GCP Wheat CI Team
• Indian Agricultural Research Institute, New Delhi
• Punjab Agricultural University, Ludhiana
• J L Neh...
Source: IWMI
Water Stress Indicator Map
GCPWheatCITeam,India
Abiotic Stresses of Wheat in India
• Wheat grown in 30 million hectares and over 85% of wheat is
irrigated
• But nearly 70...
Wheat Breeding Strategies for Drought and
Heat Tolerance
 Development of varieties which adapt to high temperatures
durin...
Centres and Responsibilities
•IARI, New Delhi MARS, MABB : Phenotyping, Genotyping
•JNKVV, Powarkheda MARS: Phenotyping
•P...
Traits targeted
Morphological traits
1. Days to flag leaf emergence
2. Days to 50% heading
3. Ground cover
4. Grain yield ...
GCPWheatCITeam,India
Per cent ground cover : Proportion of the Mean Grey Value to mean value if
the image were completely white (255). (Grey va...
VARIABLE AVERAGE MINIMUM MAXIMUM HERITABILITY
GERM 88.664 40 95 0.59
DH 66.154 50 84 0.94
YLD 179.243 66 373 0.64
TKW 31.4...
Validation of markers linked to known
QTLs
• More than two hundred microsatellite markers associated with physiological,
p...
QTL references Chromosome no. Traits targeted
Pinto et al. 2010 2B, 5A, 4A, 1B CT, Chl, NDVI,
yield
Kumar et al. 2010 7B, ...
Drought tolerance from Parent B
HI 1500
Yield traits from Parent
DBW43
z
1A 1B 2B 2D 3A 3B 3D1D 2A
Best possible individua...
FAMILY. 1
FAMILY. 4FAMILY. 3
FAMILY 2
x x
2nd Recombination
x
MARS for Accumulating Favourable Genes
F1F1
GCPWheatCITeam,I...
SSRs markers linked to QTLs for Drought
tolerance which are segregating population
HI1500 X DBW43
GWM 484(2D)
GWM 644(7B...
Breeding CrossGCPWheatCITeam,India
HI 1500 (P1)
F1
F 4 And F5 generation
F2
Phenotyping in
E1, E2 E3 and
E4 in Rainfed
and irrigated
Parental
polymorphism
13...
Traits phenotyped (HI 1500 x DBW 43)
Morphological traits
1. Days to flag leaf emergence
2. Days to 50% heading
3. Ground ...
Procedure used to select Individuals for
intercrossing
• Final selection based on AMMI ranks on multilocation yield data
(...
LOCATIONS
DELHI
LUDHIANA
POWARKHEDA
PUNE
TRAITS
CANOPY TEMPERATURE
CHLOROPHYLL CONTENT
YIELD
BIOMASS
HARVEST INDEX
GCPWhea...
**Yield in Rainfed and Restricted irrigation conditions
**Yield Among locations and **Among lines
**Canopy Temperature (CT...
HI 1500 X DBW 43
GCPWheatCITeam,India
Environment NE Em IPCAe[1] IPCAe[2]
Delhi 1 639.4 21.09613 -14.15923
Ludhiana 2 709.9 -1.32147 20.07025
Powarkheda 3 349.1...
Environment NE Em IPCAe[1] IPCAe[2]
Delhi 1 499.5 25.85725 15.92357
Ludhiana 2 624.4 -25.47537 12.53966
Powarkheda 3 277.8...
S.No Population Families selected for intercrossing
1 IARI 1 19,26,43,44,76,84,87,98,105,137,143,146,155,156
2 IARI 2 68,1...
Procedure followed for intercrossing
• In each family, 40 individuals were tagged as females and
emasculated
• Selfed seed...
ENT
NO 20 26 32 34 38 59 87 88 98 110 136 137 146 156
20 X X X X X X X X X X X X X
26 X X X X X X X X X X X X
32 X X X X X...
Few examples of cross made to accumulate QTLs
Marker inretval Traits linked Family no 20 (POSITIVE FOR 2 QTLs) x Family no...
GCPWheatCITeam,India When Richard Trethovan, our Coordinator gave us a
surprise when he directly reached the field from
Sy...
