This document summarizes the goal and objectives of pre-breeding work at the International Potato Center (CIP) focused on using crop wild relatives (CWR) to improve potato varieties. The goal is to introduce valuable genes from CWR and landraces into improved varieties to enhance sustainable farming. Objectives include identifying, combining, and documenting new traits from CWR and landraces for use in potato breeding programs. The document provides examples of traits and pre-breeding approaches used at CIP, such as introducing resistance to pests, diseases and abiotic stresses into advanced potato populations and varieties.

1. Overview of pre-breeding w/
focus on the use of CWR in
potato improvement at CIP
M. Bonierbale, E. Mihovilovich, B. Ordoñez
Global Crop Diversity Trust CWR Consultancy: Potato PreBreeding
2/22/2012, Lima, Peru

2. Pre-breeding Goal
Assure the flow of valuable genes and diversity
from CWR and landrace germplasm into
productive, improved varieties that enhance
sustainable and profitable farming systems.
Objectives
– Identify, upgrade, combine and document new sources of
needed traits for their incorporation into useful support
population and stocks.
– Support the continued utility of CIP’s broad-based tropically-
adapted potato populations for variety selection and potato
improvement.

3. General Pre-breeding Targets
& Cases for Potato at CIP
A. Traits “missing” from advanced Ø Pests, PVY, PVX, BW
populations and adapted varieties. resistance
B. Higher levels and more highly
heritable sources of needed traits. Ø PLRV resistance
C. Complementary sources to build
durable resistance to variable Ø Late blight resistance
pathogens.
D. Horizon scanning: Emerging Ø Heat, salinity and
Constraints; Climate Change. drought tolerance
E. Opportunities for new/added-value Ø Nutritional value
traits not previously attended by (micronutrient density)
breeding programs.

4. Searching for traits
1. Crop Wild Relative
2. Cultivated Landraces
3. Broad-based advanced Populations

5. Progress and prospects of Potato
Improvement by sources and traits
(CIP at a glance)
Before CIP's
40 years' CIP History Future
foundation
80 85 90 95 00 05 10
Late Blight dms ,
A B P T
Potato Cyst Nematode
(Globodera pallida)
Frost
PVY, PVX acl,
s to
PLRV
Potato Tuber Moth
Root Knot Nematode
Early Blight
Bacterial Wilt
Drought
Bioavailable Micronutrients

6. Foundation & Early Breeding
Research Contracts

7. Assets: Raw and Improved Germplasm
Adapted to Tropical Agro-ecologies
Lowland
phu
Tbr Adg tropics and
Neo dms Virus Resistant
tbr (LTVR)
Population
dms
1.2
Lesion radius (mm)
y = a+bx
Tbr 1.0
blb phu
b=LGR= 5.7
Highland
0.8
0.6
acl Neo
Adg tbr
0.4
0.2
y = a+bx
b=LGR= 3.6
tropics
0.0
3 5 7
Late Blight
Days after inoculation
Resistant (B3)
Population

8. Sustainability Population Improvement
Progenitors Incorporation Progenitors
New Cycles
Nobilisation and Genetic studies:
Introgression of • Parental value
Evaluation • Heritability
new diversity
• Variability
Support Yield, Stability
Populaciones Advanced
populations
Adaptation
Elite clones

9. Historical PreBreeding
Approaches & Results LB
BW
Virus
PLRV

10. Pre-breeding forBW + RKN Resistance
1977 -1995
BW, LB
dms × tbr (Mex germ:Atzimba) S.
sparsipilum
x S.
chacoense
tbr (earliness),
phu (CCC)x
tbr (Katahdin)
(University
of
Wisconsin)
Katahdin,
Atlantic,
DTO-­‐28 E S.
microdontum
(
CIP,
1977)
S.
phureja BW
2x
BR,
MS,
PSP,
PSW Katahdin,
Atlantic,
DTO-­‐28 E, HT
HT Stn,
phu.
Gon
tbr (heat
tolerance), (Long
adapted
Group
V
(2x)
Group
I
X
M.
tropical,
Serrana
germplasm)
X MBN
(BW
+
RKN)
CGN-­‐69.1 (1977) CGN-­‐69.1
PI/PS
bulk
X (
CIP,
1977)
(NCSU) (1980)
Group
II
‘Nobilisation”
Group
III (1979)
BW (1979-­‐80)
4x
x
2x
Group
VI
(2x)
X MBN
+
TUBEROSA
Group
IV (1983)
(1980)
raph
,
chc
(AVRDC-­‐1287.19)
Group
VII CIP
BW TD
(4x)
(1985) X LTVR
population E, HT, V
(4x)
4x
population
(1995)
Highly
heterozygote
Four
sources
of
BW
resistance
+
virus
resistance

