WESTERN REGIONAL PLANT INTRODUCTION STATION ANNUAL REPORT, CALENDAR YEAR 1999 REGIONAL PROJECT W-6: Plant Genetic Resource Conservation and Utilization

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1.  Note: changes in personnel due to retirements are listed in the Personnel Activities section.

OTHER SUPPORT PERSONNEL

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STATION ACTIVITIES and ACTIVITIES FOR RESEARCH LEADER
Richard M. Hannan

According to Dave Stout, our Seed Storage Manager, at year's end W-6 was responsible for 2,621 taxa in 376 genera (24% of the NPGS) with a total of 68,652 accessions.

In 1999 we received 1,088 new accessions from collections, plant explorations and germplasm exchanges. We also received 83 accessions from the National Seed Storage Laboratory, Ft. Collins, CO for inclusion into our Pullman collections and 10 accessions were transferred from other sites (Table RH1). Therefore, we had a total of 1,181 accessions in 132 genera added to the W-6 collections.

During the year 11,182 accessions were distributed in 16,412 packets. There were 527 separate orders filled, and of these, 404 (76.7%) were domestic and 123 (23.3%) were sent to foreign scientists (Table RH2). Within the 101 genera distributed (Table RH3) there were 722 taxa represented in the 16,412 packets sent. Within the Western Region alone, there were 47,637 accessions received by users of germplasm. Of this 34.4% was provided by the Western Regional Plant Introduction Station.

By the end of CY99 the number of PI accessions (50,676) backed up at NSSL increased to 74% of our collection. During the year a total of 1,764 accessions were sent to NSSL for back-up. The goal continued to be getting back-up samples of all of the accessions which are so difficult to propagate. As yet we have not had the resources to implement an ongoing tissue culture/ cryogenic program at the Pullman location, but will continue trying to coordinate with the cryogenic laboratory at NSSL.

Maintenance of the collections is described by the respective curators in the subsequent text. However, it should be noted that we made some significant changes to who maintains some of these collections. Although the responsibility of curating the Phaseolus, the Cool Season Food Legumes and the Beta collections remained the same, we reallocated many of the other genera. During 1999 we established Parlier, California as the National Arid Land Plant Germplasm Resource and NPGS Regeneration Unit and made it an NPGS priority site for twelve arid land genera to begin with. Similarly, after hiring a new curator for the the National Arctic Plant Germplasm Resource and NPGS Regeneration Unit Palmer, Alaska, it too has become the NPGS priority site for at least ten genera with which to begin.

The bulk of the transition, however, has fallen on the shoulders of our Hort Crops Curator, Barbara Hellier. She made the transition from being responsible for six genera to 108 genera. Her taxa now include many medicinal and ornamental species, and some taxa with, as yet, unknown utilization.

Personnel Activities

To begin with we were very proud that Leslie Elberson was named Scientific Technician of the Year for the Pacific West Area.

At the beginning of the year bill Kaiser retired after serving 28 years for WRPIS. His knowledge and skill in cleaning extremely diverse types of seed will be missed. One month later Gary Bloomfield, our Pullman Farm Manager, retired after 34 years of dedicated service to Plant Introduction. We celebrated his retirement and highlighted his three decades of contributions at a fun dinner event in Pullman.

Following a five month period with no computer support, Gwen Pentecost started in March in a temporary position as our computer specialist. During the course of her 130 day assignment she got us back on track and was hired as our permanent full time computer specialist in August.

Our previous Research Plant Pathologist, Dr. Walter Kaiser, retired in January 1998. During the later half of 1998 and early 1999 we advertized for the position and successfully recruited Dr. Frank Dugan as our new Research Plant Pathologist, and he began his work at Pullman in April.

Shari Lupien, who was working as a part time employee for the Hort Crops program, moved over into Franks's pathology program; then successfully competed, and was hired for the permanent Biological Technician position for plant pathology in October.

Following a search to replace Gary Bloomfield, in June we hired Wayne Olson as our Pullman Farm Manager.

Not long after coming on board, Wayne filled Bill Kaiser's vacant position by hiring Scott McGee in July. Scott has extensive experience both in agriculture related work and electronics.

In addition to her work in the field and greenhouse for the Hort Crops Program, Vicki Newman began a program to obtain a Masters Degree in Horticulture. Dr. Richard Hannan is Vicki's committee chair.

Dr. Eric Storlie- half-time post-doctoral researcher funded by Grain Legume Unit until Sept. 30 working on chickpea and pea BAC libraries. Currently funded by NARF.

Dr. Marie-Laure Pillet Post-doctoral research fellow from INRA, France (10-16-99 thru 4-18-00) working on analysis of Aphanomyces root rot resistance from the germplasm line PI >557500. >

Melissa McClendon-graduate student in the Department of Horticulture working on a Master of Science degree studying genetic resistance to Fusarium wilt race 1 in the germplasm line W6 12612. Funded by USDA-CSREES- Special Grants program for Cool Season Food Legumes.

Makiko Miruma- graduate student in the Department of Crop and Soil Sciences studying for a Master's of Science degree examining genetic diversity in edamame soybean funded by an IMPACT grant of Dr. Tom Lumpkin (Coyne co-P.I..).

Up north, at our Alaska project, we successfully filled the existing vacancies. First, we hired Dr. David Ianson to serve as Horticulturist/Curator of the Arctic Plant Germplasm Resource and Regeneration Unit. He came on board in June and has had the fall and winter to start this germplasm conservation effort from scratch.

Also at the Palmer site, Dr. Nancy Robertson, Research Plant Pathologist, successfully recruited Denice Knight-Slater as the plant pathology technician in July, and Denice has aggressively been learning the latest in virology research.

In addition to losing Gary and Bill, we had a few other staff members leave. Barbara Brandon, who has worked with us since 1995 left in November for a position across campus. Meg Vigil has worked for the Agronomy program (both Johnson and Bradley) since 1994 and left in September of 1999 for a permanent position with the wheat breeders at Washington State University. Lastly, Jake Wheeler, our bean technician who introduced a whole new concept in growing beans to new age music, left in November to go back home to Massachusetts where, although beans are not a major agricultural commodity, his learned skills in greenhouse plant production will come in handy.

Germplasm and Research Activities

I have been conducting greenhouse seed increases to provide virus free seed of another twelve accessions of germplasm from the National Seed Storage Laboratory that are used as part of the common virus test set for conducting virus host indicator studies. This is a service that we provide the NSSL.

In collaboration with Dr. Phil Miklas, USDA, ARS, Prosser, WA, Dr. Maria Jenderek and I conducted our part of the field trials in the second year of a study to determine the relationship of growth habit and sensitivity to high temperature growing conditions on the yield of kidney beans. A heat tolerant landrace cultivar "indeterminate Jamaica Red" was crossed with "Red Hawk" dark red kidney bean in an attempt to increase yield potential of kidney bean. Most of the dark red kidney bean lings out yielded Red Hawk across the moderate temperature environments, whereas not out yielded Red Hawk in the high temperature environments. Overall, indeterminate vine growth habit expressed greater yield potential than determinate bush growth habit, and light red kidney had greater yield potential than dark red kidney suggesting that genes which condition dark red seed coat color in kidney bean may be associated with lower yield.

We planted the third year of a four location garlic study. This study is designed to determine the gene by environment interaction on identical clones of garlic at four different locations in the United States. We are also following the adaptation of clones from one season to the next at each location to determine if genetic responses to the respective environments remain stable. This work is a collaborative effort between Phil Simon, USDA, ARS & University of Wisconsin, Ron Voss, University of California at Davis, Maria Jenderek, USDA, ARS, NPGS, Arid Land Plant Germplasm Resource Unit, Parlier, California, and me here in Pullman.

In conjunction Khalid Al-Saad, a doctoral candidate, with the Department of Chemistry at Washington State University I have continued studies on the effect of aging on the triacylglycerols (TAGs) in seed. This past year we have been exploring the use of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry and new sample preparation methods of these nonpolar analytes. We found that MALDI enables the rapid analyses of TAG standards and mixtures of whole oils. The new method provides excellent reproducibility, making quantification possible. Detection limits were in the mid femtomole range and resolution was such that TAGs could be separated by one double bond difference. We tested this on aged and fresh onion seed lipids.

Talks and Presentations for R. Hannan (unless otherwise indicated)

Travel, Invitations, Special Awards and/or Assignments for Hannan or as indicated.

January 8	Gary Bloomfield, Bill Kaiser, Kevin Miller traveled to the Washington Ag Show in Spokane, WA.
January 22	Participated in PhD prelims for Khalid Al-Saad, Department of Chemistry, Washington State University.
February 17-19	Traveled to Phoenix, AZ and visited Dr. William Feldman, Director of the Boyce Thomson South Western Arboretum. Then attended the Desert Legume Program meeting in Tucson, AZ.
March 11-13	Traveled to Parlier, CA for business.
March 26-28	Travel to Salem, OR to attend Oregon Tilth Meeting and Farmer Cooperative Genome Project (FCGP). Presented the 'The role of the National Plant Germplasm System in meeting the needs of the FCGP' to the group.
April 8	Travel to Palmer, AK. Business and interview RoseAnn Leiner who interviewed for the germplasm Curator position.
July 19-23	Rich Hannan and David Stout traveled to Aberdeen, ID to attend the Plant Germplasm Operations Committee Meeting.
July 28	Traveled to Tucson, AZ to attend DELEP meeting. Elected to be vice-chair.
September 8-9	Travel to Parlier, CA. Business.
September 15-17	Travel to Palmer, AK. Business.
October 13-15	Presented invited talk to National Garlic Symposium. Tulsa, OK
December 8-10	To Palmer, AK for business.
December 15-17	To Parlier, CA for business.

Committees, Other Assignments, Activities, and News

WSU, CAHE Greenhouse Facilities Advisory Committee (membership terminated 8/17/99)

WSU, Graduate School Representative for Ph.D. and MS defenses

WSU, Adjunct Faculty, Department of Horticulture and Landscape Architecture

Serve as Chair of Committee for Master's Degree Candidate, Vicki Newman

Serve on Committee for Doctoral Candidate, Khalid Al-Saad

Serve on Committee for Master's Degree Candidate, Barbara Hellier

ASHS, Outstanding Research, Educator Award Committee 1997-2000

W150 Regional Project

USDA, NPGS, Plant Germplasm Operations Committee (PGOC)

USDA, NPGS, PGOC, In situ Conservation Subcommittee

USDA, NPGS, PGOC, NPGS Special Recognition Award Subcommittee.

ARS, CRIS Research Project Statement Peer Review Panel

Desert Legume Program Advisory Board, Tuscon, AZ >DELEP meeting. Elected to be vice-chair

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COMPUTER RESOURCES
Gwen Pentecost

Increased connectivity via the local area network (LAN) and the World Wide Web (WWW) has accentuated compatibility issues between different ARS computers. Therefore, WRPIS focused on upgrading its hardware and software resources in 1999. The increase in technical uniformity has reduced communications problems and allows better use of up-to-date software.

The replaced computers were then redistributed. The most useful systems went to individuals and labs within the group for continued use. Other equipment was donated to local public schools and nursery schools. A high school class in computer repair received our defective equipment.

Specialized computers were also purchased, including a complete graphics system and two durable laptops. Special design features of the 'ruggedized' laptops enable them to operate under harsher conditions than normal laptops. They will be used during plant collection trips and for entry of bar-coded data in the field.

When the FTS2001 telephone contract required a change in equipment, WRPIS switched to a router system that greatly improved the speed and reliability of connections to the GRIN server in Beltsville. The improved access is available to all WRPIS employees via the Washington State University LAN.

Revision of the WRPIS WWW Home Page has begun. The update will include suggested improvements to the current web page, plus reflect changes that have occurred within the group's structure and capabilities.

The computer assistant also took on the duty of Collateral Duty Safety Officer (CDSO) for Plant Introduction just two months before a major USDA safety inspection. Parallel efforts were made to become familiar with group safety issues, identify numerous training requirements and meet increasingly rigorous documentation standards. Cooperation within the unit helped the USDA CARE Review go smoothly.

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PROGRESS REPORT OF THE PULLMAN STATION FARM AND FACILITIES MANAGER
Wayne L. Olson

Introduction

Dr. Rich Hannan contacted me in May of 1999 and asked if I would interview with the Staff of the Pullman Station to replace Gary Bloomfield who retired earlier in the year. After touring the facilities, visiting with some of the Curators and Research personnel, and interviewing with the whole Staff; I accepted the position of Farm and Facilities Manager June 15th, 1999. With me I bring eighteen (18) years experience in agribusiness providing technology transfer, promotion of agricultural products, and interpersonal communication skills. Five (5) of those years I spent in the creation and development of the Wilbur-Ellis; Genesee, Idaho retail Ag Chemical Facility. The last five (5) years with the Wilbur-Ellis Company I spent as the Palouse Regional Technical Advisor, Promotions Strategist, Crop Complaint Investigator, and Research Agronomist all rolled into one. The area I covered has been from the Canadian Border to Pendleton, Oregon; and from Grangeville, Idaho to Connell, Washington. During my time with the Wilbur-Ellis group, the least number of research sites I managed were thirty (30), and the highest number of sites was in 1997 when I established, maintained, and took data on sixty-three (63) sites within my region, managed by myself with the help one summer intern student. I have my Bachelor's Degree in Agricultural Mechanization from the University of Idaho, with heavy emphasis in Weed Science and Soils, and have passed testing and accreditation for the Regional and National Certified Crop Advisor under the American Society of Agronomy.

New Ideas and New Direction

Since I have come on board, Kurt Tetrick (Central Ferry Farm Manager) and I have worked together and purchased a mechanical transplanter that will replace planting thousands of plants by hand with the curators and technicians on hands and knees. This tool will be a major part of our plans to address the scale-up in the number of plants and number of accessions within the next three years.

Here on the Pullman Station we are blessed with hills (just like the rest of the Washington State University) to the point that we have no level fields to speak of. This last fall we planted over-winter cover crops to hold the soil from erosion for the winter season, and add organic matter back the following year. Considering the winter rains to date, this system has worked very well, both in the general fields and also between the garlic and Allium accessions.

In the seed cleaning facility we purchased a LAH Brush Machine for thrashing and preliminary cleaning of grasses primarily, but we are finding new uses every day. This machine replaces most of the thrashing that has been done in the past with the hammer mill. Since the Brush Machine uses a brush material similar to a stiff shop broom for thrashing the seed. We believe the seed leaving our facility today is in better shape for storage than using the steel hammers of the Hammer Mill for thrashing. Also, we see faster and easier cleaning of the seed by using the Brush Machine.

