09th apr 2016
DESCRIPTION
carotenoid and quality proteinTRANSCRIPT
The project has been approved on Jan 15, 2013, and was initiated the same day. From that time forward, we have received, all agreed funds (not all) towards purchase of the
project. The Progress report is due for the period of 31-36th month period. According to agreed objectives, we have met the required targets, and we are proceeding with great strength and
intended goal.
Objectives To Get To The Purpose
Period of
Study
(In Months)
Achievable Targets
(Physical and Technical)
Current
Status
0-6
1. Phenotypic identification of the diverse maize germplasm for carotenoid content
2. Donor plants carrying opaque 2 will be crossed with our NSL elite lines to generate F1
progeny
√
7-12
1. Identification of donors with superior levels of carotenoids, to use in the crossing program
and transfer superior carotenoid alleles into elite lines.
2. F1 progeny for QPM will be backcrossed with elite recurrent parent to generate BC1F1
√
13-18
1. Standard PCR based cloning techniques, genome walking, sequencing, and data analysis
using bio-informatics tools
2. BC1F1 will be self-pollinated to produce BC1F2 and the same will undergo for foreground
selection for opaque 2 using gene specific markers
3. Lines carrying opaque 2 will be screened for background selection using recurrent parent √
Stacking Traits….Along with Profit.
13-183. Lines carrying opaque 2 will be screened for background selection using recurrent parent
specific SSR markers.
4. Donor plants carrying superior carotenoid alleles will be crossed with elite lines to
generate F1 progeny
√
19-24
1. F1 progeny carrying superior carotenoid alleles will be backcrossed with elite recurrent
parent to generate BC1F1, while performing biochemical and molecular analysis.
2. BC1F2 carrying opaque2 will be self-pollinated to generate BC1F3 and will undergo for
foreground and background selection
√
25-30
1. BC1F1 carrying superior carotenoid alleles will be self-pollinated, to generate BC1F2, while
performing biochemical and molecular analysis.
2. BC1F3 carrying opaque2 will be self-pollinated to generate BC1F4 and will undergo for
foreground and background selection
√
31-36
1. BC1F4 carrying opaque2 will be self-pollinated to generate BC1F5 and will undergo for
foreground and background selection
2. BC1F2 carrying superior carotenoid alleles will be self-pollinated, to generate BC1F3, while
performing biochemical and molecular analysis.
In
Process
37-42
1. BC1F5 carrying opaque2 will be self-pollinated to generate BC1F6 (QPM Fixed elite line)
and will undergo for foreground and background selection
2. BC1F3 carrying superior carotenoid alleles will be self-pollinated, to generate BC1F4, while
performing biochemical and molecular analysis.
In
Process
43-481. Fixed QPM elite line will be crossed with BC1F4, carrying superior carotenoid alleles, to
generate F1 carrying both traits of interest.
49-54 1. F1 progeny will be self-pollinated to generate F2
55-60
1. F2 population seed will be subjected to both morphological, biochemical, and molecular
analysis for both traits of interest.
2. Selected lines will be further self-pollinated and continued to fix the traits.
and was initiated the same day. From that time forward, we have received, all agreed funds (not all) towards purchase of the intended equipment, consumables related to
month period. According to agreed objectives, we have met the required targets, and we are proceeding with great strength and full speed towards achieving the
Objectives To Get To The Purpose
Current
Status
√
√
√
Stacking Traits….Along with Profit.
Uniq
ue S
elli
ng P
oin
t (U
SP
)
and S
ocia
l
sib
ility
√
√
√
In
Process
In
Process
Uniq
ue S
elli
ng P
oin
t (U
SP
)
Com
merc
ial V
entu
reand S
ocia
l
Corp
ora
te r
esponsib
Back Cross scheme, using conventional Genetic component Genetic System Conventional breeding
o2 The recessive mutant allele of the o2 gene Phenotypic observation in F2
Hardness modifiers The endosperm hardness modifier genes/QTL Through light box screening
Amino acid modifiers The amino acid modifier genes/QTL influencing amino acid content in the endosperm
Through estimation of Lysine and Tryptophan
SELFING
PopulationPopulation
SELFING
X
Foreground selection and limited
background selection
Determination of optimum population size
for BC2
Foreground selection and limited
background selection
Determination of optimum population size
for BC2
Progeny
F1
Elite LineElite LineX
BC1F1
Population
Foreground selection
background selection of recurrent parent
Identification of individuals with highest
recurrent genome content
Foreground selection
background selection of recurrent parent
Identification of individuals with highest
recurrent genome content
PopulationBC1F2
SELFING
al and MAS to convert normal to QPM lines Conventional breeding Molecular breeding Current Status
Phenotypic observation in F2 Using SSRs with in o2 in BC and F2
Standardized the method is currently in use
Through light box screening Markers??? Standardized the way and is currently in use
Through estimation of Lysine and Tryptophan
Markers??? In progress
SELFING
SELFING
PopulationBC1F3
Population
SELFING
Select agronomicallyand biochemically
superior BC2F3 line
BC1F4 Population
Select agronomicallyand biochemically
superior BC2F3 line
--
Foreground selection and limited
background selection
Determination of optimum population size
Foreground selection and limited
background selection
Determination of optimum population size
Line
Foreground selection
Phenotypic selection for improved
agronomic characters
Phenotypic selection for kernel
modification and tryptophan conc.
