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

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

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

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

History of significant events and personnel additions at PGRU, Geneva, NY  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; 1 st propagations of Malus and Vitis  1984 – Curator hired; campus building started  1986 – CR building occupancy and dedication; 1 st orchard and vineyard plantings; merger with NERPIS  1987 – S. Kresovich hired as 1 st 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; 1 st 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

Vegetatively-propagated crops at PGRU  Apple 3909 accessions  Grape1175 accessions  Sour Cherry 87 accessions  Total5171 accessions

Major Seed-Propagated Crops Conserved at Geneva Larry Robertson: Curator/Geneticist Joanne Labate: Molecular Biologist

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)

The Geneva® Apple Rootstock Breeding Program Gennaro Fazio, USDA/ARS, PGRU in cooperation with Cornell University

Staff for clonal collections at PGRU  SYs FTE - P. Forsline1.0 Horticulturist/Curator - C. Simon0.85 Research Geneticist / RL - H. Schwaninger 1.0 Molec. Biol. / Support Sci.  Technical FTE - 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)  Database - R. Nearpass 0.40 Supervisory IT specialist - D. Dellefave 1.0 Germplasm Program Assistant

Current CRIS Project: Conservation and Utilization of the Genetic Resources of Apples, Grapes, and Tart Cherries  Objective 1: Acquire and maintain genetic resources and associated information of apples, cold-hardy grapes, and tart cherries and refine the collection based on morphological and molecular characterization and distribute germplasm 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 systematic characterization and evaluation for important morphological and horticultural traits.

Acquisition and Maintenance of Clonal Collection

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

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

Digital imaging of Vitis clones PI ‘Vignoles’ PI ‘Catawba’ PI ‘V. coignetiae’ PI ‘V. riparia’’

Prunus (tetraploid cherry) collection at Plant Genetic Resources Unit Geneva, 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

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

Cherry digital imaging MontmorencyBalaton DanubeSchatten Morello

Malus collection at Plant Genetic Resources Unit Geneva, NY, U.S.A.  2376 clones - all maintained as duplicate field plantings M.x domestica; 329 hybrid; 685 (54 M. sp) backed up in cryogenic storage at NCGRP, Ft Collins, CO * 436 also in cryogenic storage on-site at PGRU 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 seedlings under evaluation from 340 of these acc.  897 of wild acc. are Malus sieversii from Central Asia  A total of 3909 accessions

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

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

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

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

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

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

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

Codes for fire blight, shoot Y (natural) of apple Code DefinitionNo. of Accessions 1 Very resistant- no occurrence596 2 Moderately resistant - only light rating127 3 Intermediate - light to medium rating174 4 Moderately susceptible - medium to heavy rating363 5 Very susceptible - very heavy rating Y Cumulative records 1990 – 2003 for entire ‘clonal collection’

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

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

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

Pilot Project: Forsline, P.L., C. Stushnoff, L.E. Towill, J.W. Waddell, W.F. Lamboy and J. R. McFerson Recovery and longevity of cryopreserved apple buds. J. Amer. Soc. Hort. Sci. 123: 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

Pilot project to determine protocol for cryogenic storage of dormant buds: % bud recovery of 84 apple accessions (processed ); tested after up to 8 years of storage in liquid nitrogen Treatment Recovery % Desiccated Control85.3 a Z Storage one month63.0 b Storage one year64.2 b Storage two years66.5 b Storage four years68.6 b Storage after eight Y years68.3 b Z Separation 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

Annual cryopreservation of Malus accessions at NCGRP following the 4-yr pilot project YearM. x domesticaHybridsOther speciesTotal Total

Successful cryopresevation for > 90% of accessions stored at NCGRP: those with < 30% viability will be reprocessed No. of accessions in storage 171 or 9% of total were unsuccessful 1760 or 91% were successful

Successful cryopresevation for > 95% of accessions stored at PGRU: those with < 30% viability will be reprocessed No. of accessions in storage 18 or 4% of total were unsuccessful 418 or 96% were successful