Throughout the summer and fall I heard about and investigated the slow growth of the grasses when planted in the Bubblehouse in the spring. The practice through 1999 was to mix soil and peatmoss in 30 gallon garbage cans up at the soils bins; take the cans to the autoclave and place the cans weighing 250 lbs. inside and steaming them to sterilize the soil. Then the cans were taken out of the autoclave reloaded onto a truck and taken to the farm for planting grasses. Soil testing of this soil revealed toxic levels of zinc, manganese, and boron which were not present before placement in the autoclave. The conclusion was that we were removing the galvanized material from the garbage cans during the sterilization process and leached into our planting soils, thus slowing plant development.

This whole soil sterilization system is being replaced with Stainless Steel tote bins with lids that will be placed in the autoclave with a forklift. After sterilization, the greenhouse peat mix will be added to the bins and the soil in the tote boxes mixed by the rotation on the forklift. The boxes will then be set back on the truck for transfer to the farm for use in planting. Our estimate is that we will need to turn eight tons of soil through the autoclave system within two weeks this year to meet our planting expectations. This year we should easily meet our soil needs with the new system in place. With the planned projections, in two years we will be looking at double the soil amount to sixteen tons of soil that will be needed to meet the planting needs as we scale up production and keep producing quality seed for storage and distribution.

Safety through New BioFuels

One of the issues we resolved this year was a safety issue in one of our headhouses that also house offices for our station staff. Headhouse 109B is also home to one of our diesel Kabota tractors that we use to load and transport soil around the facility. The diesel smoke from the tractor is usually picked up in the air system and is noticeable throughout the facility. This fall and winter we switched to BioDiesel for the Kabota tractor made from canola oil and synthesized with grain alcohol. The switch has been good for everyone affiliated with the facility. The only smell now is warm cooling oil, and health wise better for the curators, researchers, and the support personnel.

New Equipment Yet To Come

We have plans to upgrade the single row "Cone Planter" with the conversion to a Hege Distribution Cone instead of the "dribble cone". This will help us from doubling seeds when planting and give us even seed placement for the whole plot length for better quality and yield. It will also keep our plots to a standard length for insect exclusion (tenting) so we can reuse a standard size material and better grade of material year after year over the accessions.

I have designed and plan to build a new piece of equipment for planting multiple rows of beets at the same time, something that has not been done before. By this time next year I look to have the beet planting equipment modified to put down a starter band 2" below the beets at planting time.

Solid Technical Support at Pullman

The field production and seed cleaning portion of the Pullman WRPIS unit has a solid base with two technicians, Kevin Miller and Scott McGee. Kevin continues to be an excellent source of information and knowledge to blend the past operation of the Unit with new ideas and techniques that are emerging throughout the year. Scott joined the unit this summer replacing Bill Kaiser when he retired earlier this year. Scott's vast background in farming methods and harvesting equipment and methods has added new dimension to the overall performance of the production and the seed cleaning at the station.

The Future

The thought and design of the planting, harvesting, and seed cleaning equipment this year I see as only a starting point as we look to automate the planting and harvesting of future crops, not only for the 2000 year, but beyond. Curators and Researchers of the Pullman Station have been "brainstorming" on new ways we can develop our practices and procedures to increase numbers of accessions, solidify genetic diversity of accessions, and raise the quality of our accessions in both the near and distant future.

Meetings of 1999

June 21-22	W-6 Annual Meetings, Pullman, Washington I attended the annual W-6 meetings held in Pullman, Washington. The meetings were an excellent insight into the workings and understanding the business and the group I had just became employed with; not only here at Washington State University, but Nationally and Internationally in depth and scope.
June 24-25	Pioneer Seed, Moscow, Idaho; Potlatch, Idaho; Grangeville, Idaho; Dayton, Washington; Reardon, Washington. Pioneer Seed requested my presence on their canola plot tour throughout the Pacific Northwest. These plots I had put in while previously employed with PACER Corporation. The representatives had many questions on production methods with spring and winter canola in this region since our weather patterns are very different from the canola growing regions of Canada. They also asked many questions in regards to my insight as to grower thought patterns and habits with canola production in the Palouse Region. I was able to help them in many areas drawing on my twenty (20) years of experience with the crop; four (4) of which were with the InterMountain Canola group.
December 7-9	First Aid Training, Pullman, Washington. Both Scott McGee and I would like to commend Washington State University on their First Aid Training for use within the group. Over the years we have taken many different First Aid courses, but the Washington State University course we would recommend to anyone based on content, focus, and usable knowledge.
December 14-16	Far West Fertilizer Meeting, Spokane, Washington. Meetings at the Far West Fertilizer Conference were very good and very diverse as always. Meetings included, but were not limited to: FQPA: Science and Public Perception; Soil Health; Watershed Water Quality; New (Chemical) Products and Practices on the Horizon; How CCA's Can Help Implement GWMA; FQPA: What It Means to Fieldmen; Genetic Engineering; Climatology; Phosphate Issues; Multiple Chemical Sensitivity; Chemical Drift Training; Plant Disease Detection; IPM Use of Seed Treatments.

I did conduct some side meetings with different Chemical Representatives to seek out new chemicals we can use in our greenhouses here at Pullman, Washington for resistant insects and diseases. The information I received has been very valuable and we have already brought in some new (safer) chemistries to rotate with our existing chemical use to decrease resistant pest populations and to keep from developing new resistance to the pests we work with today.

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PROGRESS REPORT OF CENTRAL FERRY RESEARCH FARM
Kurt Tetrick

It was a very busy year at the Central Ferry Farm in 1999. Farming activity for the WRPIS curators and researchers included the following:

1. Al Hodgdon had a .5 acre Beet nursery for seed increase and reported that it did fairly well. There was a one week long cold snap during the spring which killed a few of the more southern latitude adapted accessions. The beet crops required 4 applications of insecticides last summer for mite and aphid control.

2. Vicki Bradley had a 2.5 acre safflower nursery, which was half seed increase and half for ornamental evaluations. The seed increase portion of the crop used a new idea implemented in 1999. We installed a trellis to suspend caging material over in order to create a pollinator exclusion situation. The only problem we had was the material we used for caging was not strong enough for the heavy winds we receive at this location, resulting in only a few lines getting harvested. The safflower crop only required one application of insecticide.

Vicki's perennial grass increase nurseries, which take 32 acres of the farm, had very little problems other than weeding, and cultivator blight. We did one application at a half rate of tilt fungicide for rust and powdery mildew.

3. Clare Coyne had a 2 acre plot of peas for descriptor data. We applied one application of Dimethoate for leaf weevil. Clare also had a 3 acre nursery of chickpeas for descriptor data and a second look at the core collection.

4. Molly Welsh had a small, 2 acre, bean crop for increase and to see if we put BCMV free beans in the field would they increase and remain virus free. There were no insecticide applications to this nursery.

5. Barbara Hellier and Rich Hannan planted 2 X 350-foot rows of garlic for virus screening before introducing them to the collection. Barbara has a perennial nursery of wild Allium species for seed increase that is about an acre. This nursery needs to be watched close for onion thrips, which affect seed quality and quantity. I applied four applications of insecticides on this plot of land. Along with the wild Allium there is the leek nursery which needs a similar care and attention.

6. Steve Clement had two entomology research plots on the farm. One was a study in conjunction with the Dr. Nilsa Bosque-Perez, University of Idaho Extension Entomologist, which is a study for Russian wheat aphid and hessian fly resistance on different varieties of spring soft white wheat. The second was some perennial wheat and wheat grass nursery with Dr. Steve Jones, Washington State University Winter Wheat Breeding, for resistance studies of Russian wheat aphid and hessian fly.

Other research projects from Washington State University and Extension included :

1.   WSU Hop pathology and breeding research. This two (2) acre plot is a remote yard for the research done at the WSU, Irrigated Agriculture and Extension Center in Prosser, WA. The study of powdery mildew done at Prosser was a great concern to local growers in that area, fearing the spread of mildew into their fields. The studies done at Central Ferry give researchers an area of isolation so that the study of possible fungicides and resistant crops is in a more controlled atmosphere.

The continuing problem of not being able to deliver enough continuous water to this plot, to produce young susceptible growth, will be solved this year with the remodel of the irrigation system.

2.   A study by Dr. Dave Bragg, WSU Extension Entomologist, John Burns, WSU Extension Agronomist, and me on Russian Wheat Aphid and Hessian fly research in our buffer strips of winter wheat was conducted on a 15 acre research plot. It studies different insecticide methods for economic control of Russian wheat aphid and hessian fly on soft white spring and winter wheat, spring and winter feed barley, and winter and spring canola. Results from 1999 were not as conclusive because of a decrease in insects populations.

3.   USDA Wheat Genetics (Club Wheat) Dr. Kim Campbell planted a 3 acre club wheat-breeding plot which is an early look at higher elevation plantings in other areas. This planting can be up to four weeks earlier because our 550 ft. elevation at this farm. This can give an early look at diseases, rusts, and blights. I have been told that the Central Ferry Farm is very important to their study.

4.   The WSU Wheat Breeding (Winter Wheat) plots put in by Dr. Steve Jones comprised a 2 acre plot, and was planted for the same reasons as Dr. Campbell's. For this group there was a bonus result. Their nurseries were frozen out during the 98-99 winter in higher elevations. They were able to harvest plots at this location that had been lost at other location. As usual, they were very pleased with their early results in soft white winter wheat.

5.   WSU Spring Wheat Breeding program led by Dr. Kim Kidwell established a plot to study new varieties for Russian wheat aphid and hessian fly resistance.

6.   Mark Minten, a WSU Botany PHD candidate, set up a 2 acre plot to conduct a plant competition study.

Central Ferry Facilities Report:

Buildings had the standard maintenance with a start to a major project in one building. The equipment shed known as the "open shed" had a face-lift by removing the equipment and laying gravel on the dirt floor. Eventually I plan, provided funding exists, to install sliding doors and dedicate a 20 by 50 stall for shelving and manifold storage.

The improvements on grounds were mainly done for ease of access to buildings and to open the grounds for safe travel and reduce the chance of accidents in the building areas. We mainly moved fences away from buildings, graded, and laid gravel this gave us an area to park equipment and to have ease of travel around the buildings.

We removed some fences that were used as pasture many years ago; to begin expansion of the farm, to improve isolation of our germplasm.

For the past four years I have been working on changing the irrigation system. After many discussions and changes, the project was open for bid at the end of the year. The drip system Dr. Hannan and I had gone over and over in 99 should be able to do everything that is required for the CF Farm.

The drip system requirements are:

1.   Be able to drip as well as run sprinklers on different nurseries at the same time.

2.   Be able to use the existing risers for drip as well as sprinklers.

3.   Be able to schedule drip intervals to individual crops through automatic valves, to prevent over watering or under watering from seat of my pant scheduling.

4.   To install a complete weather station so that I can schedule irrigation, archive daily weather, and report any pesticide applications or labor done to a specific crop.

Major purchases for the farm in 99 included:

The weather station, which I explained earlier.

A flatbed trailer to pull behind pickups to haul equipment and materials.

A plastic mulch layer to go with the drip system. The use on this farm will be to control weeds, harvest original seed (eliminate volunteer in a line of germplasm), to reduce the amount of moisture on plants which reduces the risks of mildews and disease, and to control the moisture in the soil.

I had two people employed in 1999. Jonathan Bragg worked from April to October, 40 hrs./ week. He did weed control, irrigation work, equipment maintenance, building maintenance and other farm labor activities. Sue Nelson worked staggered part time hours, doing the same general farm labor April to August.

Future plans for 2000 and beyond:

To establish and begin using the remodeled irrigation system.

To catalog and establish a reporting system for each crops grown on the farm.

To begin the steps of expanding the size of the farm; to improve the isolation and reduce the risk of out crossing.

Year 2000 PI crops:

3000 lines of chick peas

36 lines of beans

480 lines of grass- 1st and 2nd year grass

125 lines of increase safflower.

80 lines of ornamental safflower

Carryover from last year:

Leeks

Alliums

Perennial wheat

Other research activities for the upcoming season at the Central Ferry farm include: WSU Extension, USDA Wheat Genetics, WSU Winter Wheat Breeding, WSU spring wheat Breeding, WSU Hops Pathology, and WSU Potato Research, Mark Menten, Botany Project.

In conclusion I feel The Central Ferry Farm did well last year, and kept pace with the demands put on us. I look forward to the future changes and challenges for the year 2000.

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PROGRESS REPORT OF THE STATION RESEARCH ENTOMOLOGIST
Stephen L. Clement

Activities/Research

Pollination Research

Lesquerella. A detailed glasshouse experiment to quantify self- and cross-pollination in individual plants of three species of Lesquerella was terminated when two-spotted spider mites colonized experimental plants, thereby precluding attempts to maintain pest-free plants in the experiment. The likely source of mites were infested plants in a nearby glasshouse bay or infested nursery plants directly outside the glasshouse. After evaluating the need for this research to supplement earlier findings from a 3-year field study, Dr. Clement decided in December 1999 that the research is not required and no attempts will be made to repeat it. This joint research with Dr. R.C. Johnson, W6 Research Agronomist, is now complete.

Seed Storage and Endophyte Survival

This 8-year study to determine the effect of different seed storage temperatures on seed germination and Neotyphodium endophyte survival was started by Drs. Kaiser and Bruehl before they retired. Dr. Clement, with help from Nadeer Youssef, a graduate student at the University of Idaho, has taken over this project. The tall fescue portion of this project was completed in 1998, the results of which were summarized in last year's report. Here we report the results with perennial ryegrass seed germination and Neotyphodium survival at four storage temperatures (Table SC1). Clearly, storage temperatures of 4^o, -10^o, and -196^oC are optimal for seed germination and endophyte viability.

Table SC1.   Effect of seed storage temperature on seed germination and endophyte viability after 8 years, perennial ryegrass (PI462339).

Tall Fescue-Neotyphodium Diversity-Aphid Interaction Research

This research aims to characterize fungal endophyte diversity in Mediterranean tall fescue in the Pullman seedbank and learn more about the complexity of grass-endophyte-insect relationships by characterizing the differential effects of different tall fescue-endophyte associations on aphid reproduction and survival (Bird Cherry-Oat Aphid and Rose Grass Aphid). Previously planned work on Neotyphodium diversity in W6 tall fescue accessions has been completed and summarized for publication. Completed in 1999 was a full range of experiments involving tall fescue accessions from Sardinia, Tunisia, and Morocco, some Neotyphodium-infected and some Neotyphodium-free, and aphid assays to measure susceptibility of these plants to aphid attack and reproduction. In short, and contrary to previous findings by Dr. Clement and other investigators, Neotyphodium infection in tall fescue did not always confer resistance to Bird Cherry-Oat Aphid reproduction and population growth. Table SC2 depicts the results from an assay involving the Bird Cherry-Oat Aphid and selected infected and uninfected plants originating from Tunisia (results from other assays are not shown), which shows that some infected plants were highly susceptible to the aphid. This research has been summarized, statistically analyzed, and prepared for publication.