Foreground selection
Phenotypic selection for improved
agronomic characters
Phenotypic selection for kernel
modification and tryptophan conc.
Foreground selection
background selection of recurrent parent
Identification of individuals with highest
recurrent genome content
Foreground selection
background selection of recurrent parent
Identification of individuals with highest
recurrent genome content
PopulationQPM version
of elite line
Foreground Selection of BC1F2 progenies for
P1 P2 O O O O H H O H H H H H O N
N H N O H N N N
P1 P2 H H H H O O H H H H H O H N
H O H H O H N N
P1 P2 O H H H O H H H H H H O H H
H O H H H H H O
P P H H H N N N N H N N H O H H P1 P2 H H H N N N N H N N H O H H
N N H O N N O N
P1 P2 O N O N N N N H H H H H H H
O H O N N H H O
P1 P2 N H H O H H H H H H H O O N
H H O H O H O H
P1 P2 H H H O O H O H H H O H O H
H H N H O O O
Foreground Selection of BC1F2 progenies for opaque2 allele
O O O O H H O H H H H H O N
H H H H O O H H H H H O H N
H H H N N N N H N N H O H H H H H N N N N H N N H O H H
O N O N N N N H H H H H H H
N H H O H H H H H H H O O N
H H H O O H O H H H O H O H
Elite parents selected for the
work
3001 3003 M183
1 O2o2 [Hetero] + Elite type 46 40 22
2 o2o2only 24 35 11
3 O2o2 [Hetero] only 66 20 18
4 Normal 40 45 50
III III
Hardness of endosperm - Scoring of Endosper
IV V
A212 B114
Type (Modification Score) I: Not opaque
Type (Modification Score) II: 25% opaque
Type (Modification Score) III: 50% opaque
Type (Modification Score) IV: 75% opaque
erm modification through conventional light box
G270
Type (Modification Score) IV: 75% opaque
Type (Modification Score) V: 100% opaque
Estimation of Tryptophan in maize donor parent seed and recurrent parent seed
Donors Elite Cultures
BC1F3 (Bulk)
P
Estimation of Tryptophan in maize donor parent seed and recurrent parent seed
Elite Cultures
BC1F3 (Bulk)
P
A
A
A
B
B
1.01
1.02
1.03
1.04
1.05
1.06
1.0
2.01
2.02
2.03
2.04
2.05
2.0
bnlg1179
bnlg1007
bnlg1203
bnlg1811
umc2083
umc1035
umc1335
bnlg1092
bnlg1017
mmc0111
bnlg2248
mmc0231
bnlg1175
phi049
bnlg1144
bnlg1325
bnlg1447
bnlg1113
B
G
G
B
B1.07
1.09
1.08
1.10
1.11
1.12
2.05
2.06
2.07
2.08
2.09
2.10
bnlg1629
phi037
bnlg1502
umc1802
umc1737
bnlg1055
phi120
umc1605
bnlg1036
bnlg1887
bnlg2077
bnlg2144
bnlg1267
bnlg1746
mmc0381
phi1099
bnlg1904
bnlg2136
bnlg505
bnlg2242
dupssr23
bnlg1605
umc1148
bnlg1182
1
23
3.0
3.01
3.02
3.03
3.04
4.01
4.0
4.02
4.03
4.04
5.01
5.02
5.0
5.03
5.04
7.0
7.01
7.02
9.0
9.02
9.03
9.01
10.0
10.01
10.02
phi049
bnlg1144
bnlg1325
bnlg1447
bnlg1113
umc2148
bnlg1126
bnlg1162
bnlg1741
bnlg1265
mmc0321
bnlg1066
mmc0121
bnlg105
bnlg1879
bnlg1046
bnlg1902
bnlg2323
6.0
6.01
bnlg1043
bnlg1600
bnlg1432
dupssr18
bnlg1642
bnlg2132
bnlg1200
bnlg10708.01
8.0bnlg1194
bnlg2235
bnlg2289
bnlg1272
bnlg1288
bnlg1401
dupssr19
umc1191
bnlg1688
bnlg1451
mmc0501
bnlg1655
umc1863
3.05
3.06
3.07
3.08
3.09
4.05
4.06
4.07
4.08
4.09
4.11
4.10
5.05
5.06
5.07
5.08
5.09
7.03
7.04
7.05
7.06
9.04
9.06
9.07
9.08
9.05
10.03
10.04
10.05
10.06
10.07
bnlg1904
bnlg2136
bnlg505
bnlg2242
dupssr23
bnlg1605
umc1148
bnlg1182
bnlg1137
bnlg2291
dupssr34
bnlg1444
bnlg2244
dupssr28
umc1834
bnlg1237
mmc0282
mmc0481
bnlg1306
bnlg1711
6.02
6.03
6.04
6.05
6.06
6.07
6.08
dupssr18
umc1083
umc2316
bnlg1154
bnlg1617
bnlg2249
bnlg345
bnlg1740
umc2165
umc1001
bnlg1666
umc1799
phi057
phi116
8.01
8.02
8.03
8.04
8.05
8.06
8.07
8.08
8.09
umc1415
bnlg1176
bnlg1146
bnlg1031
bnlg1828
bnlg1056
dupssr14
bnlg1688
bnlg1270
umc2346
bnlg1129
bnlg1588
umc1077
bnlg1185
bnlg2190
bnlg1480
3 4
5
6
7
8
9
10
Total Markers screened:200Best Markers: 77
77 are being used to recover the
recurrent genome
Elite QPM Het. Elite QPM Het.
