Germplasm and Heterosis Breeding in Mustard
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
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CM
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06
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MS
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MS
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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