Table SC2.   Mean numbers of adult and immature Rhopalosiphum padi on endophyte-infected and uninfected tall fescue originating from Tunisia.

Other experimental research addressed the susceptibility of Neotyphodium-infected and -free wild barley to Bird Cherry-Oat Aphid and Rose Grass Aphid reproduction and survival. This research will be repeated, but there are early indications that Neotyphodium in wild barley does not impart resistance to Bird Cherry-Oat Aphid. On the other hand, the presence of Neotyphodium in some wild barleys seems to impart a measure of resistance to Rose Grass Aphid.

Dryland Wheat Production, Cereal Aphids, and Hessian Fly

For the fourth year the W6 entomology program participated in a multi-disciplinary team of scientists researching the sustainability of spring wheat production in the arid and semiarid winter wheat-fallow region of central Washington. Dr. Frank Young, ARS Pullman, is the Project Leader and the large study is situated on the farm of Curtis Hennings near Ralston, Washington. The 1999 plots were devoid of pest aphids and predacious ladybird beetles. On the other hand, Hessian fly population densities were high and economically important in no-till continuous hard red-spring wheat plots, i.e., the P5 and P6 plots in Table SC3. This suggests that a switch to no-till cropping with continuous spring wheat will exacerbate the Hessian fly problem. This hypothesis will be examined again in 2000, the final year of this study. The biotype composition of the Ralston Hessian fly population was determined, as well, by sending material to Dr. Roger Ratcliffe, ARS/Purdue University, for genetic typing. As determined by Dr. Ratcliffe, the biotype composition is 61% GP (not virulent to presently deployed resistance genes in wheat), 16% biotype E (virulent to gene H3), 13% F (virulent to H6), and 9% G (virulent to genes H3 and H6). This information is important to the future deployment of Hessian fly resistance genes in wheat in Washington, and it comes as a result of Dr. Clement's initiative.

Table SC3.   Hessian Fly Infestations, Ralston Project, 1998 and 1999.

A small plot study at Central Ferry to quantify the susceptibility of wheatgrass germplasm to cereal aphids and Hessian fly was started in 1999. This plot also included plants of perennial wheat, which is being developed by Dr. Steve Jones, WSU winter wheat breeder. Sampling was insufficient in 1999 to gain a clear understanding of levels of insect susceptibility and resistance among the entries. Nonetheless, we determined that perennial wheat is a host plant for Russian wheat aphid and Hessian fly. We plan to do a better job of sampling in 2000.

Presentations

January 4	Presented "Agent peregrinations at the Central Ferry Research Station, Eastern Washington," Pacific Northwest Insect Management Conference, Portland, Oregon.
January 4	Leslie Elberson presented "Insect pests and beneficials in small grains under conservation tillage in the semiarid wheat-fallow regions of Washington State," Pacific Northwest Insect Management Conference, Portland, Oregon.
January 25	Gave talk "Ralston Cropping Systems Research" at Garfield County Farmers' Day, Pomeroy, Washington.
February 16	Gave invitational address as Visiting Professor at Whitman College, Walla Walla, Washington. Title "Plant-microbe-insect interactions: the grass endophyte story.
February 23	Gave talk to Alpha Zeta students, Washington State University, on plant introduction and the W6 Station.
April 2	Lectured on endophytes in host-plant resistance class, University of Idaho, Moscow.
December 14	Presented the poster "Endophyte diversity in tall fescue and the issue of aphid resistance." National Conference of the Entomological Society of America, Atlanta, Georgia.

Travel, Invitations, Special Awards

January 4-6	Pacific Northwest Insect Management Conference, Portland, Oregon.
February 15-16	Whitman College, Walla Walla, Washington.
March 28-30	Corvallis, Oregon to work in Dr. Doss's lab on PCR and endophytes.
April 8	Invited to write a chapter (Maintaining crop diversity - the need to keep a diverse gene-bank) for Cambridge Encyclopedia of Biodiversity and Conservation by editor of the volume.
April 28	Leslie Elberson was the recipient of the "Scientific Technician of the Year" award for ARS Pacific West Area.
30 July	Invited by organizers of next international symposium on grass endophytes to organize and chair a session on endophyte diversity in grass germplasm, Patorborn, Germany, September 24-29 2000. (some expenses covered).
August 25	Took and passed exams to obtain Pesticide License (Laws & Regulations, Insects & Diseases), Washington State Department of Agriculture. Good till 8/2004.
October 28-29	Leslie Elberson to Seattle, WA for graphic production training.
December 12-17	Entomological Society of America, national conference, Atlanta, Georgia.
December 14	Asked to consider spending a sabbatical at ICARDA, Aleppo, Syria (with full expenses covered).
December 20	Invited to visit endophyte research group in Hamilton, Agriculture Victoria, Australia and give seminar plus initiate cooperative research. Offer came with some funding (declined for 2000 because of other commitments).

Committees, Other Assignments, Activities, and News (Selected Examples)

Chair, Arrangements Committee for 2000 meeting in Montreal, Canada, Entomological Society of America

Member, Nominations Committee, Pacific Branch, Entomological Society of America

Member, Graduate Faculties, Entomology, University of Idaho and Washington State University. Committee member of five M.S. graduate students

Faculty Advisor, Alpha Zeta, Washington State University

Reviewed 9 manuscripts for various editors and authors

Leslie Elberson reviewed a manuscript for editor of Environmental Entomology

Reviewed proposal by CIAT at request of Swiss National Centre for International Agriculture, a branch of the Swiss Government

Article appeared in ARS Agricultural Research detailing Dr. Clement's past research in biocontrol of yellow starthistle

Hosted visiting scientists and visitors from China and Italy

Received approximately 10 calls of a 'cooperative extension' nature on a variety of subjects

Acting Research Leader on multiple occasions in absence of Dr. Hannan

The Clement-Quisenberry book "Global Plant Genetic Resources for Insect-Resistant Crops" was favorably reviewed in Crop Science and was designated a "Best Seller" by CRC Press, the publisher

Leslie Elberson is a volunteer in the Palouse Area Therapeutic Horsemanship Program, affiliated with the College of Veterinary Science, WSU

Research Plans for 2000

1.   Advise and consult on insect management with Facilities Manager and curators as needed.

Initiate pilot studies with Barbara Hellier, W6 Horticulture Curator, to assess the usefulness and value of Dipteran pollinators of caged accessions of Allium and poppies.

A.   Complete research to ascertain susceptibility of endophyte-infected and uninfected plants of three wild barley species to Rose Grass Aphid and Bird Cherry-Oat Aphid. Results will be reported in Brazil in August 2000 and compared with published effects of endophyte-infected wild barley on Russian wheat aphid.
B.   Initiate laboratory, greenhouse, and field experiments to determine extent of Bird Cherry-Oat Aphid susceptibility in large number of Neotyphodium-infected tall fescue accessions from Tunisia. This research will help characterize the level of Neotyphodium diversity in Tunisian tall fescue germplasm.
C.   Complete the last part of an 8-year project to document survival of fungal endophyte in seed of grass germplasm stored under different storage temperature regimes. This research will entail determination of wild barley seed germination and endophyte survival in this seed. The tall fescue and perennial ryegrass portions of this research have been completed. This research will tell us if W6 seed storage conditions are optimal to preserve Neotyphodium microbial germplasm in grass seed.

A.   Complete second year of sampling in a plot at Central Ferry designed to determine the susceptibility of wheatgrass germplasm to pest aphid and Hessian fly attack.
B.   Complete 5^th and final year of sampling in the 'Ralston plots' for pest aphids, aphid natural enemies, and Hessian fly.
C.   Participate, as time permits, in further efforts to genetically type Hessian fly populations in the Pacific Northwest. This is cooperative work with Drs. Roger Ratcliffe, Bragg (WSU Extension), and Bosque-Perez (U. Idaho).

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PROGRESS REPORT OF THE STATION RESEARCH AGRONOMIST
Richard C. Johnson

Research Activities

1.   Comparison of Agronomic and Biochemical Traits in Balanced and Bulk Populations.

Background

Last year (1998) a paper was published on genetic changes of isozyme markers with regeneration (Crop Sci 38:851-857), which expanded the information available on regeneration of heterogenetic accessions. Although studying isozyme loci provides valuable insight into how regeneration affects genetic structure, most traits potentially important for genetic resources are agronomic and/or quantitative, and data from the isozyme loci studies may not predict genetic changes in these factors. For example, a shift toward earlier flowering and seed production per spike in perennial ryegrass (Lolium perenne L.) has been observed in work in Wales associated with inadvertent selection. Therefore, a study was initiated to examine potential changes in quantitative along with isozyme markers associated with repeated regeneration cycles. The results will help us understand and improve our regeneration program through an understanding of how regeneration is changing accessions integrity at both the agronomic and molecular levels.

Objectives

In original, balanced (equal seeds per plant combined), and bulk (seeds combined proportionally to yield per plant) populations of annual ryegrass accessions, compare genetic changes in accessions using isozyme markers and agronomic traits after three regeneration cycles.

Progress

Design and Statistical Analysis:
A third regeneration cycle of the three annual ryegrass accessions, consisting of balanced and bulk populations derived from a population of original seed, was completed under isolation (summer 1997). Balanced and bulk seed samples from the third regeneration were prepared in the fall/winter of 1997/1998 for each population. An experiment was established in the spring of 1998 consisting of an original seed population, a balanced population, and a bulk population derived from each of the three accessions. These populations (nine total) were grown in two contrasting locations (Pullman and Central Ferry) in randomized complete blocks with four replications. Each replication consisted of 10 plants spaced 0.5 m apart. Thus each population was represented by 100 plants, 50 at each location.

Isozymes-eight isozymes determined previously to have polymorphic loci in these annual ryegrass populations were examined in each population. Original, balanced, and bulk populations of each accession will be compared for changes in allele frequency, heterozygosity, alleles per locus, and genetic distance. With this data we are also able to estimate effective population size, which is perhaps the most important factor affecting genetic drift.

Agronomic factors--such as plant phenology, plant height, spikes/plant, leaf length, leaf width, leaf area, specific leaf weight, and biomass production were determined for each plant in each population in 1998. Statistical comparisons of populations showed that most of the agronomic factors showed differences attributable to location, accession, and the location by accession interaction. It would be expected that these quantitative factors would show these effects and interactions. However, only biomass estimates differed significantly among the regeneration samples. Bulk samples after the second and third regeneration cycles had higher estimates of biomass than the original population. The effect of bulk samples was minimal on all other agronomic factors. Thus for most agronomic factors measured, genetic shifts could not be attributed to bulk sampling. But since there was an effect on biomass, the question is if this is important enough to justify the high, and currently prohibitive, labor costs associated of balanced sampling.

Application

In the current year we will complete the analysis of isozyme data which will provide insight into the effect of regeneration samples on effective population size and other genetic factors. The information on using both biochemical markers and agronomic factors will be applied to improve regeneration at the Pullman Station.

2. Effective Population Size of Heterogenetic Grass Accessions

Background

Normally, in a population of N plants, not all are effectively randomly mating, so the effective population size (Ne) is usually less than N. High variation in the number of progeny per plant in a population is a major factor leading to lower Ne and a higher potential for random genetic drift. For balanced samples the variance in seeds per plant is zero, eliminating variation in fecundity as a factor lowering Ne. But seed balancing is labor intensive and therefore very costly. In previous work we found that variation in seeds per plant sample reduced the ratio of Ne to N to an average of 0.73 for annual ryegrass. Thus, in a population of 100 plants only 73 are on average randomly mating. This information is needed on a broader set of accessions and species to gain insight into how bulk sampling will affect genetic drift in our outcrossing accessions.

Objective

Determine the reduction in effective population size, Ne, associated with variation in seed production per plant in three heterogenetic grass species.

Progress

In 1997, three accessions of three perennial grass species were established at two locations within current regeneration plots at the Western Regional Plant Introduction Station. The species were diploid accessions of perennial ryegrass (Lolium perenne), meadow fescue (Festuca pratensis), and bluebunch wheatgrass (Pseudoroegneria spicata). For each of the nine entries there were two replications of 30 plants each at both locations spaced 0.5 m apart and isolated by at least 50 m distance. In 1998, harvest of seeds from individual plants was completed.

Application

Estimates of the effect of variation in seed production per plant on Ne were made. The ratio of Ne to N averaged only 0.53. This meant that there was an average reduction of 47% in the effective population size attributable to variation in seeds production per plant associated with bulk sampling. This was substantially greater than the 27% reduction estimated for annual ryegrass in previous studies. The greater than expected reduction in Ne associated with bulk sampling suggests that we must balance regeneration samples and/or increase current population sizes to compensate for the low Ne values. Since balanced sampling is at presently prohibitively expensive, we are increasing basic population sizes from 60 to 120 plants.

3. Evaluation of diverse Kentucky Bluegrass germplasm for seed production in alternative residue management systems.

Background

This project has been funded through a USDA Special Grant program for Grass Seed Cropping Systems for A Sustainable Agriculture, and is cooperative with Bill Johnston at Washington State University. The goal is to identify material and/or characteristics that give good turf quality and improved seed production under no-burn residue management systems. A three-year field study of production and turf quality factors, starting in 1997, is ongoing on the core subset and selected accessions.

Objectives

1. Using morphological and RAPD data, study methods of core collection development in Kentucky bluegrass.

2. Determine seed production potential of diverse germplasm in burn and alternative residue management systems while evaluating for turf quality.

Progress

Objective 1.    Use agronomic and RAPD data to study core subset development methods in Kentucky bluegrass.

We have evaluated most of the Kentucky bluegrass collection for 17 descriptors recommended by the Forage and Turf CGC. In addition to agronomic descriptors, plant material was collected for DNA extraction and subsequent RAPD analysis was completed on 245 accessions and check cultivars times three replications (735 total).

For agronomic data, average Euclidean distances was calculated and used in multivariate statistical analyses. For RAPD markers, the Sorenson resemblance coefficient was used to calculate the distance between each accession. Thus, we derived a 245 X 245 accession matrix for distances among accessions based on agronomic data and an analogous matrix based on RAPD data. The two matrices were subjected to correlation analysis to determine if there was any relationship between the two types of data in terms of analysis of collection diversity. We found that although the majority of the variation between the agronomic and molecular data sets was not explained, the correlation coefficient between the two matrices was highly significant (r= -0.32). This suggested that in terms of diversity analysis there was a degree of correspondence between the agronomic data and the RAPD data.

There was no evidence of duplication among the total collection of 228 accessions. For agronomic data, suspected duplicates would be those pairs of accessions with an average Euclidean distance of zero. The two accessions closest to zero were PI 349212 and 349219 with a distance of 0.28. The relatively small difference between these accessions stemmed from variation in plant height and seed yield. For the RAPD analysis, duplication would be suspected if a pair of accessions had a Dice similarity value of one. The accession pair closest to one was PI 314733 and PI 204490, which had a similarity value of 0.93.

