Twenty Years of Germplasm Management: Geneva, NY to the Forests of Central Asia, China, Russia, and Turkey

1. Twenty Years of Germplasm Management: Geneva, NY to the Forests of Central Asia, China, Russia, and Turkey Philip Forsline April 5, 2004

3. Plant Genetic Resources Unit (PGRU) located at Cornell University, Geneva, New York PGRU facilities expanding Cornell Geneva Greenhouse & screen- house for clonal collections New office wing and greenhouses

4. History of facilities: Clonal repository at PGRU Campus Bldg 1985 Campus Bldg 1985 Equipment storage, 1984 Farm preparation, 1984 Tile drainage 1984

5. Farm development 1985 to present Clonal collection with developing plantings 1990: addition of farm for seed section of PGRU 1985 prior to planting 2002 from east to west 2002 from west to east

6. 1953 – Northeast Regional PI Station (NERPIS) established for vegetables and ornamental germplasm 1982– CSRS grant to Cornell; 50 acres purchased for Clonal Repository (CR) 1983 – CR Farm Manager hired; 1st propagations of Malus and Vitis 1984– Curator hired; campus building started 1986 – CR building occupancy and dedication; 1st orchard and vineyard plantings; merger with NERPIS 1987 – S. Kresovich hired as 1st Research Leader for NERPIS and CR – new name: Plant Genetic Resources Unit (PGRU) 1988 – J. McFerson hired as curator of seed group; Cryopreservation project began; 1st of seven germplasm expeditions initiated 1989 – 40 acres purchased for seed group 1993– J. McFerson appointed RL for PGRU; PGRU scientists granted ‘Courtesy/Adjunct’ appointments with H.S. at NYSAES 1995– Sour cherry collection established at PGRU 1996 – S. Hokanson hired as post doc 1998– W. Lamboy appointed RL; USDA apple rootstock breeder hired at PGRU; L. Robertson hired as curator of seed group 1999 – Grape genetics group of PGRU initiated with hiring of grape rootstock breeder; Molecular geneticist hired for seed group 2002 – Grape scion breeder hired 2003– C. Simon hired as RL after serving 5 years as RL at Davis, CA Clonal Repository; Grape pathologist and computational biologist hired; 2004– Grape genomics specialist hired History ofsignificantevents and personnel additions at PGRU, Geneva, NY

7. Vegetatively-propagated crops at PGRU • Apple 3909 accessions • Grape 1175 accessions • Sour Cherry87 accessions • Total 5171 accessions

8. Major Seed-Propagated Crops Conserved at GenevaLarry Robertson: Curator/GeneticistJoanne Labate: Molecular Biologist

9. USDA grapevine genetics -- Geneva • 1999 rootstock breeder/geneticist (Peter Cousins) • 2002 molecular genetics/genomics of scion traits (Chris Owens) • 2003 molecular plant-microbe interactions of fungal pathogens (Lance Cadle-Davidson) • 2003 Computational biologist – also working across Clonal and Seed projects of PGRU (Angela Baldo) • 2004 evolutionary genomics/population genetics (Amanda Garris)

10. The Geneva® Apple Rootstock Breeding ProgramGennaro Fazio, USDA/ARS, PGRU in cooperation with Cornell University

11. SYsFTE - P. Forsline 1.0 Horticulturist/Curator - C. Simon 0.85 Research Geneticist / RL - H. Schwaninger 1.0 Molec. Biol. / Support Sci. TechnicalFTE - W. Srmack 1.0 Collections Manager / Supervisor - D. Beckhorn 1.0 Field Assistant III - R. Vaughan 1.0 Biol. Sci. Technician (field) - N. Lepak 1.0 Agri. Res. Sci. Technician (field) - Vacant 1.0 Biol. Sci. Technician (molecular) Staff for clonal collections at PGRU • Database - R. Nearpass 0.40 Supervisory IT specialist - D. Dellefave 1.0 Germplasm Program Assistant

12. Current CRIS Project:Conservation and Utilization of the Genetic Resources of Apples, Grapes, and Tart Cherries • Objective 1: Acquireandmaintaingenetic resources and associated information of apples, cold-hardy grapes, and tart cherries and refine the collection based on morphological and molecular characterization and distributegermplasm to the user community. • Objective 2:Enhance efficiency and effectiveness of germplasm maintenance by applying genomic sequencing and molecular marker techniques to genetically characterize germplasm and determine phylogenies. • Objective 3:Enhance value and utilization of genetic resources of apples, cold-hardy grapes, and tart cherries by systematiccharacterization and evaluationfor important morphological and horticultural traits.