QTL references Marker interval Traits linked
Pinto et al. 2010 Barc186 – Barc151
(5A)
Barc 147(3B)
CT
Kadam et al. 2012 Ba...
HD 2733 X CROSS 1 -59 (5 Spikes ) CROSS 1-59X CROSS 4 P4 CROSS 1 P 85 X CROSS 4 P4
HD 2733 X CROSS 1- 20 (5 Spikes ) CROSS...
Sl.No. Double cross(In each case 15 spikes were
emasculated and pollinated)
1 (34 Χ136) Χ (59 Χ 146)
2 (146 Χ4) Χ (20 Χ 88...
Xbarc 186 linked Canopy Temperature by olivares el al 2008
Segregation pattern in bulked families
HI 1500 203 bp
DBW 43 21...
HI 1500 DBW 43
Barc 151 Segregation pattern in
Families
F1 type
HI 1500 217 bp
DBW 43 231 bp
GCPWheatCITeam,India
Segregation pattern Xbarc 68 linked to Chlorophyll content
DBW 43 145 bp
HI 1500 137 bp
GCPWheatCITeam,India
WMC 487 linked to 1000 grain weight by son et al 2008
A1 = HI1500 216
A2 =DBW 43 216 and 222bp
GCPWheatCITeam,India
Cfd 73 segregation pattern in F4 families
A1 = HI1500 264bp
A2 =DBW 43 256bp
GCPWheatCITeam,India
Chromosome No. of
Polymorphic
markers
1A,1B,1D 58
2A,2B,2D 60
3A,3B,3D 48
4A,4B,4D 37
5A,5B,5D 46
6A,6B,6D 61
7A,7B,7D 110...
A1 = HI1500 264bp
A2 =DBW 43 256bp
Identification of new QTLs (Cfd 73) in F4
families
GCPWheatCITeam,India
A1 = HI1500
A2 =DBW 43
Identification of new QTLs (Xbarc 327) in F4
families
GCPWheatCITeam,India
Development of backcross populations
• In the last crop season crosses were developed among contrasting
parents for heat a...
S.No Backcrosses PopSize
BC1F1
Trait targeted Chr no.
1 HD 2733*1/C306 645 Yield, CT, Chl 4B,1D, 3B
2 HD 2733*1/HI 1500 51...
HD2733 X HD2888
F1 X HD 2733
BC1F1 (population size 454)
L 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
...
HD2733 X C306
F1 X HD 2733
BC1F1 (population size 645)
Kadam et al Funct Integr Genomics (2012)
HD2733 C 306
HD2733 C 306
...
Foreground selection :QTL for Stay green under drought on 7D
HD 2733 (Recurrent parent) X HI 1500 (Donor parent )
F1 X HD ...
Generation advancement, intermating
and backcrossing at Off-season
(Lahaul Spiti, Kashmir) in summer
GCPWheatCITeam,India
An opportunity created
GCP + DBT (Govt of India) for prolonged use through MAS products of
HD 2967, HD2733, GW 322, GW 366...
IndianAgriculturalResearchInstitute
 Evaluation of BC2F3 at all four locations for drought and heat
 Intercross NILs: HD...
This was a presentation on behalf of our GCP-Wheat
Challenge Initiative Network on abiotic stress tolerance
breeding
GP Si...