We then developed sets of trail core collections to determine which methods were most effective in representing the diversity in the total collection. From 228 total accessions, cores representing 10% of the collection were developed using random sampling, UPGMA hierarchal cluster analysis, and stratification by broad geographic regions. Cluster analysis of agronomic attributes provided cores with increased variances and ranges compared to cores developed without cluster analysis. Clustering resulted from more uniform sampling of accessions from the total collection. Stratification over broad geographic areas without clustering produced cores that were similar to a core selected at random. In addition, the UPGMA agronomic core (without stratification) had increased molecular diversity (lower Sorenson matrix mean) when analyzed with RAPD marker data. Thus, it provided a core with both high agronomic and molecular diversity, and among the cores developed, appeared to have the highest overall quality.

Objective 2.    Determine seed production potential of diverse germplasm in burn and alternative residue management systems while evaluating for turf quality.

Thirty-eight accessions from the collection of over 200 were selected for evaluation under different residue management systems. Nine commercial cultivars from the National Turfgrass Evaluation Program (NTEP) were included among the entries to represent the current genetic base and provide checks. The core subset developed from morphological data was included plus accessions that showed a combination of high turf potential and high seed yield in previous tests.

The seed production plots were established in the fall of 1996 and each plot consisted of five, 6-foot (1.8 m) rows spaced 7-inches (0.18 m) apart, and there was 2 feet (0.6 m) of border area between each plot. The first year seed production data was in 1997 and thus there was no residue that year. After harvest in 1997, three residue environment treatments were established: open-field burning, mechanical residue removal (bale), and no residue removal. The experiment was randomized in complete blocks with a strip-plot arrangement with three replications. The main plots are the residue treatments, arranged in strips, and the subplots are the accessions. Main plots were randomized within the blocks and accessions randomized over the main plots.

Turf plots with the same entries as the seed production plots were established in the spring of 1997 in 1 m² plots randomized in complete blocks with three replications. On these plots, turf quality factors are being evaluated (leaf texture, leaf color, spring green-up, and overall quality assessment).

Compared to open field burning, the average reduction in yield was 30% for the baled treatment and 76% when no residue was removed. For yield, a strong interaction between germplasm entry and residue treatment was found. Numerous accessions from the baled treatments maintained yields that were similar to that of the burned treatments. A few accessions in the baled treatment showed reasonably high turf quality with above average yield. Thus, it appeared that losses associated with non-thermal residue removal can be reduced through germplasm selection. As open-field burning becomes more restricted, the greater genetic potential of some genotypes to yield under non-thermal residue management, such a baling, needs to be exploited. Although finding genotypes with high turf quality and seed yield may be challenging, there did not appear to be an intrinsic reason why those two factors could not be combined since turf quality and seed yield were not correlated.

4. Genetic changes with seed aging

Background

Seed aging results in the loss of germination that eventually regeneration will be needed to preserve a given accession. Germination levels of between 60-70% germination have often been suggested as levels where regeneration is recommended. Often, for various reasons, an accession's germination will be much lower than 60% at the time of regeneration. Little, if anything, is known about the changes in allelic composition that may take place in a population as a result of seed aging. The loss of germination could be also associated with the loss of alleles so that the genetic integrity of the population is compromised. This is especially relevant for heterogenetic, cross-pollinating species, but could also be important for self pollinating accessions with a mixture of genetic material.

Objective

Determine if, and to what extent, seed ageing affects the genetic profiles of heterogenetic accessions

Progress

The plant material used above in the study "Effective Population Size of Heterogenetic Grass Accessions" will also be used for this study. Three accessions of three species of diploid grass accessions--perennial ryegrass (Lolium perenne), meadow fescue (Festuca pratensis), and bluebunch wheatgrass (Pseudoroegneria spicata)-were grown and seeds from individual plants harvested. The nine populations were grown at both Central Ferry and Pullman, and replicated twice at each location. In 1998, harvest of seeds from individual plants was completed.

The seeds from each treatment combination within a location were combined, resulting in 18 populations per location for a total of 36 experimental units. There are 36 populations (3 species x 3 accessions x 2 locations x 2 reps) so the basic design is a randomized complete block at two locations. Germination on this material was tested to give baseline values. The overall average was 98%. Each germination test followed official rules, and was done for each population.

In the summer of 1999, plants from each entry and location were grown in the greenhouse in containers. DNA from12 plants from each experimental unit was extracted, and gels to estimate the quality and quantity of DNA were completed. This required preparation of 432 samples of DNA, which were then visualized on gels. DNA quantities now need to be estimated and roughly equal amounts of DNA bulked into working stocks. Then, AFLP fingerprinting will be completed on the basic 36 experimental units.

Seed will be placed in different environments and germination tested over time. As germination declines due to aging we will examine the genetic profiles of populations to determine if, and to what extent genetic change has taken place. The genetic distance from any two populations can be calculated using a resemblance coefficient, such as the Sorenson similarity coefficient, and statistical analysis used test for differences in populations.

Germplasm Activities

With Vicki Bradley and Wayne Olsen, initiated improved regeneration standards for grasses. This involves an increase in plant populations to120 and isolation distances to 50 m.

Obtained Lygus bug resistance data on the alfalfa core collection from Dr. L. Teuber, UC-Davis. Working with Dave Stout, facilitated its entry into GRIN.

Working with Dave Stout, entry of evaluation data for the Kentucky bluegrass for 17 agronomic factors on 228 accessions was completed.

Working with staff, data was complied into a spreadsheet for over 1,700 safflower accessions for 29 evaluation factors for future entry into GRIN. The data was provided from Professor Li Dajue, China, though IPGRI support.

Provided information requested by Henry Shands, Deputy Director, ARS, concerning pollen contamination in outcrossing grasses, 3/2/99.

Ask by the International Plant Genetic Resources Institute (IPGRI) to develop a web page for safflower under the aegis of the International Safflower Germplasm Committee. Funded for $2,000, 3/16/99.

Continued work toward germplasm exchange and evaluation with G. Becker in Patagonia, Argentina, through evaluation of Argentina grasses in the W-6 collection.

Talks and Presentations

April 16	Presentation on "The Western Regional Plant Introduction System," to University of Idaho Range Reclamation Class. Requested by Dr. Jim Kingery.
June 22	Brief presentation on program activities to the W-6 Technical Committee meeting at Pullman, Washington.
November 2	Presentation "Seed production and turf quality of the USDA Kentucky bluegrass core collection," Crop Science Society of America, Salt Lake City.

Travel, Invitations, Special Awards

June 2-5	Invitation to Rome, Italy: Consultant and senior editor on the publication "Core collections for today and tomorrow," recently published by the International Plant Genetic Resources Institute (IPGRI). (The invitation was by Toby Hodgkin, IPGRI, Rome.) This involved editing, book layout, and other aspects of publication. Visit to Peter Griffee at FAO to establish International Safflower Germplasm Committee goals and a safflower Internet information site through FAO.
June 7-8	Córdoba, Spain. Meet with José Fernández-Martínez at the Instituto de Agricultura Sostenible, Spain, to establish a cooperative grow-out and evaluation program for safflower wild relatives in Spain and establish goals for the International Safflower Germplasm Committee.
July 25- August 5	Invitation to collect germplasm in Crimea, Ukraine. Along with Harold Bockelman and the Ukranian team, wild grasses (including wheat relatives) and legumes were collected on the Crimean Peninsula. Two hundred seventy accessions were collected. Prior to this trip there was hardly any representation for grasses and legumes from Crimea.
October 31-   November 4	Salt Lake City, Utah. Participated in the Crop Science Society of America meetings.
November 2	Awarded Outstanding Paper in the Plant Genetic Resources Section, 1998, for "Genetic structure of regeneration populations of annual ryegrass," Crop Science 38:851-857. Selected by a special peer review panel of nationally recognized scientists.

Committees, Other Assignments, Activities, and News

Chair, C852.12 Crop Registration Subcommittee for Sunflower, Safflower, Rapeseed, and other misc. oilseeds, 1995-2000. Responsible for reviews, revisions, and decisions on publications of approximately 12 registration manuscripts per year.

Member, C-118, Nominations Committee for Plant Genetic Resources Division (C-8), Crop Science Society of America, 1998-1999.

Served as member of Ad-hoc Committee on "Registration of mapping populations in Crop Science," by Loren Wiesner for CSSA Committee C852, Crop Registration. A procedure to register mapping populations in Crop Science was approved by the CSSA Board of Directors, 11/99.

Member of Graduate Faculty, Washington State University.

Appointed to the Organizing Committee, Vth International Safflower Conference, to be held at Williston, North Dakota, June 2002.

Member of the Technical Advisory Committee for the Grass Seed Cropping Systems for a Sustainable Agriculture Special Grant Program. The Committee reviews proposals and recommends funding. Attended meeting 8-9 Dec.,1999, Portland, OR. Reviewed and ranked 17 proposals for funding under a USDA Special Grant program

Cooperator with Bill Johnston and Jerry Sitton on project entitled "Disease control in bluegrass cropping systems without open-filed burning." Funded for 1999 at $26,786.

Acting RL: 2/17-2/19; 9/15-9/17; 4/15-4/23/99.

Graduate Studies Representative for Washington State University Graduate School, Preliminary Exam for Can Erol Ergenekan, Department of Biochemistry, 3/10/99.

Ask to serve on Graduate Committee for Grant Poole, Department of Crop and Soil Sciences, Washington State University, 8/25/99.

Peer Reviews for journals (3) and for ARS (2).

Plans for 2000

Continue to work with staff to implement plans to increase populations for regeneration to 120 plants and insolation distances to 50 m.

Compare differences among original, balanced, and bulk populations of annual ryegrass accessions using isozyme markers and agronomic traits after three regeneration cycles. Also complete analysis of the reduction in effective population size, Ne, associated with variation in seed production per plant in three heterogenic grass species. Apply this information to collection management and prepare manuscripts on these projects for review in peer reviews journals.

Continue studies of seed production potential of diverse germplasm in burn and alternative residue management systems while evaluating for turf quality. Complete data analysis for 1999 data and compare with 1997 and 1998 results.

Write a manuscript "Characterization of the USDA Kentucky bluegrass collection with RAPD analysis," for peer review.

Continue studies on the relationship between seed aging and potential genetic changes in accessions using molecular markers. Initiate studies to use molecular markers to identify duplication among accessions in the alfalfa collection.

Complete evaluation of grasses from Argentina currently in the W-6 collections to help set priorities for future germplasm exchange and collection in Argentina.

Hire and train new SY to manage and expand the use of molecular markers for improved management of germplasm collections.

Collect safflower from the Xinjiang Autonomous region of China in the extreme northwest. This is the main safflower growing area in China, but is not represented by accessions currently in our collection.

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PROGRESS REPORT OF THE STATION RESEARCH PLANT PATHOLOGIST
April 12, 1999 (hired) - December 30, 1999
Frank M. Dugan

Activities/Research

A.   Research in Support of W6 and NPGS:

Fungi endophytic to grasses and grass seeds

Rationale:
Host-endophyte records for the grasses concentrate on a few species of forage grasses and clavicipitaceous endophytes. Other grasses are non- or under-sampled with regard to infection by endophytes (symbiotic and/or quiescent fungi). With the exception of vesicular-arbuscular mycorrhizae (VAMs), non-clavicipitaceous fungi are seldom targeted in surveys of fungi endophytic to grasses. This survey will expand host-endophyte records with respect to grasses, and better document the fungi endophytic and/or quiescent in non-symptomatic, above-ground tissues of grass hosts (nodes and seeds). An expansion of such records is integral to improved hypothesis formulation, such as attended the expansion of records pertinent to VAMs or clavicipitaceous endophytes. Summary as of December, 1999: After plating of approximately 2600 surfaced-disinfested items onto agar (800 nodes and 1800 seeds) from 15 species (Aegilops cylindrica, Agropyron cristatum, Agropyron repens (Elytrigia repens), Agropyron spicatum (Pseudoroegneria spicata), Arrhenatherum elatius, Avena fatua, Bromus inermis, Bromus tectorum, Elymus cinereus (Lymus cinereus), Festuca idahoensis, Dactylis glomerata, Melica subulata, Phalaris arundinacea, Phleum pratense, and "feral" Triticum aestivum) approximately 305 strains of fungi have been recovered. The most commonly isolated genus appears to be Alternaria, a genus in strong need of taxonomic revision.

Other than Fusarium, Alternaria is the genus with the most entries in standard texts of seed pathology (Agarwal and Sinclair, 1997, Principals of Seed Pathology, 2^nd ed., CRC; Maude, 1996, Seedborne Diseases and their Control, CAB). Alternaria species occur endophytically in other plants with effects on vertebrate herbivores (Braun and Liddell, 1998, Phytopathology 88:S10) and are known producers of mycotoxins in Gramineae and other plants (Chelkowski and Grabarkiewicz-Szczesna 1991, Alternaria and their metabolites in cereal grain, in: Cereal Grain Mycotoxins, Fungi and Quality in Drying and Storage, Elsevier; Chelkowski and Visconti (eds), 1992, Alternaria Biology, Plant Diseases and Metabolites, Elsevier). Alternaria species are agents of black point, black head & other diseases of graminicolous plants. Other frequently isolated genera included Cladosporium spp. (agents of sooty molds) and Selenophoma (Pseudoseptoria) spp. (agents of halo spot). Significantly, Fusarium, Aspergillus and Penicillium (the classic mycotoxin producers) have been completely absent to rare in isolations from non-symptomatic seeds. Miscellaneous taxa of ascomycetes, hyphomycetes and coelomycetes have been isolated but identification is not complete.

Production of keys and descriptions for important fungal taxa recovered from non-symptomatic grass nodes and seeds

Rationale:
Available taxonomic treatments of the genus Alternaria are inadequate for the following reasons: Rotem (1994, The Genus Alternaria, Biology, Epidemiology and Pathogenicity) did not provide a key. Simmons (1992, Alternaria taxonomy, in Alternaria: Biology, Plant Diseases and Metabolites) produced a key to species groups (not to species), but that key omits one very common species recorded from grasses (A. tenuissima) and omits several lesser-known species as well, including some subsequently described by Simmons (see below). Ellis (1971 Dematiaceous Hyphomycetes, 1976 More Dematiaceous Hyphomycetes) produced a series of descriptions from herbarium material, but no key. The keys and descriptions from Joly (1964 Le Genre Alternaria; 1967 Plant Dis. Rep. 51:296) are incomplete because of omissions of major and minor taxa. Neergaard (1945, Danish Species of Alternaria and Stemphylium) is still well-regarded, but the keys omit one major and several minor species from grasses. Keys and descriptions of Joly and Neergaard are not based on standardized media and incubation conditions. The extensive series of articles by Simmons (Alternaria Themes and Variations, in Mycotxon, 1981-present) is the most complete set of descriptions of Alternaria species in which strains have been grown under standard conditions; however, except for the key to groups above (Simmons 1992) these descriptions have never been accompanied by a key.

