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Application of Molecular Tools for WheatQuality Improvement: Indian Scenario
H. S. Balyan
Ch. Charan Singh University,Meerut
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007
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ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007
Wheat Grain Quality1. GPC & micronutrients (Fe, Zn)
- Nutritional quality
2. Pre-harvest Sprouting (PHS)- Activates hydrolyzing enzymes
- Endosperm constituents are broken down- Decreased volume & poor crumb of bread- Lowers quality of grain for bread/feed and seed
3. GW- Aesthetic, market & industrial quality (flour recovery)
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ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007
Wheat Grain Quality4. GPC, GW, sedimentation value, kernel
hardness, amylose content- Bread, biscuit, chapati and noodle quality
5. HMW-GS and LMW-GS- Flour processing properties/dough qualitysuch as viscoelasticity & extensibility
6. Phytic acid- Reduces absorption of Fe and Zn
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Applications of Molecular Markers
- Preparation of genetic maps- QTL analysis
- High density mapping
- Validation of linked markers- Molecular marker assisted selection
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007
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RIL Populations Used
- RIL pop for GPC (HD2329 PH132)
- RIL pop for PHST (WL711 SPR8198)
- RIL pop for GW (CS Rye Selection111)
- RIL population for GH (K65 Lerma Rojo)
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007
PAU & IARI
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Framework Molecular MapsGPC Map: 214 loci (CCSU &NCL)
GW Map:298 loci (CCSU)PHST Map:214 loci (CCSU)
GH Map: Under preparation (IARI)
Prasad et al. (2003). TAG 106:659-667
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007
M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15PH132
WL711
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7/31/2019 Session VII Balyan
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Xgwm830
Xgwm1063Xgwm164Xgwm357
2A
Xgwm359Xgwm1198
Xgwm1256
Xgwm1052Xgwm515Xgwm372
Xgwm895
Xgwm1115
C
1D
C
Xgwm957
Xgwm608
2B
Xgwm319Xgwm1249
Xgwm374
Xwmc272
Xgwm1027
Xgwm429Xgwm410
C
2D
C
Xwmc41
Xgwm1264Xgwm1204
C
3A
Xgwm1171
C
7A6B
Xgwm680
Xgwm133
Xgwm771
Xgwm889C
Xgwm456
3D
Xgwm892C
Xgwm894
Xgwm601
Xgwm397
C
4A
Xgwm940b
4B
C
Xgwm513Xgwm805
C
5D5B
Xgwm831
C
7D
C
NIL2233
Xgdm86
C
Xgwm666
2B
CXgwm429
Xgwm410
Xgwm1027
3B
C
Xgwm493
5A
C
NIL2210
Xgwm456
Xgwm493
2B
C
Xgwm429Xgwm410Xgwm319
3D
C
Xgwm1249
Xgwm374
Xwmc272
3B 4B
C
Xgwm513
C
2A
C
Xgwm830
NIL2215
Xgwm940b
2A
Xgwm940b
NIL2218
Xgwm1027
2B
Xgwm429
C
Xgwm1249
3B
C
Xgwm493Xgwm726
C
4B
C
NIL2232
1A
Validation of SSR markers Linked With QTLs for GPC in NILs (CCSU)
7/31/2019 Session VII Balyan
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ICIM mapping of QTLs for GW
using IciMapping v1.4 34 QTLs on 18 chromosomes
3 major QTLs detected
QTL on1B explained 26.0 to 82.0% andanother QTL explained 10.6- 45.7 % PV
QTL on 2B explained 11.37-32.77 % PV
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007
Kumar et al. (2006). Euphytica 151:135-144
CSSU
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QTL Analysis for PHST
CIM
7 QTLs on 1A, 2A, 2D, 3A, 3B
One major QTL each on 2A and 3A
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007 CSSU
HD2329 SPR8198 1 2 3 4
5 6 7 8 9
Kulwal et al. (2005). TAG 111: 1052-1059
