W6-MISC

A 3 year survey of the bee associates of flowering accessions of Astragalus and Onobrychis species was conducted in open-pollinated field nurseries at Central Ferry, Washington to gain insight into the potential for bee-mediated inter-accession out-crossing in these nurseries. Such nurseries are used by genebank managers at the USDA-ARS Western Regional Plant Introduction Station (WRPIS), Pullman, Washington, to multiply seed of accessions low in viability and/or supply. Fifty-two species of bees (Hymenoptera) in the Andrenidae (4 species), Apidae (18), Colletidae (1), Halictidae (5), and Megachilidae (24) were recorded. Species-rich genera were Bombus (8 species) and Osmia (16). One undescribed species of Osmia was collected. The bee fauna was more diverse on Astragalus (45 bee species) compared to Onobrychis (29 species). Among the 24 Astragalus and 13 Onobrychis species censused, bee diversity was highest on A. bungeanus Boiss. (24 bee species), A. chaborasicus Boiss. & Hausskn. (24 species), A. miser var. serotinus (A. Gray ex J.G. Cooper) Barney (14 species), A. cicer L. (13 species), A. schistosus Boiss. & Hohen. (12 species), A. onobrychis L. (11 species), A. podolobus Boiss. & Hohen. (11 species), and Onobrychis viciifolia Scop. (10 species). The presence of a rich bee fauna in WRPIS nurseries at Central Ferry, including documented pollinators of the allogamous species A. cicer and O. viciifolia, could complicate efforts to prevent hybridization and maintain genetic integrity of genebank accessions of allogamous plant species. Our results support a 1993 decision by WRPIS curators to discontinue the practice of clustering accessions of single species of Astragalus and Onobrychis in open-pollinated nurseries to regenerate seed.

Central Ferry farm picture.

An Excel spread sheet of Clement's data from Central Ferry farm in Washington state(.xls).

Images of plants grown for seed increase at the WRPIS Farm at Central Ferry, WA. Nursery established 5/9 to 5/17/2012 using seedlings started in the greenhouse. Seed were started between 3/22/2012 and 4/23/2012.

Collection pictures from the Richard C. Johnson, Harold Bockelman, Korzhenevsky, V., Boguslavsky, R. collection in 1999 from Krym, Ukraine.

Collection pictures taken in 2000 from the Kazakstan trip made by Rich Hannan, Stephanie Greene, A. Khusainov, A. Afonin, and N. Dzyubenko.

Collection pictures taken in 2001 from the Faroe Island trip made by Stoney Wright

Collection pictures taken in 2003 from the Greenland trip made by Stoney Wright

Collection pictures taken in 2003 from Norway, including Spitzbergen, trip made by Stoney Wright

Collection pictures from Fred Meuhlbauer and Walt Kaiser 2004 trip to Georgia.

For a Powerpoint view of 24 pictures of the Georgia collection click here. It is 7793 KB in size.

Collection pictures taken in 2006 from the Tajikistan trip by Barbara Hellier, Kenneth Street, Zebuniso Muminshoeva, Farkhod Kosimov, Shakhlo Safarzoda, John Sheppard, Natalya Rukhkyan and Sergey Shuvalov.

Tajikistan collection trip map.

Collection pictures from the country of Georgia in 2007 taken by Marina Mosulishvili.

Pictures taken from plants growing in the Pullman nursery in 2000

Images of seedling were taken in 2012 from seed obtained from the National Germplasm System (NPGS) and also of many other native forbs of the Inland Northwest. You can find the Forb Seedling Identication Guide here .

Ploidy level and cellular DNA content were determined by flow cytometry (DeLaat et al. 1987) using the CyStain UV Precise P or the CyStain UV Absolute P kits (Partec). For ploidy level determination, small amounts (0.5 cm2 maximum) of leaf tissue were cut into segments and placed in 0.5 ml nuclei extraction buffer for 1 min. The suspension was passed through a 50 μm CellTrics disposable filter (Partec) and stained for 2 min with 2 ml 4′,6-diamidino-2-phenylindole (DAPI) staining buffer. Samples were then excited by UV irradiation from a mercury arc lamp and analysed in the blue fluorescence channel of a flow cytometer (Partec-ROBBY). O. viciifolia accession 1127, corresponding to PI 212241 in Kidambi et al. (1990), which is known to be tetraploid, was used as a control. The cellular DNA content for O. viciifolia 1001 was estimated by DAPI staining as described above (GC content dependent quantification), and also by staining with propidium iodide after ribonuclease treatment, followed by analysis in the red fluorescence channel of the flow cytometer (GC content independent quantification). A range of plant species with known 2C values (Doležel et al. 2007) were assessed as standards for measuring the DNA content of O. viciifolia. Zea mays L. CE-777 (2C = 5.43 pg) was identified as the most appropriate of these references.

Somatic chromosome counts were carried out with conventional squashes of root tip cells. Seeds were placed between moist filter paper at 20 °C and young roots harvested when approximately 1 cm in length, usually after three or four days. The excised roots were immediately placed in 2 mM hydroxyquinoline for 4-5 h at room temperature. The roots were then fixed overnight at 5 °C in Carnoy’s solution (60 % ethanol, 30 % chloroform, 10 % glacial acetic acid). Hydrolysis was carried out in 1 M HCl at 60 °C for 8-9 min, and the roots stained for 30 min in Feulgen solution (leuco basic fuchsin). The meristematic tips were then removed, placed on a slide in a drop of 45 % acetic acid, and observed with a phase contrast microscope (Carl Zeiss Axioskop 40) equipped with a camera and image analysis software (Improvision OpenLab 4.0.2).

Papaver bracteatum grown in irrigated field at our Central Ferry farm which is in eastern Washington, United States along the Snake river, elevation about 200 meters. Seeds were planted in a greenhouse in 2007 and transplanted in the field to the field in 2007. Plants flowered, were caged, set seed and were harvested in the summer of 2008. Flower color data was taken on May 23, 2008.

Images of plants grown for seed increase at the Observatory Hill field at WSU, Pullman, WA. Nursery was established 5/25 to 5/31/2011 using seedlings started in the greenhouse.

Images of pods and/or seed taken from the original seed packet. Seed used for planting.

Images of plants grown for seed increase at the Tukey Orchard plot at WSU, Pullman, WA. Nursery was established 5/19/2010 except for the Purshia which was established in 2009. Seedlings started in the greenhouse were planted for all accessions.

A picture/image of the accession.

A picture/image of the accession.

100 seed weight at the Western Regional Plant Introduction Station, Pullman Washington, USA