Species of graminicolous Selenophoma (Pseudoseptoria) are well monographed from the perspective of the herbarium mycologist or pathologist focusing on characters in planta (Sprague 1950, Diseases of Cereals and Grasses of North America; Sprague and Johnson 1950, Species of Selenophoma on North American Grasses; Sutton 1980, The Coelomycetes) but cultural characters are inadequately described (Brokenshire and Cooke, 1975, Trans. Br. mycol. Soc. 64:201); an Aureobasium-like state and another synanamorph of Selenophoma lack descriptions. Selenophoma can be seed borne (Punithalingam and Waller, 1973, CMI Descriptions) and was recovered from seed in this study. Summary as of December, 1999: Permits have been obtained or requested from APHIS and reference cultures of Alternaria and Selenophoma have been ordered from culture collections. A preliminary key has been constructed for graminicolous Alternaria to serve as a hypothesis and materials have been obtained for growth of strains under standardized conditions. Preliminary descriptions and drawings of synanamorphs have been produced for several strains of Selenophoma isolated from grasses during this project.

Location of Neotyphodium in tissues of wild barley seedlings

Rationale:
Wild Hordeum species have been shown to contain Neotyphodium endophytes with effects on plant-insect interactions (Clement et. al. 1997, Entomologia Experimentalis et Applicata 82:275-282). The precise location of such endophytes within host tissues is therefore of interest. Summary as of December, 1999: Seedlings of one accession of wild Hordeum brevisubulatum subsp. violaceum known to be colonized by Neotyphodium have been subjected to clearing and staining techniques, and the position of fungus within tissues has been documented photographically in sections and whole mounts. Identity of the colonizing fungus from the same leaves used for sections and whole mounts has been confirmed by recovery of the fungus into pure culture. The experiment was repeated once. In collaboration with Nadeer Youssef (University of Idaho).

Fungi endophytic in asymptomatic grape berries and dormant buds

Rationale:
Endophytic and/or quiescent fungi have been demonstrated in several horticultural crops, including grapes, in which Botrytis is of special interest (Coley-Smith, Verhoeff and Jarvis 1980, The Biology of Botrytis; Nair et al. 1995, Australian J. Exp. Agri. 35:1177). Botrytis is a primary target of standard management practices in central Washington and the Pacific Northwest (Pscheidt, 1997, An on-line guide to plant disease control, www.orst.edu/dept/botany/epp/guide; Grove, 1998, Bunch rot of grape, http://disease.tfrec.wsu.edu). Our objective is to ascertain the degree to which Botrytis or other fungi might persist as quiescent infections in non-symptomatic grape tissues in central Washington. Taxa recovered should be tested with Koch's postulates to see which are pathogenic to ripe fruit. Summary as of December, 1999: 1000 berries and 80 buds have been sampled to date with recovery of approximately 180 isolates. Alternaria, Cladosporium, Ulocladium and Penicillium appear to predominate; Aureobasidium-like strains and lesser numbers of ascomycetes and coelomycetes were recovered. Botryis has not yet been recovered. Note: Pre-existing fungicide trials, planned and executed by WSU's G.G. Grove near Prosser, WA., presented the possibility of cooperative research; no labor, monies or materials are expended by WRPIS for establishing plots or conduct of the trials, but sampling is available to W6 in the control replications for recovery of endophytic fungi. This effort is projected at approximately one quarter of the effort devoted to the floristic survey of grasses.

B. Germplasm Activities in Support of W6

Pathogens Report

The cooperation of the pathologist was elicited in the following instances:

Identification of pathogens or potential pathogens

Lettuce drop was a problem at Whitlow farm. Curator Barbara Hellier has provided samples of diseased lettuce from which sclerotia were extracted. Symptoms of drop, white mycelium at the root collar, and morphology of sclerotia are congruent with Sclerotinia. Growth in culture had characters of S. sclerotiorum.

A soil fungus in the greenhouses was submitted for identification by Technician B. Brandon; after the departure of Ms. Brandon, Technician J. Thayer submitted an additional sample. The fungus was identified as the Chromelosporium state of Peziza ostracoderma; it was growing abundantly on soil surfaces. The fungus can occasionally become a problem on plants (Ellis, 1976, More Dematiaceous Hyphomycetes), but the only host listed in Farr et al. (1989, Fungi on Plants and Plant Products in the United States) is colewort.

Curator Barbara Hellier submitted for identification several fungi isolated from leafspots of Allium. With the exception of Botrytis cinerea, all were common saprophytes (Cladosporium herbarum, Ulocladium sp., Stemphylium botryosum, and small-spored Alternaria (not A. porri). Botrytis allii was not isolated. With the exception of B. cinerea, none of the fungi isolated was among the serious pathogens listed by the APS Compendium of Onion and Garlic Diseases.

Seedlings of Phaesolus vulgaris PI 197683 were submitted by Technician Paula Rose for identification of the pathogen producing lesions on hypocotyls. Rhizoctonia was repeatedly recovered from surface-disinfested sections of hypocotyl. Suggestions were relayed to Curator Molly Welsh for improvements in the techniques of testing accessions for seed-borne pathogens.

Provision of information

Documentation was furnished to Dave Stout regarding the 1) presence of Ditylenchus dipsaci (alfalfa stem nematode) on alfalfa in Washington state, host range, its transmission and persistence in seed, and eradicative seed treatments; 2) absence of seed transmission of Beet Necrotic Yellow Vein virus.

Documentation was furnished to Curator Dave Stout regarding common names of diseases caused by Mycoleptodiscus terrestris. A URL was provided which can be consulted for common names of plant diseases; the site is searchable by key word.

Documentation was furnished to Resarch Leader Rich Hannan regarding references to seed borne nematode infection of carrot. Only one, somewhat ambiguous, record (for Ditylenchus dipsaci, in Agarwal and Sinclair, 1997) was found in searches of the on-line Agricola electronic database and standard references.

Provision of strains

Packets of strains of Ascochyta fabae and Ascochtya rabiei were distributed to Antonio Trapera Casas in Spain and to A. Darie in Iran, respectively.

An agreement was reached with Research Leader John Kraft and Curator Clare Coyne for the provision of storage of isolates from Dr. Kraft's collection of pea pathogens. Liquid nitrogen space is available; strains and documentation will also be forwarded to other culture collections.

Talks and Presentations

Manuscripts Reviewed

Bailey, B.A., P.C. Apel-Birkhold, N.R. O'Neill, J.P. Plaskowitz, S. Alavi, J.C. Jennings, and J.D. Anderson. Evaluation of infection processes and resulting disease caused by Dendryphion penicillatum and Pleospora papveracea on Papaver somniferum. Sent for review by Senior Editor D. Weller (Phytopathology).

Clement, S.L. Maintaining crop diversity - the need to keep a diverse gene-bank, in: The Cambridge Encyclopedia of Biodiversity and Conservation, Cambridge University Press. Pre-submission review.

Croan, S.C. Lyophilization of hyphal-forming tropical wood-inhabiting basidiomycotina. Sent for review by Associate Editor Albert Torzilli (Mycologia).

Kaiser, W.J., F. Coca. W., and S. Vega O., First Report of Ascochyta blight of chickpea in Latin America. Pre-submission review for Disease Notes in Plant Disease.

Pimental, G., T.L. Peever and L.M. Carris. Genetic variation among natural populations of Tilletia controversa and T. bromi. Pre-submission review for Phytopathology.

Ju, Yu-Ming, and J.D. Rogers. Xylariaceae of Taiwan. Pre-submission review for Mycotaxon.

Whalley, M.A., Y.M.-Ju & J.D. Rogers, and A.J.S. Whalley. New Xylariaceous fungi from Malasia. Pre-submission review for Mycotaxon.

Proposals Reviewed

'Evaluation of the Chickpea (Cicer arietinum L.) Core Collection for Resistance to Pea Enation Mosaic Enamovirus' submitted by Dr. Richard Larsen to the Cool Season Legume Crop Germplasm Committee.

'Systematics of Biocontrol Fungi: Trichoderma and its Hypocrea Sexual States' submitted by Dr. Gary Samuels for ARS Merit Review.

Off-site Contact Visits

On November 2, I visited with Dr. Stephen Peterson, the curator of the ARS Aspergillus and Penicillium collections, in his laboratory at the USDA-ARS National Center for Agricultural Utilization Research in Peoria, Illinois. The Peoria facility is the site for the largest ARS collection of fungi of importance to agriculture. Dr. Peterson and several of the fungi stored at the site are featured in the November issue of Agricultural Research.

Outreach

Fusarium and Phoma were recovered from roots of pea plants brought in by Mike Devoe of Moscow Idaho Seed Company. Pea plants had discolored roots and yellowed tops. The Fusarium keyed to F. oxysporum on the basis of morphology (as opposed to F. solani, which usually has longer phialides); of the Phoma and Ascochyta species listed in the APS Compendium of Pea Diseases, the Phoma most resembled A. pinoides on the basis of morphology. Because both Fusarium and Phoma are well documented in root disorders, I suggested that Mike send samples to the diagnostic lab in Prosser.

Tests for recovery of fungi and bacteria from chickpea seed were performed for Randy Lunning of the USDA Federal Grain Inspection Service in Moscow, Idaho. Although several fungi (but no bacteria) were recovered from surface-disinfested seeds, there was no correlation between recovery of fungi and symptom formation on the seeds. A written report was submitted to Mr. Lunning.

Literature citations were provided to Tom Lumpkin (Crop & Soils Sciences Chair) for several photographic guides to nutritional deficiencies in plants and to the URL for Washington State plant disease diagnostic centers in Prosser and Puyallup.

Committees, Other Assignments, Activities, and News

American Phytopathological Society (APS)

Mycological Society of America (MSA)

American Institute for Biological Sciences

United States Federation for Culture Collections (USFCC)

Department of Plant Pathology (Courtesy Appointment) Washington State University.

Chair, Mycology Committee, APS

Biodiversity Liaison, MSA

Member-at-Large, Executive Board, USFCC.

Plans for 2000

Plans for floristic survey of fungi endophytic to grasses:

Sampling of seeds and nodes of field materials and seeds in the W6 collection will proceed for an additional year for incorporation of more genera, species and populations into the study. Heavy emphasis will be given to sampling of W6 seed accessions over the winter. Assignment of species names in Alternaria, Cladosporium, Selenophoma and other genera, and to frequently isolated Aureobasidium-like and Phoma-like taxa is projected, as are new host records for a number of infrequently isolated taxa. A draft manuscript is anticipated by the end of 2000.

Plans for keys and descriptions for important fungal taxa recovered from non-symptomatic grass nodes and seeds:

Initiate simultaneous culturing of reference cultures and selected strains isolated during this project; record and analyze characters, develop keys, write descriptions, produce illustrations. Once culture-based keys are constructed, names can be assigned to recovered taxa with increased confidence. Contingent on timely receipt of reference cultures, manuscript review and documentation in the Resource Management Information System (RMIS) is projected for the end of August 2000.

Plans for location of Neotyphodium in tissues of wild barley seedlings:

Writing of results has been initiated; inner negatives will be produced from color slides. Manuscript review and documentation in RMIS is anticipated during or before June 2000.

Plans for fungi endophytic in asymptomatic grape berries and dormant buds:

In year 2000 are projected one set of dormant season samples of buds and three additional sets of growing season samples of berries. For most pertinent taxa, Koch's postulates will be performed on grapes in fall of 2000. A final sample of dormant buds is projected for later winter or early spring 2001. Dependent on results of field samples, dormant buds of cuttings can be sampled from germplasm at Davis and/or Geneva. Projected efforts for this project are approximately a fifth to a quarter of those for the grass endophyte project (above).

Long-Term Goals

The primary long term goal is to establish a reputation for this facility as a center for taxonomic expertise in fungi associated with stored seeds. The primary means to this goal is the production of culture-based keys and descriptions of the pertinent fungal taxa. A subsidiary means to this goal is the introduction of molecular techniques to the taxonomic work. Attainment of the primary goal is the means to the subsidiary (but very important) goal of securing substantial external funding. The production of the Alternaria key mentioned above, together with a culture-based key to Cladosporium (published in 1999) will constitute significant steps to attainment of the long term goals.

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PROGRESS REPORT OF THE STATION AGRONOMY CURATOR
Vicki L. Bradley

Although curatorial reassignments were finalized in December the Agronomy Regeneration Program at the Western Regional Plant Introduction Station (WRPIS) maintained responsibility for cool-season forage and turf grass, safflower, jojoba, guayule, Lesquerella, and a number of forage legumes and miscellaneous species for the 1999 growing season.

Activities/Research

Grass Collection:

The grass collection consists of 17,031 accessions and represents almost one quarter of the WRPIS holdings. Although there are 95 grass genera, Festuca, Elymus, Dactylis, Eragrostis, Lolium, and Bromus, combined, represent nearly 55% of the entire grass collection. We planted 600 accessions for regeneration in 1999. We received 518 new accessions, and we distributed 4,810 seed packets. The most requested grass genera were Lolium (739 packets), Eragrostis (687), Festuca (585), and Phalaris (429).

In 1999 we completed a project to collect inventory data on more than 2,300 W6-numbered grass accessions. We have begun to clean these accessions, as needed, so they are ready for planting and/or to be placed in -20°C storage. We also began a program to identify mis-identified or unidentified grass accessions. Herbarium samples of twelve accessions were prepared and the contract botanist, Sandra Saufferer, arranged for their identification.

We increased the isolation distance within each of 11 planting strips (4 rows wide) in the 1999 nursery at the Central Ferry, WA research farm from 25 meters to 50 meters. It is our goal to increase isolation distance to 50 meters in all directions, as well as to increase the number of plants from 60 to 120. We may have to decrease the number of accessions regenerated in order to make the changes necessary to improve the quality of our regenerated grass seed. Acquiring more land for nurseries may be possible, but other resources (greenhouse and lathhouse space for growing transplants, planting flats, etc.) are limited at this time.

Safflower:

The Carthamus collection accounts for 3.5% of the total number of accessions at the WRPIS. This collection includes 2,287 accessions of Carthamus tinctorius (safflower) as well as accessions of 6 other species, most of which are weedy. Carthamus oxyacanthus (90 accessions) is a noxious weed in the United States. There was a good deal of curatorial activity in the Carthamus collection this year. We planted a regeneration nursery of 130 accessions at Central Ferry and sent seed of 56 other accessions for increase in cages at Parlier.