4 60 27 14 30 72 10 17
24 52 26 22 23 72 15 13
11 54 24 24 17 70 14 16
6 61 23 16 12 67 15.4 17.6
1 56.6 21.1 22.3 27 63 20 17
9 57 18 24 2 62.3 12 25.7
32 54 17 29 26 74 10 16
13 58.1 16.2 25.7 28 73 12 16
25 33.7 28.9 37.4 22 71 13 17
21 57 24 18 31 70 7.9 22
20 56 21 23 5 69 12 19
10 58 21 21 19 69 15 16
27 63 20 17 8 68 14 19
33 67 17 16 33 67 17 16
18 54.3 17.2 28.5 15 63 13 24
3 54.95 17.03 17.02 16 63 14 23
QP
M P
lan
ts i
n L
OT
-A
Plant
No.
%of genome
ELI
TE
Pla
nts
in
LO
T-A
Plant
No.
%of genome
Selection of Best Elite and QPM counterparts for [F1] Hybridization program (pre
Elite QPM Het. Elite QPM Het.
2 37.7 23.4 38.9 1 70.0 9.4 20.6
3 54.2 16.7 29.2 5 61.0 15.7 23.3
4 57.7 20.8 21.4 12 58.6 16.0 25.3
7 47.7 20.1 32.2 13 58.4 13.7 28.0
8 42.3 23.3 34.4 9 57.6 18.8 23.6
9 57.6 18.8 23.6 14 43.6 13.3 43.0
12 58.6 16 25.3 17 53.0 16.7 30.4
15 45 25.4 29.6 10 75.4 15.0 9.6
16 50 18.7 31.3 19 74.6 10.1 15.4
17 53 16.7 30.4 18 64.3 18.5 17.3
18 64.3 18.5 17.3 4 57.7 20.8 21.4
20 50.6 16.3 33.1 3 54.2 16.7 29.2
21 53.6 12.7 33.7
Plant
No.
%of genome
QP
M P
lan
ts i
n L
OT
-G
Plant
No.
%of genome
ELI
TE
Pla
nts
in
LO
T-G
Elite QPM Het. Elite QPM Het.
17 17 70 14 27 73 11 25
1 14 68 18 4 63 18 27
10 18 67 17 8 60 12 36
23 61 26 17 3 59 20 30
7 53 23 30 18 57 14 29
16 46 21 35 7 53 23 30
3 59 20 30 19 54 15 40
5 52 19 36 24 57 13 26
9 48 18 36 21 46 15 41
6 46 33 26 29 58 12 31
28 50 32 26 23 61 26 17
2 41 30 35 22 61 18 28
15 55 28 26 15 55 28 26
12 44 24 39 11 55 18 35
13 45 24 33 14 55 21 23
14 55 21 33 5 52 19 36
QP
M P
lan
ts i
n L
OT
-B
Plant
No.
%of genome
ELI
TE
Pla
nts
in
LO
T-B
Plant
No.
%of genome
Selection of Best Elite and QPM counterparts for [F1] Hybridization program (pre-commercial activity)
Elite QPM Het. Elite QPM Het.
27 42.7 25.2 32.1 1 63.6 20.7 15.7
5 50.7 22.9 26.4 2 61.6 13.7 24.7
10 52.1 20.8 27.1 3 67.4 5.9 26.7
1 63.6 20.7 15.7 6 58.1 10.1 31.8
26 57.0 18.3 24.6 7 68.7 6.8 24.5
14 58.1 18.2 23.6 9 65.5 9.5 25.0
22 55.8 15.9 28.3 11 63.1 14.1 22.8
13 54.5 14.5 31.0 12 58.8 8.8 32.4
19 57.9 14.5 27.6 15 67.2 10.2 22.6
11 63.1 14.1 22.8 16 73.3 10.4 16.3
16 73.3 10.4 16.3 17 62.1 13.1 24.8
23 60.2 26.6 13.3 18 62.2 10.4 27.4
2 61.6 13.7 24.7 20 71.2 9.4 19.4
17 62.1 13.1 24.8 21 78.8 6.8 14.4
28 58.7 13.0 28.3 23 60.2 26.6 13.3
24 54.7 12.8 32.4 25 67.1 7.7 25.2
18 62.2 10.4 27.4 28 58.7 13.0 28.3
15 67.2 10.2 22.6 29 66.7 8.3 25.0
Plant
No.
%of genome
QP
M P
lan
ts i
n L
OT
-F
Plant
No.