Distribution of Germplasm

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

Characterization and Evaluation

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

Collaborative evaluation with Specific Cooperative Agreements (SCA) of clonal collections in process or recently completed  NPGS-funded projects in Malus evaluation 1)Preservation of alleles from wild collections: Volk, Walters, Richards 2)Antioxidants in Malus collection: Stushnoff 3)Evaluation of wild apple species for disease resistance: Aldwinckle 4)Evaluation of elite M. sieversii and some of its hybrids for apple scab resistance genes: Aldwinckle, Luby, Gardiner and Bus 5)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 1)Evaluation of Vitis for susceptibility to crown gall: Burr 2)Evaluation of Vitis for resistance to Phomopsis viticola and powdery mildew: Wilcox and Reisch 3)Evaluation of Russian tetraploid cherry selections for cherry leaf spot resistance: Iezzoni 4)Evaluation and rescue of sour cherry germplasm for use as sweet cherry rootstocks: Iezzoni

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)

New Acquisitions

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

Germplasm collections to add mostly wild Malus species to the PGRU collection YearCountrySpeciesPersonnel 1987 – 1988 Western U.S. & Canada Eastern U.S. & Canada Malus fusca, M. ioensis, M. coronaria & M. angustifolia Weeden, Dickson 1989Uzbekistan, Tajikistan, Kazakhstan Malus sieversiiAldwinckle, Dickson, Sperling 1993Kazakhstan, Kyrgyzstan Malus sieversii, Vitis vinifera, misc. Forsline, Dickson, Mink, New Zealand scientist 1995KazakhstanMalus sieversii, Vitis vinifera, misc. Forsline, Dickson, Luby, S. African scientists 1996KazakhstanMalus sieversii, Vitis vinifera, misc. Forsline, Hokanson, Unruh, Pellett 1997China (Sichuan)Malus species (7 endemic to Sichuan) Forsline, Aldwinckle. Benson 1998RussiaMalus orientalis, Prunus cerasus and misc Prunus spp. Forsline, Iezzoni, Karle, German scientist 1999TurkeyMalus orientalis, selected local Malus cultivars Forsline, Aldwinckle

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

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

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

1995 USA and S. African collection team Dickson Britz Forsline Luby Human Arrival at site 9 Meals in yurt: site 5 Housing in sanitarium at HQ in Almaty Gala dinner with hosts in Almaty: return to USA Seed extraction at hotel: site 9

1996 USA collection team Hokanson Unruh Forsline Pellett Site 5 Site 9 Site 12 HQ in Almaty

SITE INFORMATION RECORDED  Latitude (GPS)  Longitude (GPS)  Elevation  Slope  Aspect  Light/Shade  Assoc. Tree spp.  Assoc. Shrub spp.  Assoc. Herb. spp.  Population Abun./Dis.  Climate  Soil

Site descriptions in Central Asia Country/RegionSite Lat o N / Long o EElevation ( m )Rainfall ( mm ) Tajikistan / / Uzbekistan/ / Kazakstan/Zailisky 3 43 / Kazakstan/Djungarsky 4 45 / / Kazakstan/Karatau 6 43 / / Kyrgyzstan/ / Kazakstan/Tarbagatai 9 48 / Kazakstan/ Ketmen10 44 / Kazakstan/Talasky12 42 /

Kazakhstan site 3: ’89, ’93, ’95, ‘96 Fruit from 30 randomly-collected trees Depleted by dachas Habitat 1940 Habitat present Studies by Professor A. Djangaliev

Kazakhstan site 4, ’93, ’95, ‘96 Scab infected tree

Kazakhstan site 5: ’93, ’95, ‘96 Forestry camp at 1200 m headquarters in ’93, ’95 & ‘ yr old M. sieversii Bear scat w/ many apple seeds Apple forest: 1800 m 1100 m First collection morning (Sept. 1993) starting out at 600 m with hike to 1800 m

Kazakhstan site 6: ’93 & ’95 Camp at 600 m: headquarters Standing at 900 m M. sieversii at 900 m Fruit from 30 randomly-collected trees at 900 m