Thank you
GCP
and “Special” Thanks to
Prof. R. Trethowan
Dr. Xavier Delanay
GCPWheatCITeam,India
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GRM 2013: Molecular Breeding and Selection Strategies to Combine and Validate QTLs for Improving WUE and Heat Tolerance of Wheat in India -- KV Prabhu

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GRM 2013: Molecular Breeding and Selection Strategies to Combine and Validate QTLs for Improving WUE and Heat Tolerance of Wheat in India -- KV Prabhu

  1. 1. GCP Wheat CI Team • Indian Agricultural Research Institute, New Delhi • Punjab Agricultural University, Ludhiana • J L Nehru University of Agricultural Sciences, Powarkheda • Agharkar Research Institute, Pune • National Research Centre on Plant Biotechnology, New Delhi India kvinodprabhu@rediffmail.com Molecular Breeding and Selection Strategies to Combine and Validate QTLs for Improving WUE and Heat Tolerance of Wheat in India GCPWheatCITeam,India Generation Challenge Programme : GRM 2013, Lisbon September 27-30, 2013
  2. 2. Source: IWMI Water Stress Indicator Map GCPWheatCITeam,India
  3. 3. Abiotic Stresses of Wheat in India • Wheat grown in 30 million hectares and over 85% of wheat is irrigated • But nearly 70% of irrigated area does not receive all 6 recommended irrigations • Wheat suffers losses due to abiotic stresses such as heat (terminal, early, tillering stages), drought and salinity ranging from 10-40% (yield and reduced quality) • Minor genes or QTLs based tolerance is likely to provide abiotic stress tolerance in wheat and very few varieties available for this purpose • Options such as MARS, MABB and AM are expected to pick up/utilize tolerance imparting QTLs distributed along the complex wheat genome for their deployment in varietal backgrounds GCPWheatCITeam,India
  4. 4. Wheat Breeding Strategies for Drought and Heat Tolerance  Development of varieties which adapt to high temperatures during early, mid and terminal stages (resilience)  Varieties suited to change in planting dates to avoid terminal heat stress  Varieties with high WUE, suited to low water availability  Varieties suited to altered crop rotations  Development of varieties for new agricultural areas resulting due to shift in climatic pattern GCPWheatCITeam,India
  5. 5. Centres and Responsibilities •IARI, New Delhi MARS, MABB : Phenotyping, Genotyping •JNKVV, Powarkheda MARS: Phenotyping •PAU, Ludhiana MARS, MABB : Phenotyping, Genotyping •ARI, Pune MARS: Phenotyping •NRCPB, New Delhi Genotyping GCPWheatCITeam,India
  6. 6. Traits targeted Morphological traits 1. Days to flag leaf emergence 2. Days to 50% heading 3. Ground cover 4. Grain yield (g/plot) 5. Harvest index 6. Above ground biomass (g/plot) 7. Test weight (g) 8. Grain filling duration Physiological traits 1. Stay green duration(Using NDVI) 2. Canopy temperature (CT) using Infrared thermometer 3. Flag leaf area 4. Chlorophyll content 5. Chlorophyll florescence GCPWheatCITeam,India
  7. 7. GCPWheatCITeam,India
  8. 8. Per cent ground cover : Proportion of the Mean Grey Value to mean value if the image were completely white (255). (Grey value of a completely black image is 0 ) Digital estimate of per cent ground cover for early vigour evaluation GCPWheatCITeam,India