%of genome
ELI
TE
Pla
nts
in
LO
T-F
1 3 6 10 14 18 32 5 7 8 11 12 17 22 23 26 31 33
3 X X X X
7
8
18 X X X
20
21 X X X X X X
23 X X
24 II
29
2 X X
5 X X
6 X
LOT A QPM LINES
LOT
B L
ine
s
LOT
F L
ine
s
Cross Hybridization program (pre-commercial activity) keeping quality and quantity in harmony
6 X
9 II X X X X
12 X X X X X II
13 X X
15 X II X X X X
16 X X X X X X X X
22 X
25
26 II X X X X X
28 X X X
Selected for Elite line based on genome recovery data
Selected for QPM line based on genomerecovery data
selected for both Elite & QPM nature based on genome recovery data
LOT
B L
ine
s
LOT
F L
ine
s
1 2 4 5 6 7 8 10 10 11 11 12 13 13 13 14 15 16 16 17 17 18 19 19 21 21
1 X X X X X X X X X
2 X X X X
3 II X X X X X X
4 X X X X X X
5 X X X X X X X X X
6 X X X X II X X X X
7 X X X X X X X X X X
8 X X X X X X X X X
9 X X X X X X X X X X X X
10 X X X X X X II X X X X
11 X X X II X
12 X II II II X II II X X X X
13 X X X
14 X X X X X X X X
15 X X
LOT G QPM LINES
LOT
F L
ine
s
commercial activity) keeping quality and quantity in harmony
16 X X X X
17 X X X
18 X
19 X X X X X X X X X
20 X X X X X X X
21 X X X X X X X X X
22 X X X X X X
23 X X X
24 X X X X X X X X
25 X X X X
26 X X X X X X X X X
27 X X X X X X
28 X X X X
29 X X X X X
Selected for Elite line based on genome recovery data
Selected for QPM line based on genomerecovery data
selected for Elite & QPM both natures based on genome recovery data
LOT
F L
ine
s
TSS
ATG
MGLSGATISAPLGCCVLRCGAVGGGKALKADAERWRRAGWSRRVGGPKVRCVATEKHDETA
AVGAAVGVDFADEEDYRKGGGGELLYVQMQSTKPMESQSKIASKLSPISDENTVLDLVIIGCGP
AGLSLASESAKKGLTVGLIGPDLPFTNNYGVWEDEFKDLGLESCIEHVWKDTIVYLDNNKPILIG
RSYGRVHRDLLHEELLKRCYEAGVTYLNSKVDKIIESPDGHRVVCCDKGREIICRLAIVASGAAS
GRLLEYEVGGPRVCVQTAYGVEVEVENNPYDPSLMVFMDYRDCFKEEFSHTEQENPTFLYAM
PMSPTRVFFEETCLASKDAMSFDLLKKRLMYRLNAMGIRILKVYEEEWSYIPVGGSLPNTDQK
NLAFGAAASMVHPATGYSVVRSLSEAPRYASVISDILGNRVPAEYMLGNSQNYSPSMLAWRTL
WPQERKRQRSFFLFGLALIIQLNNEGIQTFFEAFFRVPRWMWRGFLGSTLSSVDLILFSFYMFAI
APNQLRMNLVRHLLSDPTGSSMIKTYLTL
Zeamays
Complete
Introduction of carotenoid superior alleles controlling the flux towards
AU
0.000
0.010
0.020
Minutes
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00
MAAAMTSFVAKNPLLAAAARRRAPPLAGRALPFSPLASTRAPRRTVTCFVPQDTAAPAAPVPALDEE
ARAAAARRVAEKEARKRSERRTYLVAAVMSSLGVTSMAVAAVYYRFSWQMEGGEVPVIETLGTFAL
SVGAAVGMEFWARWAHRALWHASLWHMHESHHRPREGPFELNDVFAIVNAAPAISLLAYGFFHRG
IVPGLCFGAGLGITLFGMAYMFVHDGLVHRRFPVGPIADVPYFRRVAASHKIHHMDKFGGVPYGLFL
GPKELEEVGGLDELVSSPVSEATDTEDAGEEKTRPVVCVVRTSVFMGQSVPNEF
ATG
Zeamays
Complete
High β-carotene accumulating donor maize line
E1 E3E2 E4 E5 E7E6 E8 E9 E10 E11 E12 E13
354 60 207 126 57 156
181 88 152 197 98TAG
MGLSGATISAPLGCCVLRCGAVGGGKALKADAERWRRAGWSRRVGGPKVRCVATEKHDETA
AVGAAVGVDFADEEDYRKGGGGELLYVQMQSTKPMESQSKIASKLSPISDENTVLDLVIIGCGP
AGLSLASESAKKGLTVGLIGPDLPFTNNYGVWEDEFKDLGLESCIEHVWKDTIVYLDNNKPILIG
RSYGRVHRDLLHEELLKRCYEAGVTYLNSKVDKIIESPDGHRVVCCDKGREIICRLAIVASGAAS
GRLLEYEVGGPRVCVQTAYGVEVEVENNPYDPSLMVFMDYRDCFKEEFSHTEQENPTFLYAM
PMSPTRVFFEETCLASKDAMSFDLLKKRLMYRLNAMGIRILKVYEEEWSYIPVGGSLPNTDQK
