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Name of speaker : Patel Satishkumar
Reg. no. : 04-1313-2010
Major advisor : Dr. J.A. Patel
Date : 14/03/2012
Time : 1600 hrs
 Introduction
 Mechanism of Drought Tolerance
 Screening Methods
 Case studies on Drought Tolerance
Physiological
Variability and Correlation
Biochemical
 Biotechnological Studies
 Breeding approaches
 Achievements
 Limitations
 Conclusion
 Future thrust 2
INTRODUCTION
Botanical Name: Synonyms: Pennisetum glaucum (L.) R. Br.
Pennisetum typhoides (Burm. F.) Stapf. and Hubbard
Pennisetum typhoideum Rich.,
Pennisetum americanum L. Leeke
Common Names:- Bajra, Bulrush millet, Spiked millet, Cat tail millet
Family: - Poeaceae Sub family: Panicoideae Tribe: Paniceae
Origin :- Sahel zone of West Africa
Chromosome No.: 2n=14
Uses:- Feed: Fodder, Fuel, Fencing,
 Cross pollinated Crop Spices due to its protogynous flowering nature
 Annual C4 crop species.
 Stable diet for the vast majority of poor farmers
3
Table:-1 Area, Production And Productivity (2010-11)
Area
(million
hectare)
Production
(million tonns)
Productivity
(kg /hectare)
India 8.75 8.89 1015
Gujarat 0.92 1.31 1365
Source: Directorate of Economics and Statistics, Department of Agriculture and Cooperation.
4
State wise Bajra production (2010-11)
Rajasthan
31%
Uttar
Pradesh
22%
Haryana
14%
Gujarat
13%
MH
12%
MP
4%
Karnataka
2%
Tamil
Nadu
1%
Andhra
Pradesh
1%
State
Production
(Million Tonnes)
Productivity
(kg/ha)
Rajasthan
2.03 394
Uttar Pradesh 1.39 1638
Haryana 0.93 1593
Gujarat 0.92 1365
Maharashtra 0.77 741
Madhya Pradesh 0.25 1495
Karnataka 0.15 502
Tamil Nadu 0.08 1513
Andhra Pradesh 0.05 1178
Source: Directorate of Economics and Statistics, Department of Agriculture and Cooperation.
5
Drought
 Drought is event which implies the absence of a period
of time, long enough to cause moisture-depletion in soil
and water deficit with decrease of water potential in
plant tissues.
 Drought is highly heterogenous in time, space, degree
of stress, growth stage and time of stress exposure, and
it is unpredictable.
6
Types of Drought
• Meteorological Drought:- It is related to deficiencies in
rainfall compared to the average mean seasonal rainfall in
an area.
• Agricultural Drought:- Deficit rainfall over cropped
areas during their growth cycle can destroy crop or lead
to poor crop yields.
• Hydrological Drought:- It is a deficiency in surface and
sub-surface water supply. It is measured as stream flows
and also as lake, reservoir and groundwater levels.
7
Drought affected area in the world
The major bajra growing countries are Senegal, Mali, Niger, Nigeria, Sudan and India.
1.3 Billions people are under drought-prone areas (India/Africa)
8
Source: www.milletindia.org Source : http://www.mapsofindia.com
Major Bajra Growing Regions of India
Severely Affected
Moderately Affected
States affected by Drought
Bajra is a major cereals in northwestern zone as it represents approximately 25 % of the total
acreage of the crop in the country.
The chronically drought-prone areas around 33 % -receive less than 750 mm of rainfall, while
35 % classified as “drought-prone ” receive rainfall of 750-1,125 mm (in India). 9
About 36% of the land area constitutes arid and semi arid zones, arid and
semi arid areas are more prone to drought.
Drought leads to reduction in both yield and quality of economic product in
crop plants. It has adverse effect on plant growth and development.
Drought damages chloroplasts and lowers photosynthetic output.
There is an increase in proline level in the leaves of plants which are
subjected to all stresses.
Drought resistance is a genetically controlled physiological property of plant
species.
Breeding for drought tolerance is a major objective in arid and semiarid
regions of the world due to inadequate precipitation, shortage of irrigation
water and high water demand for crop evapotranspiration in such climates
Main features of drought
10
• Drought resistance : Ability of a plant to live, grow and yield
satisfactorily with limited water supply or under periodic water
deficits.
• Drought escape : Ability of plant to mature before water stress
becomes a serious limiting factor.
• Drought avoidance: Ability of a plant to withstand water deficit as
measured by degree and duration of low plant water potential.
water savers-closing of stomata
water spenders- extract more water from soil
• Drought tolerance: Ability of a plant to recover from a dry period
by producing new leaves from buds, and those were able to survive the
dry spell.
Gupta et al., 1986
11
Symptoms of Drought
• Reduced leaf area
• Early senescence of older leaves
• Effect on flowering, largely delay in flowering (Cause abscission
of flowers.)
Injury Mechanism
• Water stress directly affects cellular processes, membrane structures
and structure of macromolecules.
• Cause severe embolism formation in the xylem vessels.
12
Morphological traits Physiological factors
Higher rate of photosynthesis
Lower rate of transpiration
Higher leaf turgidity
Higher osmotic concentration
Earliness
Stomatal characters :-
Shrunken type, small size, less number per unit
area, rapid closing nature
Leaf character:- Waxy leaves, small
thick leaves, hairiness
Root characters:- Root length, root
density, R/S ratio
Growth habit:- Indeterminate
Biochemical factors
Proline content
ABA content in Leaf
13
Drought avoiding plant must maintain
High water potential
Thick and highly impermeable cuticle
Closure of stomata
More waxier leaves
Higher root -shoot ratio
14
Measurement of drought tolerance
1. Change in growth patterns
2. Change in seed production
3. Electrolyte leakage from leaf
segments
4. Leaf wilting
5. Relative leaf water content
6. Change in the transcriptome
Traits investigated
in pearl millet References
Grain and stover yield
and quality
Ibrahim et al. (1985), Kumari S (1988).
Bidinger et al. (1987, 2007), Singh and
Singh (1995), van Oosterom et al (1996),
Nepolean et al. (2006), Yadav et al.
(I999a,b, 2002, 2003. 2004) Serraj et al.
(2005)
ABA accumulation Henson et al. (1981). Henson (1983).
