Phone: +33-4-67149353 David K. Davis Fax No.: +33-4-67143637 Superintendent Lotus japonicus. Genetics, Molecular Biology, University of Missouri Agricultural Microbiology. Experiment Station entry last revised Oct 28 2003 Forage Systems Research Center and Thompson Farm 21262 Genoa Rd Pedro Díaz Gadea Linneus, MO 64653 Assistant Professor U.S.A. Facultad de Agronomía [email protected] Departamento de Biología Vegetal, Bioquímica http://www.aes.missouri.edu/fsrc Av. Garzón 780, Montevideo http://www.aes.missouri.edu/thompson Uruguay Phone: +1-660-895-5121 [email protected] Fax No.: +1-660-895-5122 http://www.fagro.edu.uy/bioquimica Lotus corniculatus. Physiology, forage, seed. Phone: +598-2-3540229 Commercial production and sale of Lotus. Fax No.: +598-2-3543004 entry last revised Nov 30 2003 Lotus corniculatus, Lotus glaber, Lotus uliginosus, Lotus japonicus, Lotus filicaulis. Biology, Physiology, Tissue Culture, José Pedro De Battista Molecular Biology. Forage researcher entry last revised Nov 18 2003 INTA EEA Concepción del Uruguay Area de Investigación en Producción Animal C.C. Nº 6, Concepción del Uruguay Allan Downie (3260) Entre Ríos Professor Argentina John Innes Centre [email protected] Colney Lane http://www.inta.gov.ar Norwich NR4 7UH Phone: +54-3442-425561 UK Fax No.: +54-3442-425578 [email protected] http://www.jic.ac.uk/staff/allan-downie/ Phone: +44-1603-450207
Lotus researchers. 87
Fax No.: +44-1603-450045 Cooperative Research Centre for Plant–based Lotus japonicus. Genetics, Molecular Biology, Management of Dryland Salinity Microbiology, Nodulation signalling. 35 Stirling Hwy entry last revised Oct 21 2003 Crawley Western Australia 6009
Australia Jeffrey S. Dukes [email protected] Assistant Professor http://www.crcsalinity.com Department of Biology Phone: +61-9-64881876 University of Massachusetts Fax No.: +61-9-648811400 100 Morrissey Blvd. Lotus corniculatus. Diversity in the Genus. Boston, MA 02125 Ecology, Germplasm, Breeding, Rhizobiology, U.S.A. Seed Production. [email protected] entry last revised Jan 13 2004 http://globalecology.stanford.edu/DGE/Duk es/Dukes.html Phone: +1-617-287-6614 Marcelo Francisco Eseiza General interest in Lotus. Plant ecology, Jefe de Trabajos Prácticos, Botánica Agrícola I community ecology, ecosystem ecology. Facultad de Agronomía, Universidad Nacional entry last revised Aug 24 2004 del Centro de la Provincia de Buenos Aires (UNCPBA)
Ciencias Básicas Agronómicas y Biológicas Nancy J. Ehlke Av. República Italia 780, C.C. 47 Department of Agronomy & Plant Genetics (7300) Azul, Provincia de Buenos Aires University of Minnesota Argentina 1991 Buford Circle [email protected] St. Paul MN 55108 http://www.faa.unicen.edu.ar U.S.A. Phone: +54-2281-433291 to 93 [email protected] Fax No.: +54-2281-433291 to 93 Phone: +1-612-625-1791 Lotus glaber, Lotus corniculatus. Biology, Fax No.: +1-612-625-1268 Genetics, Tissue Culture, seed. Lotus corniculatus. Genetics, breeding. entry last revised Nov 26 2003 entry last revised Nov. 30 2003
Osvaldo Néstor Fernández James T. English Associate Professor University of Missouri Universidad Nacional de Mar del Plata 116 Waters Facultad de Ciencias Agrarias – Group Columbia, MO 65211 Agroecology U.S.A. C.C. 276 [email protected] (CP7620) Balcarce Phone: +1-573-882-1472 Argentina Fax No.: +1-573-882-0588 [email protected] Pathology. Soilborne pathogen Phytophthora. Phone: +54-2266-439100/05 No current research with Lotus. entry last revised Oct 14 2003 Fax No.: +54-2266-439105 Lotus glaber. Population ecology and forage production. Mike Ewing entry last revised Nov 10 2003 Associate Professor, Deputy CEO
University of Western Australia
88 Lotus researchers.
Emmanouil Flemetakis Kirsten Gausing Lecturer IMSB-University of Aarhus Agricultural University of Athens C.F. Møllers Alle Department of Agricultural Biotechnology Building 130 Iera Odos 75 DK-8000 Aarhus C 118 55 Botanikos Denmark Athens [email protected] Greece Fax No.: +45 86 19 65 00 Phone: +30-210-5294347 Molecular biology. Mainly barley. [email protected] entry last revised June 18 2004 Lotus japonicus, Lotus corniculatus.
Biochemistry, Molecular Biology, Symbiotic José Manuel González García Nitrogen fixation, Carbon Metabolism. entry last revised Sep 2 2004 Ing.Agrónomo – Mejoramiento genético INTA- EEA Balcarce Departamento de Agronomia Henk Franssen C.C. 276- 7620 Balcarce, Bs. As. Assistant Professor Argentina Wageningen University mggarcí[email protected] Molecular Biology Phone: +54 2266-439103 Dreijenlaan 3 Fax No.: +54 2266-439101 6703HA Wageningen Lotus glaber. Breeding, Germplasm, Forage The Netherlands production, Utilization, seed production. [email protected] entry last revised Dec 2 2003 Phone: +31-317-483264
Fax No.: +31-317-483264 William F. Grant Lotus japonicus. Genetics, Molecular Biology, Emeritus Professor developmental program of nodule formation, McGill University nodule evolution. entry last revised Oct 24 2003 Plant Science MacDonald Campus, McGill University P.O. Box 4000 Gabino Garcia de Los Santos Ste. Anne de Bellevue Head of Seed Production Department 21,111 Lakeshore Rd. Instituto de Recursos Genéticos y Quebec H9X 3V9 Productividad Canada Colegio de Postgraduados. [email protected] Carretera Mexico-Texcoco Km. 36.5 http://www.mcgill.ca/plant/faculty/grant/ Texcoco Web page for publications: EDO. de Mexico 56230 http://eqb- Mexico dqe.cciw.ca/eman/ecotools/botanists/Grant [email protected] WF.html Phone: 595 952 02 62 Phone: +1-514-3987851, EXT. 7863 Fax No.: 34-85-381511 Fax No.: +1-514-398-7897 Lotus corniculatus. Physiology, ecology, Lotus corniculatus, Lotus glaber, Lotus spp. breeding, taxonomy. Germplasm management, Genetics; taxonomy; germplasm. Genetics of adaptation, germplasm screening for forage Lotus, especially those involved with the and seed production and breeding. evolution of Lotus corniculatus. entry last revised Nov 21 2003 entry last revised Jun 30 2004
Lotus researchers. 89
Alicia Grassano physiological and biochemical processes. Prof. Titular, Química Analítica. Specifically we have used promoter trapping, Universidad Nacional de La Pampa transfer of the Arabidopsis ethylene Facultad de Ciencias Exactas y Naturales insensitivity (via AtEtrl-l) and non-nodulation Departamento de Química mutants. Uruguay 151 entry last revised Oct 13 2003 (6300) Santa Rosa, La Pampa
Argentina Mette Grønlund [email protected] Post.doc Phone: +54-2954-436787 University of Aarhus Fax No.: +54-2954-432535 Lab. of Gene Expression Lotus glaber. Microbiology, Ecology. entry last revised Oct 21 2003 Science Park Gustav Wieds Vej 10 C DK-8000 Aarhus C Stephanie Greene Denmark Geneticist/Curator [email protected] USDA-ARS Phone: +45-89425008 National Temperate Forage Legume Fax No.: +45-86123178 Germplasm Resources Unit Lotus japonicus. Genetics, Biology, 24106 North Bunn Road Physiology, Pathology, Tissue culture, Prosser, WA 99350 Molecular biology. U.S.A. entry last revised Oct 15 2003 [email protected] http://www.forage.prosser.wsu.edu Ana María de Haro Phone: +1-509-7869265 Professor Fax No.: +1-509-7869370 Facultad de Agronomía All Lotus spp. Germplasm curator for Lotus Universidad Nacional del Centro de la collection in USDA National Plant Germplasm Provincia de Buenos Aires System. entry last revised Nov 21 2003 Departamento de Producción Vegetal Av. República Italia 780 C.C. 47 Peter Gresshoff (7300) Azul Professor/Director Provincia de Buenos Aires The University of Queensland and ARC Centre Argentina of Integrative Legume Research [email protected] ARC Centre of Excellence for Integrative http://www.faa.unicen.edu.ar/ Legume Research Phone: +54-2281-433292 John Hines Plant Science Building Fax No.: +54-2281-433292 The University of Queensland Lotus glaber. Entomology. St Lucia, Qld 4067 entry last revised Apr 23 2004 Australia
[email protected] Nate Hartwig http://www.legumecentre.cilr.uq.edu.au Pennsylvania State University Phone: +61-7-33653550 Department of Agronomy Fax No.: +61-7-33653559 116 ASI Bldg. Lotus japonicus. We conduct research into University Park, PA 16802 Rhizobium-induced nodulation and utilize U.S.A. mutagenesis and transgenics to create the [email protected] necessary diversity needed to determine