NLAFGAAASMVHPATGYSVVRSLSEAPRYASVISDILGNRVPAEYMLGNSQNYSPSMLAWRTL
WPQERKRQRSFFLFGLALIIQLNNEGIQTFFEAFFRVPRWMWRGFLGSTLSSVDLILFSFYMFAI
APNQLRMNLVRHLLSDPTGSSMIKTYLTL
LCYE Gene Structure 5393bp (1778bp)
Zeamays ε–Lycopene cyclase (lcyE) mRNA, lcyEW22 allele
Complete cds GenBank EU924262.1 (1778bp; 537aa)
Introduction of carotenoid superior alleles controlling the flux towards β-carotene
24.00 26.00 28.00 30.00 32.00 34.00 36.00 38.00 40.00 42.00 44.00 5732
MAAAMTSFVAKNPLLAAAARRRAPPLAGRALPFSPLASTRAPRRTVTCFVPQDTAAPAAPVPALDEE
ARAAAARRVAEKEARKRSERRTYLVAAVMSSLGVTSMAVAAVYYRFSWQMEGGEVPVIETLGTFAL
SVGAAVGMEFWARWAHRALWHASLWHMHESHHRPREGPFELNDVFAIVNAAPAISLLAYGFFHRG
IVPGLCFGAGLGITLFGMAYMFVHDGLVHRRFPVGPIADVPYFRRVAASHKIHHMDKFGGVPYGLFL
GPKELEEVGGLDELVSSPVSEATDTEDAGEEKTRPVVCVVRTSVFMGQSVPNEF
TAG
HYD3 gene structure 4990bp (1246)
Zeamays β-carotene hydroxylase (HYD3) mRNA , B73 allele
Complete cds; GenBank: AY844958 1246bp (319aa)
carotene accumulating donor maize line
AU
0.000
0.010
0.020
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.003001
AU
0.010
0.020
0.030
0.040
3003
To Transform NSL Corn Crop Commercial lines with Unique Selling Proposition (USP)
0.000
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.003003
AU
0.00
0.05
0.10
0.15
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00M183
Lutein Zeaxanthin β-carotene
NSL Corn Crop Commercial lines used in its NMH Hybrid breeding program
Minutes
20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00 36.00 38.00 40.00 42.00 44.00
To Transform NSL Corn Crop Commercial lines with Unique Selling Proposition (USP)
Minutes
20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00 36.00 38.00 40.00 42.00 44.00
Minutes
20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00 36.00 38.00 40.00 42.00 44.00
carotene
NSL Corn Crop Commercial lines used in its NMH Hybrid breeding program
D D D D O R R R R R R R R R
Details of ε–LCY gene introgression for Marker
D D D D O R R R R R R R R R
NQMPL's 5730 5732 5723 5721 5734 3001 3003 5896
Polymorphism D D D D O R R R
A 5' indel 2 2 2 2 3 2 - 2
B 3' indel 8 8 8 0 0 0 0 0
C 5' indel 1 2 2 2 1 1 1 1
D 3' indel 1 1 1 1 3 2 4 1
E DEL4 0 12 12 12 0 0 0 0
Class
B: 8= 8bp insertion (140bp); 0=132bp
LCY
HYD
Gene
A: 2= 250bp; 3=250 + 380bp C: 1=800bp; 2=600bp
B
AA
LCY gene introgression for Marker-Assisted Breeding
F2 R2250bp(A)
LCYE 5’ indel (2)
F2 R2
LCYE 5’ indel (3)
F1
D D D D O R R R R R R R R R
250bp + 380bp(A )
8
140bp(B)
LCYE 3’ indel (8)
R2F2
5891 5881 M199 M183 M111 M184
R R R R R R
- - 2 2 2 - 2 STS-Indel Agarose 2.5
0 0 0 8 0 0 8 STS-Indel Agarose 3
1 1 1 1 1 1 2 STS-Indel Agarose 2.5
2 1 2 3 2 2 1 STS-Indel Agarose 2.5
0 0 0 0 0 0 12 STS-Indel Agarose 3
D: 1= 543bp; 2 = 296bp; 3 = 296 + 1221bp; 4= 296 +875bp
Marker Type PCR-Assay Gel %Favourable
Alleles
E: 0= 117bp; 12=12bp insertion (129bp)
C
D D D D O R R R R R R R R R
D
D D D D O R R R R R R R R R
Details of β–HYD3 gene introgression for Marker
D D D D O R R R R R R R R R
ENon Specific Bands