Our Central Ferry safflower nursery did not do well. We used a new system for caging, and the covering material did not stand-up to the combination of wind, irrigation, and plant spines. Many of the accessions were not self-pollinated and will not be used for distribution. Environmental factors also contributed to low quality seed in this nursery. The Parlier safflower nursery was very successful. We received seed from 55 accessions. The seed harvested was of excellent quantity and quality, and we will continue to utilize this site for safflower regeneration with the cooperation of M. Jenderek.

Dr. Richard Johnson arranged for Dr. Jose´ Fernandez Martinez of the Instituto De Agricultura Sostenible in Cordoba, Spain to increase weedy species of Carthamus. Twenty five accessions of Carthamus oxyacanthus have been regenerated by Dr. Martinez.

Evaluation of the WRPIS safflower (Carthamus tinctorius) collection for ornamental potential was continued this year. We planted 80 accessions, chosen by evaluating GRIN narrative and descriptor data, in four - row plots, one and a half meters long at Central Ferry in March. Descriptor data were collected and flower samples were taken for evaluation as fresh-cut flowers. Photos were taken of each accession in the field. Photos of cut heads representing the diversity within each accession were also taken. One hundred and forty photos have been scanned into image files, edited for size and clarity, and will soon be loaded into GRIN.

Other Agronomic Collections:

In 1999, we planted 40 accessions of various broadleaf genera at Pullman, and took 20 to the Prosser station to be included in their cage regeneration nursery. In a cooperative effort with B. Hellier, cuttings from the established jojoba nursery at Maricopa, Arizona were rooted and sent to Parlier for establishment in a perennial nursery. We received regenerated seed of 9 Lesquerella, 9 Limnanthes, and 16 miscellaneous accessions from Parlier, California. B. Hellier and V. Bradley selected 31 accessions of various taxa that were sent to Palmer, Alaska for regeneration.

Related Activities:

The WRPIS purchased a mechanical transplanter in the fall. Preliminary tests were conducted at Central Ferry and at Pullman and indicate the transplanter will work well for grass transplanting. We will use it to plant the year 2000 grass nurseries.

A lap-top computer and bar-code scanning wand for use in field and greenhouse nurseries has also been purchased by the unit. We are evaluating several labeling systems that will allow us to use bar codes in plots for data collection. Various materials, including vinyl siding, plastic "credit cards", Rolodex ® card protectors, and transparency film, are being overwintered in the perennial garden in order to determine the suitability for use in next year's regeneration nurseries. We are also investigating the practicality of replacing our metal stakes with plastic ones.

Personnel

Robert Guenthner has been the Agronomic Regeneration Program Technician since October, 1997. Three full-time temporary workers help us to maintain and harvest field and greenhouse plantings from mid-May through mid-August.

Talks, Presentations and Reports

Travel, Invitations, Special Awards

Training

Committees, Other Assignments, Activities, and News

Ex-officio member of the Forage and Turf Grass Crop Germplasm Committee.

Ex-officio member of the Clover and Special Purpose Legume Crop Germplasm Committee.

Ex-officio member of the New Crops Crop Germplasm Committee.

Member of the Association for the Advancement of Industrial Crops.

Maintained close contact with the Turfgrass Breeder's Association.

Prepared crop curator statements for 2 plant exploration proposals, and reviewed 13 PI determination forms.

Trained 24 employees on Worker's Protection Standards.

Plans for 2000

1.   Regenerate 550 grass accessions and 125 safflower accessions.

2.   Increase the distance between grass accessions in all directions in the grass regeneration nurseries.

3.   Continue screening the safflower collection for ornamental potential and continue photographing safflower accessions for loading into GRIN.

4.   Begin to collect selected descriptor data on grass accessions.

5.   Utilize bar-code scanning in regeneration nurseries.

6.   Plant 20 grass accessions for identification.

7.   Find cooperators to evaluate ornamental safflower in home landscapes.

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PROGRESS REPORT FOR COOL SEASON FOOD LEGUMES
Clarice J. Coyne

Germplasm Activities

Seed increase and descriptor data:

The goal for each accession grow-out will be to (1) fill the container in the WRPIS Seed Storage facility for medium-term storage and distribution samples (2) send backup seed to National Seed Storage Laboratory for long-term storage (1500 for self-pollinating species, 3000 for cross-pollinating species), (3) collect complete descriptor data, and in the case of peas, (4) test for virus. Pea seed virus testing: 295 pea accessions were tested for virus in 1999 by Robert Yarbrough. Of these, 28 accessions had plants that tested positive. Seed of plants testing positive for virus were harvested separately and placed into seed storage separately.

Table CC1.   Status of the Cool Season Food Legumes germplasm collection held by the Western Region Plant Introduction Unit at Pullman, WA.

Genus species (number of species)	Total	Total Available	PI Total	PI Available	Non-PI Total	PI Backup NSSL	% PI Backup
Cicer arietinum	4433	4063	4000	3941	433	3879	88
Cicer species (20)	176	117	133	113	43	104	59
Lathyrus sativus	247	222	231	213	16	192	78
Lathyrus species (45)	458	214	244	196	206	89	19
Lens culinaris	2781	2631	2643	2623	138	2601	98
Lens species (3)	62	88	55	88	7	0	0
Lupinus albus	321	124	164	123	157	64	20
Lupinus species (56)	762	318	562	312	163	153	20
P. sativum	3504	2609	2936	2568	568	2382	81
P. fulvum	49	22	31	19	18	0	0
Trigonella foenum-graecum	174	163	156	151	18	155	89
Trigonella species (23)	71	49	52	48	18	25	35
Vicia faba	551	364	403	359	148	316	57
Vicia sativum	664	560	570	539	94	496	75
Vicia species (41)	786					73	9
Total	15039					10539	70

Significant seed was harvested from perennial species of Cicer from the four perennial gardens in the Pullman area in 1998. Plans are being made to move and expand the perennial gardens to increase our efficiency in the summer field season. Additionally, drip irrigation will be installed in 2000 to improve plant growth, and increase yields. Complete maps were made in 1999 of the four gardens to identify dead accessions and identify plants without NPGS accession numbers.

Field increase of Lupinus in Parlier, CA location was conducted by Dr. Maria Jenderek of the species: L. albus, L. luteus, and L. mutabilis. 55 accessions of Lupinus albus were sent for fall 1999 planting. We switched from direct seeding to transplanting seedlings in 1999 to increase the final stand count.

A field increase was conducted of 15 accessions of Vicia sativa in cages with pollinating insects in Prosser, WA by Dr. Stephanie Greene in 1998 and this continued with 30 cages of food legume accessions in1999.

Other seed increases for fall 1998-winter 1999 greenhouse planting focused on (1) accessions requested for which we have no distribution samples (2) Cicer (W6 assignments) collected from Russia by Sperling in the early 1990's for descriptors and seed increase for distribution (3) Lens (W6 assignments) collected from Turkey by Kaiser and Muehlbauer in 1989 for descriptor data and seed increase for distribution (4) collected Pisum subspecies in need of descriptors, virus testing, and seed for distribution.

New descriptor data entered into GRIN in 1999:

Pisum reaction to Fusarium root rot from John Kraft, USDA-ARS, Prosser, WA;

Pisum reaction to Fusarium wilt race 1, from Fred Muehlbauer, USDA-ARS, Pullman, WA.

Pisum reaction to Fusarium wilt race 2, from Fred Muehlbauer, USDA-ARS, Pullman, WA.

Plant germplasm conservation research 1999:

(1)   Since pea is vulnerable to an aphid-transmitted seed-borne virus, all increases are limited to the greenhouse and screen houses in pot culture. Previously, seeds were sown 4/2 gal pot. A replicated plant density study was conducted comparing seed yield with 4, 3, 2, and 1 plant per pot. This study concluded that the highest yield per plant was obtained by planting 2/2 gal pot.

(2)   Suspected duplicates of Pisum sativum cultivars in the collection were grown in the field for morphological and molecular evaluation to reduce redundancy from multiple donations to NPGS of the same genotype.

(3)   A joint project with Richard Hannan and Dave Stout under way is to evaluate the Cicer arietinum core collection to reduce the size and increase the diversity represented by including material from recent collections. The Cicer core was planted at WRPIS Central Ferry farm for evaluation and photo-documentation for entry into GRIN.

(4)   A species identification project was initiated to identify to species accessions currently only identified to genera by planting and collecting of herbarium samples to distribute to NPGS botanists and other experts.

(5)   Vicia faba experiment: 1998 field increase of 30 Vicia faba under a screen type netting to prevent cross-pollination was successful. In 1999, the netting system was improved by using a lighter material, commonly used as floating row covers in high value crops, over 60 Vicia faba accessions. The Vicia faba collection will now be increased using floating row covers to prevent cross-pollination between accessions. The following program will be implemented in the 2000 field increase of Vicia faba

a.   All Vicia faba (537) accessions will be increased from original or early increase seed from storage over the next few years.

b.   All increases will be protected from cross pollination.

(6)   Vicia faba pollination/irrigation experiment. A preliminary field trial was planted at Pullman and Prosser WA to compare seed yield with cross-pollinators present vs. absent, under irrigation vs. dryland production. Based on the preliminary results (Table CC2), a replicated trial is planned for the 2000 field season.

Table CC2.

Accession number	Genus species	Seed yield in grams per plot
		Dryland No Bees	Irrigated No Bees	Irrigated Bees
PI 275641	Vicia faba	276.4	377.8	672.4
PI 358267	Vicia faba	260.8	510.7	581.7
PI 415035	Vicia faba	36.4	351	525.5
PI 458604	Vicia faba	245.7	254.3	194.9
PI 469140	Vicia faba	789.3	486.7	760.1
PI 469163	Vicia faba	233.7	339.2	875
PI 469197	Vicia faba	802.8	584.4	1336.4
PI 567887	Vicia faba	369.9	509.1	1083.7
PI 577723	Vicia faba	830.4	908.2	1507.6
PI 577732	Vicia faba	599.6	634.2	974.5

1999 annual field planting:

Complete descriptor data was collected on the annual plantings, as many as possible with available resources. Leaves were harvested from the Pisum-duplicate cultivar trial and the Cicer core collection for use in molecular analyses. My first field season was used to set up systems to streamline data collection and increase seed yields to reduce accession grow-outs.

Cental Ferry Farm

Cicer-core collection, 505 accessions Pisum-duplicate cultivars, 234 accessions

Whitlow Farm

Vicia faba-60 accessions Vicia species-60 accessions Lens culinaris -72 W6-numbered accessions

Screen house, East and West

Pisum sativum-1999 was the first time of two cycles were completed through both screen houses.

Greenhouse, 109B-East A, B, C sections

Pisum-three cycles, continuing new planting fall 1999 Vicia-12 accessions of unidentified species for herbarium samples

Research Activities

The Cool Season Food Legume program consists of the following genera: Pisum, Cicer, Lens, Lupinus, Vicia, Lathyrus and Trigonella. These genera are advised by two Crop Germplasm Committees: Pisum CGC meeting bi-annually at the National Pea Improvement Association (Calgary, Canada, November, 1999) and the Cool Season Food Legume CGC which meets annually in conjunction with the Crop Science Society America (November, 1999).

Summary:   Develop genomic approaches to germplasm characterization, utilization, and development using pea as the model system for analyses using automated genotyping with AFLP DNA markers and chickpeas as a model system for STMS DNA markers. Visiting Scientists and their projects in the Western Regional Plant Introduction Germplasm Laboratory:

Dr. Eric Storlie - half-time post-doctoral researcher working on chickpea and pea bacterial artificial chromosome (BAC) libraries.
Dr. Marie-Laure Pilet - Post-doctoral research fellow from INRA, France (10-16-99 thru 4-18-00) working on analysis of Aphanomyces root rot resistance from the germplasm line PI 557500. Aphanomyces root rot is the greatest yield constraint world-wide on pea and we are looking for efficient methods to identify resistance in the NPGS Pisum germplasm collection.

Other Projects and Collaborations

Project Title: DNA marker for fusarium wilt race 5

Project Objective: Locate DNA marker for fusarium wilt race 5 for use in marker assisted selection. Completing post-doctoral research project initiated with the USDA-ARS Grain Legumes unit.
Collaboration: Coyne, Inglis,, McClendon and Muehlbauer.
Manuscript in preparation.

Project Title: DNA marker for fusarium wilt race 2

Project Objective: Locate DNA marker for fusarium wilt race 2 for use in marker assisted selection. Completing post-doctoral research project initiated with the USDA-ARS Grain Legumes unit.
Collaboration: Coyne, Inglis and Muehlbauer.
Manuscript in preparation.

Project Title: DNA marker for fusarium wilt race 1

Project Objective: Locate DNA marker for fusarium wilt race 1 for use in marker assisted selection. M.S. thesis project of Melissa McClendon.
Collaboration: McClendon Coyne, Inglis, and McPhee.

Project Title: Chickpea BAC library

Project Objective: Positional cloning of the two genes conferring resistance to ascochyta blight in chickpea using a BAC library.
Collaboration: Meksem, A.N. Rajesh, Tekeoglu, Coyne, Lightfoot and Muehlbauer.
Library completed. Manuscript in preparation.

Project Title: Pea BAC library

Project Objective: Positional cloning of the gene conferring resistance to pea seed-borne mosaic virus in pea using a BAC library.
Collaboration:: Meksem, Coyne, Keller, Storlie, Steber, Martin and Lightfoot.
Library 20% complete.

Project Title: Microsatellite fingerprints of Japanese vegetable soybean cultivars.

Project Objective: M.S. thesis project of Makiko Miruma,.
Collaboration: Miruma, Bickle, Lumpkin and Coyne.

Presentations

January 18	Presented, "DNA Marker for Fusarium Wilt Race 5 of Pea. International Plant and Animal Genome VII Conference, San Diego, CA.
March 12	Presented, "Quantitative trait loci in plant breeding and host plant resistance to insects and pathogens. Guest lecture: March 12, 1999 Ent 446/546, University of Idaho.
November 1	Presented, "Report of the Cool Season Food Legume curator" to CSFL CGC, ASA, CSSA, SSSA Annual Meeting, Salt Lake City, UT.
November 1	Presented, "BAC library development for the positional cloning of PSBMV and fusarium wilt race 1 resistance genes. National Pea Improvement Association Biannual Meeting, Calgary, Canada.
November 3	Presented, "Plant ready transformation BAC library of pea: progress in positional cloning of sbm-1." C-07 Cell Biology & Molecular Genetics. ASA, CSSA, SSSA Annual Meeting, Salt Lake City, UT.
November 7	Presented, "Report of the Pisum curator." Pisum Crop Germplasm Committee. National Pea Improvement Association Biannual Meeting, Calgary, Canada.
November 8	Presented, "Utilization of a pea BAC library for genetic resource development and management." C8, Plant Genetic Resources. ASA, CSSA, SSSA Annual Meeting, Salt Lake City, UT.
November 8	Presented, "Aphanomyces root rot resistance QTL: Fine mapping" prospects using automated AFLPs" National Pea Improvement Association Biannual Meeting, Calgary, Canada.
November 8	Presented, "Conservation and Availability of Critical USDA-ARS Fungal Germplasm Pathogenic to Pea (Pisum sativum L.)" National Pea Improvement Association Biannual Meeting, Calgary, Canada.
November 8	Presented, "Progress in locating DNA markers for resistance to fusarium wilt races 1, 2, and 5 in pea." National Pea Improvement Association Biannual Meeting, Calgary, Canada.

