Germplasm and Heterosis

Breeding in Mustard

Breeding of oilseed mustard

Germplasm, In vitro protocols, breeding strategy

Two major Pollination control Molecular mapping Germplasm for

gene pools methods disease resistance

1st generation hybrids DMH-1, DMH-11

2nd generation hybrids

3rd generation hybrids

4th generation hybrids

WGS Quality Disease QTLs

resistance

Cluster B

Similarity Coefficient (F value)

0.82 0.86 0.91 0.95 1.00

Varuna

Pusa Jai Kisan

RH 30 Pusa Bold

Kranti

TM 4

RLM 198 Chinensis-nigra

Sej-II

PNMB Hanzahang Gaoyou Cai

Shi Yian Kuyou Cai

D3A 2

AS 1

D1S 1.3

D3S 33

PS 6 TS 2

ATC 94395

Acc. No. 404 Donskaja IV

Zem1

Skorospieka II

Cutlass

Domo IV

Malopoloska

Acc. No. 2548

Secus

Acc. No. 409 Heera

Cluster A

Cluster C

Two major gene pools in B. juncea

Indian

East European

Srivastava et al (2000) TAG

Agronomic trait values of Indian and East European

B. juncea germplasm

Agronomic traits Indian gene

pool

Exotic gene

Pool

Heritability

D G L

Plant Height (Plht) 177.1 7.9 267.5 53.3 90.0 83.7 67.0

No. of primary branches (Pbr) 5.6 0.4 10.4 3.3 89.6 46.4 40.6

No of secondary branches (Sbr) 12.7 3.1 22.2 3.8 65.4 44.2 38.4

Main shoot length (Msl) 79.6 6.4 38.1 21.5 89.2 70.3 59.9

Siliqua per main shoot (Sqms) 50.4 5.2 37.1 9.6 71.7 54.3 58.0

Siliqua density (Sqdy) 0.63 0.06 1.13 0.31 81.6 53.2 76.4

Siliqua per plant (Sqp) 462.5 96.1 753.0 116.6 44.5 24.9 37.2

Siliqua length (Sql) 6.2 0.8 4.5 0.4 86.3 89.6 80.5

Seeds per siliqua (Ssq) 13.5 0.8 13.9 2.7 69.2 75.1 57.5

Test weight (Tsw) 5.2 1.0 2.3 0.4 94.0 92.7 75.2

Days to flower (Df) 52.3 1.4 101.36 25.2 93.5 96.1 65.2

Pradhan et al. (1995) Euphytica

Ramchiary et al. (2007) Theor. Appl. Genet.

Microspore development in normal

and ‘126-1’CMS anthers

To

ka

ma

su

CM

S B

..n

ap

us H

18

12

6-1

B.

jun

ce

a P

us

a b

old

Mo

rica

nd

ia C

MS

iP

usa

bo

ld

12

6-1

B.

na

pu

s IS

N7

06

‘og

u B

. ju

nce

a P

usa

bo

ld

‘Oxy B

.ju

nce

a P

usa

bo

ld

Dip

lota

xis

CM

S p

usa

bo

ld

‘To

ur

CM

S p

usa

bo

ld

Pu

sa

bo

ld p

are

nt

‘Og

u C

MS

B. n

ap

us H

18

‘Oxy’ C

MS

B.

na

pu

s H

18

‘To

ur’

CM

S B

. n

ap

us

‘12

6-1

CM

S B

. n

ap

us H

18

‘B. n

ap

us p

are

nt H

18

DNA fingerprinting of 126-1 CMS

Sodhi et al. (2006) TAG

Type of trials Zone / Year Number of

locations

Average increase

over checks (%)

Local National

ICAR Zone II

Zone III

13

7

13

20

29

35

Demonstration 2004 - 05

2005 - 06

12

27

31

29

-

-

Minikit 2005 - 06 395 18 -

Farmers field

Alwar

Sriganganagar

Jaisalmer

Bikaner

2007 - 08

2007 - 08

2007 - 08

2007 - 08

25

24

4

2

43

59

47

23

-

-

-

-

Yield performance of mustard hybrid DMH-1

Location: Agra

Check (Rohini) Hybrid (DMH-1)

Male sterile

barnase line

Normal male

fertile line

2004-05 2005-06

Days to

flower

Mean

yield

(kg/h)

% heterosis

Days to

flower

Mean

yield

(kg/h)

% heterosis

Varuna 55 1713 52 1886

DMH-11 56 2488 45.3

55 2415 28.0

Yield and percentage of heterosis of the

transgenic hybrid DMH-11

Erucic acid

No. of

Genes

Indian Gene

Pool

EE Gene

Pool

Glucosinolates

Seed coat color

Oil content

Seed size

Pod density

Height

+

B

+

+

+

+

-

-

Linolenic content

Linoleic content

-

-

+

Y

-

-

2r

5r

2r

7 loci

4 loci

?