D D D D O R R R R R R R R R
NQMPL's 5730 5732 5723 5721 5734 3001 3003 5896
Polymorphism D D D D O R R R
A 5' indel 2 2 2 2 3 2 - 2
B 3' indel 8 8 8 0 0 0 0 0
C 5' indel 1 2 2 2 1 1 1 1
D 3' indel 1 1 1 1 3 2 4 1
E DEL4 0 12 12 12 0 0 0 0
Class
B: 8= 8bp insertion (140bp); 0=132bp
LCY
HYD
Gene
A: 2= 250bp; 3=250 + 380bp C: 1=800bp; 2=600bp
C
D
HYD3 gene introgression for Marker-Assisted Breeding
H1 H2
HYD3 5’ indel(C)
(397bp insertion)
800bp(1)
(206bp insertion)
H1 H2
600bp(2)
HYD3 3’ indel (D)
L1 R2
190bp 350bp1
L1 R2
325bp4
1250bp
E
5891 5881 M199 M183 M111 M184
R R R R R R
- - 2 2 2 - 2 STS-Indel Agarose 2.5
0 0 0 8 0 0 8 STS-Indel Agarose 3
1 1 1 1 1 1 2 STS-Indel Agarose 2.5
2 1 2 3 2 2 1 STS-Indel Agarose 2.5
0 0 0 0 0 0 12 STS-Indel Agarose 3
D: 1= 543bp; 2 = 296bp; 3 = 296 + 1221bp; 4= 296 +875bp
Marker Type PCR-Assay Gel %Favourable
Alleles
E: 0= 117bp; 12=12bp insertion (129bp)
L1 R2
L1 R2R1
1250bp3
D4F D4R
12bp insertion
I
D4F D4R
D
HYD3 DEL 4 (E)
Is it possible to increase more
Introgress β-carotene accumulating lines into High AU
0.00
0.02
0.04
0.06
0.08
0.10
0.12
0.14
0.16
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00
0.08
0.10
3876
3113
AU
0.00
0.02
0.04
0.06
0.08
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00
AU
0.000
0.010
0.020
0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00
3113
5732
Line Development - NSL Corn Crop Commercial lines used in its NMH Hybrid breeding program
Is it possible to increase more β-carotene?
carotene accumulating lines into High Zeaxanthin maize lines
Minutes
20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00 36.00 38.00 40.00 42.00 44.00
Minutes
20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00 36.00 38.00 40.00 42.00 44.00
Minutes
20.00 22.00 24.00 26.00 28.00 30.00 32.00 34.00 36.00 38.00 40.00 42.00 44.00
NSL Corn Crop Commercial lines used in its NMH Hybrid breeding program
Hyd5'TE Hyd3'TE Del4 Lcye3'TE
HOMO 47 34.0 0.0 34.0 31.9
HETERO 47 17.0 0.0 17.0 12.8
HOMO 44 0.0 22.7 0.0 31.8
HETERO 44 0.0 13.6 0.0 4.5
HOMO 41 29.3 14.6 26.8 31.7
HETERO 41 19.5 29.3 22.0 17.1
HOMO 46 0.0 0.0 0.0 41.3
HETERO 46 0.0 0.0 0.0 0.0
HOMO 40 0.0 2.5 0.0 32.5
HETERO 40 0.0 40.0 0.0 27.5
HOMO 40 0.0 37.5 0.0 32.5
HETERO 40 0.0 20.0 0.0 27.5
HOMO 49 20.4 29.4 12.2 24.5
HETERO 49 34.7 28.6 42.9 28.6
HOMO 38 28.9 23.7 31.6 23.7
HETERO 38 7.9 10.5 10.5 13.2
HOMO 45 31.1 0.0 31.1 22.2
1107X5732
1077X5730
1065X5723
1112X5716
1079X5730
1082X5730
S.NoRabi 2016
field code
1
2
3
4
5
6
7
8
Total%
nature
R1516-139
TO 141
R1516-172
TO 174
R1516-181
R1516-182
TO 185
cross
R1516-134
R1516-186
1120X5732
1064X5723
R1516-215
TO 218
R1516-203
R1516-135
Elite Line development with carotenoid superior
HOMO 45 31.1 0.0 31.1 22.2
HETERO 45 8.9 0.0 8.9 22.2
HOMO 43 0.0 7.0 0.0 20.9
HETERO 43 0.0 32.6 0.0 25.6
HOMO 42 0.0 23.8 0.0 16.7
HETERO 42 0.0 9.5 0.0 16.7
HOMO 48 16.7 4.2 25.0 22.9
HETERO 48 20.8 29.2 4.2 12.5
HOMO 40 32.5 25.0 25.0 25.0
HETERO 40 10.0 12.5 20.0 12.5
HOMO 50 18.0 22.2 16.0 24.0