  9. 9. VARIABLE AVERAGE MINIMUM MAXIMUM HERITABILITY GERM 88.664 40 95 0.59 DH 66.154 50 84 0.94 YLD 179.243 66 373 0.64 TKW 31.404 20 44 0.76 DM 102.822 80 125 0.81 CT 28.259 20.6 31.2 - CTD 4.373 1.8 12.1 0.28 NDVI 0.747 0.58 0.88 0.33 SPAD 53.588 43.7 62.3 0.51 DH DM YLD TKW CT CTD NDVI DM 0.84** YLD -0.22** -0.11 TKW 0.20** 0.19** 0.20** CT -0.07 -0.16** -0.36** -0.09 CTD 0.08 0.19** 0.38** 0.04 -0.74** NDVI 0.39** 0.46** 0.24** -0.07 -0.34** 0.28** SPAD -0.04 -0.03 0.02 0.20** -0.06 -0.03 -0.12* GCPWheatCITeam,India Variability profile of the different populations: An example
  10. 10. Validation of markers linked to known QTLs • More than two hundred microsatellite markers associated with physiological, phenological and agronomic traits were selected. • These microsatellite markers associated with specific traits on the parents of our backcross generations to Introgress the QTLs (16 back cross BC1F1, now BC2F1 with 600-800 size generated). • • Polymorphic markers to target five physiological traits covering 17 chromosomes for identification of known QTls in our backgrounds. • • These include Canopy temperature QTLs on 15 Chromosomes (1A, 1B, 2A, 2B, 3A, 3B, 4A,4B, 5A, 5B, 6A, 6B, 6D, 7A,7B), NDVI on 3 chrs (2A, 2B and 4A), Chlorophyll on15 chrs (1A,1B,1D, 2A, 2B, 3A, 3B, 4B, 5A, 5B, 6A, 6B, 6D, 7A and 7B), WSC on 17 chrs (1A,1B,1D, 2A, 2B, 2D, 3A, 3B, 4A, 4B, 4D, 5A, 5B, 6B, 6D, 7A and 7B), stay-green on 3chrs (1A,3B and 7D). GCPWheatCITeam,India
  11. 11. QTL references Chromosome no. Traits targeted Pinto et al. 2010 2B, 5A, 4A, 1B CT, Chl, NDVI, yield Kumar et al. 2010 7B, 7D, 3B Stay-green trait Mohammadi et al. 2008a and b 2D, 1B, 5B and 7B Yield and GFD under heat Kumar et al. 2012 1D, 3B CT, Chl Olivares et al. 2007 2A NDVI Kadam et al. 2012 4B, 3D Yield under drought and root traits Significant QTLs reported and utilized in our populations GCPWheatCITeam,India
  12. 12. Drought tolerance from Parent B HI 1500 Yield traits from Parent DBW43 z 1A 1B 2B 2D 3A 3B 3D1D 2A Best possible individual by modeling 6A 6B 7A 7D7B 5A GCPWheatCITeam,India
  13. 13. FAMILY. 1 FAMILY. 4FAMILY. 3 FAMILY 2 x x 2nd Recombination x MARS for Accumulating Favourable Genes F1F1 GCPWheatCITeam,India
  14. 14. SSRs markers linked to QTLs for Drought tolerance which are segregating population HI1500 X DBW43 GWM 484(2D) GWM 644(7B) GWM 156 (5A) GDM 132(6D) GWM 383(3D) GWM 165(4B) GWM 368(4B) GWM273(1B) GWM 582(1B) BARC 147(3B) GWM 566 (3B) GCPWheatCITeam,India
  15. 15. Breeding CrossGCPWheatCITeam,India
  16. 16. HI 1500 (P1) F1 F 4 And F5 generation F2 Phenotyping in E1, E2 E3 and E4 in Rainfed and irrigated Parental polymorphism 130 SSR markers Genotyping and QTL validation Selection of lines for intercrossing based on multilocation AMMI Analyses of phenotype (RF + RI) and QTLs genotyping F1 F1F1 F1 F2 F2 F2F2 .1432Fam.1 54 7…6 Selection of homozygous plant BDW 43(P2) Double cross Double cross GCPWheatCITeam,India
