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

1
Twenty Years of Germplasm Management Geneva, NY
to the Forests of Central Asia, China, Russia,
and Turkey

• Philip Forsline
• April 5, 2004

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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
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History of facilities Clonal repository at PGRU
Campus Bldg 1985
Campus Bldg 1985
Equipment storage, 1984
Farm preparation, 1984
Tile drainage 1984
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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
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History of significant events and personnel
additions at PGRU, Geneva, NY

• 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

• 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

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Vegetatively-propagated crops at PGRU

• Apple 3909 accessions
• Grape 1175 accessions
• Sour Cherry 87 accessions
• Total 5171 accessions

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Major Seed-Propagated Crops Conserved at
GenevaLarry Robertson Curator/GeneticistJoanne
Labate Molecular Biologist
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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)

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The Geneva Apple Rootstock Breeding
ProgramGennaro Fazio, USDA/ARS, PGRU in
cooperation with Cornell University
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Staff for clonal collections at PGRU

• SYs FTE
• - P. Forsline 1.0
• Horticulturist/Curator
• - C. Simon 0.85
• 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

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Current CRIS ProjectConservation 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.

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Acquisition and Maintenance of Clonal Collection
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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

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Vitis collection at PGRU
Early / Late leaf fall
Vineyard (1045 clones)
Diversity Among clusters
Diversity among clones
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Digital imaging of Vitis clones
PI 588058 V. coignetiae
PI 181481 Vignoles
PI 588054 V. riparia
PI 588070 Catawba
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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

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Cherry collection at PGRU
P. cerasus / P. fruticosa
Collection (87 acc.)
spreading
Early / late bloom
upright
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Cherry digital imaging
Montmorency
Balaton
Danube
Schatten Morello

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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

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Diversity in Malus
Fruit
Bloom
Late leaf fall
Early leaf fall
Bloom M. baccata
Dormant Kansas 14
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Malus / seedling collection at PGRU
Early stage after planting 1986
More advanced stage
Apples/seedling
vineyard
Tree removal after repropagation on EMLA 7
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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
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Digital imaging of Malus clones
Gala
Belle de Boskoop
Geneva
Malus hupehensis
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Malus core collection at PGRU
Planting in 2000
Label system M. transitoria
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
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Malus / EMLA 7 collection at PGRU
Planting in 2000
Establishment 2001
Apples/seedling
Extension orchard, 2002
Apples/EMLA 7
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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
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Codes for fire blight, shootY (natural) of
apple Code Definition No. of
Accessions 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
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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

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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
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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
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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. 123365-370.

• 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

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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 eightY
years 68.3 b ZSeparation of grand means of
84 accessions at P lt 0.01 by test for
differences between two proportions (LSD
7.2) Y Fifteen year test in process 2004 to 2007
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Annual cryopreservation of Malus accessions at
NCGRP following the 4-yr pilot project 1988-1992
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Successful cryopresevation for gt 90 of
accessions stored at NCGRP those with lt 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
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Successful cryopresevation for gt 95 of
accessions stored at PGRU those with lt 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
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Distribution of Germplasm
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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 (mean89) Apple orders/yr (range 30-163Z)
Grape total no. 11,765 Grape total orders 782
(mean46) Grape orders/yr (range 14-72Z )
Cherry distributions started in 1998 191
accessions to 21 orders
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Characterization and Evaluation
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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

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Collaborative evaluation with Specific
Cooperative Agreements (SCA) of clonal
collections in process or recently completed

• 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

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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)

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New Acquisitions
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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


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Germplasm collections to add mostly wild Malus
species to the PGRU collection
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Collection team for 1989 Central Asian expedition
to Uzbekistan, Tajikistan and Kazakhstan (site 3)
The late Calvin Sperling
Herb Aldwinckle
Elizabeth Dickson
Uzbekistan
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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
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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
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1995 USA and S. African collection team
Dickson Britz Forsline Luby Human
Arrival at site 9
Seed extraction at hotel site 9
Meals in yurt site 5
Gala dinner with hosts in Almaty return to USA
Housing in sanitarium at HQ in Almaty
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1996 USA collection team
Hokanson Unruh Forsline Pellett
HQ in Almaty
Site 9
Site 12
Site 5
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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