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Two-Locus QTL AnalysisQTLMapper
PVE due to M-QTLs:GPC=7.22 to 7.22%; GW=28.11%; PHST=37 to 48%
PVE due to QQ:GPC= 2.68 to 6.04%; GW=42.12%; PHST=27 to 28.73%
PVE due to QE:GPC= 25.00 to 47.99% ; GW= nil; PHST= 3.24%
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007CSSU
7/31/2019 Session VII Balyan
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1A
1B
1D
2A
2B
2D
3A
3B
3D
4A
4B
4D
5A
5B
5D
6A
6B6D
7A
7B
7D
: a : a + ae: ae : aa : aae : aa + aae : centromere
Q Q and Q E Interactions Resolved Through Two-Locus
QTL Analysis for GPC in Bread Wheat
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007 CSSU
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GPC: gwm1264, gwm608, gwm1171, gwm508, gwm193,
gwm361, gwm644, PCR-CAPS marker, ASA marker,uhw89
PHST:cdo795, psr115, bcd1431.1, bcd1431.2,
bcd1380, fbb370, bcd907, fbb283, ATPase.2,wmc104,Mst101,wmc153,gwm155,
GW: wmc333, gwm419,E35M4714, E35M4718Leaf rust: Lr9(SCS5550); Lr24(SCS73719);Lr28(SCS421570); Lr19(SCS265512); (SCS253736)
Linked Markers and Their Validation
CSSU
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Strategy for MAB Three step selection during backcrossing
- Foreground selection for alleles for donor-parent markers flanking the QTL
- Background selection for the recipient-parent-alleles on loci of individualchromosome carrying the QTL
- Background selection for the recipient-parent-alleles for the whole-genome
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007
CSSU
7/31/2019 Session VII Balyan
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MAB of GPC-B1,a high GPC QTL, into 10 Indian WheatGenotypes
Yecora Rojo
F1s
Foreground & background selections
BC3F1s (85-100% genomic similarity)
BC3F2sSelection of individuals homozygous for GPC-B1a
Indian cultivars
Respective RP
BC1F1S
10 Recipient Parents (RP) High GPC Donor Parent (DP)
Xuhw89
DP RP BC3F2 progenies
* ** * * * *
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007Asterisk indicate individuals homozygous for the GPC-B1 allele
CSSU
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10
11
12
13
Proteincontent(%)
HD2687
HD2329
PBW343
PBW
343(L
r19)
PBW
343(L
r9)
PBW
343(L
r24)
K9107
PBW
373
Raj3765
HI977
Performance of BC3F1 Containing GPC-B1 QTL Allele
(Change in GPC by -0.93 to 1.72%)
CSSU
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Summary of MAS for Gpc-B1
Key long--2 0 0 4F1
2 0 0 7
2 0 0 6 - 0 7
2 0 0 6 - 0 7
2 0 0 5 - 0 6
2 0 0 4 - 0 5
Year
PAU, Ludh iana2 5~ 5 0F4
PAU, Ludh iana7 41 6 7BC1F1
PAU, Ludh iana1 6 83 0 0BC2F2
Key longI n
p rog res s
DNA
iso la t i on
BC2F3/ F5
Key long1 1 03 3 0BC2F1
Locat ion+ v e f orASA
To t a l p l an t sana lysed
Genera t ion
Recipient Parents- BW3652, C273, C518, DBW16, DBW17, BW4406,BW4426, PBW509, PBW554, PBW563, PBW564, PBW568, PBW571
PAU
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20
15
10
5
0
Grain Protein content of F4s progenies of the crosses of
GluPro with wheat
%GPC
PBW3
43
Glupro
13.0 to 14.3%
11.3%
15.6%
Marker assisted selection for HGPC geneGpcB1 using allele specific amplification marker (ASA)
PAU
7/31/2019 Session VII Balyan
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PH
Sscale
Parents and Progeny0
3
6
9
PHS scaleSusceptible =9
Resistance =1
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007 CSSU
7/31/2019 Session VII Balyan
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Gr aph ica l geno t yp es o f PBW 34 3 / SPR5in t r og r ession l ines