  17. 17. Traits phenotyped (HI 1500 x DBW 43) Morphological traits 1. Days to flag leaf emergence 2. Days to 50% heading 3. Ground cover 4. Grain yield (g/plot) 5. Harvest index 6. Above ground biomass (g/plot) 7. Test weight (g) Physiological traits 1. Stay green duration 2. Canopy temperature (CT) using Infrared thermometer 3. Flag leaf area 4. Chlorophyll content 5. Leaf area index (0nly in 2011- 12) 6. Chlorophyll flouresence (0nly in 2011-12) 7. NDVI index( only in 2011-12) GCPWheatCITeam,India
  18. 18. Procedure used to select Individuals for intercrossing • Final selection based on AMMI ranks on multilocation yield data (common individuals in top 30 ranks in all four location) • AMMI analysis was done over the location for yield data. Rain fed and restricted irrigation data was combined as two treatments and for each location two replications were included • Separate AMMI analysis was done separately for rain fed data and Four best lines were also included in selection. • Four best performing lines from the least favorable environment were also added • Homozygous lines for favourable QTLs are selected for intercrossing GCPWheatCITeam,India
  19. 19. LOCATIONS DELHI LUDHIANA POWARKHEDA PUNE TRAITS CANOPY TEMPERATURE CHLOROPHYLL CONTENT YIELD BIOMASS HARVEST INDEX GCPWheatCITeam,India
  20. 20. **Yield in Rainfed and Restricted irrigation conditions **Yield Among locations and **Among lines **Canopy Temperature (CT) at vegetative and reproductive stages **Chlorophyll content at vegetative stage HI 1500 X DBW 43GCPWheatCITeam,India
  21. 21. HI 1500 X DBW 43 GCPWheatCITeam,India
  22. 22. Environment NE Em IPCAe[1] IPCAe[2] Delhi 1 639.4 21.09613 -14.15923 Ludhiana 2 709.9 -1.32147 20.07025 Powarkheda 3 349.1 - 22.06025 -13.91356 Pune 4 191.6 2.28559 8.00254 Number Environment Mean Score 1 2 3 4 1 Delhi 639.4 21.10 G28 G86 G34 G84 4 Pune 191.6 2.29 G87 G34 G149 G137 2 Ludhiana 709.9 -1.32 G109 G87 G14 G149 3 Powarkheda 349.1 -22.06 G76 G88 G93 G136 HI 1500 X DBW 43 GCPWheatCITeam,India
  23. 23. Environment NE Em IPCAe[1] IPCAe[2] Delhi 1 499.5 25.85725 15.92357 Ludhiana 2 624.4 -25.47537 12.53966 Powarkheda 3 277.8 3.47501 -27.55992 Pune 4 114.0 -3.85689 -0.90331 Number Environment Mean Score 1 2 3 4 1 Delhi 499.5 25.86 G88 G133 G32 G84 3 Powarkheda 277.8 3.48 G121 G76 G93 G88 4 Pune 114.0 -3.86 G156 G76 G121 G136 2 Ludhiana 624.4 -25.48 G138 G151 G142 G73 FINAL SELECTIONS (RF + RI) Entries: 19,26,43,44,76,84,87,98,105,137,143,146,155,156 AMMI for RAINFED CONDITIONSGCPWheatCITeam,India
  24. 24. S.No Population Families selected for intercrossing 1 IARI 1 19,26,43,44,76,84,87,98,105,137,143,146,155,156 2 IARI 2 68,111,46,7,72,107,101,68 3 PAU 1 51,5,43,113,105,95,49,23,104,84,48 4 PAU 2 8,11,74,97,80,28,44,112,153,118,52,4. Families selected for intercrossing in Winter 2013 GCPWheatCITeam,India
  25. 25. Procedure followed for intercrossing • In each family, 40 individuals were tagged as females and emasculated • Selfed seeds of each female and male plant were collected • Five spikes were pollinated in each cross • Crossed seed from each spike collected separately • Crossed seed were planted along with selfed male and female plants and F2 + Selfed seed harvested GCPWheatCITeam,India
