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Improvement of drought Improvement of drought tolerant variety, IR57514 , for tolerant variety, IR57514 , for
grain aroma and cooking quality grain aroma and cooking quality using marker-assisted selectionusing marker-assisted selection
Ubon Ratchathani University
IR57514 is an elite drought tolerant and high yielding rice IR57514 is an elite drought tolerant and high yielding rice variety developed by IRRI which is well adapted to rainfed variety developed by IRRI which is well adapted to rainfed lowland in Mekong region where the areas are mostly limited lowland in Mekong region where the areas are mostly limited by abiotic stresses with unpredictable rainfall patterns. by abiotic stresses with unpredictable rainfall patterns.
Problem ?
BUT it was not
successfully adopted by farmers due to
unsatisfied of grain qualities.
Breeding IR57514 for grain aroma and good cooking quality traits will greatly enhance both quality and productivity of rice in the rainfed lowland areas in Mekong region.
IR57514 +aroma +
good cooking quality
The research was aimed to improve grain aroma and cooking quality of IR57514. Marker-assisted selection (MAS) in backcross breeding will be used to transfer favorable alleles of genes / QTL of grain aroma and good cooking qualities traits from KDML105 into genetic background of IR57514.
Objective
• The marker-assisted backcross breeding was conducted using IR57514 as the recipient parent. • KD571-77, the backcross introgression line of KDML105 carrying Xa21 for bacterial leaf blight resistance and Sub1A, Sub1B and Sub1C for submergence tolerance, was used as a donor parent for grain aroma and good cooking quality traits. • The backcross procedure for introgression of the target QTLs was conducted at Ubon Rice Research Center and Kasetsart University, Kamphangsean Campus, Nakorn Pathom.
Plant materialsPlant materials
Breeding scheme:
IR57514 x KD571-77Recipient parent Donor parent
F1 x IR57514
BC1F1
IR57514 x Selected BC1F1
BC2F1 (~6 line, ~27 seed)
~188 BC1F2MAS
IR57514 x selected BC1F2 (~18 plant)
90 BC2F1
MAS
IR57514 x selected BC2F1 (~14 plant)
BC3F1
11 selected BC3F1BC3F2 (1,640 plants)
436 Selected BC3F2 plants from 11 families
selected BC3F2 (33 plants)
MAS
Nov. 2007
July 2005
May, 2007
Jan. 2006
MAS
2,595 BC3F3
MAS
370 Selected BC3F3
May, 2008
MAS
Initial submergence tolerance testing (370 BC3F4 lines) Initial drought tolerance testing (319 BC3F4 lines)
173 selected lines
- YT in aerobic condition in KPS (173 BC3F4 lines)- YT in anaerobic condition in KPS (173 BC3F4 lines, stimulated flooding condition)
Yield trial(56 BC3F5 lines)
Trait validation (56 BC3F5 lines)- submergence tolerance- drought tolerance
Grain aroma and cooking quality testing
(56 BC3F5 lines)
August, 2008June, 2008
January, 2009
Preliminary grain aroma and cooking quality validation
56 selected lines
IR57514 x KD571-77Recipient parent Donor parent
F1 x IR57514
BC1F1
IR57514 x Selected BC1F1
BC2F1 (~6 line, ~27 seed)
~188 BC1F2MAS
IR57514 x selected BC1F2 (~18 plant)
90 BC2F1
MAS
IR57514 x selected BC2F1 (~14 plant)
BC3F1
11 selected BC3F1BC3F2 (1,640 plants)
436 Selected BC3F2 plants from 11 families
selected BC3F2 (33 plants)
MAS
Nov. 2007
July 2005
May, 2007
Jan. 2006
MAS
2,595 BC3F3
MAS
370 Selected BC3F3
May, 2008
MAS
Initial submergence tolerance testing (370 BC3F4 lines) Initial drought tolerance testing (319 BC3F4 lines)
173 selected lines
- YT in aerobic condition in KPS (173 BC3F4 lines)- YT in anaerobic condition in KPS (173 BC3F4 lines, stimulated flooding condition)
Yield trial(56 BC3F5 lines)
Trait validation (56 BC3F5 lines)- submergence tolerance- drought tolerance
Grain aroma and cooking quality testing
(56 BC3F5 lines)
August, 2008June, 2008
January, 2009
Preliminary grain aroma and cooking quality validation
56 selected lines
• 352 BC3F1 were produced from the crosses between the selected 14 BC2F1 and IR57514.