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Site descriptions in Central Asia
Country/Region Site Lat oN / Long
oE Elevation (m) Rainfall (mm) Tajikistan /
--- 1 39 / 68 ---
--- Uzbekistan/ --- 2 41 / 69
--- --- Kazakstan/Zailisky 3 43
/ 77 1170-1690 700 Kazakstan/Djungarsky 4
45 / 80 1170-1760 800 5 46
/ 81 1190-1360 850 Kazakstan/Karatau 6
43 / 70 600-910 250 11 42
/ 70 780-1230 250 Kyrgyzstan/ --- 7
41 / 73 1300-1500 1300 Kazakstan/Tarbagatai
9 48 / 82 870-1120 450 Kazakstan/
Ketmen 10 44 / 80 1600-1700
650 Kazakstan/Talasky 12 42
/ 71 1000-1025 320
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Kazakhstan site 3 89, 93, 95, 96
Studies by Professor A. Djangaliev
Habitat 1940
Habitat present
Fruit from 30 randomly-collected trees
Depleted by dachas
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Kazakhstan site 4, 93, 95, 96
Scab infected tree
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Kazakhstan site 5 93, 95, 96
Forestry camp at 1200 m headquarters in 93, 95
96
Apple forest 1800 m 1100 m
Bear scat w/ many apple seeds
300 yr old M. sieversii
First collection morning (Sept. 1993) starting
out at 600 m with hike to 1800 m
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Kazakhstan site 6 93 95
Camp at 600 m headquarters
Standing at 900 m
Fruit from 30 randomly-collected trees at 900 m
M. sieversii at 900 m
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Kyrgyzstan site 7 1993
Apple and walnut forests as seen from helicopter
Heavy grazing in M. sieversii areas
Village in Kyrgyzstan near collection sites
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Kazakhstan site 9 95 96
Grazed areas M. sieversii
on slopes
Diverse, elite M. sieversii
M. sieversii
Super-elite M. sieversii
Heavy grazing of habitat
Fruit from 30 randomly-collected M. sieversii
trees
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Kazakhstan site 10 1996
Breakdowns common occurrence
M. niedzwetzkyana
Uygur tribes tradition of bread baking
Fruit from 20 randomly-collected trees
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Kazakhstan site 11 1996
Elite Yellow transparent-type M. sieversii
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Kazakhstan site 12 1996
Canyon 400 m deep w/ M. sieversii on the N-facing
wall
Bottom of canyon M. sieversii on this side
Trail to bottom
Fruit from 10 randomly- collected M.
sieversii trees
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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/InsectsLeaves/Fruit
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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) 2 10 65 42
60-74 32-50 5 / (96) 14
54 58 35 54-65 25-49 9 / (95/96)
57 150 56 43
46-72 28-62 11 / (96) 13
40 55 42 44-76 29-63 10 / (96)
2 20 49 40
47-51 29-51 5 / (95) 14
60 46 36 37-56 27-48 3 /
(95/96) 11 80 45 34
32-56 26-49 4 / (95/96)
10 87 44 34 32-55 28-44
6 / (95) 25 64 42 41
33-46 28-54 Totals 148 565
51 38 32-76 25-63
Note difference in fruit size observed in
1995 and 1996 at Site 5
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Summary of Central Asian M. sieversii collection,
distribution and storage
Accessions Seeds Seeds
Storage Storage Group Obtained
Obtained Distributed at PGRU at NSSL
1989 and 1993 179 33,000 9200
13,900 10,900 Collections 1995
and 1996 148z 67,000 16,000
29,200 21,800 Elites 1995 and 1996
565 30,000 4100 19,100
6800x Random populations Totals 892
130,000 28,300Y
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)
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Seed storage at -200 C
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Evaluation of New Material
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PGRU / Cornell cooperative evaluation of Malus
sieversii
Lab Evaluation Type
No. of Seedlings H. Aldwinckle Disease
resistance 5124Z PGRU Horticultural
/ Molecular part of 5124 - P.
Forsline - S. Hokanson - W. Lamboy - G. Fazio
- L. Benson N. Weeden Isozyme / Molecular S.
Mehlenbacher / W.
Reissig Insect resistance Aldwinckle /
Korea Rosellinia / Helicobasidium 400 S.
Brown Genetic dwarf 250 I.
Merwin Nematode resistance 200 Z 2108 of this
group of screened seedlings were sent to OH, NJ,
WA, MN and WI for further evaluation
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PGRU / SAES cooperative evaluation of M. sieversii
State / Lab Evaluation type
No. of seedlings MN / Luby Hort,
Disease, Cold H. 1498Y NJ / Goffreda Hort,
Disease, Molecular, Elite clones 1899Y CO /
Stushnoff Hort, Cold H., Antioxidants 720 WA
/ Barritt Hort, Sunburn, Cold H. 624Y OHZ
/ Lynd, Miller Hort, Disease, Late Bloom, Elite
clones 950Y WI / Smith Hort, Disease, Cold
H. 655Y AR / Rom Hort, Disease, Late
Bloom 400 AK / McBeath Hort, Disease, Cold
H. 346 IL / Korban Hort, Disease,
Molecular 200 Z Seedlings planted at Dawes
Arboretum, Newark, OH c/o D. Miller Y Some
seedlings at _at_ of these sites screened for apple
scab, fire blight and cedar-apple rust at
Cornell, Geneva by Dr. Aldwinckle
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PGRU / International cooperative evaluation of
Malus sieversii
No. of Country / Lab
Evaluation type seedlings New
Zealand / Bus, Noiton, Gardiner Disease,
Hort 4426 Germany / Buttner, Geibel, Höfer
Disease, Hort 1367 Norway /
Roen Disease, Hort 692 N.S.,Canada /
Deslauriers, Embree Disease, Hort 1155 Man.,
Canada / Davidson Disease, Hort, Cold H.
169 B.C., Canada / Quamme, Hampson Disease,
Hort 325 N.B., Canada / Hunter Hort
120 Japan / Bessho Disease, Hort
100 UK, Reading /
Farrel Molecular 300 UK, Oxford
/ Juniper Molecular 150 S.
Africa / Human, Britz Dis., Insect, Sunburn,
Chilling ?Z Netherlands / Kemp Elite
clones -- Italy / Sansivini Elite
clones -- Z Parallel collections were made in
1995 expedition in Kazakhstan