11. Selections from 1990’s Series Show Reduced
BW Severity Under High Pressure

12. Products of Collaborative BW Resistance
PreBreeding Program
391930.1 394904.17
BW, LB, PVX,
BW,
High
high concentration concentration
Zinc and Vit C Iron and Zinc
394908.13
395443.103
BW, PVX,
high concentration BW, LB –HR, PVX,
Zinc high concentration
Zinc and Vit C
395446.1
BW, PVX, PVY,
High concentration Iron and Zinc

13. Highly Heterozygous Nematode Resistant Variety
2x Breeding follow by 2x-4x Hybridization
vrn 2488-A (C.P.C. collection)
82M124.27
HT Bulk - stn-phu (NCSU)
84.28.42 stn-phu (NCSU)
378908.43
381343.40 (MBN) S. spl
Bulk Kura CIP 760147.7
85.27.13 (2x) (Pi 1230502)
(CIP PTL 80M (virus)
591099.13) MI-7.10 stn-phu (NCSU)
BW
84.194.11 S. spl
(MBN)
Nemared CIP 760147.7
MI-49.10 (Pi 1230502)
387559.3
released in
1994
AQUILA [tbr x (tbr x phu)]
LT-1
KATAHDIN (tbr)
LT-8 (4x)
PVXY Bk. (adg, neo-tbr)

14. Strategic Broadening of the Late Blight
Resistance of Advanced PopB3
1.2
Lesion radius (mm)
y = a+bx
1.0
b=LGR= 5.7
B3 0.8
0.6
Population 0.4
y = a+bx
0.2 b=LGR= 3.6
S bulbocastanum S. .tuberosum 0.0
S phureja 2x=48 3 5 7
2x=24
S acaule Days after inoculation
2x=24
4x=48 S. demissum
2x=48
S. tuberosum 25
S. andigena
Number of families
2x=24 2x=48 20
15
10
B3C3
5
B3C0
0
AUDPC (Family Means)

15. 1. Determine the Architecture of LB
Resistance in B3 Population
chr1 chr2 chr3 chr4 chr5 chr6
EatgMcag80 0.0 STI0024f 0.0 EaacMcac205 0.0 STMGX55 0.0 STI0032f 0.0 EatgMacg249 0.0
STM5127f 4.2 STG0006f 7.5 EacaMcag51 1.3
STG0020 10.5 EaaaMacg340 6.7 T0892_COS6f
STM1008 9.8 9.9
STI0034 10.8 STM3016 11.4 At5g14320 11.8 At3g10920f 10.2
STM5114f 10.7 EaacMcag243 16.2 EaacMcac167 12.3 STM0019 16.0
STI0043f 17.7 STI0029f 20.3 At3g58490f 20.8
STI0036f 21.0 At5g51110f 26.5 STM5140 21.8 STM5119 21.4
At1g29520 23.4 At5g12370 28.3 EaaaMcag328 25.8
EaacMcac244 At1g44575f 28.2
EaacMcac560 32.0 29.0
STG0033 32.7 At1g02140 At1g44760 30.2
STM1029f 33.1 30.3
STI0031f 34.2 EaaaMacg246 32.0 At1g52200 36.7
T0869_COS2.1 38.6 STM0020f 39.4
STI0013f1 35.7 STM1100 40.2
GP25b1895 36.7 STG0021 48.8
dPI02
STI0013f2 37.8 T1169f 48.2
EagaMcat391 50.2 STI0049f 50.4 EacaMcag96 51.8
STG0016f 53.4 At5g24120f2 38.8 STPOAC58b86 58.5
At1g51160f STG0008 54.7 STI0004f 53.9
EatgMacg528 54.7 STM0038 57.4 39.6 EaaaMacg191 59.6
STM1009f2 59.9 At5g53490 42.3 EaaaMacg86 61.8
At3g02910f 49.1 STM5149 65.1
At5g41040f 50.9 At5g26360 66.2
At1g07020 54.5
STM3011 77.1 STI0050f 57.1
STG0022 61.1 STI0012 80.0
STM5115 75.5
STM2030 88.4 At1g16080 76.8
EacaMacg829 89.5 GP198b1572 86.1
STM1049 90.6 STG0010 92.4
chr7 chr8 chr9 chr10 chr11 chr12
STLSIG239
dPI07
0.0 STM0024 0.0 STM1102f 0.0 STM5132f 0.0 STM0037 0.0 STM5121 0.0
STM3009f 3.1 LEUZIP119 2.6 STI0002f 2.4 STI0023f 1.4 STG0004 1.1
dPI09a
STM0028 4.2 STI0038 3.9
STM1051 STM1106b 10.4
4.6
STM1057f1 14.6 STI0057f 7.9
STI0019f 21.7 STMIP26f STM3012 10.4
23.1
STM2013f 22.5 STM1057f2 EagaMcat323 20.0
27.1 STI0028f 25.8 STM0030f
STM0010 29.8 27.2
STI0027f 27.8 STM5109 29.9
STM0017_172 31.6 STI0030f 31.4
At3g26060 35.9 STI0003 36.4 STG0015 STM2005f
32.8 35.5
EaacMcac382 37.1 STI0022b127 37.7 EagaMcat196 35.0
STI0044b310 39.0 EaacMcac700 43.0
STM0031f 46.8 At4g11600 47.1
EaaaMacg451 47.8 STI0046b547 51.2
STI0040f 52.2 STWAX-2f 53.5
STGBSS-1f STINHWI228 64.4 STM0025f 53.1
EagaMacg582 55.5 54.7 STMIR63 57.2
STM1104f 58.1 STI0014 66.8
STM1021f 68.2 STMCL13 63.5
dPI09b
At2g24270 65.5
dPI12
STINHWI277 69.2 STG0007 66.3
At3g24160f2 76.2 EaacMcac368 71.4
DMC42152bf 77.6 STM0003f 72.8
dPI09c
PI09
DMC42144af 78.8 STI0051f 77.7
Rpi_svnt1_367 81.0
At3g24160f1 85.6