•14 BC3F1 were selected based on
MAS:• BADH for grain aroma, • waxy for amylose content & gel consistency,• SNP 2341 for gelatinization temperature• PB7-8 for Xa21 bacterial leaf blight resistance
1st workshop (May 21 – 30, 2007)1st workshop (May 21 – 30, 2007)IR57514 x KD571-77
Recipient parent Donor parent
F1 x IR57514
BC1F1
IR57514 x Selected BC1F1
~188 BC1F2
MAS
IR57514 x selected BC1F2 (~18 plant)
90 BC2F1
MAS
IR57514 x selected BC2F1 (~14 plant)
352 BC3F1
11 selected BC3F1
MAS
July 2005
1st workshop ( 21 -30 May, 2007)
Jan. 2006
MAS
IR57514 x KD571-77Recipient parent Donor parent
F1 x IR57514
BC1F1
IR57514 x Selected BC1F1
~188 BC1F2
MAS
IR57514 x selected BC1F2 (~18 plant)
90 BC2F1
MAS
IR57514 x selected BC2F1 (~14 plant)
352 BC3F1
11 selected BC3F1
MAS
July 2005
1st workshop ( 21 -30 May, 2007)
Jan. 2006
MAS
• 1640 BC3F2 plants were generated from 11 BC3F1
• The 436 BC3F2 progeny were selected from 1640 BC3F2 based on phenotype.
• 33 lines were selected using MAS:
• BADH for grain aroma, • waxy for amylose content & gel consistency,• SNP 2341 for gelatinization temperature• PB7-8 for Xa21 bacterial leaf blight resistance characters.
2nd workshop (Nov 10 – 24, 2007)2nd workshop (Nov 10 – 24, 2007)
BC3F2 (1,640 plants)
436 Selected BC3F2 plants
selected BC3F2 (33 plants)
MAS
Preliminary quality validation
11 selected BC3F1
IR57514 x KD571-77Recipient parent Donor parent
F1 x IR57514
BC1F1
IR57514 x Selected BC1F1
~188 BC1F2
MAS
IR57514 x selected BC1F2 (~18 plant)
90 BC2F1
MAS
IR57514 x selected BC2F1 (~14 plant)
352 BC3F1
MAS
July 2005
Jan. 2006
MAS
May, 2007
BC3F2 (1,640 plants)
436 Selected BC3F2 plants
selected BC3F2 (33 plants)
MAS
Preliminary quality validation
11 selected BC3F1
IR57514 x KD571-77Recipient parent Donor parent
F1 x IR57514
BC1F1
IR57514 x Selected BC1F1
~188 BC1F2
MAS
IR57514 x selected BC1F2 (~18 plant)
90 BC2F1
MAS
IR57514 x selected BC2F1 (~14 plant)
352 BC3F1
MAS
July 2005
Jan. 2006
MAS
May, 2007
2nd workshop
• These 33 families of BC3F2 were selfing to generate 2,595 lines of BC3F3 progeny.
• Marker-assisted selection (MAS):
• BADH for aroma • waxy for amylose content and gel consistency• SNP2341 for gelatinization temperature and • PB 7-8 for Xa21 bacterial left blight resistance
• 370 BC3F3 progeny were selected based on genotype (260 lines) and based on phenotype (110 lines).
• The selected 370 BC3F3 progeny will be evaluated for the traits of interest comparing to the recipient variety, IR57514 .
