Session VII Balyan

7/31/2019 Session VII Balyan

1/31

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


2/31


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)


3/31


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


4/31

Applications of Molecular Markers

- Preparation of genetic maps- QTL analysis

- High density mapping

- Validation of linked markers- Molecular marker assisted selection



5/31

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)


PAU & IARI


6/31

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


M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15PH132

WL711


7/31


8/31

Xgwm830

Xgwm1063Xgwm164Xgwm357

2A

Xgwm359Xgwm1198

Xgwm1256


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


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)


9/31

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


Kumar et al. (2006). Euphytica 151:135-144

CSSU


10/31

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


11/31

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


12/31

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



13/31

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


14/31

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


CSSU


15/31

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


16/31

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


17/31

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


18/31

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


19/31

PH

Sscale

Parents and Progeny0

3

6

9

PHS scaleSusceptible =9

Resistance =1



20/31

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


21/31

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


22/31

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


23/31

Chinese Spring (PhI)F1 CS (PhI) XAe. kotschyi Ae.kotschyi

Spikes of Parents and F1 of wide crosses



24/31

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



25/31

Sterile

Colchicine treatment

Synthetic amphiploid

AABBDDUUSS

Chinese Spring (PhI) Ae.kotschyiF1 CS (PhI) XAe. kotschyi

UUSSABDUSAABBDDSynthetic

amphiploids

IARI, ARI, PAU, IITR


26/31

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)



27/31

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



28/31

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



29/31

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


31/31

Thank You

Documents

Session VII Balyan