Henson et al. (1983). Henson(1984)
Water potential Henson (1982)
Osmotic potential Henson (1982)
Osmolytes Patil et al. (2005), Kholova et al. (2008)
Antioxidative enzymes Patil et al. (2005), Kholova et al. (2008)
Photosynthetic
pigments
Ibrahim et al. (1985), Ashraf et al. (2001)
Transpiration related
traits
Ibrahim et al. (1985), Squire (1979), Black
and Squire (1979), Henson et al. (1981),
Henson (1984), Kholova et al. (2008, 2010
a, b.c)
Canopy temperature Singh and Kanemasu (1983)
15
Drought
Tolerance
Physiolo
gical
processes
Expression
Profiling
Reverse
Genetics
Allele
Mining for
natural
Variation
MAS/Tran
sformation
Map
Based
Cloning
Forward
Cloning
Forward
Genetics
Comparative
mapping
Fig. 1 Drought tolerance improvement tools and processes
16
Screening Criteria
17
Screening Criteria
The selection criteria primarily based on morphological characters could be
selection of parents as well as desirable segregants followed by hybridization.
During selection, characters have high heritabilities and high correlation with
yield under stress across the environments.
Grain yield under stress conditions is usually the primary traits for
selection.
A suitable secondary traits should have (Edmeades et al. 2001).
1) Genetically association with grain yield under drought,
2) High heritability,
3) Stable and feasible to measure,
4) Lack of association with yield loss under ideal growing conditions.
18
Screening methods for Drought
tolerance in Pearl millet
19
Laboratory method
 In Laboratory method to identify genotypic difference in germinability, osmotic
solutions like polyethylene glycol (PEG) is used.
 The osmotic effect of drought are known to be comparable to true drought effects
Field method:
 The field is uniformly irrigated with overhead system using perforated pipes.
 Also used sprinkler method.
 The percentage of seedling that emerge is computed.
Tested Material
Tested Material
Line source irrigation method
Increasewaterstress
Water
Source/chennel
20
Case Studies
21
Table 2 :-Effect of osmotic stress on seedling traits of pearl millet genotype during drought induced by PEG in
Vitro condition.
ICRISAT (A. P.) Govindaraj et al. (2010) 22
Genotype Germination % Root length
(cm)
Seedling height
(cm)
Dry mass of
seedling (g)
Vigor index
Normal Stress Normal Stress Normal Stress Normal Stress Normal Stress
X7 99.5 98.0 10.89 6.53 17.90 10.71 0.058 0.074 1781.3 1049.6
X6 97.0 96.0 10.23 6.17 17.48 10.65 0.070 0.080 1695.6 1022.4
Co7 98.3 95.5 9.28 5.58 16.17 9.03 0.073 0.094 1589.5 862.4
WC-C75 99.8 98.3 11.18 4.64 18.43 7.75 0.093 0.101 1839.3 761.8
CD
(P<0.05 %)
T 0.636** 0.650** 0.755** 0.0065** 72.133**
V 0.900** 0.919 (NS) 1.068** 0.0092* 102.002 (NS)
T X V 1.272
(NS)
1.300* 1.510** 0.0131 (NS) 144.268 (NS)
Table 3 :- Germination and physiological parameters under normal (N) and
induced stress (PEG) (S) treatments (T) in pearl millet cultivars (V)
Vijayalakhsmi et al. (2000)
Coimbatore (T.N.)
23
Treatment Plant
height
(cm)
Plants/m Tillers/m Effective
tillers/m
Length
of ear
(cm)
Weight
of ear
(g)
1000
Grain
weight
(g)
Average
Yield
(kg/ha)
Percent
Reduction
T1, control
154 5.0 15.9 8.1 24.2 30.4 9.45 2101 -
T2, Rain out
during crop
establishment
149 4.8 17.5 6.2 22.2 28.8 9.27 1852 11.80
T3, Rainout during
tillering stage 144 4.8 11.8 5.8 20.4 26.3 8.89 1629 22.50
T4, Rainout during
earing and
flowering stage
139 5.2 11.6 6.3 21.2 27.8 8.67 1707 18.80
T5, Rainout during
grain-filling and
maturity stage
152 4.7 16.2 5.8 20.5 28.2 7.75 1970 6.20
CD (P < 0.05 %) NS NS 2.23 0.30 0.78 NS 0.25 - -
Table 4:-Average growth and yield attributes of pearl millet as affected by different
treatments
Agra (U.P.) Prakash et al. (2008) 24
WW (36 %) MS(21 %) SS (9 %)
Ethiopia Yalew and Yemane (2011)
WW (36 %) MS(21 %) SS (9 %)
Fig. 2 Response of pearl millet cultivars (Dadda and Shella ) to post-flowering
drought stress.
WW= Well Water
MS= Moderately Stress
SW= Severely Stress
WW (36 %) MS(21 %) SS (9 %)
25
Yalew and Yemane (2011)Ethiopia
Fig.3 Potential quantum yield of two cultivars of Pearl millet subjected to
three soil moisture levels.
WW= Well Water (36 %)
MS= Moderately Stress ( 21 %)
SW= Severely Stress ( 9 %)
Dadda
Shella
26
Table 5:- Grain yield and physiological parameters as influenced by terminal
moisture stress in B-line and inbreds
Pearl
millet
lines
Days to
flower
Grain
yield
(kg/ha)
Harvest
index
Threshing Relative
Water
content (%)
Drought
susceptibility
Index
81B 70.8 136.1 4.7 22.0 74.9 1.196 + 0.06
218B 65.7 234.4 8.7 28.4 63.4 1.259 + 0.12
89111B 59.3 263.3 11.2 26.0 77.4 1.104 + 0.12
95444B 58.7 628.9 22.4 47.7 74.2 0.812 + 0.06
J-108 57.8 648.9 18.2 40.0 72.0 0.846 + 0.10
J-998 62.8 353.3 9.3 29.7 69.0 1.057 + 0.04
J-2290 68.0 375.0 8.2 31.7 75.7 1.099 + 0.09
J-2296 59.2 376.7 15.4 37.7 75.0 0.880 + 0.08
J-2340 58.8 806.7 18.1 44.4 77.8 0.761 + 0.18
LSD
(P=0.05)
4.7 304.2 5.2 13.5 6.5 -
CV % 3.9 39.7 36.4 19.0 7.1 -
Joshi et al. (2005)Jamnagar (Gujarat) 27
Table 6:- Physiological parameters in pearl millet hybrids as influenced by
high temperature and receding soil moisture at seedling stage
Entries/
Hybrids
Root
Dry
mass
(mg/plant)
Shoot
dry
mass
(mg/plant)
Total
Dry
Mass
(mg/plant)
Root
/shoot
ratio
(dry
wt.