Kyrgyzstan site 7: 1993 Apple and walnut forests as seen from helicopter Heavy grazing in M. sieversii areas Village in Kyrgyzstan near collection sites

Kazakhstan site 9: ’95 & ‘96 Grazed areas – M. sieversii on slopes Diverse, elite M. sieversii Super-elite M. sieversii Heavy grazing of habitatFruit from 30 randomly-collected M. sieversii trees M. sieversii

Kazakhstan site 10: 1996 Fruit from 20 randomly-collected trees Breakdowns; common occurrence Uygur tribes tradition of bread baking M. niedzwetzkyana

Kazakhstan site 11: 1996 Elite ‘Yellow transparent’-type M. sieversii

Kazakhstan site 12: 1996 Canyon 400 m deep w/ M. sieversii on the N-facing wall Fruit from 10 randomly- collected M. sieversii trees Bottom of canyon; M. sieversii on this side Trail to bottom

ACCESSION INFORMATION RECORDED + Fruit Over Color/Intensity+ Fruit Size + Fruit Ground Color+ Fruit Texture + Fruit Russet+ Fruit Flavor + Fruit Shape+ Harvest Season + Stem Character + Tree Habit + Diseases/Insects:Leaves/Fruit

Variability of fruit size of Malus sieversii among sites in Central Asia No. collected Mean size ( mm ) Size range ( mm ) Site / Yr(s) Elite Random Elite Random Elite Random 12 / (‘96) * 5 / (‘96) / (‘95/’96) / (‘96) / (‘96) * 5 / (‘95) / (‘95/’96) / (‘95/’96) / (‘95) Totals * Note difference in fruit size observed in 1995 and 1996 at Site 5

Summary of Central Asian M. sieversii collection, distribution and storage Accessions Seeds Seeds Storage Storage Group Obtained ObtainedDistributed at PGRU at NSSL 1989 and , ,900 10,900 Collections 1995 and z 67,00016,000 29,200 21,800 Elites 1995 and , , x Random populations Totals ,00028,300 Y 62,200 39,500 z 44 of best accessions also obtained as clones Y Distributed to 24 evaluators x Stored as 19 bulked populations (each population includes seeds from 10 to 60 trees with an average of 30 trees / population)

Seed storage at C

Evaluation of New Material

PGRU / Cornell cooperative evaluation of Malus sieversii Lab Evaluation Type No. of Seedlings H. AldwinckleDisease resistance 5124 Z PGRUHorticultural / Molecular part of P. Forsline - S. Hokanson - W. Lamboy - G. Fazio - L. Benson N. WeedenIsozyme / Molecular S. Mehlenbacher “ / “ W. ReissigInsect resistance Aldwinckle / KoreaRosellinia / Helicobasidium 400 S. BrownGenetic dwarf 250 I. MerwinNematode resistance200 Z 2108 of this group of screened seedlings were sent to OH, NJ, WA, MN and WI for further evaluation

PGRU / SAES cooperative evaluation of M. sieversii State / Lab Evaluation type No. of seedlings MN / LubyHort, Disease, Cold H.1498 Y NJ / GoffredaHort, Disease, Molecular, Elite clones1899 Y CO / StushnoffHort, Cold H., Antioxidants 720 WA / BarrittHort, Sunburn, Cold H. 624 Y OH Z / Lynd, MillerHort, Disease, Late Bloom, Elite clones 950 Y WI / SmithHort, Disease, Cold H. 655 Y AR / RomHort, Disease, Late Bloom~400 AK / McBeathHort, Disease, Cold H. 346 IL / KorbanHort, Disease, Molecular ~200 Z Seedlings planted at Dawes Arboretum, Newark, OH; c/o D. Miller Y Some seedlings of these sites screened for apple scab, fire blight and cedar-apple rust at Cornell, Geneva by Dr. Aldwinckle