13. Acquisition and Maintenance of Clonal Collection

14. Vitis collection at Plant Genetic Resources UnitGeneva, NY, U.S.A. • 1045 clones - all maintained as duplicate field plantings - 650 hybrid; 395 (23 Vitis sp) - cryogenic storage (?) - 138 are part of a core collection • 130 accessions of wild Vitis sp (seed lots and/or seedling populations) - 420 seedlings under evaluation from 30 of these acc. • A total of 1175 accessions at PGRU* * Remainder of Vitis collection at Davis, CA - 814 hybrids; 1838 (45 Vitis sp) * 3827 accessions of Vitis in NPGS

15. Vitis collection at PGRU Early / Late leaf fall Vineyard (1045 clones) Diversity Among clusters Diversity among clones

16. Digital imaging of Vitis clones PI 588058 ‘V. coignetiae’ PI 181481 ‘Vignoles’ PI 588054 ‘V. riparia’’ PI 588070 ‘Catawba’

17. Prunus (tetraploid cherry) collection at Plant Genetic Resources UnitGeneva, NY, U.S.A. • 87 clones - all maintained as duplicate field plantings - 57 Prunus cerasus (sour cherry) - 12 P. fruticosa (wild progenitor of sour cherry) - 7 interspecific hybrids - 12 other (misc. Prunus sp) - 52* backed up in cryogenic storage at NCGRP * 8 also in cryogenic storage on-site at PGRU

18. Cherry collection at PGRU P. cerasus / P. fruticosa Collection (87 acc.) spreading Early / late bloom upright

19. Cherry digital imaging Montmorency Balaton Danube Schatten Morello

20. Malus collection at Plant Genetic Resources UnitGeneva, NY, U.S.A. • 2376 clones - all maintained as duplicate field plantings - 1362 M.x domestica; 329 hybrid; 685 (54 M. sp) - 2146 backed up in cryogenic storage at NCGRP, Ft Collins, CO * 436 also in cryogenic storage on-site at PGRU - 195 are part of a core collection (multi-state plantings) * 60 new additions to core in 2003 • 1533 accessions of wild M. spp. (stored as seed) from World centers of origin - 3300 seedlings under evaluation from 340 of these acc. • 897 of wild acc. are Malus sieversii from Central Asia • A total of 3909 accessions

21. Diversity in Malus Fruit Bloom Late leaf fall Early leaf fall Bloom M. baccata Dormant “Kansas 14”

22. Malus / seedling collection at PGRU Early stage after planting 1986 More advanced stage Apples/seedling vineyard Tree removal after repropagation on EMLA 7

23. Malus / EMLA 9 collection at PGRU Initial plantings after establishment in 1986 Established plantings summer Established plantings at bloom Tree removal for conversion to EMLA 7

24. Digital imaging of Malus clones Gala Belle de Boskoop Geneva Malus hupehensis

25. Malus core collection at PGRU Label system M. transitoria Planting in 2000 Core collection in bloom: 2001 Apples/seedling clean infected infected Extension orchard 02 Apples/EMLA7 Core collection fruiting: 2001 Success w/ core collection in discovering new virus indicator

26. Malus / EMLA 7 collection at PGRU Planting in 2000 Establishment 2001 Apples/seedling Extension orchard, 2002 Apples/EMLA 7

27. Fire blight challenges and control in the Malus collection at PGRU Heavy FB on blossoms; 2000 Severe FB epidemic; 1996 425 trees removed No ‘Apogee’ treatment ‘Apogee’ treatment ‘Apogee’

28. Codes for fire blight, shootY (natural) of apple Code Definition No. ofAccessions 1 Very resistant - no occurrence 596 2 Moderately resistant -only light rating 127 3 Intermediate - light to medium rating 174 4 Moderately susceptible - medium to heavy rating 363 5 Very susceptible - very heavy rating 1091 2351 YCumulative records 1990 – 2003 for entire ‘clonal collection’

29. Measures being used to minimize fire blight in the PGRU apple collection • Cryogenic storage of dormant buds; as a result, ~ 90 accessions that have died (fire blight) have been rescued by direct grafting of cryopreserved buds • Repropagation of collection to EMLA 7 rootstock: Replacing collection previously held on seedling and EMLA 9 rootstock • Application of ‘Apogee’ post-bloom to control vegetative growth reducing shoot blight • Removal of all lesions in dormant season; timely annual copper spray; and corrective pruning of infected shoots throughout the growing season • Continued use of antibiotics under defined protocols to minimize blossom blight

30. Cryopreservation of MalusA back up collection at $1/accession/yr! Base collection (2146 acc.) at NCGRP, Ft. Collins, CO Processing 35 mm bud segments Hundreds of seedlings budded with cryopreserved buds testing viability Recovery of accessions by bud grafting Active collection (436 acc.) at PGRU Geneva, NY

31. Cryopreservation of Prunus Base collection (52 acc.) at NCGRP, Ft. Collins, CO Injury to primary bud following LN exposure Active collection (8 acc.) at PGRU Geneva, NY Typical recovery scenario from axillary bud