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M. sieversii seedling grow outs in Germany and
New Zealand
Dresden, Germany
Vincent Bus Havelock, North, New Zealand
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Disease Resistance Screening
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Screening young M. sieversii seedlings for apple
scab (Venturia inaequalis)
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Codes used in rating apple scab resistanceH. S.
Alwinckle and H. L. Gustafson

• 1 Pits, small pin-prick marks (similar to Vm
  gene) - RESISTANT
• 2 Chlorotic lesions (similar to Vf 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 Vf gene) -
  RESISTANT
• 9 Stellate (star shaped) necrotic (similar to
  Vr gene) - RESISTANT
• A Usually has some cupping and or chlorosis
  but may have little or no symptoms
• (Characteristic of Vf gene) ( numbers 2,5,7,
  8) - RESISTANT
• B Stellate necrotic (Characteristic of Vr
  gene) (number 9) - RESISTANT
• Other resistant reactions
• - LN Large necrotic lesions - RESISTANT
• - N Necrotic lesions - RESISTANT
• - P Pits ( number 1 ) - RESISTANT

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Apple scab resistance of Malus sieversii
populations from Central Asia H. Aldwinckle, P.
Forsline, H. Gustafson and S. Hokanson
Site No. Seedlings Inoculated
Resistant 4 405
50 6 705
37 5
1224 29 9
1325 28 7 383
25 2 101
24
11 244
23 3 460
17 12 133
14 10 123
6
1 21
5 Totals 5124
28
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Wild Malus sieversii seedling plantings
1989 collection
1995/96 collections (1200 sdgs)
M. niedzwetzkyana form of M. sieversii
Early stage after planting 1986
More advanced stage
1993 collection
M. sieversii in bloom
M. sieversii fruiting
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Summary of scab resistance in grow-out of
1188 Malus sieversii seedlings in Geneva, New York
80
Cedar apple rust resistance in M. sieversii
seedlings from 8 different sites in Kazakhstan
of population resistant
9 6 4 3 5 11 10
12
Total
81
Natural incidence of fire blight on shoots of the
Malus sieversii grow-out in Geneva, New York
Fire blight
82
Natural occurrence of fire blight in M. sieversii
seedlings from 7 different sites in Kazakhstan
9
of trees in each category
9
6
6
6
Codes 1 2
Codes 4 5
Fire Blight Severity
83
Horticultural Evaluations
84
Descriptors used to characterize M. sieversii at
http//www.ars-grin.gov/cgi-bin/npgs/html/crop.p
l?115

• 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

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Wild Malus sieversii clones and seedlings in
grow-out plantings
PI 613972 clone
in bloom
PI 613975 clones fruiting
PI 613978
More advanced stage
Fruit from half-sib seedlings of some of the M.
sieversii clones
13 different M. sieversii clones
vineyard
Empire
Gala
Empire
Gala
86
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
87
Harvest season and soluble solids of M. sieversii
fruit from grow-outs in yrs 5 - 7 (2001 - 2003)
in Geneva, New York in addition the no. of trees
to characterize in 2004 (yr 8 or 7)
88
Fruit shape in M. sieversii seedling populations
from 6 sites in Kazakhstan
of seedlings in each category
89
Digital imaging of Malus sieversii seedlings