Xwmc153 and Xgwm155 suitable for marker assisted
mobilization of PHST to elite wheat backgroundsICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007 PAU
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Transfer of new GluAxand GluAysubunits to
bread wheatX
BC2F3(2006-07)
Half seed analysis
X
X
X
Half seed analysis
Half seed analysisHalf seed analysis
Half seed analysis
T. durum-T. dicoccoidesPBW343
F1 PBW343
BC1F1 PBW343
BC2F1 PBW343
BC3F1BC2F2
BC3F2(2006-07)
~300 half seeds analysedand 8 homozygous and 11
segregating progeniesidentified PAU
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01020
304050607080
90100
Ae. kots
chyi3573
Ae. kots
chyi3774
Ae. pere
grina3791
Ae. pere
grina13772
Ae. lo
ngissima
3770
Ae. lo
ngissima
3507
T.dic
occoides
4640
T.dic
occoides
477
T.ae
stivu
mPBW
34
T.du
rumPDW
27
Accessions
IronandZinc(mg/kg)
Iron (mg/kg) Zinc (mg/kg)
Iron and zinc of wild relatives as
compared with cultivars
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007 IARI, ARI, PAU, IITR
7/31/2019 Session VII Balyan
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Chinese Spring (PhI)F1 CS (PhI) XAe. kotschyi Ae.kotschyi
Spikes of Parents and F1 of wide crosses
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007 IARI, ARI, PAU, IITR
7/31/2019 Session VII Balyan
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a b
dc
fe
Meiotic chromosome pairing in F1 of wide crosses
CS (PhI) XAegilops kotschyi3573
(a) 2II (b) 4II
CS (PhI) X
Aegilops kotschyi396(c) 4II (d) 6II
CS (PhI) X
Aegilops kotschyi3790),(e) 3II;,1III (f)2II
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007 IARI, ARI, PAU, IITR
7/31/2019 Session VII Balyan
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Sterile
Colchicine treatment
Synthetic amphiploid
AABBDDUUSS
Chinese Spring (PhI) Ae.kotschyiF1 CS (PhI) XAe. kotschyi
UUSSABDUSAABBDDSynthetic
amphiploids
IARI, ARI, PAU, IITR
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Grain Iron and Zinc of
Amphiploids
41.6774.55Ae. kotschyi 3790
17.2330.87WL 711
19.6130.84CS (Phi)
32.7577.27CS(Phi) x Kot 379033.1370.00WL711 x Kot 391
33.6870.73CS(Phi) x Kot 396
25.2071.96WL711 x Kot 3774
51.0777.00Ae. kotschyi 396
41.2282.67Ae. kotschyi 3774
Parents/ Amphiploid Fe (ppm) Zn (ppm)
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007 IARI, ARI, PAU, IITR
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0
20
40
60
80
100
120
140
160
180
a b c d e f g h i j k l m n o
Recipients, F1 hybrids, Donors
Ironcontent(mg/kg)
Recepient F1 Hybrid Donor
0
10
20
30
40
50
60
70
a b c d e f g h i j k l m n o
Recipients, F1 Hybrids, Donors
Zinccontent
(mg/kg)
Recipient F1 Donor
Leaf iron and zinc content in sterile F1 hybridsbetween wheat and Aegilopsdonors
ICAR-ACIAR Workshop on Molecular Marker Assisted Breeding In Wheat, October 12, 2007 IARI, ARI, PAU, IITR
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Future Research Programmes to Identify
Novel QTLs & Linked Markers
AB-QTL analysis LD and association analysis
Identification of novel positive alleles
Transciptome analysis
Expression QTL (e-QTL) analysis
Genetic mapping of newly identified genesequences including regulatory sequences
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Participating Institutions CCSU, Meerut
IARI, New Delhi
PAU, Ludhiana
DWR, Karnal ARI, Pune
NCL Pune
IIT, Roorkee
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ACKNOWLEDGEMENTS
Funds provided by DBT and NATP are
gratefully acknowledged
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Thank You