32. Pilot Project: Forsline, P.L., C. Stushnoff, L.E. Towill, J.W. Waddell, W.F. Lamboy and J. R. McFerson. 1998. Recovery and longevity of cryopreserved apple buds. J. Amer. Soc. Hort. Sci. 123:365-370. 84 accessions were processed periodically over 4-yr period • 12/12/1988 – 8 accessions • 02/07/1989 – 8 accessions • 12/12/1989 – 6 accessions • 01/19/1990 – 9 accessions • 12/14/1990 – 12 accessions • 01/15/1991– 13 accessions • 12/18/1991– 13 accessions • 01/18/1992– 15 accessions

33. Pilot project to determine protocol for cryogenic storage of dormant buds: % bud recovery of 84 apple accessions (processed 1989-1992); tested after up to 8 years of storage in liquid nitrogen Treatment Recovery % Desiccated Control 85.3 aZ Storage one month 63.0 b Storage one year 64.2 b Storage two years 66.5 b Storage four years 68.6 b Storage after eightYyears 68.3 b ZSeparation of grand means of 84 accessions at P< 0.01 by test for differences between two proportions (LSD = 7.2) Y Fifteen year testin process 2004 to 2007

34. Annual cryopreservationof Malusaccessions at NCGRP following the 4-yr pilot project 1988-1992

35. Successful cryopresevation for > 90% of accessions stored at NCGRP: those with < 30% viability will be reprocessed 1760 or 91% were successful 171 or 9% of total were unsuccessful No. of accessions in storage 0-10 20-30 40-50 60-70 80-90 100

36. Successful cryopresevation for > 95% of accessions stored at PGRU: those with < 30% viability will be reprocessed 418 or 96% were successful 18 or 4% of total were unsuccessful No. of accessions in storage 0-10 20-30 40-50 60-70 80-90 100

37. Distribution of Germplasm

38. Annual distribution of germplasm from 1988 to 2004 Z mean = 2418 No. of accessions annually Z mean = 692 Apple total no. = 41,111 Apple total orders = 1514 (mean:89) Apple orders/yr (range: 30-163Z) Grape total no. = 11,765 Grape total orders = 782 (mean:46) Grape orders/yr (range = 14-72Z ) Cherry distributions started in 1998: 191 accessions to 21 orders

39. Characterization and Evaluation

40. Characterization of collections at Plant Genetic Resources Unit, Geneva, NY • Malus Clones - 1150 acc. with 33 descriptors - 900 digital images • Malus species characterization at origin site - 1088 with 25 descriptors plus other passport records • Malus species grow-out - 465 seedlings with 33 descriptors - 375 digital images • Vitis clones - 850 acc. with 16 descriptors - 414 digital images • Prunus clones - 50 acc. with 8 descriptors & 50 digital images

41. NPGS-funded projects in Malus evaluation Preservation of alleles from wild collections: Volk, Walters, Richards Antioxidants in Malus collection: Stushnoff Evaluation of wild apple species for disease resistance: Aldwinckle Evaluation of elite M. sieversii and some of its hybrids for apple scab resistance genes: Aldwinckle, Luby, Gardiner and Bus Molecular characterization of seedling populations of 10 Malus species with 90 mapped microsatellite markers in relation to apple rootstocks: Fazio, Baldo NPGS-funded projects in Vitis and Prunus evaluation Evaluation of Vitis for susceptibility to crown gall: Burr Evaluation of Vitis for resistance to Phomopsis viticola and powdery mildew: Wilcox and Reisch Evaluation of Russian tetraploid cherry selections for cherry leaf spot resistance: Iezzoni Evaluation and rescue of sour cherry germplasm for use as sweet cherry rootstocks: Iezzoni Collaborative evaluation with Specific Cooperative Agreements (SCA) of clonal collections in process or recently completed

42. Molecular studies on clonal germplasm at PGRU: past & present • Apple core collection – Hokanson • Wild apple germplasm – Benson and Dickson (PhD projects) • Grape genotyping – Lamboy • Grape phylogeny – Schwaninger • Overall coordination presently – Simon (Research leader / molecular geneticist)

43. New Acquisitions

45. The origin of the cultivated apple Ancient Malus species of China: bird disseminated to Central Asia M. sieversii of Central Asia Dr. B. Juniper: Theory on early and recent evolution of the cultivated apple Mammal disseminated North America became a secondary center of origin: ‘Red Delicious’, ‘Golden Delicious’, etc. Johnny Appleseed

46. Germplasm collections to add mostly wild Malus species to the PGRU collection

48. Collection team for 1989 Central Asian expedition to Uzbekistan, Tajikistan and Kazakhstan (site 3) ‘The late’ Calvin Sperling Herb Aldwinckle Elizabeth Dickson Uzbekistan

49. Professor Djangaliev Geneva: making plans, 1992 Organizing expedition: first morning, Sept. 1993; ready to leave to board helicopter Collection admiration, 1995 One of frequent lectures; 1996 Local Kazak media highlighting American & S. African invasion 1995

50. 1993 collection team: USA & New Zealand Mink Noiton Dickson Transfer in Moscow – hotel and visit to Red Square Preparation for departure Significant travel by helicopter Moscow to Alma Ata via ‘Aeroflot’ Team in Alma Ata with Kazakh hosts