3rd workshop (May 1 – 15, 2008)3rd workshop (May 1 – 15, 2008)
BC3F2 (1,640 plants)
436 Selected BC3F2 plants
selected BC3F2 (33 plants)
MASPreliminary quality
validation
11 selected BC3F1
IR57514 x KD571-77Recipient parent Donor parent
BC1F1
IR57514 x Selected BC1F1
90 BC2F1
MAS
IR57514 x selected BC2F1 (~14 plant)
352 BC3F1
MAS
Jan. 2006
MAS
May, 2007
2,595 BC3F3
MAS
370 Selected BC3F3
BC3F2 (1,640 plants)
436 Selected BC3F2 plants
selected BC3F2 (33 plants)
MASPreliminary quality
validation
11 selected BC3F1
IR57514 x KD571-77Recipient parent Donor parent
BC1F1
IR57514 x Selected BC1F1
90 BC2F1
MAS
IR57514 x selected BC2F1 (~14 plant)
352 BC3F1
MAS
Jan. 2006
MAS
May, 2007
2,595 BC3F3
MAS
370 Selected BC3F3
3rd workshop
Traits Validation• Grain quality: aroma, cooking quality
• BC3F2 • BC3F5
• Submergence tolerance• BC3F4• BC3F5
• Drought tolerance• BC3F4
4th workshop (May 1 – 15, 2008)4th workshop (May 1 – 15, 2008)
Grain aroma : sensory test
Amylose content : colorimetric method described by Julaino (1987)
Gelatinization temperature : Alkaline test (alkali spreading value ) as described by Little et al. (1958)
Gel consistency : described by Tan et al. (1999)
Traits Validation : Grain aroma and cooking quality in BC3F2 and BC3F5
Grain QualityGrain Quality
Traits Validation : Submergence tolerance (BC3F4 and BC3F5)Traits Validation : Submergence tolerance (BC3F4 and BC3F5)
Data recording ;
•Percent of plant survival (PPS)
• Percent of plant elongation (PPE)
• Leaf senescence
(Siangliw et al., 2003).
IR57514IR57514KDML105KDML105
FR13AFR13A
RD6RD6
Results 370 BC3F4 173 BC3F4
PPS
56 BC3F5 20 BC3F5
Location : Kasetsart University,
kampangsean campus,
Nakorn Pathom
PPS, LS, PPE
Percentage of plant survival
-5.000.005.00
10.0015.0020.0025.0030.0035.0040.0045.0050.0055.0060.0065.0070.0075.0080.0085.0090.0095.00
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55 57 59
Lines
Pe
rce
nta
ge
of
pla
nt
su
rviv
al
RD
6K
DM
L105
IR57514
KD
571-7
7
Percentage of plant survival
-5.000.005.00
10.0015.0020.0025.0030.0035.0040.0045.0050.0055.0060.0065.0070.0075.0080.0085.0090.0095.00
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55 57 59
Lines
Pe
rce
nta
ge
of
pla
nt
su
rviv
al
RD
6K
DM
L105
IR57514
IR57514
KD
571-7
7K
D571-7
7
Leaf greenness
0.00
10.00
20.00
30.00
40.00
50.00
1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46 49 52 55 58
Lines
SPAD
val
ue
IR57
514
KD57
1-7
7RD
6KD
ML1
05
Leaf greenness
0.00
10.00
20.00
30.00
40.00
50.00
1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46 49 52 55 58
Lines
SPAD
val
ue
IR57
514
KD57
1-7
7RD
6KD
ML1
05
Percent of plant elongation
0.00
20.00
40.00
60.00
80.00
100.00
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55 57 59
Lines
perc
ent o
f pla
nt
elon
gatio
n KD57
1-7
7
RD6
KDM
L105
IR57
514
Percent of plant elongation
0.00
20.00
40.00
60.00
80.00
100.00
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55 57 59
Lines
perc
ent o
f pla
nt
elon
gatio
n KD57
1-7
7
RD6
KDM
L105
IR57
514
IR57
514
Submergence tolerance validation in 56 BC3F5 lines. Submergence tolerance validation in 56 BC3F5 lines.
Validation : Drought tolerance (BC3F4) Validation : Drought tolerance (BC3F4)
Data recording
GY (Grain yield) PH (Plant height)
PN (Panicle number) TN (Tiller number)
FWD (Flowering date)
Experiment Design : Randomized Complete Block Design in CPA
Augmented Design in RGDU
Check varieties: IR57514 Tolerance check
KDML105, KD571-77 Susceptible check
Population size: 319 BC3F4 lines Chumpae Rice Research Center
173 BC3F4 lines Kasetsart University, Kampangsean campus
Grain aroma and cooking quality testing (BC3F5 progeny)Grain aroma and cooking quality testing (BC3F5 progeny)
Grain quality genotyping and validation
No.