90 Lotus researchers.
Lotus corniculatus. Forage, utilization, Mostly Lotus corniculatus. Biology, Molecular management. Cover crops, living mulches in Biology, Genetics corn and soybean. entry last revised June 22 2004 entry last revised July 20 2004
Alberto A. Iglesias Makoto Hayashi Profesor Asociado UNL Assistant Professor Investigador Principal CONICET Osaka University Grupo de Enzimología Molecular Department of Biotechnology, Graduate Bioquímica Básica de Macromoléculas School of Engineering Facultad de Bioquímica y Ciencias Biológicas 2-1, Yamada-Oka Universidad Nacional del Litoral Suita, Osaka 565-0871 Paraje "El Pozo", CC 242 Japan S3000ZAA Santa Fe [email protected] Argentina Phone: +81-6-6879-7417 [email protected] Fax No.: +81-6-6879-7418 entry last revised Nov 30 2003 Lotus japonicus. Symbiosis, Genetics,
Physiology entry last revised July 1 2004 Pedro Insausti Profesor Adjunto Institute for Agricultural Plant Physiology and José A. Herrera-Cervera Ecology Assistant Research Professor CONICET – Faculty of Agronomy (University Facultad de Ciencias. University of Granada of Buenos Aires – Argentina) Departamento de Fisiología Vegetal Av. San Martín 4453 Campus de FuenteNueva s/n. 18071 1417 Buenos Aires Granada Argentina Spain [email protected] [email protected] http://www.ifeva.edu.ar/ Phone: +34-958243382 Phone: +54-11-4524-8070 to 71 Ext 8126 Fax No.: +34-958248995 Fax No.: +54-11-4514-8730 Lotus japonicus. Genetics, Physiology, Lotus glaber and Lotus corniculatus. Pathology, Tissue Culture, Molecular Biology, Ecophysiology. Microbiology. entry last revised Mar. 17 2004 entry last revised Oct 14 2003
Sachiko Isobe Ann M. Hirsch National Agricultural Research Center for Professor Hokkaido Region UCLA Forage Legume Breeding Lab. Molecular, Cell and Developmental Biology Hitsujigaoka 1 405 Hilgard Avenue, Los Angeles, CA 90095- Toyohira 1606 Sapporo, 062-8555 USA Japan [email protected] [email protected] http://www.mcdb.ucla.edu/Research/Hirsc http://cryo.naro.affrc.go.jp/sakumotu/mame h/ ka/etop.htm Phone: +1-310-206-8673 Phone: +81-11-8579272 Fax No.: +1-310-206-5413 Lotus japonicus. Breeding, Genetics. entry last revised Oct 17 2003
Lotus researchers. 91
Euan James Research Scientist Sangho Jeong University of Dundee Postdoc School of Life Sciences University of California, San Diego (UCSD) Centre for High Resolution Imaging & Yanofsky Lab Processing(CHIPs) Section of Cell and Developmental Biology MSI/WTB Complex La Jolla, CA 92093-0116 Dundee DD1 5EH USA UK Phone: +1-858-534-7298 [email protected] Fax No.: +1-858-822-1772 Phone: +44-1382-344741 [email protected] Fax No.: +44-1382-345893 Lotus japonicus. Development, Genetics, Lotus corniculatus, Lotus uliginosus, Lotus Molecular Biology. entry last revised Jan 7 2004 japonicus. Physiology, structure (especially of nodules). entry last revised Oct 18 2003 Qunyi Jiang Laboratory manager Martín Jaurena The University of Queensland Research Assistance ARC Centre of Excellence for Integrative Ministry of Livestock Agriculture and Legume Research Fisheries John Hines Building Department of Soil Microbiology Brisbane Uruguay Qld 4072 Phone: ++598 2 2038152 Australia Fax No.: ++598 2 2038152 [email protected] [email protected] www.legumecentre.cilr.uq.edu.au http://fp.chasque.net:8081/microlab/LMSCI Phone: +61-7-33657227 /LMSCI.htm Fax No.: +61-7-33653556 Lotus corniculatus, Lotus glaber, Lotus Lotus japonicus. Genetics, genomics and molecular biology. uliginosus. Genetics, Germplasm, Taxonomy, entry last revised Nov 21 2003 Ecology, Microbiology. entry last revised Aug 2 2004 Bjarne Jochimsen
Associate professor Erik Østergaard Jensen IMSB-Lab. of Gene Expression Associate Professor University of Aarhus University of Aarhus Department of Molecular Biology Dept. Molecular Biology. Lab. of Gene C.F. Møllers Alle Expression Gustav Wieds Vej 10C Science Park DK-8000 Aarhus C Gustav Wieds Vej 10 C Denmark DK-8000 Aarhus C [email protected] Denmark Phone: +45-89-42-50-39 [email protected] Fax.no.: +45-86-12-31-78 http://130.225.13.27/~eoj/ Lotus japonicus. Seed development. Phone: +45-89425014 Entry last revised Jan 7 2004 Lotus japonicus. Molecular biology, physiology, organ development. entry last revised Nov. 7 2003
92 Lotus researchers.
Bodil Jørgensen Iera Odos 75 Senior scientist 11855 Athens Danish Institute of Agricultural Science Greece Depart. Plant Biology, Biotechnology Group [email protected] Thorvaldsensvej 40, opg. 8, 2 sal Phone: +30-210-5294314 1871 Frederiksberg C Fax No.: +30-210-5294314 Denmark Lotus japonicus. Molecular Biology, [email protected] Biochemistry, Physiology. Phone: +45-35282578 entry last revised Oct 22 2003
Fax No.: +45-35282589
Lotus japonicus. Tissue culture, Tony Kavanagh transformation, Molecular biology, Professor, Head of Department biochemistry. entry last revised Nov 25 2003 Trinity College Dublin Smurfit Institute Department of Genetics Dublin 2 Adem Kamalak Ireland Assistant Professor [email protected] Kahramanmaras Sutcu Imam University Phone: +353-1-6081035 Faculty of Agriculture Fax No.: +353-1-6714968 Department of Animal Science Lotus japonicus. Genetics. Kahramanmaras, 46040 entry last revised Oct 14 2003 Turkey [email protected] Masayoshi Kawaguchi Phone: +903442237666 Associate Professor Fax: +903442230048 University of Tokyo Lotus corniculatus. Forage Production, Graduate School of Sciences Utilization. entry last revised Jul 28 2004 Department of Biological Sciences Hongo Bunkyo-ku Norihito Kanamori Tokyo 113-0033 National Food Research Institute Japan Food Engineering Division [email protected] Kannondai 2-1-12 Fax No.: +81-3-58414458 Tsukuba-city Lotus japonicus, Lotus burttii. Molecular basis Ibaraki 305-8642 of symbiosis and systemic regulation of Japan nodule development [email protected] entry last revised Oct 21 2003 Phone: +81-29-8388047
Fax No.: +81-29-8391552 Walter Kelman Lotus japonicus. Breeding and Molecular Research Scientist Biology. entry last revised Nov 5 2003 CSIRO Division of Plant Industry Institute of Plant Production and Processing GPO Box 1600 Panagiotis Katinakis Canberra ACT 2601 Professor Australia Agricultural University of Athens [email protected] Botanikos Phone: +61-2-62465083
Lotus researchers. 93
Fax No.: +61-2-62465255 Lotus uliginosus, Lotus corniculatus. Cultivar Tatiana E. Kramina breeding and genetic analysis of agronomic Scientific Researcher traits. Moscow State University entry last revised Nov 7 2003 Biological Faculty
Higher Plants Department
Vorobyevy Gory, 1, building 12, Peter Kemp 199992 Moscow Associate Professor Russia Institute of Natural Resources [email protected] Massey University Phone: +7-095-9391603 Private Bag 11 222 Fax No.: +7-095-9391827 Massey University Section Lotus, Lotus corniculatus group. Palmerston North Taxonomy. Natural hybridization. New Zealand entry last revised Oct 13 2003 [email protected] Lotus corniculatus. Ecology, forage production, grazing management, animal Lene Krusell responses. Max Planck Institute of Molecular Plant entry last revised Aug 4 2004 Physiology
MPI-MP
Am Mühlenberg 1 Kyung-Nam Kim 14476 Golm Assistant Professor Germany Sejong University [email protected] Department of Molecular Biology Phone: +49-331-5678150 98 Gunja-Dong Lotus japonicus. Gwangjin-Gu entry last revised Oct 16 2003 Seoul 143-747 Korea [email protected] Alejandro La Manna Phone: +82-2-34083647 Researcher Fax No.: +82-2-34083661 Instituto Nacional de Investigacion Arabidopsis thaliana. Lotus japonicus. Agropecuaria (INIA) Molecular Biology. Dairy Department entry last revised Nov 20 2003 Ruta 50 km 11 CC 39173
70000 Colonia
Uruguay Joseph H. Kirkbride [email protected] USDA-Agricultural Research Service http://www.inia.org.uy/ Systematic Botany & Mycology Laboratory Phone: +598-574-8000 Bldg 011A, Rm 304 Fax No.: +598-574-8012 BARC-West Lotus corniculatus, Lotus uliginosus. Animal Beltsville, MD 20705-2350 Nutrition, nutrient management and U.S.A. environmental issues. Ecology, Biology, [email protected] Forage Production, Utilization, Seed Phone: +1-310-5049447 Production. Fax No.: +1-310-5045810 entry last revised March 24 2004 All Lotus spp. Taxonomy. Systematics of Lotus spp. entry last revised Oct 9 2003
94 Lotus researchers.
Pedro Laterra Av. Gral Paz entre Albarellos y Constituyentes Universidad Nacional de Mar del Plata INTI, Edif. 24 (Colectora Gral Paz 5445) Facultad de Ciencias Agrarias (1650) San Martín CC 276, 7620 Balcarce Buenos Aires Prov. Buenos Aires Argentina Argentina [email protected] [email protected] http://www.iib.unsam.edu.ar http://www.mdp.edu.ar Lotus glaber, Lotus japonicus. Microbiology. Phone: +54-2266-439100 entry last revised Nov 7 2003 Fax No.: +54-2266-439101
Lotus glaber. Ecology. entry last revised Oct 29 2003 Lina A.C. Lett Assitant Professor Facultad de Agronomía W.H. (Bill) Leakey Universidad Nacional del Centro de la Deer Creek Seed Provincia de Buenos Aires Box 105 Ciencias Básicas Agronómicas y Biológicas Ashland, WI 54806 Av. República Italia 780 U.S.A. C.C. 47, (7300) Azul [email protected] Provincia de Buenos Aires. Phone: +1-715-2783200 Argentina Fax No.: +1-715-2783209 [email protected] Lotus corniculatus, Lotus glaber, Lotus http://www.faa.unicen.edu.ar uliginosus. Seed. Commercial production and Phone : +54-2281-433291 to 93 sale of Lotus. Fax No.: +54-2281-433292 entry last revised Oct 28 2003 Lotus glaber, Lotus corniculatus.
Microbiology, Molecular Biology, Forage
production, Genetics. Richard H. Leep entry last revised Nov 10 2003 Michigan State University Crop and Soil Sciences W. K. Kellogg Biological Station Dasharath Prasad Lohar 3700 E. Gull Lake Drive Research Associate Hickory Corners, MI 49060 University of Minnesota, St. Paul U.S.A. Department of Plant Biology [email protected] 1445 Gortner Ave. http://www.msue.msu.edu/fis/ 250 Biological Sciences Building Phone: +1-269-671-2323 Saint Paul, MN 55108 Fax No.: +1-269-671-2104 U.S.A. Lotus corniculatus. Forage Production, [email protected] physiology. Phone: +1-612-6249230 entry last revised Mar 3 2004 Fax No.: +1-612-6251738
Lotus japonicus. Molecular biology,
microbiology, tissue culture, biology, genetics. Viviana Lepek entry last revised Oct 17 2003 CONICET researcher and Assistant Professor of Universidad de Gral San Martín Instituto de Investigaciones Biotecnológicas W. L. (Bill) Lowther IIB-UNSAM Scientist Departamento de Microbiología (Interacción AgResearch Rhizobium-leguminosas) Plant Breeding and Genomics
Lotus researchers. 95
Invermay Agricultural Centre Jim H. McAdam Private Bag Principle Scientific Officer Mosgiel Agriculture and Food Science Centre New Zealand Applied Plant Science Research Division [email protected] Newforge Lane http://www.agresearch.co.nz Belfast BT9 5PX Phone: +64-3-4899053 Northern Ireland Fax No.: +64-3-4893739 [email protected] Lotus uliginosus, Lotus corniculatus. Ecology, Phone: +44-28-90255275 utililization, microbiology, inoculation, Fax No.: +44-28-90255003 rhizobia. Lotus uliginosus. Ecology, Utilisation. entry last revised Oct 23 2003 entry last revised Nov 21 2003
Antonio J. Marquez Mark McCaslin Profesor Titular Forage Genetics Departamento de Bioquímica Vegetal y N5292 Gills Coulee Rd. Biología Molecular West Salem WI 54669 Facultad de Química U.S.A. Apartado 553 [email protected] 41080-Sevilla Phone: +1-608-7862121 Spain Fax No.: +1-608-7862193 [email protected] Lotus corniculatus. Forage, seed. Phone: +34-95-4557145 entry last revised Nov 5 2003 Fax No.: +34-95-4626853
Lotus japonicus. Biochemistry, Molecular Robert L. McGraw biology, Biotechnology, Nitrogen assimilation, Associate Professor of Agronomy Enzymology, Mutagenesis entry last revised Oct 7 2003 University of Missouri Department of Agronomy 208 Waters Gabriela Martinoia Columbia, MO 65211 Research Assitant U.S.A. Facultad de Agronomía [email protected] Universidad Nacional del Centro de la Phone: +1-314-882-6608 Provincia de Buenos Aires Fax No.: +1-314-882-1467 Departamento de Producción Vegetal Lotus corniculatus. Forage, utilization, Av. República Italia 780 physiology. Management strategies to improve C.C. 47 performance and persistence. (7300) Azul entry last revised Nov 30 2003 Provincia de Buenos Aires
Argentina Mariana Melchiorre [email protected] Post-doc student http://www.faa.unicen.edu.ar/ INTA Phone: +54-2281-433292 Instituto de Fitopatologia y Fisiologia Vegetal Fax No.: +54-2281-433292 Córdoba Lotus glaber. Entomology. Argentina entry last revised Apr 23 2004 [email protected] http://www.inta.gov.ar/iffive/ Phone: +54-351-4974343