2 loci

3 loci

3 loci

Molecular map of B. juncea genome

By 2003

(TAG, 106:607-614)

By 2007

1. Total no. of markers 1029 1808

2. Types of markers

a) AFLP 996 1282

b) RFLP 33 70

c) SSR 0 70

d) Gene markers

e) Intron spanning markers

0

0

13

373

3. Total length (cM) 1628.7 1850.2

4. Assigning LGs to A and

B genomes

- 10 A genome LGs

8 B genome LGs

5. Traits tagged

a) Erucic acid - Candidate gene

b) Linoleic acid - Candidate gene

c) Seed coat colour

d) Glucosinolates

-

-

SSR marker

Candidate gene

Aliphatic glucosinolates biosynthesis in Brassica species

Methionine

Propyls Butyls Pentyls

Methylthiopropyl Methylthiobutyl Methylthiopentyl

Methylsulphinyl-

propyl

Methylsulphinyl-

butyl

Methylsulphinyl-

pentyl

Propenyl

Butenyl Pentenyl

Hydroxybutenyl Hydroxypentenyl

Side chain

elongation

Development of

glucone moiety

Side chain

modification

Gsl-elong

Gsl-Alk Gsl-Alk Gsl-Alk

Gsl-Oxid Gsl-Oxid

Gsl-Oxid

Gsl-elong

Gsl-elong

Gsl-OH Gsl-OH

Gsl-pro

Cys

Myb 28, myb 29

Development of Low GSL Varuna through DH

Varuna X Heera No. of low GSL plants Year

F1 DH Expected - 50% Actual - 44%

7/ 752 1999 - 2000

X Varuna

BC1 DH

11 / 1263 2000 - 01

Expected - 75% Actual - 61%

BC2 DH

Expected - 87.5% Actual - 75%

8 / 728 2001 - 02

X Varuna

X Varuna

BC3 DH Expected - 93.7% Actual - 82%

13 / 1380 2002 - 03

X Varuna

BC4 DH

22/3479

2003 - 04 27/ 3550

Expected - 96.8% Actual - 86%

BC5DH 2004 - 05

x Varuna

Expected - 96.8% Actual - 86%*

*No further increase in Varuna genome due to linkage drag

BjG

sl1

B

jGsl2

B

jGsl6

B

jGsl3

A2

A3

A9

At5g05920 2.4

At5g13190a 12.8

At5g53920 31.9

At1g74640a 48.5

At5g27740b 77.6

GER1 90.1

At5g61970 95.4

At4g35530 6.9

At4g21105 18.3

At1g74640b 27.8

GER4/5 29.9

At3g54670b 37.1

At3g10572b 50.6

At4g01897a 62.5

GAR1 64.2

At2g30200a 77.5

GAR2 0.6

At5g27560 8.6

At5g63905 21.5

At2g14170a 34.2

At2g19260 44.9

At3g46560 57.9

At2g03060 68.3

At1g22860 78.5

At3g55430a 90.6

At3g62620 104.4

At2g20490b 118.0

At2g22370 129.7

Msl

Sqm

s

Tsw

S

ql

Pbr

BjG

sl4

B

jGsl5

B1

B4 At4g13720 0.0

At3g52990 11.2

At3g63420a 23.5

At3g57650 33.5

At2g21620b 60.6

At1g15370 68.0

At2g32080 74.0

At5g65220 85.2

At5g63760 92.2

At1g47570 101.6

At3g06483 8.8

At3g10572a 19.1

At3g20920 30.1

At5g47890 44.6

At5g27830 47.2

At5g66290 51.2

At1g58220a 55.3

At3g23980 61.6

At3g14850 74.3

At3g12260b 84.9

At3g13062 100.4

Ramchiary et al. (2007) TAG

Mapping of GSL-elong, GSL-alk, GSL-pro and Myb 28 genes

in B. juncea

p58m71h154 0.0

ec3f1 7.7

p65m47v155*v 15.1

e46t65h723 26.5