basis)
Survival
(%) at
19-22
DALI*
Leaf
elongatio
n
Rate
(cm/day)
index at
15 DALI*
Chlorophyll
Stability
GHB-558 26.3 30.9 57.1 0.9 44.9 0.35 0.111 + 0.016
GHB-559 29.5 26.4 55.7 1.2 68.4 0.48 0.079 + 0.007
GHB-316 26.9 25.9 52.8 1.1 61.3 0.47 0.096 + 0.019
GHB-526 38.7 37.4 76.1 1.1 76.4 0.59 0.053 + 0.016
GHB-538 32.8 34.9 67.8 1.0 72.9 0.55 0.058 + 0.014
LSD
(P=0.05)
NS 5.3 NS NS 16.4 0.09 -
CV (%) 10.5 15.1 10.8 17.9 9.1 17.3 -
* DALI – Days after last Irrigation, NS- Non significant
Joshi et al. (2005)Jamnagar (Gujarat) 28
29
Character GCV % PCV % h2 (B.S.) % Genetic
Advance
GA % of
Mean
E % 12.83 13.52 90.1 20.783 25.09
FSL 12.21 12.85 90.3 2.806 23.89
FRL 12.57 13.65 98.7 4.987 25.73
FSW 16.32 16.60 96.6 0.02 33.05
FRW 21.36 21.91 95.0 0.015 42.87
DSW 22.48 23.14 94.4 0.002 44.99
DRW 24.99 25.32 97.4 0.002 50.79
R/S 22.87 24.03 90.6 0.203 44.83
Table 7:- Estimates on GCV, PCV, heritability in broad sense(h2 B.S.), Genetic
advance as percentage of mean (GA %) of seedling traits in 63 pearl millet
genotypes
Coimbatore (T.N.) Arulselvi and Selvi (2009)
E %=Emergence %, EI= Emergence index, ERI= Emergence rate, FSL=Fresh Shoot Length,
FRL= Fresh Root Length, FSW=Fresh Shoot Weight, FSL= Fresh Root Weight,
DSW= Dry Shoot Weight, DRW= Dry Root Weight.
30
Table 8:-Character contribution towards genetic divergence
Sr. No. Character Contribution (%)
1 Emergence % 4.86
2 Fresh Shoot Length 2.66
3 Fresh Root Length 39.63
4 Fresh Shoot Weight 26.73
5 Fresh Root Weight 2.05
6 Dry Shoot Weight 7.32
7 Dry Root Weight 16.28
8 Root/Shoot 0.36
Coimbatore (T.N.) Arulselvi and Selvi (2009) 31
Source df E % EI ERI FSL FRL FSW FRW DSW DRW R/S
Replications 1 6.2222 0.002 0.000007 0.0287 0.0229 0.00001 0.000059 0.000002 0.000001 0.0178
Genotypes 62 238.3574** 0.0069 0.000027 4.3309** 11.9518** 0.000201** 0.000107** 0.000002** 0.000001** 0.0226**
Error 62 12.4158 0.0035 0.000014 0.2211 0.0778 0.000003 0.000003 0.0000001 0.0000001 0.0011
SE 2.4717 0.0414 0.0027 0.3298 0.1956 0.0013 0.0012 0.0002 0.0001 0.0234
CD @ 5% 4.9409 0.0828 0.0053 0.6593 0.3911 0.0026 0.0023 0.0004 0.0001 0.0468
Mean 82.8254 1.0956 0.0139 11.7437 19.3833 0.0609 0.0338 0.0047 0.0021 0.4528
Table 9:- Mean Square from analysis of variance for seedling traits conferring drought
tolerance in pearl millet genotypes.
** Significant at 0.01 probability level
Coimbatore (T.N.) Arulselvi and Selvi (2009)
E %=Emergence %, EI= Emergence index, ERI= Emergence rate, FSL=Fresh Shoot Length, FRL= Fresh
Root Length, FSW=Fresh Shoot Weight, FSL= Fresh Root Weight, DSW= Dry Shoot Weight, DRW= Dry
Root Weight.
32
Traits FSL FRL FSW FRW DSW DRW R/S
E %
0.2782 0.4332** 0.4555** 0.5009** 0.4479** 0.3204** -0.0508
FSL 0.4688** 0.7518** 0.5672** 0.5909** 0.5542** 0.0606
FRL 0.5536** 0.4769** 0.5793** 0.5411** 0.0506
FSW 0.7682** 0.8105** 0.6978** -0.0060
FRW 0.5633** 0.8800** 0.4457**
DSW
0.5176** -0.3404**
DRW 0.6158**
Table 10:- Simple Correlation coefficients Between seedling traits
(conferring Drought Tolerance ) among 63 pearl millet genotypes.
Arulselvi and Selvi (2009)Coimbatore (T.N.) ** Significant at 0.01 probability level
E %=Emergence %, EI= Emergence index, ERI= Emergence rate, FSL=Fresh Shoot Length, FRL=
Fresh Root Length, FSW=Fresh Shoot Weight, FSL= Fresh Root Weight, DSW= Dry ShootWeight,
DRW= Dry Root Weight.
33
Table 11:-Means and F ratios of genotypes for growth and yield components
measured in the irrigated control (c) and drought stress (s) treatments.