96 Lotus researchers.
Fax No.: +54-351-4974330 physiology, tissue culture, molecular biology, Lotus japonicus. Physiology, Molecular microbiology. biology. entry last revised Nov 18 2003 entry last revised Jun 22 2004
Ken Moore Rodolfo Mendoza Professor Senior Research Iowa State University CONICET Department of Agronomy CEFYBO 1567 Agronomy Hall Serrano 669 Ames Iowa 50011 1414 Buenos Aires U.S.A. Argentina [email protected] [email protected] http://www.public.iastate.edu/~kjmoore/ Phone: +54-1148570847 Phone: +1-515-2943160 Fax No.: +54-1148570847 Fax No.: +1-515-2943163 Lotus corniculatus, Lotus glaber. Ecology, Lotus corniculatus. Forage, utilization, Biology, Physiology, Forage Production, ecology, physiology. Utilization, Microbiology. entry last revised Oct 27 2003 entry last revised Jul 1 2004
Jorge A. Mosjidis Réal Michaud Professor Agriculture et Agroalimentaire Canada / Auburn University Agriculture and Agri-Food Canada Department of Agronomy and Soils Centre de recherche et de développement sur 202 Funchess Hall les sols et les grandes Cultures / Soils and Auburn, AL 36849-5412 Crops Research and Development Centre U.S.A. 2560 Hochelaga Blvd. [email protected] Ste-Foy Québec G1V 2J3 http://www.ag.auburn.edu/ay/mosjidis Canada Phone: +1-334-8443976 [email protected] Fax No.: +1-334-8443945 Phone: +1-418-6577980 ext. 261 Genetics. Fax No.: +1-418-6482402 entry last revised Nov 6 2003 Lotus corniculatus. Forage, utilization. entry last revised Oct 29 2003 John Mundy Professor Jorge Monza Institute of Molecular Biology Professor Oester Farimagsgade 2A Facultad de Agronomía 1353 Copenhagen K Departmento de Biología Vegetal, Bioquímica Denmark Av. Garzón 780 [email protected] Montevideo http://www.biobase.dk/~mundy Uruguay Phone: +45-35322131 [email protected] Fax No.: +45-35322128 http://www.fagro.edu.uy/bioquim/web Lotus japonicus. Phone: +598-2-3540229 entry last revised Oct 16 2003
Fax No.: +598-2-3543004
Lotus corniculatus, Lotus glaber, Lotus uliginosus, Lotus japonicus. Biology,
Lotus researchers. 97
Jeremy Murray Columbia, MO 65211 Post Doctoral Fellow U.S.A. Southern Crop Protection and Food Research [email protected] Centre (SCPFRC), Agriculture Canada Phone: +1-314-882-2801 1391 Sandford St. Fax No.: +1-341-882-1467 London Ont N5V 4T3 Lotus corniculatus. Physiology, forage. Canada entry last revised Jul 15 2004 [email protected]
Phone: +1-519-457-1470 Ext 643 Minoru Niizeki Fax No.: +1-519-457-3997 Professor Lotus japonicus, Lotus filicaulis. Genetics of Hirosaki University nodulation, molecular biology. entry last revised Jan 5 2004 Faculty of Agriculture and Life Science Plant Breeding and Genetics Hirosaki P.S. Nagar Aomori-ken 036-8561 Coordinator, IMPLANT, Scientist Japan IMPLANT [email protected] Department of Biosciences Phone: +81-172-393776 Saurashtra University Fax No.: +81-172-393750 Rajkot – 360005 Lotus corniculatus, Lotus japonicus. Genetics, India breeding and molecular biology. Plant [email protected] breeding and genetics by using somatic cell http://wwwsaurashtrauniversity.edu/ hybridization and somaclonal variation. Phone: +91-281-2571154 entry last revised Oct 18 2003
Lotus corniculatus var. minor, Lotus corniculatus, Lotus garcini. Taxonomy, Nazhat-Ezzamane Noureddine ccology, forage production, tissue culture, Teacher/Searcher molecular biology. entry last revised Dec 31 2003 Faculté des Sciences Biologiques Laboratoire de Biologie du Sol USTHB - BP 32 El Alia - Bab Ezzouar Valeria Negri 16111 - Alger Associate Professor Algeria Universita' Degli Studi Di Perugia [email protected] Biologia Vegetale Biotecnologie Phone: +213-21-24-79-50 to 64 ext. 936 Agroambientali Fax No.: +213-21-24-72-17 Borgo XX Giugno 74 Lotus conimbricensis, Lotus creticus, Lotus 06100 Perugia roudairei, Lotus jolyi, Lotus glinoides, Lotus Italy palustris (all species present in Algeria). [email protected] Microbiology, symbiosis, biological nitrogen http://www.agr.unipg.it/dbvba fixation, taxonomy, seed, utilization. Phone: +39-075-5856218 entry last revised Mar 5 2004
Fax No.: +39-075-5856224
Lotus curator. Germplasm, genetics. entry last revised Nov 25 2003 Takuji Ohyama Professor Faculty of Agriculture C. Jerry Nelson Niigata University University of Missouri Department of Applied Biological Chemistry 210 Waters Hall 2-8050 Ikarashi
98 Lotus researchers.
Niigata, 950-2181 [email protected] Japan http://www.ibvf.cartuja.csic.es/ [email protected] Phone: +34-95-4489526 Phone: +81-25-2626643 Fax No. : +34-95-4460065 Fax No.: +81-25-2626643 Lotus japonicus. Physiology, nitrogen Lotus japonicus. Genetics, physiology, assimilation. symbiosis, molecular biology, microbiology. entry last revised Oct 31 2003 entry last revised Nov11 2003
María Cristina de Pablo Felicia Oliva Tejera Profesor Adjunto Doctoral Student Universidad Nacional del Centro de la Cabildo Insular de Gran Canaria Provincia de Buenos Aires (UNCPBA) Jardín Botánico Canario “Viera y Clavijo” Facultad de Agronomía Apartado de Correos, 14. Laboratorio Regional de Análisis de Semillas y Tarija Alta Granos 35017 Las Palmas de Gran Canaria Av. República Italia 780, C.C. 47 Gran Canaria, Islas Canarias (7300) Azul Spain Provincia de Buenos Aires [email protected] Argentina Phone: +34-928219580 [email protected] Fax No.: +34-928219581 http://www.faa.unicen.edu.ar Lotus sect. Pedrosia. Taxonomy, ecology, Phone: +54-2281-427566 molecular biology. Fax No.: +54-2281-433292 entry last revised Jun 2 2004 Lotus glaber, Lotus corniculatus. Seed
production and quality. entry last revised Nov 4 2003 Fernando Olmos Assistant Researcher Instituto Nacional Investigación Agropecuaria Cristina Pacios-Bras - INIA Postdoctoral position Pasture Ecology Leiden University Ruta 5, Km. 386 Laboratoire des Proteines du Cytosquelette Tacuarembo 41, Rue Jules Horowitz Uruguay F-38027 Grenoble Cedex 1 [email protected] France Phone: +598-63-22407 [email protected] Fax No.: +598-63-23969 Phone: +33-4-38789219 Lotus corniculatus; Lotus subbiflorus; Lotus Fax No.: +33-4-38785494 uliginosus. Germplasm, ecology, physiology, Former Lotus japonicus research. forage production. entry last revised Oct 31 2003 entry last revised Nov 5 2003
Peter Paľove-Balang Alicia Orea Researcher Postdoctoral Position Institute of Botany SAV Instituto de Bioquímica Vegetal y Fotosíntesis Department of Plant Physiology CSIC-Universidad de Sevilla Dúbravská cesta 14 Avda. Américo Vespucio, s/n Bratislava 41092-Seville Slovak Republic, 845 23 Spain [email protected]
Lotus researchers. 99
Phone: +421-2-59426127 Phone: +49-551-3914430 Fax No.: +421-2-54771948 Fax No.: +49-551-395749 Lotus japonicus. Physiology, molecular Lotus japonicus. Physiology, utilization, tissue biology. culture, molecular biology. entry last revised Oct 11 2003 entry last revised Jun 29 2004
Yousef A. Papadopoulos Andrea Pedrosa-Harand Agriculture and Agri-Food Canada PostDoc 14 Fundy Drive Department of Cell Biology and Genetics Truro, NS B2N 5Z3 Institute of Botany Canada University of Vienna [email protected] Rennweg 14 Phone: +1-902-8960400 A-1030 Vienna Fax No.: +1-902-8960200 Austria Lotus corniculatus. Genetics, breeding, forage. [email protected] Developing germplasm for seedling vigor and Phone: +43-1-4277-54026 competitive ability for cultivar development. Fax No.: + 43-1-4277-9541 entry last revised Oct 22 2003 Lotus japonicus, Lotus filicaulis, Lotus burttii. Genetics (cytogenetics, physical mapping,
comparative genomics), taxonomy. Currently Martin Parniske no research activities with Lotus. Professor entry last revised Aug 17 2004 University of Munich Department Biology I - Genetics Maria Ward Strasse 1a Carsten Poulsen D-80639 Muenchen University of Aarhus Germany Dep.Mol.Biol. - Lab.Gene Expression [email protected] Science Park http://www.jic.bbsrc.ac.uk/science/sl/smp.h Gustav Wieds Vej 10 C tm DK-8000 Aarhus C Phone: +49-89-21806150 Denmark Fax No.: +49-89-1785633 [email protected] Lotus japonicus, Lotus filicaulis. Genetic and http://130.225.13.27/~chp/ physical mapping of plant genes. Molecular Phone: +45-89425007 genetic analysis of pathogenic and symbiotic Fax.no.: +45-86123178 plant-microbe interactions. Nitrogen fixing root Lotus japonicus. Genetics, molecular biology, nodule and arbuscular mycorrhiza symbiosis. microbiology. entry last revised Aug 4 2004 entry last revised Oct 16 2003
Katharina Pawlowski Braj Nandan Prasad Group Leader Professor & Coordinator,Programme in Göttingen University Biotechnology Plant Biochemistry Department Tribhuvan University Justus-von-Liebig-Weg 11 Programme in Biotechnology C/O Central 37077 Göttingen Department of Botany Germany Kirtipru [email protected] P.O.Box 9782 http://www.plant-biochem.uni- Kathmandu goettingen.de/pawlowski.html Nepal