p62t78v278*v 36.1

p48t94v336*v 47.7

ec2c12 58.9

p80m48v270 66.9

p33t83h171 76.7 p58m35h184 82.5

GSLelonR1/R2 84.2 e34m43v260 86.8 A75 88.3

e34m43h335 91.3

A2

GSLalkR2 0.0

e44m50h155 6.3

e34m43h360 13.1

e31m60h440 18.6

e58m44v202 27.2

p33t83v197 37.1

DGAT1 43.1

e50t66v251 49.1

e48m39v773 60.4

Dfr-R 65.0

p58m71v952 78.9

e34m62v390*h 87.7

e34m43v310 95.7

p53m47h350 102.1

e46m59v304 110.4

e48t78h132 120.5

p48t94h261 129.2

A9

e35t69h736 0.0

e31m50h174 10.8

e34m62v219 25.3

p64t66h390 34.5

FAD6.2 43.5

e51t78v450 53.9

e47t64h233 62.4

e34m43h350 71.5 e31m62v199 75.3 N17 79.8 GSLalkN1.2 80.9 N19 83.3

e31m60h223 90.7

N183 102.7

p53t76v392 110.5

e62t65h147 121.6

N89 130.6

B4

WR 0.0

p62m59h197 9.5

e62m32h183 16.4

e47m43h330 23.2

e31m58v101 33.4 e48m39v100 35.3 GSLelonR3 37.0 ec4c11 40.0

p58t68v455 47.1

A131 54.1 p63t78v400 57.2

A4

e50m32v292 0.0

p33t66v200 16.7

e32t66h147 24.5

wg3g9a 32.8

e31m50h335 40.3

e35t63h280 45.6

p64m32h258 63.9

p53m42v139 74.2

p62m59v465 82.1

p33m38v260 97.4

e44m39v89 104.0

e34m49v163 113.5 p48m67h295 115.5 GSLelonN2 117.5 N15 120.

6 e62m32v179 124.5

e44m50h105 137.2

p63t78h267 148.2

B8

e32t70v718 0.0

p61m57v318 8.1

e45m34h420*v 15.4 e31m61v99 24.0 R42.1 27.2 R187 27.9 GSLelonR4/R5

myb28a 28.4

31.6 ec2d8 34.3

wg6d6 44.4

e47t74v415 53.1 p33m32h332 58.7 e31m47h153 63.6

GSLalkR1.1 64.2 e62m32h170 67.9 e48m39v160 71.4

p53m47h173*h 81.3

A1 90.2

e39t74v736 99.1

p33m38h274*h 104.9

e34m62h290 117.0

A3

102.1

e42t70h733

e34t74h719 0.0

e39m55h425

p32m66v901

e58m40h85

91.7 p33m32h148

p53m42h188 77.3

58.0 e44m39h131

e45m34h101

8.4

16.7

24.1

41.8

B1

wg1g6b 0.0

p48t67v370 9.1

p63m50h215 18.1

p62t78v265 29.5

p53t91h252 36.1

e34m50v205 46.4

e62m33h241 55.6

N135 63.0

p87m38h176 72.6

e31m62h288 77.7

GsLelonN1 89.3

B6

Gsl-pro

myb28b

Ann.locus

At5g05920 2.4

At5g13190a 12.8

At5g53920 31.9

At1g74640a 48.5

At5g27740b 77.6

ELONG R1 90.1

At5g61970 95.4

Msl S

qm

s

At4g35530 6.9

At4g21105 18.3

At1g74640b 27.8 ELONG R4/Myb28a 29.9

At3g54670b 37.1

At3g10572b 50.6

At4g01897a 62.5 ALK R1 64.2

At2g30200a 77.5 Tsw

S

ql

Pbr

ALK R2 0.6 At5g27560 8.6

At5g63905 21.5

At2g14170a 34.2

At2g19260 44.9

At3g46560 57.9

At2g03060 68.3

At1g22860 78.5

At3g55430a 90.6

At3g62620 104.4

At2g20490b 118.0

At2g22370 129.7

At3g06483 8.8

At3g10572a 19.1

At3g20920 30.1

At5g47890 44.6 At5g27830

46.2

At5g66290 53.2

At1g58220a 55.3

At3g23980 61.6

At3g14850 74.3

At3g12260b 84.9

At3g13062 100.4

A2 A3 A9 B1

117

75

25

118

12

123

4-5

63520

ELONG R1

61970

?