From site 5 in Kazakhstan
From site 6 in Kazakhstan
From site 9 in Kazakhstan
From site 11 in Kazakhstan
90
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
91
Elite clone (PI 613971) and progeny (3 half-sibs)
Clone
Seedling .a
Seedling .b
Seedling .h
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Elite clone (PI 613976) and progeny (3 half-sibs)
Seedling .a
Clone
Seedling .k
Seedling .n
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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

94
Other Recent Collections
95
1997 Sichuan collection team
Crew collecting minutes before monkey thievery
Geneva, 11/96 Prof. Li plans
Hotel in Chengdu Prof. Li wife H B
Aldwinckle
monkey
Laura Benson
Crew at 3500 m pass in mountains
Post expedition conference with Yunnan and
Guizhou researchers in Chonqing
96
Malus sp. collections in Sichuan, China
Malus kansuensis
M. hupehensis
M. prattii
M. toringoides
97
1997 collections of Malus spp. in Sichuan,
ChinaPhil Forsline, Laura Benson and Herb
Aldwinckle
Species Site(s)
No. of accessions No. of seeds Malus
prattii 01, 05, 06 22 1475 Malus
transitoria 02 9 650 Malus hupehensisZ 03,
04, 07 23 1100 Malus toringoides 03, 04 16
900 Malus kansuensis 03 13 800 Malus
sieboldiiZ 05 7 375 Malus
zhaojiaoensis 06 11 1900 Total --
101 7200 Z Likely to be apomictic
98
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 eval
uation
Many of these sdgs distributed to 10 arboreta for
preservation
99
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 (0304), 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
100
Variability of fire blight resistance in M.
hupehensis and M. toringoides collected from
different sites in Sichuan, China
of sdgs fire blight resistant
of sdgs fire blight resistant
3
4
7
4
3
M. hupehensis
M. toringoides
101
Apple scab resistance in 5 Malus species
collected in Sichuan, China in 2002 by M. Geibel
of seedlings scab resistant
toringoides
kansuensis
hupehensis
transitoria
sieboldii
Fire blight and cedar apple rust screen is in
process
102
Malus orientalis in Russian Caucasus
Vavilov Inst. St. Petersburg
103
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
104
Malus orientalis in Turkey
Forsline, Aldwinckle 6 Turks
Local type Seker Elmasii Sugar apple
Q 42413
105
Screening populations of M. orientalis from
Russian Caucsus, Turkey and Armenia for fire
blight, apple scab and cedar apple rust
No. of sdgs screened
Russia Turkey Armenia Scab
238 786 191 F.B. 206
565 77 C.A.R. 129 191
TBD
of seedlings resistant
A, B, P-type 312 ratio
Mostly A-type
A/B 21
27 populations
62 populations
7 populations
106
Screening 5 populations of European wild
crabapple (M. sylvestris) received from gene bank
in Dresden, Germany
of seedlings resistant
107
Expedition to Russia to exchange sour cherry and
cherry rootstock for crop improvement, July
10-30, 1998A. Iezzoni, R. Karle, P. Forsline and
M. Fischer
Cherries in St. Petersburg
Cherry accession in Orel
A. Iezzoni
M. Fischer R. Karle
Prof. Yushev
Krymsk Prunus nursery
Breakdown Orel
Michurinsk
G. V. Eremin M. Fischer
108
Expedition to Russia to Exchange sour cherry,
cherry rootstock, and Malus germplasm for crop
improvementJuly 10-30, 1998A. Iezzoni, R.
Karle, P. Forsline and M. Fischer