100 Lotus researchers. [email protected] http://www.inia.org.uy/sitios/lnl/ Phone: +977-1-330582 Phone: +598-574-8000 Ext 1479 Fax No: +977-1-4331964 Fax No.: +598-574-8012 Lotus corniculatus, Lotus japonicus. Lotus corniculatus, Lotus uliginosus, Lotus Physiology and molecular biology. subbiflorus. Breeding, genetics, Lotus curator, entry last revised Oct 18 2003 Lotus Newsletter editor. entry last revised Oct 2 2003
Daniel Real Senior Plant Breeder Kevin Reed University of Western Australia Department of Primary Industries University Field Station Pastoral and Veterinary Institute 1 Underwood Avenue PB 105 Shenton Park Hamilton, Victoria 3300 Perth, WA 6008 Australia Australia [email protected] [email protected] Phone: +61-55-730911 Phone: +61-8-92-872831 Fax No.: +61-55-711523 Fax: +61-8-93-839907 Lotus uliginosus, Lotus corniculatus. Ecology, Lotus corniculatus, Lotus japonicus, Lotus forage, utilization, germplasm. Plant glaber, Lotus uliginosus, Lotus creticus, Lotus introduction, cultivar evaluation, animal cytisoides, Lotus marrocanus, Lotus australis, production. Lotus cruentus. Breeding and genetics. entry last revised Oct 12 2003 entry last revised Mar 4 2004
J.S. Grant Reid Daniel Real Professor Emeritus Senior Plant Breeder University of Stirling National Institute of Agricultural Research Department of Plant Biochemistry INIA Tacuarembó Stirling FK9 4LA Ruta 5 km 386 Scotland C.C. 78086 C.P. 45000 United Kingdom Tacuarembó [email protected] Uruguay Phone: +44-1786-467762 [email protected] Fax No.: +44-1786-464994 Phone: +598-63-22407 Lotus japonicus. Physiology, molecular Lotus ornithopodioides, Lotus angustissimus, biology. Lotus conimbricensis, Lotus shoelleri. entry last revised Nov 30 2003
Breeding and genetics. entry last revised Mar 4 2004 Joel Reynaud Laboratoire de Botanique Mónica Rebuffo Faculté de Pharmacie Senior Researcher Avenue Rockefeller National Institute of Agricultural Research 69373 Lyon Cedex 08 INIA “La Estanzuela” France Ruta 50, km 11 [email protected] C.P. 70000 Colonia http://ispb.univ-lyon1.fr/cours/botanique Uruguay Teaching. No current research activity with [email protected] Lotus. http://www.inia.org.uy entry last revised Nov 30 2003
Lotus researchers. 101
Fax No.: +44-1970-823242 Ken Richards Lotus corniculatus, Lotus japonicus. Genetics, Research Manager biology, physiology, molecular biology. Plant Gene Resources of Canada entry last revised Jul 2 2004 Agriculture and Agri-Food Canada
Saskatoon Research Centre Craig A. Roberts 107 Science Place Associate Professor Saskatoon University of Missouri Saskatchewan S7N0X2 Agronomy Department Canada 214 Waters Hall [email protected] Columbia, MO 65211 http://agr.gc.ca/pgrc-rpc U.S.A. http://agr.gc.ca/rpc-pgrc [email protected] Phone: +1-306-956-7641 http://www.psu.missouri.edu/roberts/ Fax No.: +1-306-956-7246 Phone: +1-573-8822801 Germplasm collection with many species of Fax No.: +1-573-8844317 Lotus housed at PGRC. Germplasm Mainly Lotus corniculatus, but other species as conservation, taxonomy, seed production, well. Germplasm screening, forage quality, genetics, breeding. entry last revised Sep 30 2004 biochemical defense. entry last revised Oct 22 2003
Diego F. Risso William John Rogers Senior Researcher Profesor Titular National Institute of Agricultural Research Facultad de Agronomía INIA “Tacuarembó” Universidad Nacional del Centro de la Ruta 5 km 386 Provincia de Buenos Aires C.C. 78086 C.P. 45000 Ciencias Básicas Agronómicas y Biológicas Tacuarembó Av. República Italia 780, C.C. 47 Uruguay (7300) Azul [email protected] Provincia de Buenos Aires. http://www.inia.org.uy Argentina Phone: +598-63-22407 [email protected] Lotus corniculatus, Lotus uliginosus, Lotus http://www.faa.unicen.edu.ar subbiflorus. Forage management and Phone: +54-2281-433292 utilization under grazing, physiology. entry last revised Mar 8 2004 Fax No.: +54-2281-433292 Lotus corniculatus, Lotus glaber. Genetics, breeding, molecular biology. Mark P. Robbins entry last revised Nov 25 2003 Researcher
Institute of Grassland and Environmental Clive Ronson Research Chair in Genetics Plant, Animal and Microbial Sciences University of Otago Aberystwyth Research Centre Microbiology Department Plas Gogerddan P.O. Box 56 Aberystwyth Dunedin Ceredigion New Zealand United Kingdom [email protected] [email protected] Phone: +44-1970-823113
102 Lotus researchers. http://microbes.otago.ac.nz/dept/STAFF/pr ofile-ronsonc.html Niels Sandal Fax No.: +64-4798540 Permanent academic staff Lotus japonicus, Lotus corniculatus. University of Aarhus Microbiology and genetics of the Department of Molecular Biology-Lab. of microsymbiont Mesorhizobium loti. Gene Expression entry last revised Oct 9 2003 Gustav Wieds Vej 10 C DK-8000 Aarhus C
Denmark Oscar Adolfo Ruiz [email protected] Researcher (CONICET) and Professor Phone: +45-89-425006 (UNSAM) Fax No.: +45-86-123178 Instituto de Investigaciones Biotecnológicas- Lotus japonicus, Lotus filicaulis. Genetics, Instituto Tecnológico de Chascomús (IIB- molecular biology, map based cloning, INTECh) recombinant inbred lines, symbiosis. Unidad de Biotecnología 1 IIB-INTECh entry last revised Mar 23 2004 Casilla de Correo 164 Camino de circunvalación Km 5 (B7130 IWA), Chascomús Matt A. Sanderson Provincia de Buenos Aires Research Agronomist Argentina USDA-ARS [email protected] Pasture Lab Building [email protected] Curtin Road http://www.iib.unsam.edu.ar University Park Phone: +54-2241-424049/430323 PA 16802-3702 Fax No.: +54-2241-424048 U.S.A. Lotus corniculatus, Lotus glaber, Lotus [email protected] japonicus. Biology, physiology, molecular General interest in Lotus. Production, biology, microbiology utilization, grazing land ecologist. entry last revised Oct 6 2003 entry last revised Aug 17 2004
Federico Sanchez Graeme Sandral Professor Research Officer (Plant ecology and genetics) Universidad Nacional Autónoma de México University of Western Australia and NSW Instituto de Biotecnología Agriculture Plant Molecular Biology Department University Field Station Av. Universidad 2001 1 Underwood Drive, Shenton Park Col. Chamilpa. 62210 Perth, WA 6009 Cuernavaca, Morelos Australia México [email protected] [email protected] [email protected] Fax No.: +52-73-172388 http://www1.crcsalinity.com/ Lotus japonicus, Lotus corniculatus. Molecular Phone: +61-8-92-872-851 and cell biology, cell imaging, actin Fax No.: +61-8-93-839-907 cystoskeleton, cell signaling, nodule Mobile: + 61-040-922-6235 development, Rhizobium-legume interactions. Perennial Lotus species known to the entry last revised Nov 30 2003 Macaronesian Islands (Azores, Madeira’s,
Canary Islands and Cape Verde) and the more cultivated species such as Lotus corniculatus
Lotus researchers. 103 and Lotus glaber. Plant breeding, Science Park phylogenetics, taxonomy, ecology. Gustav Wieds Vej 10 C entry last revised Mar 24 2004 DK-8000 Aarhus C
Denmark
[email protected] Juan Sanjuán Fax No.: +45 86 12 31 78 Research Scientist Consejo Superior de Investigaciones Científicas (CSIC) Simone Meredith Scheffer-Basso Estación Experimental del Zaidin Professor Departamento de Microbiologia del Suelo y Universidade de Passo Fundo Sistemas Simbióticos Instituto de Ciências Biológicas Prof. Albareda 1 Departamento Biologia E-18008 Granada Rua Silva Jardim, 303 apto. 701 Spain 99010-240 Passo Fundo [email protected] Rio Grande do Sul http://www.eez.csic.es/inves/grupos/D3.htm Brazil Phone: +34-958-181600 Ext 259 [email protected] Fax No.: +34-958-129600 http://www.upf.tche.br Lotus japonicus, Lotus corniculatus, Lotus Phone: +55-316-8100 Ext. 8326 glaber, Lotus uliginosus. Microbiology. Lotus corniculatus. Morphophysiology, entry last revised Oct 13 2003 evaluation of forage production (cutting- grazing tolerance, growth habit, nutritive value, Sindhu Sareen competition, mixtures with grasses). Indian Grassland and Fodder Research Institute entry last revised Oct 16 2003
CSKHPKV Campus Palampur 176062 (H.P.) Ms Anne Schneider India Executive Secretary Delegate [email protected] AEP, European Association for Grain Legume entry last revised Oct 9 2003 Research Executive Secretariat Shusei Sato 12 Avenue George V Researcher 75 008 Paris Kazusa DNA Research Institute France Plant gene research group [email protected] 2-6-7 Kazusa-Kamatari http://www.grainlegumes.com Kisarazu, 292-0818 Phone: +33-1-40694909 Japan Fax No.: +33-1-47235872 [email protected] The AEP is an international multidisciplinary http://www.kazusa.or.jp/lotus/ network of scientists and end-users concerned Phone: +81-438-523935 with grain legumes. The AEP office is Fax No.: +81-438-523934 interested in being informed about the progress Lotus japonicus. Genomics, molecular in Lotus investigations for establishing genetics. connection with other legumes research entry last revised Nov 24 2003 activities. entry last revised Nov 20 2003