Characters 1988 (34 Genotypes) 1989 (34 Genotypes) 1990 (32 Genotype)
Treatment Means F ratio Means F ratio Means F ratio
Time to flowering (days) Control 64 5.80** 66 5.92** 67 11.90**
Stress 62 5.40** 65 5.99** 68 15.60**
Biomass (g m-2) Control 621 3.02** 858 1.79** 662 2.49**
Stress 437 2.49** 585 3.28** 598 2.76**
Stover (g m-2) Control 398 3.29** 482 1.91** 431 6.50**
Stress 300 4.38** 359 4.57** 423 3.91**
Panicle (g m-2) Control 223 2.54** 378 1.77** 230 0.97
Stress 137 2.05** 228 2.19** 175 1.41
Grain yield (g m-2) Control 156 2.30** 271 1.66** 158 1.07
Stress 83 2.48** 140 2.64** 121 0.78
Panicle No. m m-2 Control 9.8 1.58* 11.5 3.61** 10.3 1.50*
Stress 8.2 1.47 10.0 2.33** 8.4 1.51*
Panicle yield (g) Control 16.2 4.02** 23.7 3.81** 15.0 2.46**
Stress 9.8 3.00** 13.7 3.23** 14.2 0.72
No. Grains panicle-1 Control 2440 4.47** 3090 3.75** 2260 1.96**
Stress 1840 1.90** 2300 3.16** 2330 0.72
Grain mass (g 100-1) Control 0.67 10.39** 0.77 3.96** 0.66 5.90**
Stress 0.53 7.30** 0.53 2.86** 0.61 2.55**
No. Grains m-2 (*103) Control 2.306 3.34** 35.3 1.93** 23.5 1.16
Stress 15.4 1.88** 23.3 2.43** 19.6 0.81
Harvest index Control 25.4 4.04** 32.0 3.20** 23.0 2.01**
Stress 18.6 4.38** 24.0 3.81** 20.0 0.65
Threshing Percentage Control 70 3.61** 71 1.90** 67 2.30**
Stress 58 3.44** 60 3.76** 69 0.61
Sadore Peter (1992) 34
Characters
Correlation Coefficients
Flowering DRI
1988 1989 1990 1988 1989 1990
Time to flowering (days) --- ----- --- -0.07 0.07 -0.02
Biomass (G m-2) 0.53** 0.47** 0.56** 0.56** 0.59** 0.20
Stover (G m-2) 0.76** 0.63** 0.77** 0.26. 0.37** 0.06
Panicle (G m-2) -0.31 -0.21 -0.35* 0.73** 0.74** 0.31
Grain yield (G m-2) -0.46** -0.35* -0.28 0.69** 0.71** 083**
Panicle No. M-2 -0.55** -0.47** -066** 050** 0.11 0.10
Panicle yield (g) -0.29 -0.16 0.06 0.62** 0.78** 0.88**
No. Grains per panicle 0.13 0.22 0.04 0.55** 0.63** 0.92**
Grain mass (G) -0.47** -0.57** 0.04 0.28 0.36* -0.01
No. Grains m-2 (x103) -0.22 -0.08 -0.28 0.65** 0.62** 0.85**
Harvest index -0.74** -069** -044* 0.44* 0.25 0.82**
Threshing % -0.60** -057** -0.09 0.48**
0.44*
0.44* 0.91**
Table 12 :- Correlations of yield parameters in the drought stress treatment to
time to flowering under drought stress and Drought Response Index.
** P<0.01 * P<0.05
Peter (1992)Sadore 35
Character
Days to 50
per cent
Flowering
Days to
Maturity
Ear
head
length
(cm)
Ear
head
girth
(cm)
No. of
produc-
tive
tillers
per
plant
Ear
head
weight
per
plant
(g)
Total
biomass
accumulati
on per
plant (g)
Grain
yield per
plant (g)
Harvest
index
Panicle
harvest
index
Test
weight
(g)
Root
length
per
plant
(cm0
Proline
content
(µ g g-1 f.
w.)
Drought
response
index
(DRI)
Days to 50 %
Flowering
1.00
Days to
Maturity
0.14** 1.00
Ear head
length
-0.22** -0.07 1.00
Ear head
Girth
-0.06 0.07 0.07 1.00
No of
Productive
tillers per plant
-0.06 0.04 0.03 -0.18** 1.00
Ear head
weight per
plant
-0.08 -0.08 -0.01 -0.16** 0.13* 1.00
Total biomass
accumulation
per plant
-0.05 -0.08 -0.10 -0.06 0.20** 0.24** 1.00
Grain Yield per
plant
-0.06 -0.12 0.13* -0.20** 0.44** 0.48** 0.19** 1.00
Harvest
index
-0.18** -0.15** 0.27** -0.08 0.22** 0.11 -0.26** 0.65** 1.00
Panicle harvest
index
-0.14** -0.08 0.16** -0.08 0.39** -0.21** 0.03 0.74** 0.66** 1.00
Test
weight
-0.14** -0.15** 0.14** -0.12 0.23** 0.13* 0.09 0.59** 0.42** 0.55** 1.00
Root length per
plant
-0.00 0.03 0.17** 0.11 0.21** 0.04 0.09 0.35** 0.26** 0.36** 0.33** 1.00
Proline
Content
0.04 -0.00 0.04 -0.09 0.36** 0.10 0.13* 0.49** 0.31** 0.45** 0.50** 0.37** 1.00
Drought
response index
-0.01 0.00 -0.11 -0.30** 0.35** 0.53** 0.01 0.74** 0.28** 0.41** 0.30** 0.17** 0.35** 1.00
Table 13:-Phenotypic correlation coefficient for different characters under terminal
drought condition in pearl millet.
*, ** Significant at 5 and 1 percent level of probability, respectively
AAU (Anand) Patil and Jadeja (2005) 36
Transpiration Rate
37
Fig 4:-Water Conserving mechanisms with the terminal drought tolerance of
pearl millet
Kholova et al. ( 2010)ICRISAT (A.P.)
Drought Tolerant
PRTL 2/89-33
863B-P2
Drought Sensitive
H 77/833-2
ICMB-841-P3
38
ABA content in Leaf
39
Fig. 5:- ABA content in pearl millet test cross hybrids (Drought Tolerant Drought Sensitive)
in well-water (WW) and water stress (WS) condition
Vegetative Stage Reproductive Stage
NIL-QTLs Drought Tolerant
PRTL 2/89-33
863B-P2
Drought Sensitive
H 77/833-2
ICMB-841-P3
Kholova et al. ( 2010)
ICRISAT (A.P.) 40
Drought Tolerance QTL under salt stress
41
0
2
4
6
8
10
12
14
16
843A X H 77/833-
2 (Drought
sensitiveparent)
843A X 01029
(QTL-NIL)
843A X PRTL
2/89-33 (Drought
tolerant parent)
pH 8.5
pH 9.0
pH 9.4
LeafNa(mg/gdrywt.)
FIG. 6:- Na+ accumulation in the leaves of drought- sensitive and drought-tolerant
parents, at three Alkalinity levels.
Sharma et al. (2010)CSSRI (Karnal)
42
LeafNa(mg/gdrywt.)