  26. 26. ENT NO 20 26 32 34 38 59 87 88 98 110 136 137 146 156 20 X X X X X X X X X X X X X 26 X X X X X X X X X X X X 32 X X X X X X X X X X X 34 X X X X X X X X X X 38 X X X X X X X X X 59 X X X X X X X X 87 X X X X X X X 88 X X X X X X 98 X X X X X 110 X X X X 136 X X X 137 X X 146 X 156 INTERCROSSES MADE BETWWEN SELECTED FAMILIES DURING 2011-12 IN CROSS HI 1500 X DBW 43 In each cross 5 spikes are emasculated and from male and female plants selfed seeds were collected and planted along with crosses in lahoul spiti now in all above mentioned cross F2 seeds are available GCPWheatCITeam,India
  27. 27. Few examples of cross made to accumulate QTLs Marker inretval Traits linked Family no 20 (POSITIVE FOR 2 QTLs) x Family no 59 ( POSITIVE FOR 6 QTLs) Barc 186 (5A) CT Homo for DBW 43 (-/-) Homo for HI1500 (+/+) Barc 147(3B) CT Homo for DBW 43 (-/-) Homo for HI1500 (+/+) Gwm 273() CT Homo forHI1500 (+/+) Homo for DBW 43 (-/-) Barc68-barc 101(3B) Fv/Fm, chl Homo for DBW 43 (-/-) Homo for HI1500 (+/+) Barc 170 (4A) Fv/Fm Hetero (+/-) Homo for HI1500 (+/+) Gwm 533() GFD Homo for DBW 43 (-/-) Homo for HI1500 (+/+) Wmc 487 (6B) 1000 Gn wt Homo for DBW 43 (-/-) Homo for HI1500 (+/+) Gwm 368-barc20 (4B) Yld (drought),root biomass Homo for HI1500 (+/+) Homo for DBW 43 (-/-) Marker Traits linked Family no 38 ( POSITIVE FOR 4 QTLs) x Family no 59 ( POSITIVE FOR 6 QTLs) Barc 186 (5A) CT Homozygous for HI1500 (+/+) Homozygous for HI1500 (+/+) Barc 147(3B) CT Homozygous for DBW 43(-/-) Homozygous for HI1500 (+/+) Gwm 273 () CT Homozygous for HI1500 (+/+) Homozygous for DBW 43 (-/-) Barc68-barc 101(3B) Fv/Fm, chl Homozygous for HI1500 (+/+) Homozygous for HI1500 (+/+) Gwm 533(3B) GFD Homo for DBW 43 (-/-) Homo for HI1500 (+/+) Barc 170 (4A) Fv/Fm Homozygous for DWB43(-/-) Homozygous for HI1500 (+/+) Wmc 487 (6B) 1000 Gn wt Homo for DBW 43 (-/-) Homo for HI1500 (+/+) Gwm 368-barc20 Yld (drought),root biomass Homozygous for HI1500 (+/+) Homozygous for DWB43 (-/-) marker Traits linked Family no 34 (POSITIVE FOR 2 QTLs) Family no 20 (POSITIVE FOR 2 QTLs) ) Barc 186 (5A) CT Homozygous for HI1500 (+/+) Homozygous for DBW 43 (-/-) Barc 68-barc 101 (3B) Fv/Fm, chl Homozygous for HI1500 (+/+) Homozygous for DBW 43 (-/-) Gwm 368-barc 20 (4B) Yld (drought),root biomass Heterozygous (+/-) Homozygous for HI1500 (+/+) Barc 170(4A) Fv/Fm Heterozygous (+/-) Heterozygous (+/-) Gwm 273() CT Homozygous for DBW 43 (-/-) Homo forHI1500 (+/+) GCPWheatCITeam,India
  28. 28. GCPWheatCITeam,India When Richard Trethovan, our Coordinator gave us a surprise when he directly reached the field from Sydney to watch the intermating in operation!!
  29. 29. QTL references Marker interval Traits linked Pinto et al. 2010 Barc186 – Barc151 (5A) Barc 147(3B) CT Kadam et al. 2012 Barc20-gwm368(4B) grain yield per plant, harvest index, and root biomass under drought. Wang et al. 2009 gdm 132-cfd42 (6D) Barc113-gwm533(1A) GFD Kumar et al. 2012 Barc68-barc101(3B) Xgwm369 (3A) CT, Chl, Fv/Fm Son et al. 2007 Wmc 487(6B) 1000 grain wt Significant QTLs reported and utilized in our populationsGCPWheatCITeam,India