Visiting Scientists

August 1- December 31	Dr. Eric Storlie- post-doctoral researcher working on chickpea and pea BAC libraries. Flip 84-92C is an ascochyta blight resistant germplasm release from ICARDA.
October 16- April 18	Dr. Marie-Laure Pillet Post-doctoral research fellow from INRA, Rennes, France working on analysis of Aphanomyces root rot resistance from the germplasm line PI 557500.

Travel

January 17-21	Traveled to Plant and Animal Genome VII, presented research poster.
March 14- April 3	Traveled to Southern Illinois University, BAC library construction.
July 14	Traveled to Mount Vernon, WA presented talk at Twilight Pea Tour.
October 31- November 3	Traveled to ASA-CSSA-SSSA meeting-presented two posters and one talk.
November 7-9	Traveled to Calgary, Canada for the National Pea Improvement Association, presented two talks and three posters.
December 1	USA Dry Peas & Lentil Council, Grower Division, Annual Meeting.
December 6-13	Traveled to Southern Illinois University, BAC library construction.

Committees, Other Assignments, Activities

Appointed, Washington State University, Graduate School Faculty.

Member, Cool Season Food Legume Crop Germplasm Committee.

Member, Pisum Crop Germplasm Committee.

Member, American Association for the Advancement of Science

Member, Crop Science Society of America

Member, National Pea Improvement Association

Graduate Major Advisor

Melissa McClendon
Master of Science, Department of Horticulture, WSU.

Graduate Major Co-Advisor.

Makito Mimura
Master of Science, Department of Crop and Soil Science, WSU.

Member, Graduate Advisory Committees

Jeff Shultz
Master of Science, Department of Crop and Soil Science, WSU

Jena Lewinsohn
Master of Science, Department of Botany, WSU

Competitive Grants funded in 1999

1.	USDA-CREES-Special Grants program for Cool Season Food Legumes. Inglis, Coyne and McPhee: Novel DNA Marker for Fusarium wilt race 1 resistance gene. $31,200.
2.	Northwest Agriculture Research Foundation: Coyne and Inglis. DNA Marker for Fusarium wilt race 2. $7900.
3.	IMPACT CONTINUING PROJECT PROPOSAL, 2000 TITLE: Multi-disciplinary Approach to the Production and Marketing of East Asian Crops in Washington State Principal Investigators: Dr. Thomas A. Lumpkin, Chair, Crop and Soil Sciences Co-P.l.s. Crop and Soil Sciences: Dr. Bill Schillinger; Dr. Clarice Coyne; Dr. Carol Miles, WSU Coop. Ext. Ag. Syst; Ag. Res. Tech, Carrie Bickle; Marie Bumback and Makiko Mimura, grad students; Dr. Gavin Sills, Sweetbriar Development Inc. $65,000.

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PROGRESS REPORT OF THE STATION HORTICULTURE CROPS CURATOR
Barbara Hellier

The WRPIS Horticulture Crops Program is responsible for 177 genera (comprising 4,976 accessions) maintained by at the Pullman station. The major collections (those genera with 40 or more accessions) in this program are Lactuca and Lactuca relatives (1,315 accessions), Allium(874 accessions), Astragalus (692 accessions), Onobrychis (599 accession), Papaver (305 accessions), Sanguisorba (120 accessions), Scorpiurus (80 accessions), Anthyllis (72 accessions), Hedysarum (77 accessions), Plantago (46 accessions), Osteospermum (43 accessions), and Salvia (40 accessions). The Horticulture Crops program began 1999 with 119 genera. An additional 58 genera were assigned to the program at the end of 1999. Although this program includes a wide range of genera with a wide range of uses, the major focus is on medicinal and ornamental species.

Activities/Research

Lactuca and Lactuca relatives collection:

Lactuca, 6 accessions of Sonchus, and one accession each of Scariola and Mycelis. This year there were 68 accessions increased in the field (~30 plants per accession), and 8 accessions grown in the greenhouse. Any accession with low germination, very low seed supply or is a crisphead type is grown in the greenhouse. Three of the 76 accessions grown tested positive for LMV. Infection was 2% for all three lines. Positive plants were eliminated. We are continuing to slowly work through increasing the wild Lactuca species. These accessions are greenhouse increased only so progress is slow due to space limitations. We increased 15 wild Lactuca accessions in 1999.

To date, we have 1144 PI numbers and 171 W6 numbers in the collection. There are 951(83%) PI and 14(8%) W6 accessions available for distribution. 1009(88%) accessions are backed-up at NSSL. From January 1 to December 1999 there were 373 accessions distributed from the Pullman PI station and 20 PI accessions from the Salinas, CA location.

We have slightly modified the tentative Core Collection by adding several crisphead types, which were under represented, and 8 lines used extensively in breeding programs increasing the Lactuca sativa Core to 138 accessions. Except for the new additions, head type and country of origin were the two criteria used to select the core. For a particular head type, an accession was selected at random from each country represented. If there were more than 15 accessions from a particular country for a particular head type, 10% of the accessions from that country were selected at random. Only lines for which we had head type data were used in selecting the tentative core.

Allium collection:

There are currently 874 accessions (431 PIs and 443 W6 numbers) in 103 species extant in the Allium collection at WRPIS. From January 1 to December 31, 1999 we received 75 new Allium accessions. During the 1998-99 season, increase plots of 356 accessions were established at the Pullman location, and 157 at Central Ferry. There are 422 accessions available for distribution, and 26% of the non-clonal species are backed up at NSSL. 955 samples were distributed to researchers, domestic and foreign (Table 1).

Many of the older, unavailable A. ampeloprasum accessions, receive from Geneva, NY, have very low germination rates. We are currently working to increase these lines. We are trying to identify optimal germination and greenhouse conditions for growth because we have had difficulty with field increasing accessions with poor vigor. There are approximately 78 older, low germination A. ampeloprasum accessions.

Both Dr. Loren Wiesner and Dr. Leigh Towill at NSSL are interested in pursuing the long-term storage of A. sativum in liquid nitrogen. Dr. Towill has expressed interest in continuing the experimentation necessary to perfect cryopreservation techniques of garlic meristems. Dr. Wiesner has indicated he is would like to begin back-up storage of the garlic collection.

Table 1

Species	Total number of accessions	Total available accessions	Number of accessions backed-up at NSSL	Number of samples distributed
sativum and longicuspis	264	206	0	872
ampeloprasum	189	56	53	7
schoenoprasum	32	22	19	0
all other species	447	147	94	76

Allium in situ research:

We concluded the current in situ study this summer. Data from one additional site for both A. fibrillum and A. columbianum were collected. On preliminary analysis of the study, in situ seed collection appears to be a viable increase method for Allium species native to North America. We are proceeding with plans to expand this program.

All other species collections:

There are currently 175 genera and 2987 accessions in these collections. In 1999, 152 new accessions were received and 491 seed packets were distributed to scientists, foreign and domestic.

The curatorial focus for the non-Allium/Lactuca collections in 1999 was to establish increase priorities and inventory the collections. Other activities related to these collections were as follows.

The Rheum collection was significantly increased in 1999. With the help of Mr. Dale Marshall, retired USDA-ARS Agricultural Engineer and the staff of the Clarksville Hort. Farm, 75 clonal accessions were received from the Michigan State University rhubarb cultivar evaluation nursery. These accessions were transplanted to plots at our Pullman farm.

Three accessions of Guizotia and two of Nigella were increased at Central Ferry in 1999. We experimented with transplanting greenhouse-started plants. Seed increase was marginal for the Guizotia and good for Nigella.

An updated inventory of the established plantings at Central Ferry was made. These plots were also trimmed with dead plants removed. Genera in these plantings include Iris, Astragalus, Thymus, Artemisia, Amorpha, and Hemerocallis.

Seed was harvested from newest plantings at Central Ferry and in Pullman. These plots of Glycyrrhiza and Amorpha were established by Vicki Bradley.

Vicki Bradley had also sent several accessions to Parlier, CA for increase. Increase for a majority of these lines was good to excellent. Genera sent to Parlier included Rogeria, Plantago, and Glycyrrhiza.

Seed of several accessions was also sent to the Palmer, AK station for increase. Genera sent included Crepis (four species), Hedysarum, Polemonium, and Arctostaphylos.

Presentations

February 7	Tour of seed storage, GH 44, and garlic storage facilities. University of Idaho class-Host plant resistance to insects and pathogens.
April 6	Tour of garlic storage facilities and Pullman garlic nursery with Rob Mercer and associates from Mercer Farms, WA.
June 22	Western Region 6-Technical Advisory Committee meeting: Horticulture Crops Program overview, Central Ferry, WA.
June 25	Washington State University Extension Othello Garlic Field Day: NPGS garlic collection. Othello, WA.
June 23	Tour of garlic storage facilities and Pullman garlic nursery with Greg Anthony and family from Irish Eyes and a Hint of Garlic, Ellensburg, WA.
August 31	Environmental Science/Regional Planning 593 seminar: A National Plant Germplasm System: exploring in situ conservation at Pullman, WA.
September 22	Visit by Ron Voss, University of California Extension. Tour of garlic storage facilities and Pullman garlic nursery along with discussion on entering garlic data collected by Ron Voss into GRIN.

Travel

February 23-25	University of Arizona Maricopa Agriculture Center, Maricopa, Arizona. Took cutting from the PI jojoba nursery for reestablishing nursery at the Parlier, CA station.
June 25	Washington State University Extension Othello Garlic Field Day. Othello, WA.
July 26-30	Root and Bulb Vegetables and the Leafy Vegetable CGC meetings held in conjunctions with the American Society of Horticultural Science meetings. Minneapolis, Minnesota.
Oct. 18-21	Attended the New Crops CGC meeting and visited Territorial Seed Company. Eugene and Cottage Grove, OR.
October 31- November 6	Visited Dr. K. Shierenbeck's lab at Chico State University to learn RAPD protocol and work on RAPDs for in situ project. Chico, CA.

Committees

WSU Plant Growth Facilities Advisory Committee member (membership expired in September).

WSU Plant Growth Facilities Manager search committee member (Jan. to Sept.).

ARS Pacific West Area awards review panel member (March).

Ex-officio member of the Root and Bulb Crop Germplasm Committee.

Ex-officio member of the Leafy Vegetable Crop Germplasm Committee.

Ex-officio member of the New Crops Germplasm Committee.

Training

Pesticide Recertification Training, Feb 3-4, Moscow, ID.

Adult CPR and First Aid, Dec 7 & 9, Pullman, WA.

Plans for 2000

Lactuca and Lactuca relatives collection:

Continue field and greenhouse increase programs with an emphasis on wild species and crisphead types for increase in the greenhouse.

Expand lettuce nursery locations to include Central Ferry and the bubble greenhouse.

Continue work on the low germination accessions. Include sterile culture in test protocols.

Allium collection:

Continue in situ program.

Continue work on the low germination A. ampeloprasum accessions. Include sterile culture in test protocols.

Continue field increase on new and unavailable accessions.

In conjunction with Steve Clement and the Entomology program, conduct preliminary experiments to determine optimal pollinator populations needed for controlled pollination of A. ampeloprasum and other wild Allium species.

All other species collections:

Genera chosen for increase:

Plantago-newest accessions

Glycyrrhiza-unavailable accessions plus "species" lines

Papaver-"species" lines

Guizotia-to be increased at Parlier, CA

Cyphomandra

Hedysarum-newest material

Sanguisorba-unavailable accessions

Lycium

Astragalus- several species. For increase in Pullman, at the Central Ferry farm, and Prosser

Borago

Salvia hispanica -unavailable accessions

Investigate efficacy of controlled pollination of Papaver bracteatum using house flies as pollinators.

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PROGRESS REPORT OF THE BEET (Beta) PROGRAM
Alan Hodgdon

Increase Activity

Thirty-nine accessions harvested at W-6 in 1999 have been cleaned and weighed. Of these, thirty-six were increased in the greenhouses. Three of these accessions will have to be redone. Only three accessions were successfully increased in the field due to very bad growing conditions. Twenty-five plots froze during the winter, and several surviving plots did not flower well during the heat of the summer. Fifty-four accessions were started at W-6 in 1999. In all, 110 accessions are in the process of increase. The increase priority list has 448 accession as of the end of 1999. Twelve accessions are being increased by seed companies in the U.S. This help is greatly appreciated. Field increases at W-6 have been a problem with poor plant numbers, poor quality seed, and low seed yield.

In the future we will try artificial vernalization, and then spring plant in at our Pullman site. This could solve the problem of plot freeze-out and provide a cooler weather grow-out site. I am not optimistic about this solution since our growing environment is a poor match with that of the wild beets, particularly Beta maritima. With the greenhouse increases, we have had good seed yield and quality, but progress has been slowed by deinduction of flowering especially with the wild beets. We are working on changing the post-vernalization conditions to improve flowering.

Seed Germination

One hundred-five Beta seed samples were tested for germination in 1999. The seed lab is using a dry germinator which gives better results. No specific seed germination data is available now. However, no sample had less than 20% germination, and most samples were higher than 50%.

Seed Storage Activity

W-6 distributed 643 samples from the Beta collection in 1999, and we acquired thirty-one new accessions. There was no new backup activity for the Beta collection.

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PROGRESS REPORT OF THE PHASEOLUS CURATOR
Molly Welsh

Activities/Research

During the year 1999 approximately 2.5% of the collection was increased. All accessions in the increase cycle were tested for the presence of BCMV. Part of the Phaseolus germplasm project includes determining presence of the virus and the clean-up of accessions that test positive. Presently 13.5% of the collection is labeled 'tested virus-free'. During the year 12 accessions, testing positive, were run through the virus cleanup program and are now labeled 'tested virus-free'.

The "BCMV clean-up" of accessions which remained as a backlog from previous years production programs has been completed and the "BCMV clean-up" program is now limited to accessions identified in the current years production program.

The BCMV files have been completely re-organized and brought up to date. They now include all past available records along with those generated in the ongoing program. The lab procedures for the BCMV testing has been reviewed and modified, and a protocol manual of the current lab procedures for the BCMV testing is being produced. This manual is available upon request and will be upgraded as needed.