Leif Schauser IMSB-Lab. of Gene Expression University of Aarhus
104 Lotus researchers.
Philippe Seguin pink-flowered species. Cladistic analyses. Assistant Professor Flower development in Lotus corniculatus. McGill University, Macdonald Campus entry last revised Oct 9 2003
Department of Plant Science
21,111 Lakeshore Rd. Gary Stacey Ste. Anne de Bellevue Professor Quebec H9X 3V9 University of Missouri Canada Plant Microbiology and Pathology [email protected] 108 Waters Hall http://www.mcgill.ca/plant/faculty/seguin/ Columbia, MO 65203 Phone: +1-514-3987851 U.S.A. Fax No.: +1-514-3987897 [email protected] Lotus corniculatus. Forage production, utilization. http://psu.missouri.edu/staceylab/index entry last revised Nov 21 2003 .htm Phone: +1-573-884-4752 Fax No.: +1-573-882-0588 Richard R. Smith Lotus japonicus. Symbiotic nitrogen fixation, Retired Research Geneticist genetics, microbiology, pathology. USDA-ARS, University of Wisconsin entry last revised Mar 23 2004 US Dairy Forage Research Center 1925 Linden R. West Madison WI 53706 Jens Stougaard U.S.A. Lecturer, Group leader [email protected] University of Aarhus Phone: +1-608-2645240 IMSB-Lab. of Gene Expression Fax No.: +1-608-2645147 Department of Molecular Biology Lotus corniculatus. Genetics, breeding, forage, Science Park cultivar evaluation. Selection for improved Gustav Wieds Vej 10 C seedling establishment and improved DK-8000 Aarhus C persistence Denmark entry last revised Nov 11 2003 [email protected] Phone: +45-89-425011 Fax No.: +45-86-21222 Dmitry Sokoloff Lotus japonicus. Plant molecular genetics and Lecturer general molecular biology. Genetic mapping, Higher Plants Department map-based cloning, gene isolation, gene Biological Faculty characterisation, expression studies. Moscow State University entry last revised Oct 15 2003 119992 Moscow Russia [email protected] Martina Stromvik http://www.herba.msu.ru/ Assistant professor Phone: +7-095-9391603 McGill University Fax No.: +7-095-9392777 Department of Plant Science Lotus australis and related taxa, Lotus Macdonald campus arabicus and related taxa, Lotus discolor and 21,111 Lakeshore Rd related taxa, Lotus creticus and related taxa. Ste.-Anne-de-Bellevue Taxonomy. Generic limits of Lotus, taxonomy QC, H9X 3V9 of its segregate genera, sectional system of Canada Lotus, taxonomy of some white-, red- and [email protected]
Lotus researchers. 105 http://www.mcgill.ca/plant/faculty/stromvik Kagawa University Phone: +1-514-398-8627 Miki-cho, Kita-gun Fax No.: +1-514-398-7897 Kagawa 761-0795 Soybean (For the moment not studying Japan Lotus). Bioinformatics, Genomics, [email protected] Molecular Biology. Phone: +81-87-8913129 entry last revised Aug 26 2004 Fax.no.: +81-87-8913021 Lotus japonicus. Genetics, molecular biology. entry last revised Oct 17 2003 Norio Suganuma Professor Aichi University of Education Nancy Terryn Department of Life Science Researcher Kariya University Gent Aichi 448-8542 Instititute Plant Biotechnology for Developing Japan Countries [email protected] K.L. Ledeganckstraat 35 Phone: +81-566-262647 9000 Gent Fax No.: +81-566-262310 Belgium Lotus japonicus. Genetics, physiology. [email protected] entry last revised Nov 5 2003 http://www.ipbo.ugent.be/ Phone: +32-9-2645201 Fax No.: +32-9-2648795 John Sullivan Lotus japonicus, Phaseolus, Lathyrus. Research fellow Transformation, seed development, stress University of Otago response, molecular biology of Grain legumes Department of Microbiology such as Phaseolus, cowpea, Lathyrus for which New Zealand there is little genomic data so we look at model [email protected] systems Lotus japonicus. Phone: +64-3-4798373 entry last revised Oct 20 2003 Fax No.: +64-3-4798540 Lotus corniculatus, Lotus japonicus. Molecular biology, microbiology. Toshiki Uchiumi entry last revised Nov 20 2003 Associate Professor Kagoshima University Faculty of Science Krzysztof Szczyglowski Department of Chemistry and BioScience Agriculture & Agri-Food Canada 1-21-35 Korimoto 1391 Sandford Street Kagoshima 890-0065 London, ON N5V 4T3 Japan Canada [email protected] [email protected] Phone: +81-99-2858164 Phone: +1-519-457-1470 ext. 273 Fax No.: +81-99-2858163 Fax.no.: +1-519-457-3997 Lotus japonicus. Physiology, molecular Lotus japonicus biology, microbiology, symbiosis. entry last revised Oct 02 2003 entry last revised Oct 21 2003
Shigeyuki Tajima Professor Lab. Molecular Plant Nutrition Faculty of Agriculture
106 Lotus researchers.
Michael Udvardi Lotus corniculatus, Lotus glaber. Germplasm, Group Leader ecology, biology, physiology, forage Max-Planck-Institut fur Molekulare production, utilization, seed production. Pflanzenphysiologie entry last revised Nov 9 2003
Molecular Plant Nutrition
Am Mühlenberg 1 Jeffrey J. Volenec 14476 Golm Professor Deutchland Purdue University [email protected] Department of Agronomy Phone: +49-331-5678149 Lilly Hall of Life Sciences Fax No.: +49-331-5678250 915 West State St. Lotus japonicus. Genetics, biochemistry, West Lafayette IN 47907-2054 molecular biology, functional genomics U.S.A. (transcriptomics, proteomics, metabolics). [email protected] Symbiotic nitrogen fixation and molecular http://www.agry.purdue.edu/staffbio/jjvbio. plant nutrition. entry last revised Oct 22 2003 htm Phone: +1-765-494-8071 Fax No.: +1-765-496-2926 Don R. Viands Lotus corniculatus. Physiology, utilization, Associate Dean and Director of Academic molecular biology, biochemistry. Programs, Professor of Plant Breeding entry last revised Oct 27 2003 Dept. of Plant Breeding and Biometry
151 Roberts Hall Trevor L. Wang Ithaca, NY 14853 Group Leader U.S.A. Metabolic Biology Department [email protected] John Innes Centre Phone: +1-607-255-3081 Norwich Research Park Fax No.: +1607-254-4613 Norwich NR4 7UH Lotus corniculatus. Breeding, pathology. United Kingdom Breeding for resistance to crown-rot and to [email protected] Fusarium wilt caused by Fusarium oxysporum. entry last revised Oct 7 2003 http://www.jic.bbsrc.ac.uk/staff/trevor- wang/index.htm Phone: +44-1603-450283 Osvaldo Ramón Vignolio Fax No.: +44-1603-450014 Professor Lotus japonicus. Genetics, biology, Universidad Nacional de Mar del Plata- EEA physiology, molecular biology, metabolism. Facultad de Ciencias Agrarias entry last revised Nov 4 2003 INTA Balcarce
Producción Vegetal Judith Webb Ruta 226, Km 73.5 Research Scientist CC 276 (7620) Balcarce Institute of Grassland and Environmental Argentina Research [email protected] Plas Gogerddan [email protected] Aberystwyth [email protected] Ceredigion Phone: +54-2266-439100 SY23 3EB Wales Fax No.: +54-2266-439101 United Kingdom [email protected]
Lotus researchers. 107
Phone: +44-1970-823124 Fax No.: +44-1970-823243 Lotus japonicus, Lotus corniculatus. Genetics, biology, physiology, tissue culture, molecular biology, microbiology. entry last revised Nov 9 2003
Lotus Newsletter (2004) Volume 34, 108 – 118.
Lotus literature Database provided by Lotus Newsletter recipients, last updated September 30 2004
BORISOV A.Y., MADSEN L.H., TSYGANOV V.E., UMEHARA Y., VOROSHILOVA V.A.., BATAGOV A.O., SANDAL A. FREDERIKSEN N., SCHAUSER L., ELLIS N., TIKHONOVICH I.A. and STOUGAARD J. 2003. The Sym35 gene required for root nodule development in Pisum sativum is is an orthologue of Nin gene from Lotus japonicus. Plant Physiology, 131, 1009-1017. Comparative phenotypic analysis of pea (Pisum sativum) sym35 mutants and Lotus japonicus nin mutants suggested a similar function for the PsSym35 and LjNin genes in early stages of root nodule formation. Both the pea and L. japonicus mutants are non-nodulating but normal in their arbuscular mycorrhizal association. Both are characterized by excessive root hair curling in response to the bacterial microsymbiont, lack of infection thread initiation, and absence of cortical cell divisions. To investigate the molecular basis for the similarity, we cloned and sequenced the PsNin gene, taking advantage of sequence information from the previously cloned LjNin gene. An RFLP analysis on recombinant inbred lines mapped PsNin to the same chromosome arm as the PsSym35 locus and direct evidence demonstrating that PsNin is the PsSym35 gene was subsequently obtained by cosegregation analysis and sequencing of three independent Pssym35 mutant alleles. L. japonicus and pea root nodules develop through different organogenic pathways, so it was of interest to compare the expression of the two orthologous genes during nodule formation. Overall, a similar developmental regulation of the PsNin and LjNin genes was shown by the transcriptional activation in root nodules of L. japonicus and pea. In the indeterminate pea nodules, PsNin is highly expressed in the meristematic cells of zone I and in the cells of infection zone II, corroborating expression of LjNin in determinate nodule primordia. At the protein level, seven domains, including the putative DNA binding/dimerization RWP-RK motif and the PB1 heterodimerization domain, are conserved between the LjNIN and PsNIN proteins. Full text at http://www.plantphysiol.org/cgi/content/full/131/3/1009
BONFANTE P., GENRE A., FACCIO A., MARTINI I., SCHAUSER L., STOUGAARD J., WEBB J. and PARNISKE M. 2000. The Lotus japonicus LjSym4 gene is required for the successful symbiotic infection of root epidermal cells. Molecular Plant Microbe Interactions, 13, 1109-1120. The role of the Lotus japonicus LjSym4 gene during the symbiotic interaction with Mesorhizobium loti and arbuscular mycorrhizal (AM) fungi was analyzed with two mutant alleles conferring phenotypes of different strength. Ljsym4-1 and Ljsym4-2 mutants do not form nodules with M. loti. Normal root hair curling and infection threads are not observed, while a nodC-dependent deformation of root hair tips indicates that nodulation factors are still perceived by Ljsym4 mutants. Fungal infection attempts on the mutants generally abort within the epidermis, but Ljsym4-1 mutants allow rare, successful, infection events, leading to delayed arbuscule formation. On roots of mutants homozygous for the Ljsym4-2 allele, arbuscule formation was never observed upon inoculation with either of the two AM fungi, Glomus intraradices or Gigaspora margarita. The strategy of epidermal penetration by G. margarita was identical for Ljsym4-2 mutants and the parental line, with appressoria, hyphae growing between two epidermal cells, penetration of epidermal cells through their anticlinal wall. These observations define a novel, genetically controlled step in AM colonization. Although rhizobia penetrate the tip of root hairs and AM fungi access an entry site near the base of epidermal cells, the
108 109 Lotus Literature
LjSym4 gene is necessary for the appropriate response of this cell type to both microsymbionts. We propose that LjSym4 is required for the initiation or coordinated expression of the host plant cell's accommodation program, allowing the passage of both microsymbionts through the epidermis layer.