27740b

74640b

ELONGR4

Myb28a

14045

05755 05590 54670b

?

Myb28b

Tagging of loci involved in glucosinolate

biosynthesis in B. juncea

?

Negative yield QTL from low GSL parent

Pyramiding the desired recombinants

A3 A2 B1 A9 Activity Schedule Identification

of desired

recombinants

2006-07

311-1 (BC5DH)

gsl3

570-497 (BC4DH) 402-23 (BC5DH) 411-166 (BC4DH)

gsl5

gsl1

gsl2 QTL-NIL

identification

and crosses

2006-07

F1

X

gsl3

Size of BC4DH and BC5DH population genotyped: 1595; No. of makers: 18

gsl1 gsl5

X

gsl2 Three loci

pyramiding 2007, Leh

gsl1/+

gsl2/+

gsl3/+

Desired

2007-08 Four loci

pyramiding

Desired gsl1/+

gsl3/+

gsl5/+

X

gsl2

Desired heterozygotes were identified

through MAS after screening 100

individuals in each cross

MAS – DH and four loci

low GSL lines DH and three loci

low GSL lines 2008-09

gsl1/gsl1, gsl2/gsl2, gsl3/gsl3 gsl1/gsl1, gsl2/gsl2, gsl3/gsl3, gsl5/gsl5

Heera

Varuna

White rust

In B. juncea

J4

At3g63420b*(3g150) 0.0 At3g54650(3g66) 2.4 At3g58500a*(3g165) At3g55440b(3C-18) 4.7 p62m59h197 5.5 At5g37480(5gAJ30k) At1g77550(174)

At4g13940*b(4C3) WR3 At5g37580(288)

6.4

At5g37630(5g59) 7.3 At5g37850(5gAJ31) 8.2 WR17 9.2 WR22 9.9 At5g41560(5gAJ35) 10.6 At1g66410(254d) At2g21150(2g135) WR26 11.8

At5g36880*(5g57v) At2g22000a(2g136) 12.5 At2g22640(2g137) 13.3 At2g24765a*(2C-9) 15.8 At2g25737(2C-10) p53m47h285 17.4 At2g28880b*(2C-15) 22.8 At1g69980(1gLP78) 25.2 e47m43h330 At2g30130*(AJ-2g11) 26.2 At2g31490a(2g59a) 29.8 At2g33040(417) At2g07440b*(2g105) At2g32520(2g61) 32.5

e31m58v101 36.4 GE-R3 At2g35790(2g66) 39.8 At2g36530c*(2C-22) 42.1 At2g37040*(ec4c11) 43.1 At2g37150*(wg4a4) 43.7 At2g40690*(wg5b2) At2g40765b(2g74b) 53.3 At1g76540(183b) 55.3 At2g42040(2g76) 56.6 p63t78v400 62.2 At2g44520(2g167) 74.9

Wh

ite

rust

Varuna-Heera

WR locus:7-17cM

TM4-Donskaja

WR locus:18-24cM

J5

Wh

ite

rust

At2g46390(2g82) 0.0

2g472.2 15.1 2g428 20.9 e47t70v258 e51t78t290 2g337 p32t77d380 27.3

2g415(b) e55t73t240 29.1 2g415(a) e50t66t380 29.6 e31m42v144 e35t63v351 e48t78v225 msa21 e34m62v89 e31m58v430 e31m61v136 e34m62v157 e31m49v336 e34m62t160 2g55(NCB)

32.6

e31m49v295 34.4 e31m50t98 43.9

p32t77v301 p32t77d310 56.9 3g14 57.5

2g385 64.5 e34m62d143 66.5 5g218 68.6 e35t63v240 70.4

NDDB / DOFCO

Farming Communities Funding and Production of Seeds

DBT Funding

ICAR

Technology Development and trials UDSC

Trials

SFCI & NSC Seed production

Collaborations

Dr. Arundhati Mukhopadhyay Prof. Akshay Pradhan

Dr. Vibha Gupta Dr. Pradeep Burma

Dr. Y. S. Sodhi Dr. Naveen Bisht

Dr. N. Arumugam Ms. Indira Sivaraman

Dr. Arun Jagannath Mr. N. Ramchiary

Mr. Jagdish Verma Dr Priya Panjabi

Mr B. S. Yadav Dr. Atika Chandra