0
5
10
15
20
25
843A X
H77/833-2
(Drought
Sensitive
parent)
843A X 01029
(QTL-NIL)
834A X PRTL
2/89-33
(Drought
tolerant parent)
EC 2 ds/m
EC 9 ds/m
EC 12 ds/m
FIG. 7:- Na+ accumulation in the leaves of drought- sensitive and drought-tolerant
parents, at three salinity levels.
CSSRI (Karnal) Sharma et al. (2010)
43
Table 14:- Correlation analysis between enzymatic activities and pigments contents
and ratios under well water and water stress condition.
Water stress
Chlorophyll a Chlorophyll b Carotenoids Chl a/Chl b Chl/Car
APX (Ascorbic peroxidase)
APX 2 ns ns ns ns 0.9344*
APX 4 ns ns ns ns 0.9037*/0.9881*
APX 8 ns ns -0.9104* ns ns
APX 9 ns ns ns ns 0.8929*
SOD (Superoxide dismutase)
Mn-SOD2 ns ns ns ns 0.969**
Mn-SOD3 ns ns ns ns 0.9516*
CAT (Catalse)
CAT1 ns ns ns ns 0.9439*
CAT2 ns ns ns ns 0.8911*
Well-watered
Chlorophyll a Chlorophyll b Carotenoids Chl a/Chl b Chl/Car
APX (Ascorbic peroxidase)
APX5 ns ns ns ns -0.8821*
APX 6 ns ns ns ns -0.8972*
APX Sum ns ns ns ns -0.9417*
Kholova et al. ( 2011)ICRISAT (A.P) 44
Fig. 8:- Gene networks involved in drought stress response and
tolerance
Function in stress tolerance Signal transduction and gene expression
Shinozaki and Shinozaki (2007)Japan
45
Breeding Strategies
46
Lines of pearl millet used in crosses to developed genetic
maps
Lines Characteristics
1 H 77/833-2 Elite male parent of grain hybrids in
north-western India, Susceptible to
downy mildew but with seedling
thermotolerance, high tillering capacity
and earliness
2 PRLT 2/89-33 Inbred 33 in ICRISAT potential R- line
Trail conducted in 1989; derived by
selfing in the ICRISAT bold seeded Early
Composite, low tillering, large seeds,
drought tolerant which is largely based
on lniadi landrace germplasm from West
Africa.
47
Fig. 9:- Strategy for development of Genetic linkage map to identify
QTLs linked to traits
Yadav et al. 2010ICRISAT
48
Fig:-10 Genetic map
Genetic map of a pearl millet population that segregates for drought tolerance showing the distribution
of molecular marker on the different linkage group. The highlighted regions indicate parts of the
genome controlling grain yield, and its components, during drought stress.
49
Linkage
Group
(QTL on)
Drought Response
for Grain yield
Genetic Background References
LG2 Up to 32 % H 77/833-2 x PRTL 2/89-33 Yadav et al., 1999,2002
LG 3 & 4 11.6-17.3 % ICMB 841 x 863 B Bidinger et al., 2007
LG 5 14.8 % ICMB 841 x 863 B Yadav et al., 2004
LG 6 & 1 QTL has Pleiotropic to
decreased panicle
number
H 77/833-2 x PRTL 2/833 Yadav et al., 2010
Table:- 15 QTLs associated with drought tolerance of grain yield
50
Fig:-11 Marker Assisted Backcross Breeding for Drought Tolerance
51
Fig. 12:- Fine mapping population or High Resolution Cross
Yadav et al. 2010
Objective of HRC
1. To fine map the DT-QTL interval
on LG 2
2. To Pyramid this DT-QTL with
the Downy-mildew resistance
QTLs on LG 1 & 4
52ICRISAT
Table 16:-Comparison of the DT-QTL based and Field performance-
based Hybrids.
Moisture Environment
Crop Trait
QTL topcross Hybrids Field topcross hybrids LSD (P=0.05)
Non- Stress
Flowering (d) 39.1 41.3 0.19
Biomass (g /m2) 777 845 15.6
Harvest Index (%) 49.6 45.9 0.55
Grain Yield (g/m2) 381 393 7.7
Terminal Stress
Flowering (d) 41.1 43.5 0.15
Biomass (g /m2) 581 619 11.4
Harvest Index (%) 41.7 38.3 0.60
Grain Yield (g/m2) 245 239 5.6
Line Source
Flowering (d) 35.1 38.1 0.12
Biomass (g /m2) 537 562 7.9
Harvest Index (%) 49.5 43.8 0.5
Grain Yield (g/m2) 268 255 5.1
Serraj et al.(2005)ICRISAT 53
Fig. 13:-Marker Assisted Selection
1. Marker-assisted selection. Genetic composition at the drought tolerance QTL
( to constitute a MAS TCP)
2. Phenotypic selection. Field performance (best 16) in the drought trials used to
identify QTLs (to constitute a phenotype TCP)
3. A Random control. A random sample from within the mapping population
(to constitute a random TCP).
Bidinger et al. (2005)ICRISAT
54
55
The mapped progeny were phenotyped as testcross hybrids
rather than as the skeleton-mapped F2 plants.
• To restore heterotic vigour to partially inbred mapping progeny that might
otherwise be too weak for effective screening under stress conditions
(Inbreeding depression)
• To reduced variation in flowering time among the test units, in order to
focus the mapping on specific drought tolerance traits rather than traits or
responses associated with variation on capacity for drought escape
• To have test units that approximate the genetic structure of the F1 hybrids
grown by farmers rather than partially inbred F3 or F4 lines.
56
Framework of an integrated strategy for genetic enhancement of crop grain yield
(GY) and its components under water-limited conditions at ICRISAT.
TR=total plant water transpired; TE=transpiration efficiency; HI= harvest index.
57
Breeding Approaches
 Breeding under optimum (water-stress free)
condition
 Breeding under actual drought condition
 Breeding under artificially created
environment
 Incorporation of drought tolerance
58
1. Introduction (PRLT 2/89-33, lniadi landrace germplasm from West Africa.)
2. Interspecific and intergeneric hybridization
Single backcross
Three way cross (Gene pyramiding)
3. Pedigree selection
4. Back cross breeding
5. Mutation breeding
6. Ideotype breeding : Breeding activity aimed at producing
new genotypes with novel morpho-physiological features that fit a
pre-defined architecture thought to be advantageous based on
experimental physiology and/or modelling.
Breeding Methods
59
7. Marker assisted breeding
8. Marker-assisted backcrossing (MABC): Repeated
backcrossing of the F1’s to reconstitute the
recipient genome without losing the desirable
gene.