PGRU / International cooperative evaluation of Malus sieversii No. of Country / Lab Evaluation typeseedlings New Zealand / Bus, Noiton, GardinerDisease, Hort4426 Germany / Buttner, Geibel, Höfer Disease, Hort 1367 Norway / Roen Disease, Hort 692 N.S.,Canada / Deslauriers, EmbreeDisease, Hort1155 Man., Canada / Davidson Disease, Hort, Cold H. 169 B.C., Canada / Quamme, HampsonDisease, Hort 325 N.B., Canada / HunterHort ~120 Japan / BesshoDisease, Hort ~100 UK, Reading / FarrelMolecular ~300 UK, Oxford / JuniperMolecular ~150 S. Africa / Human, BritzDis., Insect, Sunburn, Chilling ? Z Netherlands / KempElite clones-- Italy / SansiviniElite clones-- Z Parallel collections were made in 1995 expedition in Kazakhstan

M. sieversii seedling grow outs in Germany and New Zealand Dresden, Germany Vincent Bus; Havelock, North, New Zealand

Disease Resistance Screening

Screening young M. sieversii seedlings for apple scab (Venturia inaequalis)

Codes used in rating apple scab resistance H. S. Alwinckle and H. L. Gustafson  1 = Pits, small pin-prick marks (similar to V m gene) - RESISTANT  2 = Chlorotic lesions (similar to V f gene) - RESISTANT  3 = Necrotic lesions ( brown) - RESISTANT  4 = Sporulation - SUSCEPTIBLE  5 = Nonsporulating - RESISTANT  6 = Abaxial sporulation - SUSCEPTIBLE  7 = No symptoms - RESISTANT  8 = Cupped or convoluted (similar to V f gene) - RESISTANT  9 = Stellate (star shaped) necrotic (similar to V r gene) - RESISTANT  A = Usually has some cupping and or chlorosis but may have little or no symptoms [(Characteristic of V f gene) ( numbers: 2,5,7,& 8)] - RESISTANT  B = Stellate necrotic [(Characteristic of V r gene) (number: 9)] - RESISTANT  Other resistant reactions - LN = Large necrotic lesions - RESISTANT - N = Necrotic lesions - RESISTANT - P = Pits ( number 1 ) - RESISTANT

Apple scab resistance of Malus sieversii populations from Central Asia H. Aldwinckle, P. Forsline, H. Gustafson and S. Hokanson SiteNo. Seedlings Inoculated% Resistant Totals

Wild Malus sieversii seedling plantings Early stage after planting 1986 More advanced stage 1993 collection 1995/96 collections (1200 sdgs) M. sieversii in bloom 1989 collection M. niedzwetzkyana form of M. sieversii M. sieversii fruiting

Summary of scab resistance in grow-out of 1188 Malus sieversii seedlings in Geneva, New York

Cedar apple rust resistance in M. sieversii seedlings from 8 different sites in Kazakhstan Total % of population resistant

Natural incidence of fire blight on shoots of the Malus sieversii grow-out in Geneva, New York Fire blight

Natural occurrence of fire blight in M. sieversii seedlings from 7 different sites in Kazakhstan Fire Blight Severity % of trees in each category Codes 1 & 2 Codes 4 &

Horticultural Evaluations

Descriptors used to characterize M. sieversii at:  Date collected / Harvest season  Fruit bloom  Fruit flesh color  Fruit flesh firmness  Fruit flesh texture  Fruit flesh flavor  Fruit flesh oxidation  Fruit skin overcolor  Overcolor pattern  Overcolor intensity  Fruit ground color  Fruit juiciness (sp. gravity)  Fruit size (LxW in mm)  Fruit weight (g)  Fruit russet type  Fruit russet location  Fruit russet intensity  Fruit shape  Fruit shape uniformity  Fruit size uniformity  Fruit-top shape  Stem cavity  Stem thickness  Stem length  Calyx basin  Fruit tenacity to premature abscission  Soluble solids  Bloom date  Budbreak date