FORSLUND K., MORANT M., JORGENSEN B., OLSEN C.E., ASAMIZU E., SATO S., TABATA S. and BAK S. 2004. Biosynthesis of the nitrile glucosides rhodiocyanoside A and D and the cyanogenic glucosides lotaustralin and linamarin in Lotus japonicus. Plant Physiology, 135, 71-84. Lotus japonicus was shown to contain the two nitrile glucosides rhodiocyanoside A and rhodiocyanoside D as well as the cyanogenic glucosides linamarin and lotaustralin. The content of cyanogenic and nitrile glucosides in L. japonicus depends on plant developmental stage and tissue. The cyanide potential is highest in young seedlings and in apical leaves of mature plants. Roots and seeds are acyanogenic. Biosynthetic studies using radioisotopes demonstrated that lotaustralin, rhodiocyanoside A, and rhodiocyanoside D are derived from the amino acid l-Ile, whereas linamarin is derived from Val. In silico homology searches identified two cytochromes P450 designated CYP79D3 and CYP79D4 in L. japonicus. The two cytochromes P450 are 94% identical at the amino acid level and both catalyze the conversion of Val and Ile to the corresponding aldoximes in biosynthesis of cyanogenic glucosides and nitrile glucosides in L. japonicus. CYP79D3 and CYP79D4 are differentially expressed. CYP79D3 is exclusively expressed in aerial parts and CYP79D4 in roots. Recombinantly expressed CYP79D3 and CYP79D4 in yeast cells showed higher catalytic efficiency with l-Ile as substrate than with l-Val, in agreement with lotaustralin and rhodiocyanoside A and D being the major cyanogenic and nitrile glucosides in L. japonicus. Ectopic expression of CYP79D2 from cassava (Manihot esculenta Crantz.) in L. japonicus resulted in a 5- to 20-fold increase of linamarin content, whereas the relative amounts of lotaustralin and rhodiocyanoside A/D were unaltered.
GRANT W.F. and MCDOUGALL R.B. 1995. Registration of H401-4-4-2 birdsfoot trefoil germplasm resistant to sulfonylurea. Crop Science, 35, 286-287.
GRUBER M.Y., RAY H., AUSER P., SKADHAUGE B., FALK J., THOMSEN K.K., STOUGAARD J., MUIR A., LEES G., COULMAN B., MCKERSIE B., BOWLEY S. and VON WETTSTEIN D. 1999. Genetic systems for condensed tannin biotechnology. In GROSS G.G., HEMINGWAY R. and YOSHIDA T. (Eds.) Plant Polyphenols 2: Chemistry and Biology. Plenum Press, New York. [Manual]
HAYASHI M., MIYAHARA A., SATO S., KATO T., YOSHIKAWA M., TAKETA M., HAYASHI M., PEDROSA A., ONDA R., IMAIZUMI-ANRAKU H., BACHMAIR A., SANDAL N., STOUGAARD J., MUROOKA Y., TABATA S., KAWASAKI S., KAWAGUCHI M. and HARADA K. 2001. Construction of a Genetic Linkage Map of the Model Legume Lotus japonicus Using an Intraspecific F2 Population. DNA Research, 8, 301–310. Among leguminous plants, the model legume Lotus japonicus (Regel) Larsen has many biological and genetic advantages. We have developed a genetic linkage map of L. japonicus based on amplified fragment length polymorphism (AFLP), simple sequence repeat polymorphism (SSRP) and derived cleaved amplified polymorphic sequence (dCAPS). The F2 mapping population used was derived from a cross between two L. japonicus accessions Gifu B-129 and Miyakojima MG-20. These parental accessions showed remarkable cytological differences, particularly with respect to size and morphology of chromosomes 1 and 2. Using fluorescence in situ hybridization (FISH) with BAC clones from Gifu B-129 and TAC (Transformation-competent Artificial Chromosome) clones from Lotus Literature 110
Miyakojima MG-20, a reciprocal translocation was found to be responsible for the cytological differences between chromosomes 1 and 2. The borders of the translocations were identified by FISH and by alignment toward the L. filicaulis×L. japonicus Gifu B-129 linkage map. The markers from the main translocated region were located on linkage groups 1 and 2 of the two accessions, Gifu B-129 and Miyakojima MG-20, respectively. The framework of the linkage map was constructed based on codominant markers, and then dominant markers were integrated separately in each linkage group of the parents. The resulting linkage groups correspond to the six pairs of chromosomes of L. japonicus and consist of 287 markers with 487.3 cM length in Gifu B-129 and 277 markers with 481.6 cM length in Miyakojima MG-20. The map and marker information is available through the World Wide Web at http://www.kazusa.or.jp/lotus/. Free text at http://dna-res.kazusa.or.jp/8/6/06/PDF/8_301.pdf
KADE M., PAGANI E.A. and MENDOZA R.E. 2003. Phosphorus utilization efficiency in populations of narrow leaf-birdsfoot trefoil. Communications in Soil Science and Plant Analysis, 34, 271-284. The widespread distribution of the pasture legume Lotus glaber Mill. throughout the Flooding Pampa in Argentina may be attributed to ecotypic differentiation among populations. In order to confirm such genetic variation, two morphologically distinct populations from contrasting soil types, particularly in the phosphorus (P) status, were compared in their response to a different P supply. Plants were cloned by stem cuttings and transplanted to a common soil fertilized with increasing amounts of P. Morphological characters including number of shoots, leaf length and width, as well as the leaf length/width ratio, remained different in spite of P supply. Biomass P content, tissue P concentration and the percentage of root length infected by mycorrhizae associated with P uptake, were similar between populations. However, both populations showed a different growth response to P fertilization in spite of being cultivated on the same soil. Consequently, they differed in the efficiency of P utilization (E), which is probably under genetic control. These results produce evidence in support of the existence of ecotypes for L. glaber adapted to local soil conditions.
KADE M., PAGANI E.A. and MENDOZA R.E. 2003. A morphological study of populations of Lotus glaber Mill. (Fabaceae). Agronomie, 23, 203-207. The pasture legume Lotus glaber Mill. has colonized the Flooding Pampa (Argentina) in spite of high environmental heterogeneity. Morphological characters of plants from different populations were compared to evaluate if plastic or genetic differentiation could have contributed to such a widespread geographic distribution. Plants were collected at random from six different sites and grown in a greenhouse for five months. Clonal replicates of those plants were cultivated in pots on a native and a common soil. Populations were replicated three times and each replicate (pot) had three plants from the same clone. The number of primary shoots, internode length, leaf length and width and shoot length were recorded and the significance of population mean differences determined. All characters, except for leaf width, differed between populations on each sampling in both sets of experiments. This variability is therefore best explained by genetic differentiation of the populations.
KRUSELL L., MADSEN L.H., GENUA A., SZCZYGLOWSKI K., AUBERT G., DUC G., SATO S., TABATA S., DE BRUIJN F., PAJUELO E., SANDAL N. and STOUGAARD J. 2002. Shoot control of root development and nodulation is mediated by a receptor-like kinase. Nature, 420, 422-426. In legumes, root nodule organogenesis is activated in response to morphogenic lipochitin oligosaccharides that are synthesized by bacteria, commonly known as rhizobia. Successful symbiotic interaction results in the formation of highly specialized organs called root nodules, which provide a unique environment for symbiotic nitrogen fixation. In wild-type plants the number of nodules is 111 Lotus Literature regulated by a signalling mechanism integrating environmental and developmental cues to arrest most rhizobial infections within the susceptible zone of the root. Furthermore, a feedback mechanism controls the temporal and spatial susceptibility to infection of the root system. This mechanism is referred to as autoregulation of nodulation, as earlier nodulation events inhibit nodulation of younger root tissues. Lotus japonicus plants homozygous for a mutation in the hypernodulation aberrant root (har1) locus escape this regulation and form an excessive number of nodules. Here we report the molecular cloning and expression analysis of the HAR1 gene and the pea orthologue, Pisum sativum, SYM29. HAR1 encodes a putative serine/threonine receptor kinase, which is required for shoot- controlled regulation of root growth, nodule number, and for nitrate sensitivity of symbiotic development.
LEEP R., JERANYAMA P., MIN D.H. and DIETZ T. 2002. Grazing effects on herbage mass and composition in grass-birdsfoot trefoil mixtures. Agronomy Journal, 94, 1257–1262. Grass–legume mixtures have the ability to supply more consistent forage yields across a wide range of environments throughout the grazing season than do grass monocultures. The suitability of diverse grass species in binary mixtures with birdsfoot trefoil (Lotus corniculatus L.) in rotational stocking systems has not been extensively studied. The objective of this study was to evaluate binary mixtures of five cool-season grasses with the birdsfoot trefoil cultivar Norcen for herbage mass, botanical composition, and cattle (Bos taurus) grazing preference under a rotational stocking. Experiments were established at Lake City and Chatham, MI, in 1994. Binary mixtures were grazed for 2 yr with beef or dairy cows three times yearly at predetermined periods from spring to fall. Total herbage dry mass production ranged from 3 to 10 Mg ha-1 yr-1 over two years and locations. The grass fraction in binary mixtures was 327 to 946 g kg-1 in swards over two years and locations. Perennial ryegrass (Lolium perenne L.) failed to persist at Lake City, probably due to less consistent snow cover. Birdsfoot trefoil fraction was highest in binary mixtures with smooth bromegrass (Bromus inermis Leyss) and timothy (Phleum pratense L.). Binary mixtures with orchardgrass (Dactylis glomerata L.) and tall fescue (Festuca arundinacea Schreb.) produced the highest herbage biomass but were less preferred by grazing animals while binary mixtures with timothy and smooth bromegrass were associated with the highest apparent herbage utilization at both locations (84–100%). Full text at http://agron.scijournals.org/cgi/content/full/94/6/1257
LEEP R., ANDRESEN J.R. and JERANYAMA P. 2001. Fall Dormancy and Snow Depth Effects on Winterkill of Alfalfa. Agronomy Journal, 93, 1142-1148. Received for publication December 11, 2000. The lack of a definitive method to assess winter hardiness in alfalfa (Medicago sativa L.) remains a challenge in the north-central region of the USA where winterkill of alfalfa can be severe. The reliability of fall dormancy ratings for describing alfalfa cultivar susceptibility to winter injury and the role of snow depth in moderating temperatures near the plant were investigated at Chatham, MI on a Chatham Stony loam (Typic Haplorthod). Four cultivars were selected with a range of fall dormancy ratings: ‘Nitro’, ‘Magnum IV’, ‘Saranac’, and ‘Vernal’. The cultivars were planted in 1993–1994, 1994–1995, and 1995–1996 seasons in plots over which 0-, 10-, and 20-cm winter snow depths were maintained. Temperatures were monitored for each plot, and stand counts were made each fall and spring to assess winter injury. Nitro suffered the most winterkill across snow cover treatments. The total yield range was 0 to 9 Mg ha-1 in the absence of a snow cover and 0.4 to 12 Mg ha-1 for a snow depth of at least 10 cm, except in 1996. Extreme minimum canopy- level (6 cm) temperatures for 10-cm snow depth averaged over three winter seasons were 12.1°C higher than the 0-cm snow cover treatment, which translated into higher yields. The results suggest that snow cover of 10 cm adequately protects alfalfa from winter injury. Cultivars within the same fall dormancy rating did not necessarily perform similarly, suggesting the need to develop other methods for assessing winter survival. Lotus Literature 112
Full text at http://agron.scijournals.org/cgi/content/full/93/5/1142
MADSEN L.H., COLLINS N.C., RAKWALSKA M., BACKES G., SANDAL N., KRUSELL L., JENSEN J., WATERMAN E.H., JAHOOR A., YALIFFE M., PRYOR A.J., LANGRIDGE P., SCHULTZE-LEFERT P. and STOUGAARD J. 2003. Barley disease resistance gene analoges of the NBS-LRR class – identification and mapping. Molecular Genetics and Genomics, 269, 150-161. The majority of verified plant disease resistance genes isolated to date are of the NBS-LRR class, encoding proteins with a predicted nucleotide binding site (NBS) and a leucine-rich repeat (LRR) region. We took advantage of the sequence conservation in the NBS motif to clone, by PCR, gene fragments from barley representing putative disease resistance genes of this class. Over 30 different resistance gene analogs (RGAs) were isolated from the barley cultivar Regatta. These were grouped into 13 classes based on DNA sequence similarity. Actively transcribed genes were identified from all classes but one, and cDNA clones were isolated to derive the complete NBS-LRR protein sequences. Some of the NBS-LRR genes exhibited variation with respect to whether and where particular introns were spliced, as well as frequent premature polyadenylation. DNA sequences related to the majority of the barley RGAs were identified in the recently expanded public rice genomic sequence database, indicating that the rice sequence can be used to extract a large proportion of the RGAs from barley and other cereals. Using a combination of RFLP and PCR marker techniques, representatives of all barley RGA gene classes were mapped in the barley genome, to all chromosomes except 4H. A number of the RGA loci map in the vicinity of known disease resistance loci, and the association between RGA S- 120 and the nematode resistance locus Ha2 on chromosome 2H was further tested by co-segregation analysis. Most of the RGA sequences reported here have not been described previously, and represent a useful resource as candidates or molecular markers for disease resistance genes in barley and other cereals.