9. Tissue culture (in vitro Screening by using PEG
6000)
10.Development of Transgenic
60
Achievements
61
Table:-17 Recommended drought-tolerant/drought-avoiding
hybrids/varieties of pearl millet in India
State Hybrids Varieties
Maharashtra Nandi 35,Saburi, PAC 903
PPC 6, HC 20, JBV 2, ICTP 8203,
ICMV 221, AIMP 92901
Tamil Nadu GHB 558, CoH (Cu)8, X7 Co7, ICMV 221, ICMV 155
Andhra Pradesh PB 106, GHB 558 AIMP 92901, ASP-1, ICTP 8203
Karnataka PB 106, GHB 558 ICMV 221, ICTP 8203
Rajasthan
HHB 67, RBH 121, GHB 538,
PB 180 CZP 9802, Raj 171
Gujarat
GHB 577, GHB 526, PB 172,
PB 112, ICMB 356 JBV 2, HC 20
Haryana
HHB 67, GHB 538, HHB 117,
ICMB 356 CZP 9802
ICRISAT REPORT (2007) 62
63
Limitations
• There is no single major gene, which has a remarkable effect on
the drought tolerance
• Drought tolerance is an environmental and developmental stage
specific character
• Drought reduces nutrients uptake, and is associated with
temperature stress and at higher elevation with cold. This
associations make the breeding programme more complicated.
• Most of the physiological and metabolic processes are affected by
water deficits: cell growth, stomatal regulation, photosynthesis,
translocation, etc.
• Large number of genes regulated up- or down- by drought
• Large genetic populations and replicates are required
• Even drought component traits are often complex and difficult to
screen
64
Conclusion
• Tillering stage (30-45 DAS) is most susceptible to drought; wherein 23-25
% of yield reductions occurres, followed by drought at grain filling and
maturity stage.
• The osmotic (PEG 6000) stress at the seedling stage is the most suitable
method for drought tolerance screening owing to their significant
relationship with declining the germination percentage, root and shoot length.
• Selection for seedling traits conferring drought tolerance such as root
length, root weight and root shoot ratio will be useful for identifying
genotypes with drought tolerance capacity.
• Post-flowering drought stress is one of the most important
environmental factors reducing the grain yield and yield stability of pearl
millet and increasing the incidence of crop failure in dryland production
environments
• Marker Assisted Selection is the most appropriate method to improve
drought tolerance genotype. 65
Future thrust
• Consolidation of yield grains through multiple resistance
to various abiotic and biotic stresses.
• To develop a plant ideotype by restructuring the
morphological attributes these can withstand drought
conditions.
• Need to saturate QTL areas to increase the efficiency
• Pyramiding with other QTLs
• In pearl millet, it need to be tested whether high leaf ABA
content and the lower Tr are linked or not.
• Need to develop Transgenic for Drought Tolerance,
though difficult because of polygenic inheritance.
66
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Breeding for Drought tolerance in Pearl Millet

  • 1. //o Name of speaker : Patel Satishkumar Reg. no. : 04-1313-2010 Major advisor : Dr. J.A. Patel Date : 14/03/2012 Time : 1600 hrs
  • 2.  Introduction  Mechanism of Drought Tolerance  Screening Methods  Case studies on Drought Tolerance Physiological Variability and Correlation Biochemical  Biotechnological Studies  Breeding approaches  Achievements  Limitations  Conclusion  Future thrust 2
  • 3. INTRODUCTION Botanical Name: Synonyms: Pennisetum glaucum (L.) R. Br. Pennisetum typhoides (Burm. F.) Stapf. and Hubbard Pennisetum typhoideum Rich., Pennisetum americanum L. Leeke Common Names:- Bajra, Bulrush millet, Spiked millet, Cat tail millet Family: - Poeaceae Sub family: Panicoideae Tribe: Paniceae Origin :- Sahel zone of West Africa Chromosome No.: 2n=14 Uses:- Feed: Fodder, Fuel, Fencing,  Cross pollinated Crop Spices due to its protogynous flowering nature  Annual C4 crop species.  Stable diet for the vast majority of poor farmers 3
  • 4. Table:-1 Area, Production And Productivity (2010-11) Area (million hectare) Production (million tonns) Productivity (kg /hectare) India 8.75 8.89 1015 Gujarat 0.92 1.31 1365 Source: Directorate of Economics and Statistics, Department of Agriculture and Cooperation. 4
  • 5. State wise Bajra production (2010-11) Rajasthan 31% Uttar Pradesh 22% Haryana 14% Gujarat 13% MH 12% MP 4% Karnataka 2% Tamil Nadu 1% Andhra Pradesh 1% State Production (Million Tonnes) Productivity (kg/ha) Rajasthan 2.03 394 Uttar Pradesh 1.39 1638 Haryana 0.93 1593 Gujarat 0.92 1365 Maharashtra 0.77 741 Madhya Pradesh 0.25 1495 Karnataka 0.15 502 Tamil Nadu 0.08 1513 Andhra Pradesh 0.05 1178 Source: Directorate of Economics and Statistics, Department of Agriculture and Cooperation. 5
  • 6. Drought  Drought is event which implies the absence of a period of time, long enough to cause moisture-depletion in soil and water deficit with decrease of water potential in plant tissues.  Drought is highly heterogenous in time, space, degree of stress, growth stage and time of stress exposure, and it is unpredictable. 6
  • 7. Types of Drought • Meteorological Drought:- It is related to deficiencies in rainfall compared to the average mean seasonal rainfall in an area. • Agricultural Drought:- Deficit rainfall over cropped areas during their growth cycle can destroy crop or lead to poor crop yields. • Hydrological Drought:- It is a deficiency in surface and sub-surface water supply. It is measured as stream flows and also as lake, reservoir and groundwater levels. 7
  • 8. Drought affected area in the world The major bajra growing countries are Senegal, Mali, Niger, Nigeria, Sudan and India. 1.3 Billions people are under drought-prone areas (India/Africa) 8