# Aroma GT AC GC
Lines BADH Score SNP2341 Score Waxy Score Score
KD571-77 1 KD 1 5 1 16 73
IR57514 3 IR 3 2 3 33 102
1 BC3F5 # 14-34-1-16-1 1 KD* 1 4.9 1 19 86
2 BC3F5 # 22-43-1-10-19 1 KD 1 5.0 1 16 82
3 BC3F5 # 22-43-1-10-24 1 KD* 1 5.0 1 19 109
4 BC3F5 # 22-43-1-10-27 1 KD 1 4.9 1 20 106
5 BC3F5 # 22-43-1-10-32 1 KD 1 4.9 1 17 108
6 BC3F5 # 22-43-1-10-44 1 KD 1 4.9 1 19 146
7 BC3F5 # 22-43-1-42-25 1 KD 1 4.9 1 22 104
8 BC3F5 # 85-4-1-24-1 1 KD 1 5.0 1 20 123
9 BC3F5 # 85-4-1-24-12 1 KD 1 5.0 1 21 131
10 BC3F5 # 85-4-1-24-41 1 KD 1 5.0 1 21 106
Final Lines
Submergence tolerance and drought tolerance validationSubmergence tolerance and drought tolerance validation
No.
Lines
Submergence validation (BC3F5)
Drought validation (BC3F4)
Grain yield (kg rai-1) Sterility (%) Flowering date
PPS PPE LS Irrigated Drained Irrigated Drained Irrigated Drained
KD571-77 67.98 33.81 20.14 87.68 90.46 3.61 7.18 90 88
IR57514 73.39 27.21 31.99 140.82 121.40 4.37 4.93 103 98
1 BC3F5 # 14-34-1-16-1 79.29 24.93 33.04 101.40 103.47 1.92 0.67 80 81
2 BC3F5 # 22-43-1-10-19 73.61 21.50 27.81 - - - - - -
3 BC3F5 # 22-43-1-10-24 81.35 25.96 30.81 125.89 93.61 1.11 2.39 83 92
4 BC3F5 # 22-43-1-10-27 74.48 24.89 32.05 94.05 89.64 1.85 6.95 81 81
5 BC3F5 # 22-43-1-10-32 75.99 16.45 31.98 95.90 102.62 0.93 1.43 81 84
6 BC3F5 # 22-43-1-10-44 69.37 25.53 33.50 119.60 100.76 2.04 1.86 81 91
7 BC3F5 # 22-43-1-42-25 73.29 35.57 32.04 125.83 96.38 2.02 4.65 89 90
8 BC3F5 # 85-4-1-24-1 71.82 22.84 28.08 137.80 110.51 4.29 11.23 101 100
9 BC3F5 # 85-4-1-24-12 74.89 22.49 32.78 139.04 95.27 5.64 13.76 104 102
10 BC3F5 # 85-4-1-24-41 78.15 24.97 28.34 120.49 93.91 9.77 14.89 102 100
Mean 72.86 26.90 29.46 118.63 103.69 2.55 4.17 88.14 90.02
Prob. ** ** ** ** ** ** ** ** **
LSD0.05 2.36 1.67 0.63 1.68 1.48 0.17 0.30 0.68 0.64
Final Lines
Conclusion
We aimed to improve grain aroma and cooking quality of IR57514 using MAS.
10 selected BC3F5 were finally selected for grain aroma and cooking quality traits based on genotype (MAS) and these lines were validated for grain quality, submergence tolerance and drought tolerance.
These selected lines will be on trial both in station and on farm in rice production areas in Mekong region.
Marker assisted selection (MAS) technology was an excellent tool for rice breeding program.
MAS technology jointly with classical breeding will increase the effectiveness and precision of selection process.
MAS will be shortening the breeding program and also reduce the cost of breeding program.
Breeding IR57514 for grain aroma and good cooking quality traits will greatly enhance both quality and productivity of rice in the rainfed lowland areas in Mekong region.
Acknowledgments Acknowledgments
RRT H A I L A N Drice gene discovery UNIT
Rice Gene Discovery Unit Rice Gene Discovery Unit
(RGDU)(RGDU)
National Center for Genetic National Center for Genetic
Engineering and Engineering and
Biotechnology (BIOTEC)Biotechnology (BIOTEC)
Generation Challenge
Programe (GCP)