MADSEN E.B. MADSEN L.H., RADUTOIU S., OLBRYT M., RAKWALSKA M., SZCZYGLOWSKI K., KANEKO T., SATO S., TABATA S., SANDAL N. and STOUGAARD J. 2003. A receptor- like kinase gene involved in legume perception of rhizobial signal molecules. Nature, 425, 637-640. Plants belonging to the legume family develop nitrogen-fixing root nodules in symbiosis with bacteria commonly known as rhizobia. The legume host encodes all of the functions necessary to build the specialized symbiotic organ, the nodule, but the process is elicited by the bacteria. Molecular communication initiates the interaction, and signals, usually flavones, secreted by the legume root induce the bacteria to produce a lipochitin-oligosaccharide signal molecule (Nod-factor), which in turn triggers the plant organogenic process. An important determinant of bacterial host specificity is the structure of the Nod-factor, suggesting that a plant receptor is involved in signal perception and signal transduction initiating the plant developmental response. Here we describe the cloning of a putative Nod-factor receptor kinase gene (NFR5) from Lotus japonicus. NFR5 is essential for Nod-factor perception and encodes an unusual transmembrane serine/threonine receptor-like kinase required for the earliest detectable plant responses to bacteria and Nod-factor. The extracellular domain of the putative receptor has three modules with similarity to LysM domains known from peptidoglycan- binding proteins and chitinases. Together with an atypical kinase domain structure this characterizes an unusual receptor-like kinase.
MCKENZIE D.B., PAPADOPOULOS Y.A. and MCRAE K.B. 2004. Harvest management affects yield and persistence of birdsfoot trefoil (Lotus corniculatus L.) in cool summer climates. Canadian Journal of Plant Science, 84, 525-528. 113 Lotus Literature
Two studies determined the effect of birdsfoot trefoil harvest managements on persistence, productivity, and species composition under cool summer growing conditions. Single and double cut managements were harvested at 10, 50, and 100% bloom; triple cuts and simulated grazing were also evaluated. Double cut harvest management produced considerably more dry matter yield (DMY) in the first production year, but its advantage over single cuts in the second year depended on bloom at harvest. Harvesting at 10% bloom for single (and perhaps double) cuts appears to be the best system based on DMY, trefoil content, and stand density.
NAGAR P.S. and PANDYA S.M. 2002. Stylosanthes hamatus (Linn.) Taub. (Papilionaceae): A new record to the flora of Gujarat. Bomb. Nat. Hist. Soc., 99, 363-364. (With one text figure).
NAGAR P.S. and PANDYA S.M. 2002. Addition to the flora of Saurashtra, Gujarat, India. J.Econ. Taxon.Bot., 26, 75-77.
NAGAR P.S. Floristic Diversity of Barda Hills and its surroundings. Scientific Publishers, Jodhpur. (Book, Publication in progress).
NAGAR P.S. Biodiversity of Barda Wildlife Sanctuary, GEER Foundation, Gandhinagar, Gujarat, India. (Book, Publication in progress).
NAGAR P.S. Medicinal plants of Gujarat, GEER Foundation, Gandhinagar, Gujarat, India. (Book, Publication in progress).
NIKOLIC R. and MITIC N. 2003. Morphological changes in atypical bird’s foot trefoil plants obtained during genetic transformation by Agrobacterium. Acta Biologica Jugoslavica, Series F, Genetics. (Acta Biolog. Jugosl. F G), 35, 177-185.
PACIOS-BRAS C., SCHLAMANN H.R.M., BOOT K., ADMIRAAL P., LANGERAK J.M., STOUGAARD J. and SPAINK H.P. 2003. Auxin distribution in Lotus japonicus during root nodule development. Plant Molecular Biology, 52, 1169-1180. For this work, Lotus japonicus transgenic plants were constructed expressing a fusion reporter gene consisting of the genes beta-glucuronidase (gus) and green fluorescent protein (gfp) under control of the soybean auxin-responsive promoter GH3. These plants expressed GUS and GFP in the vascular bundle of shoots, roots and leafs. Root sections showed that in mature parts of the roots GUS is mainly expressed in phloem and vascular parenchyma of the vascular cylinder. By detecting GUS activity, we describe the auxin distribution pattern in the root of the determinate nodulating legume L. japonicus during the development of nodulation and also after inoculation with purified Nod factors, N- naphthylphthalamic acid (NPA) and indoleacetic acid (IAA). Differently than white clover, which forms indeterminate nodules, L. japonicus presented a strong GUS activity at the dividing outer cortical cells during the first nodule cell divisions. This suggests different auxin distribution pattern between the determinate and indeterminate nodulating legumes that may be responsible of the differences in nodule development between these groups. By measuring of the GFP fluorescence expressed 21 days after treatment with Nod factors or bacteria we were able to quantify the differences in GH3 expression levels in single living roots. In order to correlate these data with auxin transport capacity we measured the auxin transport levels by a previously described radioactive method. At 48 h after inoculation with Nod factors, auxin transport showed to be increased in the middle root segment. Lotus Literature 114
The results obtained indicate that L. japonicus transformed lines expressing the GFP and GUS reporters under the control of the GH3 promoter are suitable for the study of auxin distribution in this legume.
PEDROSA A., SANDAL N., STOUGAARD J., SCHWEITZER D. and BACHMAIR A. 2002. Chromosomal map of the model legume Lotus japonicus. Genetics, 161, 1661-1672. Lotus japonicus is a model plant for the legume family. To facilitate map-based cloning approaches and genome analysis, we performed an extensive characterization of the chromosome complement of the species. A detailed karyotype of L. japonicus Gifu was built and plasmid and BAC clones, corresponding to genetically mapped markers (see the accompanying article by Sandal et al. 2002, this issue), were used for FISH to correlate genetic and chromosomal maps. Hybridization of DNA clones from 32 different genomic regions enabled the assignment of linkage groups to chromosomes, the comparison between genetic and physical distances throughout the genome, and the partial characterization of different repetitive sequences, including telomeric and centromeric repeats. Additional analysis of L. filicaulis and its F1 hybrid with L. japonicus demonstrated the occurrence of inversions between these closely related species, suggesting that these chromosome rearrangements are early events in speciation of this group. Free text at http://www.genetics.org/cgi/content/full/161/4/1661
PERRY J.A., WANG T.L., WELHAM T.J., GARDNER S., PIKE J.M., YOSHIDA S., PARNISKE M. 2003. A TILLING reverse genetics tool and a web-accessible collection of mutants of the legume Lotus japonicus. Plant Physiology 131, 866-871. Full text at http://www.plantphysiol.org/cgi/content/full/131/3/866
QUAEDVLIEG N.E.M., SCHLAMAN H.R.M., ADMIRAAL P.C., WIJTING S.E., STOUGAARD J. and SPAINK H.P. 1998. Fusions between green fluorescent protein and β-glucuronidase as sensitive and vital bifunctional reporters in plants. Plant Molecular Biology, 37, 715-727. By fusing the genes encoding green fluorescent protein (GFP) and beta-glucuronidase (GUS) we have created a set of bifunctional reporter constructs which are optimized for use in transient and stable expression studies in plants. This approach makes it possible to combine the advantage of GUS, its high sensitivity in histochemical staining, with the advantages of GFP as a vital marker. The fusion proteins were functional in transient expression studies in tobacco using either DNA bombardment or potato virus X as a vector, and in stably transformed Arabidopsis thaliana and Lotus japonicus plants. The results show that high level of expression does not interfere with efficient stable transformation in A. thaliana and L. japonicus. Using confocal laser scanning microscopy we show that the fusion constructs are very suitable for promoter expression studies in all organs of living plants, including root nodules. The use of these reporter constructs in the model legume L. japonicus offers exciting new possibilities for the study of the root nodulation process.
RADUTOIU S., MADSEN L.H, MADSEN E.B, FELLE H., UMEHARA Y., GRØNLUND M., KANEKO T., SATO S., TABATA S., SANDAL N. and STOUGAARD J. 2003. Plant recognition of symbiotic bacteria requires two LysM receptor-like kinases. Nature, 425, 585-592. Although most higher plants establish a symbiosis with arbuscular mycorrhizal fungi, symbiotic nitrogen fixation with rhizobia is a salient feature of legumes. Despite this host range difference, mycorrhizal and rhizobial invasion shares a common plant-specified genetic programme controlling the early host interaction. One feature distinguishing legumes is their ability to perceive rhizobial-specific signal molecules. We describe here two LysM-type serine/threonine receptor kinase genes, NFR1 and NFR5, enabling the model legume Lotus japonicus to recognize its bacterial microsymbiont 115 Lotus Literature
Mesorhizobium loti. The extracellular domains of the two transmembrane kinases resemble LysM domains of peptidoglycan- and chitin-binding proteins, suggesting that they may be involved directly in perception of the rhizobial lipochitin-oligosaccharide signal. We show that NFR1 and NFR5 are required for the earliest physiological and cellular responses to this lipochitin-oligosaccharide signal, and demonstrate their role in the mechanism establishing susceptibility of the legume root for bacterial infection.