HETERO 50 10.0 40.0 12.0 10.0
HOMO 42 0.0 14.3 0.0 23.8
HETERO 42 0.0 4.8 0.0 9.5
HOMO 24 0.0 20.8 0.0 29.2
HETERO 24 0.0 12.5 0.0 8.3
HOMO 49 0.0 0.0 22.4 24.5
HETERO 49 0.0 0.0 12.2 8.2
HOMO 49 16.3 16.3 16.3 28.6
HETERO 49 20.4 20.4 20.4 8.2
HOMO 47 0.0 6.4 0.0 21.3
HETERO 47 0.0 17.0 0.0 6.4
HOMO 50 0.0 14.0 0.0 26.0
HETERO 50 0.0 6.0 0.0 6.0
1075x5730
1111X5732
1094X5723
1090X5730
1076X5730
1091X5730
1080X5730
1071X5732
1066X5723
1162X5723
1081X5730
16
17
18
19
20
10
11
12
13
14
15
9
R1516-136
TO 137
R1516-152
TO 154
R1516-155
TO 156
R1516-170
TO 171
R1516-175
TO 178
R1516-138
R1516-187
TO 190
1106X5723
R1516-211
R1516-203
TO 205
R1516-192
TO 196
R1516-131
R1516-132
TO 133
Hyd5'TE Hyd3'TE Del4 Lcye3'TE
HOMO 40 0.0 17.5 0.0 20.0
HETERO 40 0.0 12.5 0.0 5.0
HOMO 45 0.0 13.3 0.0 0.0
HETERO 45 0.0 24.4 0.0 0.0
HOMO 45 0.0 22.2 0.0 0.0
HETERO 45 0.0 0.0 0.0 0.0
HOMO 50 0.0 18.0 0.0 0.0
HETERO 50 0.0 14.0 0.0 0.0
HOMO 44 0.0 18.2 0.0 0.0
HETERO 44 0.0 0.0 0.0 0.0
HOMO 50 0.0 0.0 0.0 26.0
HETERO 50 0.0 0.0 0.0 0.0
HOMO 45 0.0 13.3 26.7 0.0
HETERO 45 0.0 46.7 37.8 0.0
HOMO 50 0.0 16.0 0.0 0.0
HETERO 50 0.0 44.0 0.0 0.0
HOMHO 48 0.0 18.8 16.7 0.0
%S.No
Rabi 2016
field cross nature Total
1161X5730
1084X5730
1086X5730
1087X619
1088X5730
1092X5730
1069X5732
1089X5730
28
22
23
24
25
26
27
21
R1516-
142
R1516-
143 TO
R1516-
145 TO
R1516-
148
R1516-
R1516-
157 TO
R1516-
163 TO
R1516-
165 TO
R1516-
149 TO
rior alleles for back crossing into QPM fixed lines
HOMHO 48 0.0 18.8 16.7 0.0
HETERO 48 0.0 27.1 25.0 0.0
HOMO 49 0.0 0.0 0.0 18.4
HETERO 49 0.0 0.0 0.0 0.0
HOMO 46 21.7 19.6 21.7 0.0
HETERO 46 2.2 2.2 2.2 0.0
HOMO 47 0.0 0.0 0.0 27.7
HETERO 47 0.0 0.0 0.0 0.0
HOMO 40 32.5 21.5 22.5 0.0
HETERO 40 35.0 42.9 47.5 0.0
HOMO 48 35.4 31.3 33.3 0.0
HETERO 48 0.0 2.1 2.1 0.0
HOMO 50 26.0 26.0 26.0 0.0
HETERO 50 0.0 0.0 0.0 0.0
HOMO 46 32.6 21.7 26.1 0.0
HETERO 46 0.0 8.7 6.5 0.0
HOMO 37 13.5 13.5 13.5 0.0
HETERO 37 0.0 0.0 0.0 0.0
HOMO 44 0.0 0.0 31.8 0.0
HETERO 44 0.0 0.0 0.0 0.0
HOMO 53 32.1 9.4 30.2 0.0
HETERO 53 1.9 7.5 3.8 0.0
HOMO 43 30.2 0.0 25.6 0.0
HETERO 43 2.3 0.0 2.3 0.01113X5721
1164X5723
1100X5721
1101X5721
1102X5721
1103X5721
1109X5721
1098X5723
1095X5723
1097X5723
1121X5723
1123X5723
40
34
35
36
37
38
39
29
30
31
32
33
R1516-
197 TO
R1516-
199
R1516-
200 TO
R1516-
222 TO
R1516-
224 TO
R1516-
227 TO
R1516-
206 TO
R1516-
208 TO
R1516-
212 TO
R1516-
230 TO
R1516-
234 TO
R1516-
219 TO
(Part -1)
Hyd5'TE Hyd3'TE Del4 Lcye3'TE
homo 48 37.5 29.2 27.1 29.2
hetero 48 8.3 12.5 14.6 14.6
homo 48 0.0 22.9 0.0 29.2
hetero 48 0.0 10.4 0.0 12.5
homo 46 0.0 21.7 0.0 21.7
hetero 46 0.0 4.3 0.0 6.5
homo 48 0.0 0.0 0.0 29.2
hetero 48 0.0 0.0 0.0 0.0
homo 42 0.0 0.0 0.0 19.0
hetero 42 0.0 0.0 0.0 0.0
homo 48 0.0 0.0 0.0 18.8
hetero 48 0.0 0.0 0.0 0.0
homo 43 0.0 0.0 0.0 27.9
hetero 43 0.0 0.0 0.0 0.0
homo 38 34.2 34.2 34.2 26.3
hetero 38 7.9 7.9 7.9 15.8
homo 49 0.0 0.0 0.0 26.5
S.NoRabi 2016
field codecross nature Total
%
49R1516-300
M184X5730
47 R1516-299 5884X5730