• Goals of expedition
• tart cherry elite germplasm at two sites (Orel,
  and Michurinsk)
• broad spectrum Prunus spp (Krymsk)
• Germplasm in 1 2 with resistance to cherry
  leaf spot (Blumeriella jaapii) and twig brown
  rot (Monilinia laxa)
• wild apple collections (Maikop)
• Trip participants
• Amy Iezzoni and Renate Karle, Michigan State
  University
• Philip Forsline, USDA-ARS Plant Genetic Resources
  Unit
• Manfred Fischer, Genebank OBST, Dresden-Pillnitz,
  Germany
• Material collected
• Orel 25 accessions
• Michurinsk 12 accessions
• Krymsk 24 accessions
• Maikop area 28 seed lots (6500 seeds) of wild
  M. orientalis

109
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

110
Germplasm Utilization and Enhancement
111

• 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
• Vr Vm Vf Vb
• Accession No. P415B UBC562
  OPB12 CS5 UBC220
• GMAL 4326 YES YES NO NO
• GMAL 4327 YES YES YES YES
• GMAL 4331 YES YES NO NO
• GMAL 4333 YES NO NO NO
• GMAL 4334 YES YES NO NO
• These have been used as pollen parents X
  Gala (see next slide)

112
Germplasm enhancement to studygenetics of apple
scab resistance

• Gala X M. sieversii (GMAL 4335) 67 of 230
  sdgs resistant
• Gala X M. sieversii (GMAL 4448) 57 of 209
  sdgs resistant
• Gala X M. sieversii (GMAL 4455) 52 of 209
  sdgs resistant
• Gala X M. sieversii (GMAL 4331Z) 29 of 90
  sdgs resistant
• Gala X M. sieversii (GMAL 4334Z) 24 of 206
  sdgs resistant
• Gala X M. sieversii (GMAL 4333Z) 11 of 136
  sdgs resistant
• Gala X M. sieversii (GMAL 4327Z) 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
11 ratio
Z gt50 of progeny from these clones are scab
resistant clones themselves
are not Sdgs from crosses are all A-type
resistance
113
Selected publications (1 2) and media (3 4)
The origin of apples
2
1

• Horticultural Reviews, vol. 29. 2003. 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. 1-62. P.L. Forsline, H.S.
  Aldwinckle, E.E. Dickson, J. J. Luby, and S.C.
  Hokanson. 2003.
• Chapter 2 - Translation from Russian The Wild
  Apple Tree of Kazakhstan, p. 63-304. A.D.
  Djangaliev
• Chapter 3 Translation from Russian The Wild
  Fruit and Nut Plants of Kazakhstan, p. 305-371.
  A.D. Djangaliev and T.N. Salova

4
3
114
Conclusions and Future Directions

• 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)
• Grape
• Need to make additional collections in China and
  North America
• Grow out and evaluate collections made in
  Kazakhstan
• Expand cryopreservation research w/ NCGRP
• Sour Cherry
• Gradually building collection based on
  evaluations of Dr. A. Iezzoni of Michigan State
  University
• 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

115
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