Wild Malus sieversii clones and seedlings in grow-out plantings More advanced stage vineyard PI clone in bloom PI clones fruiting ‘Empire’ ‘Gala’ 13 different M. sieversii clones ‘Empire’ ‘Gala’ Fruit from half-sib seedlings of some of the M. sieversii clones PI &

Fruit size, flavor and red color of M. sieversii on-site in Kazakhstan, and as grow-outs in yrs 6 to 8 (2001 to 2003) in Geneva, New York SiteCategoryNo. of trees Mean diam ( mm / fruit ) Diam range (mm / fruit) Mean weight (g / fruit) Weight range (g / fruit) Accept -able flavor (%) % of trees w/ red fruit Red intensity mean (%) 9 Trees in Kazakhstan NA Progeny grow-out Trees in Kazakhstan NA Progeny grow-out Elites in Kazakhstan NA Elite grow-out

Harvest season and soluble solids of M. sieversii fruit from grow-outs in yrs ( ) in Geneva, New York: in addition the no. of trees to characterize in 2004 (yr 8 or 7) Site(s)CategoryNo. of trees completed Harvest season range Soluble solid range No. of trees to test in Elite clones1110 Aug-13 Sep Seedlings1598 Aug-17 Sep Seedlings1388 Aug-3 Nov Seedlings388 Aug-20 Sep Seedlings378 Aug-30 Sep Seedlings298 Aug-15 Oct Seedlings514 Aug-10 Sep Seedlings714 Aug-17 Sep Seedlings208 Aug-23 Sep ,5,6,11,12Elite clones522 Aug-10 Oct

Fruit shape in M. sieversii seedling populations from 6 sites in Kazakhstan % of seedlings in each category

Digital imaging of Malus sieversii seedlings From site 5 in Kazakhstan From site 6 in Kazakhstan From site 9 in KazakhstanFrom site 11 in Kazakhstan

Digital imaging of Malus sieversii clones From site 5 in Kazakhstan From site 6 in Kazakhstan From site 12 in Kazakhstan From site 9 in Kazakhstan

Elite clone (PI ) and progeny (3 half-sibs) Clone Seedling.a Seedling.b Seedling.h

Elite clone (PI ) and progeny (3 half-sibs) Clone Seedling.a Seedling.k Seedling.n

Seed bulk-up from flowering M. sieversii seedlings to save additional genetic diversity  In spring 2004 we will make controlled pollinations on seedlings from sites 6 and 9 in order to bulk up seeds for long term storage to supplement original seed collected in 1995  In spring 2005 the same project will be accomplished with seedlings from sites 4, 5, 10, 11, and 12

Other Recent Collections

1997 Sichuan collection team Geneva, 11/96: Prof. Li; plans Hotel in Chengdu; Prof. Li & wife; H & B Aldwinckle Crew collecting minutes before monkey thievery Crew at 3500 m pass in mountains Post expedition conference with Yunnan and Guizhou researchers in Chonqing Laura Benson monkey

Malus sp. collections in Sichuan, China M. hupehensis M. toringoides M. prattii Malus kansuensis

1997 collections of Malus spp. in Sichuan, China Phil Forsline, Laura Benson and Herb Aldwinckle Species Site(s) No. of accessions No. of seeds Malus prattii01, 05, Malus transitoria Malus hupehensis Z 03, 04, Malus toringoides03, Malus kansuensis Malus sieboldii Z Malus zhaojiaoensis Total Z Likely to be apomictic

Chinese Malus spp. screened for fire blight in greenhouse and field: replanted for horticultural evaluation Greenhouse fire blight screen Temporary field location for grad student L. Benson and field f.b. screen Digging sdgs following grad student project and f.b. screen ‘Permanent’ field location for Horticultural evaluation Many of these sdgs distributed to 10 arboreta for preservation