Previously the "BCMV clean-up" program was implemented when accessions tested greater than 10% or less than 15% virus-infected. This program has been expanded to clean up lines with a greater level of contamination. Additionally, accessions labeled as non virus-free are now regularly requested from seed storage to be entered into the "BCMV clean-up" program.

Material donated by Dr. George Freytag has been sorted and categorized. Some of this material was added to past contributions within the collection; some was new and has been entered into the collection. A program to classify systematically some previously unclassified material donated by Dr. Freytag has been initiated.

A uniform system of recording increase information, including plant character data, BCMV testing results, and harvest results was developed and is being used in the Phaseolus germplasm increase program. Collection and recording of this information is done on a regular basis.

Three accessions, previously not categorized systematically, have been identified. One of the accessions was sent to the Southern Regional PI Station (S9) [USDA, ARS, SRPIS, University of Georgia, Plant Genetic Resources Conservation Unit, 1109 Experiment Street, Griffin, Georgia 30223-1797] to be stored with the appropriate collection. Two of the accessions remain in the Phaseolus collection.

Information contained in the GRIN web-page descriptor site has been updated and continues to be monitored and changed as additional data is obtained. In particular emphasis has been placed on updating this information to keep it a current resource. A differential set for the disease Anthracnose has been entered into the collection and the information is available on GRIN.

The GH 44 project was completed with the following improvements: a new gravel base in the house, addition of a water permeable weed barrier, installation of new tables with at a more functional height, and installation of fertilizer injection system. A program for bio-control of insect pests, similar to the program in GH 109, was initiated in GH 44.

Fertilizer injection systems were also placed in GH 109 and GH CF. The system will provide the plants with needed nutrients in an evenly regulated manner.

Aphid proof netting has been installed in GH 109 and GH 44.

A laboratory for sterile work was set up in Johnson Hall (room 268). The work done here will include embryo rescue and tissue culture. We received in 1999 a few seed that had been preserved since 1920's at the Peabody Museum. This seed is all that remains of two lines developed by Mandan and Hidatsu tribes of the Dakotas. Embryo rescue will be used to attempt to save the lines.

A program to include photographic representation of genetic stocks and of plant character descriptors has been developed. A video camera was purchased for this project and methods for obtaining the best quality photographs are being researched. The pictures will be included with the information provided in the GRIN site as soon as feasible.

The field plantings at Central Ferry produced a good quantity of seed. Unfortunately, it became apparent shortly after plants began to be tested for BCMV that some of the seed planted had not been virus-free. This condition did not allow the completion of the original experiment. Therefore, it was decided to repeat the experiment in the next year.

Plans for 2000

1. Continue all on-going projects:

a)   Pursue the "BCMV clean-up" program and continue to increase the number of labeled 'non-virus free' accessions to be entered into the program

b)   Sort varied collections stored at the W6 site, reviewing them for material to be added to the collection, and organize the collections to make data and germplasm available to more clients

c)   Further study in the area of Phaseolus systematics and botany to identify previously unidentified species within the collection

d)   Keep the GRIN resource current and "user-friendly"

e)   Work with clients to obtain data from their research with the requested seed

f )   Spanish language studies

g)   Improved skills in the utilization of the GRIN database

h)   Further organization of collection to make better accessibility of accessions and related data to interested parties

i )   improve my experience with propagation of beans, both in greenhouse and in field: control of pests, nutritional requirements, etc.

j )   all of the regular increase, germplasm acquisition and distribution, and data collection programs

2.   Begin the program to "save" endangered accessions through embryo rescue work.

3.   The experiment to produce virus-free seed outdoors will be repeated in summer of 2000 at Central Ferry.

4.   Review the method of characterization and evaluation of the collection and determine that which will provide the most information and make such information most useful/accessible.

5.   Interact with professionals in similar work areas to learn how the material in their collections is organized, maintained, and distributed.

6.   Obtain a pesticide applicator license so I am able to deal more effectively with pest problems in the greenhouses.

7.   Attend classes to learn the protocol, etc. for handling the information in the GRIN database

Presentations

February 8	Presented lecture "Increasing Genetic Diversity of Soybean in the United States"; graduate student seminar series for the Department of Crop and Soil Sciences, WSU, Pullman, WA.
June 22	Spoke to W6 TAC members about beans growing at remote site, Central Ferry, WA.

Committees and other assignments

Member: Phaseolus C G C

Member: BIC Genetics Committee

Member: W150 Regional Project

Member: Seed Savers Exchange

Activities and News

January 19	Attended safety training course for chemical spills-Pullman
January 27	Attended workshop with W6 curators-Pullman, WA
February 15-20	Attended annual meetings (1999) of the Phaseolus Crop Germplasm Committee and the W150 Regional Project -Mayaguez, Puerto Rico
May 27	Attended safety training course for general chemical and pesticide handling safety-Pullman, WA.
June 21-22	Attended W6 TAC meetings-Pullman & Central Ferry, WA.
September 27	Conducted tour of W6 for 2 scientists (adzuki bean breeders) from Japan-Pullman, WA.
November 8-12	Attended annual meetings (2000) of the Phaseolus Crop Germplasm Committee and the W150 Regional Project, and bi-annual meeting of the Bean Improvement Cooperative-Calgary, Alberta CANADA.
December 17	Attended conference/workshop on BCMV-Prosser, WA.

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PROGRESS REPORT OF FORAGE LEGUME GERMPLASM UNIT, PROSSER, WA
Stephanie L. Greene

The Forage Legume Germplasm Conservation project manages and conducts research on the NPGS collections of alfalfa, perennial clover and trefoils. The FLGC collection contains over 13,000 accessions representing 72 taxa of Medicago, 132 taxa of Trifolium, and 39 taxa of Lotus. This report will focus on the activities and accomplishments of the FLGC project in 1999.

Service Activities

1.   In 1999 we regenerated 390 accessions, representing 95 different species in 10 genera. Our main emphasis was on Medicago and Trifolium accessions that were new acquisitions, old varieties with expired PVP status and Dr. Norman Taylor's University of Kentucky clover collection. We also surveyed species of the following genera to assess seed production at Prosser: Astragalus, Onybrycis, Vicia and Lathyrus.

2.   This summer we had good success eliminating the use of methyl bromide as a soil fumigant. Pathogen problems were limited to insects, and we had good success managing weeds using a combination of tillage and Glyphosate. Two of our greenhouses were completely rebuilt and are now in production. In October, we took delivery of a custom-built variable speed belt thresher.

3.   We are currently revising the GRIN descriptor list for the clover collection, and reorganizing and standardizing the data in GRIN. I developed a set of questions and answers (FAQ) to help users access information on the GRIN website. This has been posted at the main web site. Additionally, I developed a forage legume data dictionary that can also be access on the GRIN web site. We successfully uploaded approximately 4500 estimated latitude and longitude values for alfalfa and clover accessions. I assisted with the updating of the germplasm status report for alfalfa.

Research Activities

1.   In June, we completed screening 39 populations of red clover using RAPD markers. We also completed our second year of morphologically evaluating this same set of germplasm at Prosser and Pullman. This test array was part of the germplasm collected in the Caucasus Mountains, Russia and has extensive ecogeographic data. We will be comparing the relationships among the populations as reflected by the three marker systems (RAPDS, morphology, ecogeography). This research will also let us assess the usefulness of using GIS-derived data to predict which populations will add the most value to the current NPGS collection.

2.   We are currently analyzing the results of surveying 56 ex situ germplasm institutes in an attempt to assess the global conservation status for the genera Medicago and Trifolium

Presentations

January 22	Led Western Regional Plant Introduction Curator Workshop
October 1	University of Idaho Plant, Soil and Entomological Dept. Special Topic Field Trip "Conserving Forage Legume Genetic Resources"

Travel, Invitations, Special Awards

August 31	Reviewed Dr. Robert Webster's GIS project, NGRL, Beltsville, MD.
September 1	Attended the ASA Annual Meeting, Salt Lake City, Utah

Committees, Other Assignments, Activities, and News (Selected Examples)

Chair, CSPL CGC subcommittee on descriptor revision

Contributing member to the rewrite of ARS National Program 205 -Area: Plant Genetic Resources

Member, Crop Science Crop Registration Subcommittee - misc. legumes

Member of M.S. committee of Mercy Neumann

Reviewed manuscripts for various editors and authors

Plans for 2000

1.   Complete manuscripts on global survey, collaborative work with Marina Gritsenko, and Caucasus red clover project

2.   Develop a website and general information brochure for the Prosser Forage Legume project. Develop a brochure to assist users in accessing information on the forage legume collections

3.   Carry out a seed collection trip to Kazakhstan in August 2000

4.   Work out procedures and establish a lab for seed rescue of vulnerable germplasm

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PROGRESS REPORT OF THE NATIONAL ARID LAND PLANT GENETIC RESOURCES UNIT, PARLIER, CA
M. M. Jenderek

Germplasm Activities

Out of 231 plant germplasm accessions received in 1999 or carried over from 1998, 167 accessions (72.3%) were increased, 16 (6.9%) were carried over to CY 2000 and seeds of 43 (18.6%) accessions did not germinate (Table MJ1) The remaining 2.2% of accessions either did not flower or were damaged by gophers. The mass of seeds shipped to curators from the propagated material was ca. 158 kg. Additional 15 kg of perennial accession seeds were mailed to their main repositories (Table MJ2). 229 new accessions were planted or sown in fall of 1999 for propagation in year 2000 (Table MJ3). Considering the mass of seeds produced, plant vigor and the very limited, if any, number of pesticide treatment needed for crop protection, the 1999 Parlier climatic conditions were excellent for propagation of Carthamus, Crambe, Cucurbita, Lesquerella, Limnanthes, Lupinus, Lycopersicon, Raphanus, Umbelliferae and several perennial genera, and provided an exceptional good environment for development of garlic true seeds.

To identify additional accessions suitable for propagation at Parlier climatic conditions and to optimize plant propagation operation at the site, a CIMIS Weather Data Summary, a Germplasm Increase Requests Form, Tentative Dates for filing request forms and a Seed Information Sheet were distributed to all NPGS sites. The curators' interest in utilizing this site was modest; only curators who propagated material at Parlier in previous years have responded.

Literature review on economic characteristics of main genera, which are planned for transfer to Parlier as their primary germplasm conservation site, was prepared.

To conduct basic germplasm propagation operations at this site, few infrastructure units were established through reconstruction of existing, desolate farm structures. A part of an old pole barn was converted to a Seed Processing Room by building sheetrock walls, a leach drain, a drop down ceiling, connecting electric and pluming installation. A room at the farm's old homestead was renovated to set up a Tissue Culture Laboratory/Plant Growing Area. This area is used to process difficult in germination seeds and to propagate challenged germplasm accessions. Two other rooms of the homestead were adapted to a Seed Drying/Packing Room and a Screen Storage Area. The renovation and reconstruction was done by a hired handyman, students and employees of the Parlier NPGS site. All of the above infrastructure units are temporary in nature; they will be demolished when the location's Research Farm Operation Shop is approved for construction. A drip irrigation system was installed in field # 43, the site's major area for annual accession cultivation. The plan of the system was developed by a water management engineer from the USDA, WMRL group in Fresno, and carried out by the station's employees. The installed system allows for application of 80 different water capacities which is essential in cultivation of the diversified germplasm species propagated in Parlier.

Research Activities

1.   Fertility evaluation of selected Allium sativum germplasm accessions
70 Allium sativum accessions were evaluated for pollen viability and true seed development. In this number, 53 accessions developed flower stalks and 38 accessions produced true seeds. The results imply that fertility in garlic germplasm is not so rare as it was thought before, and the NPGS garlic collection contains a high number of fertile clones which maybe used by garlic industry to create genetic variability and to develop varieties propagated by seeds. Few other characteristics influencing efficiency of seed production were evaluated (e. g. easiness of bulbil removal, flowering time, male fertility, number of flowers per umbel). Several self-pollinated seed families were obtained. The material will be used in studies on inheritance pattern of selected economic traits in garlic.

2.   Multi - site garlic experiment
15 garlic clones representing major garlic types were evaluated for field and post harvest characteristics. This evaluation was a part of a study carried out in cooperation with UW in Madison, WI, USDA, ARS, WRIPS in Pullman, WA and UC in Davis, CA. Questions addressed in this experiment pertained to type/clone variability across tested environments, re-grow effects and photoperiod sensitivity of the vegetatively cultivated crop (randomized complete block design, 4 replications). This was the second evaluation year of a three-year study.

3.   Field evaluation of garlic WD germplasm collection
35 PI accessions were evaluated for morphological characteristics. The purpose of this evaluation was to compare a clone grouping based on leaf, stem and bulb characteristics with clustering pattern developed by over 200 RAPD markers analyzed previously for the same changes. Among the 35 PI accessions, no genetic or morphological duplicates were observed.

4.   Heat tolerance evaluation in beans
44 different bean varieties were evaluated for heat tolerance in the San Joaquin Valley growing conditions. In a completely randomized design (3 replications), maturity dates and yield were recorded. This experiment was carried out for Dr. Miklas from University of Washington in Prosser, WA.

5.   Selection and propagation of pea breeding material
406 G pea breeding line was selected for flower color and glossy appearance. Over 16 kg of the selected and propagated breeding material was shipped to Dr. Eigenbrode's program, University of Idaho, Moscow, ID.

The total number of different crop units evaluated or selected in the 1999 research projects was 363 and 279 new crop units were planted for CY 2000 evaluation (Table MJ4).

NPGS Parlier, CA
1999 Progress Report [status 12/31/1999]

Table MJ1a. Research crops evaluated in 1999

Genus	species	Purpose	No. of units/acc. rec.	No. of acc. establiched in field	No. of acc. harvested	Remarks
Allium	sativum	WD, DNA	35	35	35
Allium	sativum	Fertility	74	70	70
Allium	sativum	Set 1. Four Loc. Study	64	64	64	16x4
Allium	sativum	Set 2. Four Loc. Study	56	56	56	14x4
Allium	ampeloprasum	Fertility	5	5	0
Phaseolus	vulgaris	HT Evaluation, Dr. Miklas	132	132	0	Total 30,278 g
Pisum	sativum	Regen./selec.Dr.Eigenbrode	1	1	1	Total 16,279 g
TOTALS			367	363	226

Table MJ1b. Research crops planted for CY 2000 in fall 1999

Genus	Species	Purpose	No. of units planted
Allium	sativum	Set 1. Four Loc. Study	64
		Set 2. Four Loc. Study	56
		Set 3. Four Loc. Study	52
Allium	sativum	Fertility - replant	100
Allium	sp.	Fertility	7
TOTAL			279

Table MJ2. Mass of seeds of annual and perennial accessions shipped to curators