  • 9. Source: www.milletindia.org Source : http://www.mapsofindia.com Major Bajra Growing Regions of India Severely Affected Moderately Affected States affected by Drought Bajra is a major cereals in northwestern zone as it represents approximately 25 % of the total acreage of the crop in the country. The chronically drought-prone areas around 33 % -receive less than 750 mm of rainfall, while 35 % classified as “drought-prone ” receive rainfall of 750-1,125 mm (in India). 9
  • 10. About 36% of the land area constitutes arid and semi arid zones, arid and semi arid areas are more prone to drought. Drought leads to reduction in both yield and quality of economic product in crop plants. It has adverse effect on plant growth and development. Drought damages chloroplasts and lowers photosynthetic output. There is an increase in proline level in the leaves of plants which are subjected to all stresses. Drought resistance is a genetically controlled physiological property of plant species. Breeding for drought tolerance is a major objective in arid and semiarid regions of the world due to inadequate precipitation, shortage of irrigation water and high water demand for crop evapotranspiration in such climates Main features of drought 10
  • 11. • Drought resistance : Ability of a plant to live, grow and yield satisfactorily with limited water supply or under periodic water deficits. • Drought escape : Ability of plant to mature before water stress becomes a serious limiting factor. • Drought avoidance: Ability of a plant to withstand water deficit as measured by degree and duration of low plant water potential. water savers-closing of stomata water spenders- extract more water from soil • Drought tolerance: Ability of a plant to recover from a dry period by producing new leaves from buds, and those were able to survive the dry spell. Gupta et al., 1986 11
  • 12. Symptoms of Drought • Reduced leaf area • Early senescence of older leaves • Effect on flowering, largely delay in flowering (Cause abscission of flowers.) Injury Mechanism • Water stress directly affects cellular processes, membrane structures and structure of macromolecules. • Cause severe embolism formation in the xylem vessels. 12
  • 13. Morphological traits Physiological factors Higher rate of photosynthesis Lower rate of transpiration Higher leaf turgidity Higher osmotic concentration Earliness Stomatal characters :- Shrunken type, small size, less number per unit area, rapid closing nature Leaf character:- Waxy leaves, small thick leaves, hairiness Root characters:- Root length, root density, R/S ratio Growth habit:- Indeterminate Biochemical factors Proline content ABA content in Leaf 13
  • 14. Drought avoiding plant must maintain High water potential Thick and highly impermeable cuticle Closure of stomata More waxier leaves Higher root -shoot ratio 14
  • 15. Measurement of drought tolerance 1. Change in growth patterns 2. Change in seed production 3. Electrolyte leakage from leaf segments 4. Leaf wilting 5. Relative leaf water content 6. Change in the transcriptome Traits investigated in pearl millet References Grain and stover yield and quality Ibrahim et al. (1985), Kumari S (1988). Bidinger et al. (1987, 2007), Singh and Singh (1995), van Oosterom et al (1996), Nepolean et al. (2006), Yadav et al. (I999a,b, 2002, 2003. 2004) Serraj et al. (2005) ABA accumulation Henson et al. (1981). Henson (1983). Henson et al. (1983). Henson(1984) Water potential Henson (1982) Osmotic potential Henson (1982) Osmolytes Patil et al. (2005), Kholova et al. (2008) Antioxidative enzymes Patil et al. (2005), Kholova et al. (2008) Photosynthetic pigments Ibrahim et al. (1985), Ashraf et al. (2001) Transpiration related traits Ibrahim et al. (1985), Squire (1979), Black and Squire (1979), Henson et al. (1981), Henson (1984), Kholova et al. (2008, 2010 a, b.c) Canopy temperature Singh and Kanemasu (1983) 15
  • 18. Screening Criteria The selection criteria primarily based on morphological characters could be selection of parents as well as desirable segregants followed by hybridization. During selection, characters have high heritabilities and high correlation with yield under stress across the environments. Grain yield under stress conditions is usually the primary traits for selection. A suitable secondary traits should have (Edmeades et al. 2001). 1) Genetically association with grain yield under drought, 2) High heritability, 3) Stable and feasible to measure, 4) Lack of association with yield loss under ideal growing conditions. 18
  • 19. Screening methods for Drought tolerance in Pearl millet 19
  • 20. Laboratory method  In Laboratory method to identify genotypic difference in germinability, osmotic solutions like polyethylene glycol (PEG) is used.  The osmotic effect of drought are known to be comparable to true drought effects Field method:  The field is uniformly irrigated with overhead system using perforated pipes.  Also used sprinkler method.  The percentage of seedling that emerge is computed. Tested Material Tested Material Line source irrigation method Increasewaterstress Water Source/chennel 20
  • 22. Table 2 :-Effect of osmotic stress on seedling traits of pearl millet genotype during drought induced by PEG in Vitro condition. ICRISAT (A. P.) Govindaraj et al. (2010) 22