Screening 7 Malus species from Sichuan, China for fire blight, apple scab and cedar apple rust These seedlings were also screened for:  Apple scab – 385 seedlings were screened and 98% of them were resistant  M. hupehensis (03&04), M. toringoides (04), M. kansuensis, M. zhaojioensis, and M. sieboldii were mostly ‘A-type’ resistance  M. hupehensis (07), M. transitoria, M. toringoides (03) and M. yunnanensis were a mix of ‘A-type and P- type’ resistance  M. sieboldii (91% resistant) was the only species with some susceptible seedlings  Cedar apple rust – 370 sdgs were screened and 93% of them were resistant.  M. sieboldii (55% resistant) was the only species with some susceptible seedlings % of seedlings resistant

Variability of fire blight resistance in M. hupehensis and M. toringoides collected from different sites in Sichuan, China % of sdgs fire blight resistant M. hupehensis M. toringoides

Apple scab resistance in 5 Malus species collected in Sichuan, China in 2002 by M. Geibel Fire blight and cedar apple rust screen is in process % of seedlings scab resistant hupehensis kansuensis sieboldii toringoides transitoria

Malus orientalis in Russian Caucasus Vavilov Inst. St. Petersburg

Malus orientalis screening for apple scab, cedar apple rust, and fire blight Seedlings germinated Seedlings screened for apple scab Seedlings screened for fire blight Seedlings in high density orchard for horticultural evaluation

Malus orientalis in Turkey Local type ‘Seker Elmasii’ Sugar apple Q Forsline, Aldwinckle & 6 Turks

Screening populations of M. orientalis from Russian Caucsus, Turkey and Armenia for fire blight, apple scab and cedar apple rust A, B, P-type; 3:1:2 ratio A/B 2:1 Mostly A-type % of seedlings resistant No. of sdgs screened Russia Turkey Armenia Scab F.B C.A.R TBD 27 populations62 populations7 populations

Screening 5 populations of European wild crabapple (M. sylvestris) received from gene bank in Dresden, Germany % of seedlings resistant

Expedition to Russia to exchange sour cherry and cherry rootstock for crop improvement, July 10-30, 1998 A. Iezzoni, R. Karle, P. Forsline and M. Fischer Breakdown OrelMichurinsk Krymsk Prunus nursery G. & V. Eremin; M. Fischer Cherry accession in Orel A. Iezzoni Cherries in St. Petersburg Prof. Yushev M. Fischer R. Karle

Expedition to Russia to Exchange sour cherry, cherry rootstock, and Malus germplasm for crop improvement July 10-30, 1998 A. Iezzoni, R. Karle, P. Forsline and M. Fischer  Goals of expedition 1)tart cherry elite germplasm at two sites (Orel, and Michurinsk) 2)broad spectrum Prunus spp (Krymsk) 3)Germplasm in 1 & 2 with resistance to ‘cherry leaf spot’ (Blumeriella jaapii) and ‘twig brown rot’ (Monilinia laxa) 4)wild apple collections (Maikop)  Trip participants 1)Amy Iezzoni and Renate Karle, Michigan State University 2)Philip Forsline, USDA-ARS Plant Genetic Resources Unit 3)Manfred Fischer, Genebank OBST, Dresden-Pillnitz, Germany  Material collected 1)Orel – 25 accessions 2)Michurinsk – 12 accessions 3)Krymsk – 24 accessions 4)Maikop area – 28 seed lots (6500 seeds) of wild M. orientalis

Vitis collections in Kazakhstan in 1993, 1995 and 1996 in sites 6 and 11 Accessions collected 1993 – 17 populations: 8324 seeds and 10 local cultivars (cuttings) 1995 – 33 populations: 5723 seeds 1996 – 1 population: 318 seeds

Germplasm Utilization and Enhancement

Presence / absence of RAPD markers for 4 scab- resistance genes in elite clones of Malus sieversii S. Mehlenbacher and N. Weeden Scab-resistance Genes / RAPD Markers V r V m V f V b Accession No. P415B UBC562 OPB12 CS5 UBC220 GMAL 4326YESYES NO NO GMAL 4327*YESYESYESYES GMAL 4331*YESYES NO NO GMAL 4333*YES NO NO NO GMAL 4334*YESYES NO NO * These have been used as pollen parents X ‘Gala” (see next slide)