  30. 30. HD 2733 X CROSS 1 -59 (5 Spikes ) CROSS 1-59X CROSS 4 P4 CROSS 1 P 85 X CROSS 4 P4 HD 2733 X CROSS 1- 20 (5 Spikes ) CROSS 1 -59 X CROSS 4 P5 CROSS 1 P 85X CROSS 4 P5 HD 2733 X CROSS 1 -156(5 Spikes ) CROSS 1 -59 X CROSS 4 P8 CROSS 1 P 85 X CROSS 4 P8 HD 3076 X CROSS 1-98 (5 Spikes ) CROSS 1 -59X CROSS 4 P11 CROSS 1 P 85 X CROSS 4 P11 HD 3076 X CROSS 1 -86 (5 Spikes ) CROSS 1 -59 X CROSS 4 P44 CROSS 1 P 85 X CROSS 4 P44 CROSS 1-38 X CROSS 4 P4 CROSS 1 P-59X CROSS 4 P55 CROSS 1 P 85 X CROSS 4 P55 CROSS 1-38X CROSS 4 P5 CROSS 1-59 X CROSS 4 P35 CROSS 1 P 85 X CROSS 4 P35 CROSS 1 -38 X CROSS 4 P8 CROSS 1 -59X CROSS 3 P5 CROSS 1 P 85 X CROSS 3 P5 CROSS 1-38 X CROSS 4 P11 CROSS 1 -59X CROSS 3 P51 CROSS 1 P 85 X CROSS 3 P51 CROSS 1 -38 X CROSS 4 P44 CROSS 1 -86X CROSS 3 P5 CROSS 1-87X CROSS 4 P35 CROSS 1-38 X CROSS 4 P55 CROSS 1 -86 X CROSS 3 P35 CROSS 1 -87X CROSS 3 P5 CROSS 1 -38 X CROSS 4 P35 CROSS 1-86 X CROSS 3 P51 CROSS 1 -87X CROSS 3 P51 CROSS 1-38 X CROSS 3 P5 CROSS 1 P 104 X CROSS 3 P5 CROSS 1 -38 X CROSS 3 P51 CROSS 1 P 104 X CROSS 3 P35 Crosses made among selected individuals of different populations of MARS these crosses were forwarded in lahaul spiti F2 seed available in each case GCPWheatCITeam,India
  31. 31. Sl.No. Double cross(In each case 15 spikes were emasculated and pollinated) 1 (34 Χ136) Χ (59 Χ 146) 2 (146 Χ4) Χ (20 Χ 88) 3 (136 Χ 88) Χ (87 Χ 59) 4 (20 Χ 87) Χ (136 Χ 86) 5 (146 Χ 86) Χ (20 Χ 88) 6 (110 Χ 136) Χ (20 Χ 146) 7 (20 Χ 156) Χ (110 Χ 146) 8 (156 Χ 38) Χ (156 Χ 32) 9 (156 Χ32) Χ (156Χ 136) 10 (156Χ 38) Χ (34Χ 110) List of double crosses made to accumulate favourable QTLs in lahaul spiti during 2012-13 GCPWheatCITeam,India
  32. 32. Xbarc 186 linked Canopy Temperature by olivares el al 2008 Segregation pattern in bulked families HI 1500 203 bp DBW 43 211 bp GCPWheatCITeam,India
  33. 33. HI 1500 DBW 43 Barc 151 Segregation pattern in Families F1 type HI 1500 217 bp DBW 43 231 bp GCPWheatCITeam,India
  34. 34. Segregation pattern Xbarc 68 linked to Chlorophyll content DBW 43 145 bp HI 1500 137 bp GCPWheatCITeam,India
  35. 35. WMC 487 linked to 1000 grain weight by son et al 2008 A1 = HI1500 216 A2 =DBW 43 216 and 222bp GCPWheatCITeam,India
  36. 36. Cfd 73 segregation pattern in F4 families A1 = HI1500 264bp A2 =DBW 43 256bp GCPWheatCITeam,India
  37. 37. Chromosome No. of Polymorphic markers 1A,1B,1D 58 2A,2B,2D 60 3A,3B,3D 48 4A,4B,4D 37 5A,5B,5D 46 6A,6B,6D 61 7A,7B,7D 110 Hunt for new QTLs segregating in the cross Polymorphic markers among parental genotypes GCPWheatCITeam,India
  38. 38. A1 = HI1500 264bp A2 =DBW 43 256bp Identification of new QTLs (Cfd 73) in F4 families GCPWheatCITeam,India
  39. 39. A1 = HI1500 A2 =DBW 43 Identification of new QTLs (Xbarc 327) in F4 families GCPWheatCITeam,India
  40. 40. Development of backcross populations • In the last crop season crosses were developed among contrasting parents for heat and drought tolerance • F1s from twenty three crosses were advanced in the off-season nursey at Lahaul-Spiti • F1s were backcrossed with the high yielding recurrent parent to generate BC1F1 populations • These Bc1F1s are advanced in the current crop- season to generate BC2F1 populations. DNA samples were drawn from the BC1F1 for foreground selection of the target trait and recovery of the background genome through background selection. • A total of 850 microsatellite markers used and approximately hundred polymorphic markers were observed among them GCPWheatCITeam,India