16. 2. 2. Search for Novel Resistance Sources:
Evaluate Putative Novel Resistance Sources
Piurana and Tuberosa Groups
Field and Detached leaf
Assays
S. chiquidenum Comas. Oxapampa
2000-2004
S. paucissectum
S. piurae
S. cajamarquense

17. 3. Pre-breeding with Putative Novel Resistance Sources
Embryo Rescue
Technique
Prezygotic barriers Field Testing & Selection
4x × 2x (B3 clones x ER)
Field testing
2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013
Selection:
3rd
resistance +
2nd agronomic +
Project
start Crossing Blocks reproductive
1st Selected
wild 2x
Pre Breeding
populations traits.
Dihaplod Native
2x selections
PF A B C D PM
wild wild x =
x
A B C D
Diallel wild x wild cross
Hybridity test
Field Testing and selection

18. Approaches for Selecting Complementary
Resistance Sources and Genes
phu-703308 B3-303946.7
Chr11
Oxapampa2005 Avr-vnt1
Comas 2001
+ -
LOD
Avr-vnt1+
Rpi-vnt1
+ +
Rpi-vnt1
Rpi-pcs
N,
NL25,
R3,
R3a, b,
- -
NL27, R6, R7 Empty
Bs4, R10, R11 vector
- -

19. Comparative Gene Expression between
B3 and S. cajamarquense under LB Challenge
Study# 83 at
Solanaceae
Gene expression
database:
http://www.tigr.org/tdb/potato/
SGED_index2.shtml
Design
:
advanced B3 S. cajamarquense
clone (cjm)
Inoculation with isolates
POX067 and PE B3
274
Sample collection at 72h and 96h after 30
inoculation cjm
329
Hybridization to the TIGR10K potato array

20. Lesson learned
• Interspecific • Innovative approaches
reproductive barriers (Sli?)
• Lack of fertile • Selection &
recipients w/ good
tuber appearance consolidation of pool of
compatible recipients
• Dilution of resistance • Sufficient
during breeding recombination; Monitor
process w/ molecular tools.
• Poorly-suited genetic • Attention to adaptation
background of recipient germplasm

21. Lesson Learned
• Variability of • Use and monitor
pathogens complementary sources
• Accumulation of • Monitor glycoalkaloids
glycoalkaloids accumulation during
breeding process
• Large populations
and cumbersome • Rapid and affordable
screening methods screening methods
• Limited, short term • Develop and debate
project funding business models
• Capitalize products per
se? Courses?

22. Recomendations
• Develop mechanism to sustain funding for
maintenance of most-valuable prebreeding
populations and stocks
• Document, communicate and create
demand for products for use in house and
across institutions
• Develop Community of Practice for potato
pre-breeding

23. More Pre-breeding Products
112 rare LB
resistant
hybrids with 2n
pollen and
good
agronomic
performance
(ER)

24. Acknowledgements
• W.Perez, A.Salas, S.Priou, L.Gutarra,

25. Thank you