48R1516-344
TO 34810602X5723
45R1516-267
TO 2685881X5730
46 R1516-266 5891X5730
43R1516-251
TO 2521017x5730
44R1516-248
TO 2505883X5716
41R1516-242
TO 2445883x5732
42R1516-240
TO 2415883x5730
Elite Line development with carotenoid superior
homo 49 0.0 0.0 0.0 26.5
hetero 49 0.0 0.0 0.0 0.0
homo 44 0.0 0.0 0.0 25.0
hetero 44 0.0 0.0 0.0 0.0
homo 21 9.5 0.0 9.5 14.3
hetero 21 9.5 0.0 4.8 9.5
homo 22 45.5 31.8 40.9 27.3
hetero 22 9.1 13.6 13.6 31.8
homo 19 31.6 31.6 31.6 31.6
hetero 19 5.3 5.3 5.3 15.8
homo 38 21.1 13.2 13.2 28.9
hetero 38 13.2 18.4 18.4 5.3
homo 47 0.0 0.0 12.8 0.0
hetero 47 0.0 0.0 0.0 0.0
homo 49 0.0 0.0 28.6 22.4
hetero 49 0.0 0.0 8.2 6.1
homo 47 0.0 0.0 23.4 0.0
hetero 47 0.0 0.0 0.0 0.0
homo 50 0.0 0.0 12.0 22.0
hetero 50 0.0 0.0 10.0 6.0
homo 49 0.0 0.0 26.5 30.6
hetero 49 0.0 0.0 10.2 10.2
homo 37 0.0 0.0 0.0 27.0
hetero 37 0.0 0.0 0.0 0.0
59R1516-269
TO 2735881X5732
60R1516-351
TO 35210590X5716
57R1516-311
TO 3135894X5732
58R1516-314
TO 31610589X5732
55 R1516-310 M114X5732
56R1516-285
TO 287M111X5732
53R1516-289
TO 291M111X5723
54 R1516-343 10603X5723
51R1516-274
TO 2755881X5723
52R1516-253
TO 2561017X5723
49R1516-300
TO 301M184X5730
50R1516-295
TO 298M114X5730
Hyd5'TE Hyd3'TE Del4 Lcye3'TE
homo 45 0.0 0.0 0.0 22.2
hetero 45 0.0 0.0 0.0 0.0
homo 36 0.0 0.0 0.0 16.7
hetero 36 0.0 0.0 0.0 0.0
homo 31 0.0 0.0 0.0 16.1
hetero 31 0.0 0.0 0.0 0.0
homo 49 0.0 12.2 16.3 0.0
hetero 49 0.0 0.0 0.0 0.0
homo 40 0.0 0.0 0.0 17.5
hetero 40 0.0 0.0 0.0 0.0
homo 46 0.0 23.9 23.9 21.7
hetero 46 0.0 10.9 10.9 8.7
homo 45 17.8 22.2 20.0 0.0
hetero 45 0.0 0.0 2.2 0.0
homo 45 15.6 17.8 26.7 33.3
hetero 45 24.4 13.3 17.8 13.3
homo 43 0.0 14.0 20.9 20.9
S.NoRabi 2016
field cross
69R1516-
10590X5732
67R1516-
329 TO 10594X5732
68R1516-
325 TO 10603X5732
65R1516-
353 TO 10603X5716
66R1516-
337 TO 10608X5732
63R1516-
292 TO M111X5716
64R1516-
31710604X5732
61R1516-
2765881X5716
62R1516-
263 TO 5896X5716
nature Total%
rior alleles for back crossing into QPM fixed lines
homo 43 0.0 14.0 20.9 20.9
hetero 43 0.0 11.6 11.6 14.0
homo 43 0.0 11.6 0.0 34.9
hetero 43 0.0 2.3 0.0 7.0
homo 50 0.0 8.0 10.0 0.0
hetero 50 0.0 0.0 2.0 0.0
homo 43 0.0 20.9 0.0 27.9
hetero 43 0.0 16.3 0.0 11.6
homo 43 0.0 16.3 0.0 0.0
hetero 43 0.0 0.0 0.0 0.0
homo 41 0.0 12.2 17.1 0.0
hetero 41 0.0 0.0 0.0 0.0
homo 40 15.0 15.0 17.5 20.0
hetero 40 5.0 5.0 2.5 7.5
homo 48 18.8 14.6 18.8 31.3
hetero 48 10.4 0.0 10.4 8.3
homo 47 0.0 0.0 0.0 29.8
hetro 47 0.0 0.0 0.0 0.0
homo 46 0.0 0.0 0.0 19.6
hetero 46 0.0 0.0 0.0 0.0
homo 48 0.0 16.7 22.9 8.3
hetero 48 0.0 4.2 4.2 16.7
homo 48 0.0 16.7 0.0 0.0
hetero 48 0.0 0.0 0.0 0.0
79R1516-
3496192X5723
80R1516-
3506188 X 5723
77R1516-
357 TO 10601X5716
78R1516-
355 TO 10591X5716
75R1516-
321 TO 10591X5732
76R1516-
319 TO 10599X5732
73R1516-
3025894X5730
74R1516-
34210594X5723
71R1516-
33610597X5732
72R1516-
303 TO 10600X5730
69R1516-
31810590X5732
70R1516-
279 TO 5892X5730
(Part -2)
“Food security” is “
Thank You!
“National Security”