  41. 41. S.No Backcrosses PopSize BC1F1 Trait targeted Chr no. 1 HD 2733*1/C306 645 Yield, CT, Chl 4B,1D, 3B 2 HD 2733*1/HI 1500 516 NDVI, Stay- green,Yield 2B, 7D 3 HD 2733*1/HD 2888 454 NDVI, CT, Yield 1D, 7B, 4A 4 HD 2733*1/HW 2004 706 NDVI, yield 2B, 7D 5 HD 2733*1/NI 5439 462 NDVI, CT, Yield 1D, 7B, 4A 6 HD 2733*1/WH730 725 Yield, CT 1B, 7B 7 GW 322*1/HD 2888 650 NDVI, CT 2A, 2B 8 GW 322*1/HD 2987 780 Stay-green, Yield, GFD 7D, 4A, 6D 9 GW 322*1/HI 1500 540 Yield, stay-green, Chl content 4A, 7D, 4B 10 GW 322*1/NI 5439 352 Yield, Chl 4B, 3B 11 GW 322*1/RAJ 3765 392 CT, Yield 1B, 5B, 7B 12 GW 366*1/HD 2987 430 13 GW 366*1/WH 730 470 CT, Yield 1B, 5B, 7B 14 GW 366*1/RAJ 3765 540 CT, Yield 1B, 5B, 7B Traits targeted for introgression through MABB GCPWheatCITeam,India
  42. 42. HD2733 X HD2888 F1 X HD 2733 BC1F1 (population size 454) L 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 L 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 13 = HD2733(259bp) 14=HD2888(270bp) 13 = HD2733(300& 248 bp) 14=HD2888(306& 256bp) Plants positive for both cfd55 & cfd79 selected (plant No. 3 and 22) cfd55 cfd79 Kadam et al Funct Integr Genomics (2012) Foreground selection for QTL for yield under drought on 3DS GCPWheatCITeam,India
  43. 43. HD2733 X C306 F1 X HD 2733 BC1F1 (population size 645) Kadam et al Funct Integr Genomics (2012) HD2733 C 306 HD2733 C 306 Kadam et al Funct Integr Genomics (2012) xbarc20 gwm 368 GCPWheatCITeam,India
  44. 44. Foreground selection :QTL for Stay green under drought on 7D HD 2733 (Recurrent parent) X HI 1500 (Donor parent ) F1 X HD 2733 BC1F1 (population size 782) L 1 2 3 4 5 6 7 8 9 10 1 1 12 13 14 15 16 17 18 19 20 21 22 23 24 Lane 13 = HD 2733 Lane 14=HI 1500 L 1 2 3 4 5 6 7 8 9 10 1 1 12 13 14 15 16 17 18 19 20 21 22 23 24 Kumar et. al Euphytica (2010) gwm 111 gwm 437 GCPWheatCITeam,India
  45. 45. Generation advancement, intermating and backcrossing at Off-season (Lahaul Spiti, Kashmir) in summer GCPWheatCITeam,India
  46. 46. An opportunity created GCP + DBT (Govt of India) for prolonged use through MAS products of HD 2967, HD2733, GW 322, GW 366 (drought + rust resistant) HD2733 × HD2687+Lr19 HD2733 × HD2687+ Lr24 HD2733×HD2687 Yr15 F1 F1 F1HD2733 HD2733 HD2733× × × BC1 BC1 BC1HD2733 HD2733 HD2733× × × BC2 BC2BC2HD2733 HD2733 HD2733 BC2F1 BC2F1 BC2F2 BC2F2 BC2F2 Double cross F1 NIL for Lr19 NIL for Lr24 NIL for Yr15 BC3 BC3 BC3 ××× IndianAgriculturalResearchInstitute
  47. 47. IndianAgriculturalResearchInstitute  Evaluation of BC2F3 at all four locations for drought and heat  Intercross NILs: HD 2733 lines(DHT) X HD2733 (RR) MAS in F2-F4 (Foreground + carrier chromosome background) Sometime in 2016/17 Reconstitution of Leading varieties introgressed with drought and heat tolerance + Rust resistance A joint output from GCP and DBT
  48. 48. This was a presentation on behalf of our GCP-Wheat Challenge Initiative Network on abiotic stress tolerance breeding GP Singh, Neelu Jain, T Ramya, Rajbir, PK Singh, C Pandey, SC Mishra, PC Mishra, NK Singh, TR Sharma, Praveen Chhuneja, VS Sohu, GS Mavi, KV Prabhu + 16 Research Staff* Delhi Team X 4 = India GCP TeamGCPWheatCITeam,India
  49. 49. Thank you GCP and “Special” Thanks to Prof. R. Trethowan Dr. Xavier Delanay GCPWheatCITeam,India

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