Germplasm enhancement to study genetics of apple scab resistance 1.Gala X M. sieversii (GMAL 4335*) – 67% of 230 sdgs resistant 2.Gala X M. sieversii (GMAL 4448*) – 57% of 209 sdgs resistant 3.Gala X M. sieversii (GMAL 4455*) – 52% of 209 sdgs resistant 4.Gala X M. sieversii (GMAL 4331 Z ) – 29% of 90 sdgs resistant 5.Gala X M. sieversii (GMAL 4334 Z ) – 24% of 206 sdgs resistant 6.Gala X M. sieversii (GMAL 4333 Z ) – 11% of 136 sdgs resistant 7.Gala X M. sieversii (GMAL 4327 Z ) – 9% of 209 sdgs resistant Total – 38% of 1289 sdgs resistant *These clones are scab resistant; A-type vs. B-type resistance of sdgs from these crosses in ~ 1:1 ratio Z >50% of progeny from these clones are scab resistant: clones themselves are not; Sdgs from crosses are all A-type resistance

Selected publications (1 & 2) and media (3 & 4): The origin of apples Horticultural Reviews, vol “Wild Apple and Fruit Trees of Central Asia”. Wiley, New York. J. Janick, P. Forsline, E. Dickson, R. Way and M. Thompson (eds.).  Chapter 1 – “Collection, Maintenance, Characterization and Utilization of Wild Apples of Central Asia”, p P.L. Forsline, H.S. Aldwinckle, E.E. Dickson, J. J. Luby, and S.C. Hokanson  Chapter 2 - Translation from Russian: “The Wild Apple Tree of Kazakhstan”, p A.D. Djangaliev  Chapter 3 – Translation from Russian: “The Wild Fruit and Nut Plants of Kazakhstan”, p A.D. Djangaliev and T.N. Salova

Conclusions and Future Directions 1)Apple  Gene pool greatly increased with collection of wild species with passport data …..Make selections of seedlings w/ unique traits  Planning additional collections of wild species in Southwest China  Evaluate 7 populations (1300 seedlings of Gala X M. sieversii crosses) 2)Grape  Need to make additional collections in China and North America  Grow out and evaluate collections made in Kazakhstan  Expand cryopreservation research w/ NCGRP 3)Sour Cherry  Gradually building collection based on evaluations of Dr. A. Iezzoni of Michigan State University 4)Activities  Continue morphological characterization on current as well as additional descriptors  Continue digital imaging of fruit samples for all accessions and begin digital imaging of leaves, flower and tree  Molecular characterization – in house and collaboration

ACKNOWLEDGEMENTS  Professor Aimak Djangaliev, Kazakstan Academy of Sciences, Almaty, Kazakstan; Host for Expeditions  USA and international scientists who provided data on evaluation progress  NPGS Plant Exploration Office that provided funding for expeditions: C. Sperling, K. Williams, N. Garvey  NPGS administrative leadership: H. Shands, A. Stoner, P. Bretting  NPGS GRIN program personnel  USDA- ARS International Programs: R. Soper, R. Bennnet, E. Rosenquist  USDA- ARS National Center for Germplasm Resources Preservation: S. Eberhart, L. Wiesner, H. Shands, G. Volk, L. Towill, C. Stushnoff, C. Walters  Apple, Grape and Prunus Crop Germplasm Committees (CGC)  All plant exploration team members  PGRU administrative leadership: S. Kresovich, J. McFerson, W. Lamboy, C. Simon  PGRU administrative assistants: D. Emerson, T. Fisk, S. Walburn  Staff assigned to Clonal Repository of PGRU  Former CR technicians who completed cryo & virus indexing: Sheffer & Holleran  Cornell staff responsible for disease screening: H. Aldwinckle, H. Gustafson, T. Momol  Cornell and other SAES scientists with SCAs to evaluate germplasm  PGRU Grape Genetics Staff  Horticulture Sciences staff at NYSAES for early oversight of the CR establishment and continuing activities