SANDAL N., KRUSELL L., RADUTOIU S., OLBRYT M., PEDROSA A., STRACKE S., SATO S., KATO T., TABATA S., PARNISKE P., BACHMAIR A., KETELSEN T. and STOUGAARD J. 2002. A Genetic Linkage Map of the Model Legume Lotus japonicus and Strategies for Fast Mapping of New Loci. Genetics, 161, 1673-1683. A genetic map for the model legume Lotus japonicus has been developed. The F2 mapping population was established from an interspecific cross between L. japonicus and L. filicaulis. A high level of DNA polymorphism between these parents was the source of markers for linkage analysis and the map is based on a framework of amplified fragment length polymorphism (AFLP) markers. Additional markers were generated by restriction fragment length polymorphism (RFLP) and sequence-specific PCR. A total of 524 AFLP markers, 3 RAPD markers, 39 gene-specific markers, 33 microsatellite markers, and six recessive symbiotic mutant loci were mapped. This genetic map consists of six linkage groups corresponding to the six chromosomes in L. japonicus. Fluorescent in situ hybridization (FISH) with selected markers aligned the linkage groups to chromosomes as described in the accompanying article by Pedrosa et al. (2002, this issue). The length of the linkage map is 367 cM and the average marker distance is 0.6 cM. Distorted segregation of markers was found in certain sections of the map and linkage group I could be assembled only by combining colormapping and cytogenetics (FISH). A fast method to position genetic loci employing three AFLP primer combinations yielding 89 markers was developed and evaluated by mapping three symbiotic loci, Ljsym1, Ljsym5, and Ljhar1-3. Free text at http://www.genetics.org/cgi/content/full/161/4/1673
SCHAUSER L., HANDBERG K., SANDAL N., STILLER J., THYKJÆR T., PAJUELO E., NIELSEN A. and STOUGAARD J. 1998. Symbiotic mutants deficient in nodule establisment identified after T-DNA transformation of Lotus japonicus. Molecular Genetics and Genomics, 259, 414-423. Nitrogen-fixing root nodules develop on legumes as a result of an interaction between host plants and soil bacteria collectively referred to as rhizobia. The organogenic process resulting in nodule development is triggered by the bacterial microsymbiont, but genetically controlled by the host plant genome. Using T-DNA insertion as a tool to identify novel plant genes that regulate nodule ontogeny, we have identified two putatively tagged symbiotic loci, Ljsym8 and Ljsym13, in the diploid legume Lotus japonicus. The sym8 mutants are arrested during infection by the bacteria early in the developmental process. The sym13 mutants are arrested in the final stages of infection, and ineffective nodules are formed. These two plant mutant lines were identified in progeny from 1112 primary transformants obtained after Agrobacterium tumefaciens T-DNA-mediated transformation of L. japonicus and subsequent screening for defects in the symbiosis with Mesorhizobium loti. Additional nontagged mutants arrested at different developmental stages were also identified and genetic complementation tests assigned all the mutations to 16 monogenic symbiotic loci segregating recessive mutant alleles. In the screen reported here independent symbiotic loci thus appeared with a frequency of approximately 1.5%, suggesting that a relatively large set of genes is required for the symbiotic interaction.
SCHAUSER L., ROUSSIS A., STILLER J. and STOUGAARD J. 1999. A plant regulator controlling development of symbiotic root nodules. Nature, 402, 191-195. Lotus Literature 116
Symbiotic nitrogen-fixing root nodules on legumes are founded by root cortical cells that de- differentiate and restart cell division to establish nodule primordia. Bacterial microsymbionts invade these primordia through infection threads laid down by the plant and, after endocytosis, membrane- enclosed bacteroids occupy cells in the nitrogen-fixing tissue of functional nodules. The bacteria excrete lipochitin oligosaccharides, triggering a developmental process that is controlled by the plant and can be suppressed. Nodule inception initially relies on cell competence in a narrow infection zone located just behind the growing root tip. Older nodules then regulate the number of nodules on a root system by suppressing the development of nodule primordia. To identify the regulatory components that act early in nodule induction, we characterized a transposon-tagged Lotus japonicus mutant, nin (for nodule inception), arrested at the stage of bacterial recognition. We show that nin is required for the formation of infection threads and the initiation of primordia. NIN protein has regional similarity to transcription factors, and the predicted DNA-binding/dimerization domain identifies and typifies a consensus motif conserved in plant proteins with a function in nitrogen-controlled development.
STOUGAARD J. 2000. Regulators and regulation of root nodule development. Plant Physiology, 124, 531-540. [Review] Full text at http://www.plantphysiol.org/cgi/content/full/124/2/531
STOUGAARD J. 2001. Lotus japonicus. In Encyclopediae of Genetics, Academic press, pp 1121-1122. [Review]
STOUGAARD J. 2001. Genetics and genomics of root symbiosis. Current Opinion in Plant Biology, 4, 328-336. [Review]
Model genetics and genomics have been developed as tools for studying the third largest family of flowering plants, the Leguminosae, which includes important crop plants. Functional genomics strategies for the global analysis of gene expression, the elucidation of pathways and reverse genetics are established. These approaches provide new possibilities for investigating rhizobial as well as mycorrhizal endosymbiosis. Plant genes with central functions in these mutualistic interactions have been identified by positional cloning and gene tagging. With progress in Lotus japonicus genome sequencing, which was recently initiated by Japanese researchers, comparative genomics will contribute to our understanding of symbiosis, pathogenesis and the evolution of plant genome.
STOUGAARD J., SZCZYGLOWSKI K., DE BRUIJN F.J. and PARNISKE M. 1999. Genetic nomenclature guidelines for the model legume Lotus japonicus. Trends in Plant Science, 4, 300-301. [Manual]
STRACKE S., KISTNER C., YOSHIDA S., MULDER L., SATO S., KANEKO T., TABATA S., SANDAL N., STOUGAARD J., SZCZYGLOWSKI K. and PARNISKE M. 2002. A plant receptor-like kinase required for both bacterial and fungal symbiosis. Nature, 417, 959- 962. Most higher plant species can enter a root symbiosis with arbuscular mycorrhizal fungi, in which plant carbon is traded for fungal phosphate. This is an ancient symbiosis, which has been detected in fossils of early land plants. In contrast, the nitrogen-fixing root nodule symbioses of plants with bacteria evolved more recently, and are phylogenetically restricted to the rosid I clade of plants. Both symbioses rely on partially overlapping genetic programmes. We have identified the molecular basis for this convergence by cloning orthologous SYMRK ('symbiosis receptor-like kinase') genes from Lotus and pea, which are required for both fungal and bacterial recognition. SYMRK is predicted to 117 Lotus Literature have a signal peptide, an extracellular domain comprising leucine-rich repeats, a transmembrane and an intracellular protein kinase domain. Lotus SYMRK is required for a symbiotic signal transduction pathway leading from the perception of microbial signal molecules to rapid symbiosis-related gene activation. The perception of symbiotic fungi and bacteria is mediated by at least one common signalling component, which could have been recruited during the evolution of root nodule symbioses from the already existing arbuscular mycorrhiza symbiosis.
WANG T.L., DOMONEY C., HEDLEY C.L., CASEY R. and GRUSAK M.A. 2003. Can we improve the nutritional quality of legume seeds? Plant Physiology, 131, 886-891. Full text at http://www.plantphysiol.org/cgi/content/full/131/3/886
WEGEL E., SCHAUSER L., SANDAL N., STOUGAARD J. and PARNISKE M. 1998. Mycorrhiza mutants of Lotus japonicus define genetically independent steps during symbiotic infection. Molecular Plant Microbe Interactions, 11, 933-936.
WOPEREIS J., PAJUELO E., DAZZO F.B., JIANG Q., GRESSHOFF P.M., DE BRUIJN F.J., STOUGAARD J. and SZCZYGLOWSKI K. 2000. Short root mutant of Lotus japonicus with a dramatically altered symbiotic phenotype. Plant Journal, 23, 97-114. Legume plants carefully control the extent of nodulation in response to rhizobial infection. To examine the mechanism underlying this process we conducted a detailed analysis of the Lotus japonicus hypernodulating mutants, har1-1, 2 and 3 that define a new locus, HYPERNODULATION ABERRANT ROOT FORMATION (Har1), involved in root and symbiotic development. Mutations in the Har1 locus alter root architecture by inhibiting root elongation, diminishing root diameter and stimulating lateral root initiation. At the cellular level these developmental alterations are associated with changes in the position and duration of root cell growth and result in a premature differentiation of har1-1 mutant root. No significant differences between har1-1 mutant and wild-type plants were detected with respect to root growth responses to 1-aminocyclopropane1-carboxylic acid, the immediate precursor of ethylene, and auxin; however, cytokinin in the presence of AVG (aminoetoxyvinylglycine) was found to stimulate root elongation of the har1-1 mutant but not the wild- type. After inoculation with Mesorhizobium loti, the har1 mutant lines display an unusual hypernodulation (HNR) response, characterized by unrestricted nodulation (hypernodulation), and a concomitant drastic inhibition of root and shoot growth. These observations implicate a role for the Har1 locus in both symbiotic and non-symbiotic development of L. japonicus, and suggest that regulatory processes controlling nodule organogenesis and nodule number are integrated in an overall mechanism governing root growth and development. Full text at http://www.blackwell-synergy.com/links/doi/10.1046/j.1365-313x.2000.00799.x/full/
ZAGROBELNY M., BAK S., RASMUSSEN A.V., JØRGENSEN B., NAUMANN C.M., LINDBERG MØLLER B. 2004. Cyanogenic glucosides and plant-insect interactions. Phytochemistry, 65, 293-306. [Review] Cyanogenic glucosides are phytoanticipins known to be present in more than 2500 plant species. They are considered to have an important role in plant defense against herbivores due to bitter taste and release of toxic hydrogen cyanide upon tissue disruption. Some specialized herbivores, especially insects, preferentially feed on cyanogenic plants. Such herbivores have acquired the ability to metabolize cyanogenic glucosides or to sequester them for use in their predator defense. A few species of Arthropoda (within Diplopoda, Chilopoda, Insecta) are able to de novo synthesize cyanogenic glucosides and, in addition, some of these species are able to sequester cyanogenic glucosides from Lotus Literature 118 their host plant (Zygaenidae). Evolutionary aspects of these unique plant–insect interactions with focus on the enzyme systems involved in synthesis and degradation of cyanogenic glucosides are discussed.
ZHANG S., SANDAL N., POLOWICK P.L., STILLER J., STOUGAARD J. and FOBERT P.R. 2003. Proliferating Floral Organs (PFO) a Lotus japonicus gene required for specifying floral meristem determinacy and organ identity, encodes an F-box protein. Plant Journal, 33, 607-619. To study flower development in the model legume Lotus japonicus, a population of transgenic plants containing a maize transposable element (Ac) in their genome was screened for floral mutants. One mutation named proliferating floral organs (pfo) causes plants to produce a large number of sepal-like organs instead of normal flowers. It segregates as a single recessive Mendelian locus, and causes sterility. Scanning electron microscopy revealed that pfo affects the identity, number and arrangement of floral organs. Sepal-like organs form in the first whorl, and secondary floral meristems are produced in the next whorl. These in turn produce sepal-like organs in the first whorl and floral meristems in the second whorl, and the process is reiterated. Petals and stamens are absent while carpels are either absent or reduced. The pfo phenotype was correlated with the presence of an Ac insertion yielding a 1.6-kb HindIII restriction fragment on Southern blots. Both the mutant phenotype and this Ac element are unstable. Using the transposon as a tag, the Pfo gene was isolated. Conceptual translation of Pfo predicts a protein containing an F-box, with high overall similarity to the Antirrhinum FIMBRIATA, Arabidopsis UNUSUAL FLORAL ORGANS and Pisum sativum Stamina pistilloida proteins. This suggests that Pfo may regulate floral organ identity and meristem determinacy by targeting proteins for ubiquitination.