  • 23. Genotype Germination % Root length (cm) Seedling height (cm) Dry mass of seedling (g) Vigor index Normal Stress Normal Stress Normal Stress Normal Stress Normal Stress X7 99.5 98.0 10.89 6.53 17.90 10.71 0.058 0.074 1781.3 1049.6 X6 97.0 96.0 10.23 6.17 17.48 10.65 0.070 0.080 1695.6 1022.4 Co7 98.3 95.5 9.28 5.58 16.17 9.03 0.073 0.094 1589.5 862.4 WC-C75 99.8 98.3 11.18 4.64 18.43 7.75 0.093 0.101 1839.3 761.8 CD (P<0.05 %) T 0.636** 0.650** 0.755** 0.0065** 72.133** V 0.900** 0.919 (NS) 1.068** 0.0092* 102.002 (NS) T X V 1.272 (NS) 1.300* 1.510** 0.0131 (NS) 144.268 (NS) Table 3 :- Germination and physiological parameters under normal (N) and induced stress (PEG) (S) treatments (T) in pearl millet cultivars (V) Vijayalakhsmi et al. (2000) Coimbatore (T.N.) 23
  • 24. Treatment Plant height (cm) Plants/m Tillers/m Effective tillers/m Length of ear (cm) Weight of ear (g) 1000 Grain weight (g) Average Yield (kg/ha) Percent Reduction T1, control 154 5.0 15.9 8.1 24.2 30.4 9.45 2101 - T2, Rain out during crop establishment 149 4.8 17.5 6.2 22.2 28.8 9.27 1852 11.80 T3, Rainout during tillering stage 144 4.8 11.8 5.8 20.4 26.3 8.89 1629 22.50 T4, Rainout during earing and flowering stage 139 5.2 11.6 6.3 21.2 27.8 8.67 1707 18.80 T5, Rainout during grain-filling and maturity stage 152 4.7 16.2 5.8 20.5 28.2 7.75 1970 6.20 CD (P < 0.05 %) NS NS 2.23 0.30 0.78 NS 0.25 - - Table 4:-Average growth and yield attributes of pearl millet as affected by different treatments Agra (U.P.) Prakash et al. (2008) 24
  • 25. WW (36 %) MS(21 %) SS (9 %) Ethiopia Yalew and Yemane (2011) WW (36 %) MS(21 %) SS (9 %) Fig. 2 Response of pearl millet cultivars (Dadda and Shella ) to post-flowering drought stress. WW= Well Water MS= Moderately Stress SW= Severely Stress WW (36 %) MS(21 %) SS (9 %) 25
  • 26. Yalew and Yemane (2011)Ethiopia Fig.3 Potential quantum yield of two cultivars of Pearl millet subjected to three soil moisture levels. WW= Well Water (36 %) MS= Moderately Stress ( 21 %) SW= Severely Stress ( 9 %) Dadda Shella 26
  • 27. Table 5:- Grain yield and physiological parameters as influenced by terminal moisture stress in B-line and inbreds Pearl millet lines Days to flower Grain yield (kg/ha) Harvest index Threshing Relative Water content (%) Drought susceptibility Index 81B 70.8 136.1 4.7 22.0 74.9 1.196 + 0.06 218B 65.7 234.4 8.7 28.4 63.4 1.259 + 0.12 89111B 59.3 263.3 11.2 26.0 77.4 1.104 + 0.12 95444B 58.7 628.9 22.4 47.7 74.2 0.812 + 0.06 J-108 57.8 648.9 18.2 40.0 72.0 0.846 + 0.10 J-998 62.8 353.3 9.3 29.7 69.0 1.057 + 0.04 J-2290 68.0 375.0 8.2 31.7 75.7 1.099 + 0.09 J-2296 59.2 376.7 15.4 37.7 75.0 0.880 + 0.08 J-2340 58.8 806.7 18.1 44.4 77.8 0.761 + 0.18 LSD (P=0.05) 4.7 304.2 5.2 13.5 6.5 - CV % 3.9 39.7 36.4 19.0 7.1 - Joshi et al. (2005)Jamnagar (Gujarat) 27
  • 28. Table 6:- Physiological parameters in pearl millet hybrids as influenced by high temperature and receding soil moisture at seedling stage Entries/ Hybrids Root Dry mass (mg/plant) Shoot dry mass (mg/plant) Total Dry Mass (mg/plant) Root /shoot ratio (dry wt. basis) Survival (%) at 19-22 DALI* Leaf elongatio n Rate (cm/day) index at 15 DALI* Chlorophyll Stability GHB-558 26.3 30.9 57.1 0.9 44.9 0.35 0.111 + 0.016 GHB-559 29.5 26.4 55.7 1.2 68.4 0.48 0.079 + 0.007 GHB-316 26.9 25.9 52.8 1.1 61.3 0.47 0.096 + 0.019 GHB-526 38.7 37.4 76.1 1.1 76.4 0.59 0.053 + 0.016 GHB-538 32.8 34.9 67.8 1.0 72.9 0.55 0.058 + 0.014 LSD (P=0.05) NS 5.3 NS NS 16.4 0.09 - CV (%) 10.5 15.1 10.8 17.9 9.1 17.3 - * DALI – Days after last Irrigation, NS- Non significant Joshi et al. (2005)Jamnagar (Gujarat) 28
  • 29. 29
  • 30. Character GCV % PCV % h2 (B.S.) % Genetic Advance GA % of Mean E % 12.83 13.52 90.1 20.783 25.09 FSL 12.21 12.85 90.3 2.806 23.89 FRL 12.57 13.65 98.7 4.987 25.73 FSW 16.32 16.60 96.6 0.02 33.05 FRW 21.36 21.91 95.0 0.015 42.87 DSW 22.48 23.14 94.4 0.002 44.99 DRW 24.99 25.32 97.4 0.002 50.79 R/S 22.87 24.03 90.6 0.203 44.83 Table 7:- Estimates on GCV, PCV, heritability in broad sense(h2 B.S.), Genetic advance as percentage of mean (GA %) of seedling traits in 63 pearl millet genotypes Coimbatore (T.N.) Arulselvi and Selvi (2009) E %=Emergence %, EI= Emergence index, ERI= Emergence rate, FSL=Fresh Shoot Length, FRL= Fresh Root Length, FSW=Fresh Shoot Weight, FSL= Fresh Root Weight, DSW= Dry Shoot Weight, DRW= Dry Root Weight. 30
  • 31. Table 8:-Character contribution towards genetic divergence Sr. No. Character Contribution (%) 1 Emergence % 4.86 2 Fresh Shoot Length 2.66 3 Fresh Root Length 39.63 4 Fresh Shoot Weight 26.73 5 Fresh Root Weight 2.05 6 Dry Shoot Weight 7.32 7 Dry Root Weight 16.28 8 Root/Shoot 0.36 Coimbatore (T.N.) Arulselvi and Selvi (2009) 31
  • 32. Source df E % EI ERI FSL FRL FSW FRW DSW DRW R/S Replications 1 6.2222 0.002 0.000007 0.0287 0.0229 0.00001 0.000059 0.000002 0.000001 0.0178 Genotypes 62 238.3574** 0.0069 0.000027 4.3309** 11.9518** 0.000201** 0.000107** 0.000002** 0.000001** 0.0226** Error 62 12.4158 0.0035 0.000014 0.2211 0.0778 0.000003 0.000003 0.0000001 0.0000001 0.0011 SE 2.4717 0.0414 0.0027 0.3298 0.1956 0.0013 0.0012 0.0002 0.0001 0.0234 CD @ 5% 4.9409 0.0828 0.0053 0.6593 0.3911 0.0026 0.0023 0.0004 0.0001 0.0468 Mean 82.8254 1.0956 0.0139 11.7437 19.3833 0.0609 0.0338 0.0047 0.0021 0.4528 Table 9:- Mean Square from analysis of variance for seedling traits conferring drought tolerance in pearl millet genotypes. ** Significant at 0.01 probability level Coimbatore (T.N.) Arulselvi and Selvi (2009) E %=Emergence %, EI= Emergence index, ERI= Emergence rate, FSL=Fresh Shoot Length, FRL= Fresh Root Length, FSW=Fresh Shoot Weight, FSL= Fresh Root Weight, DSW= Dry Shoot Weight, DRW= Dry Root Weight. 32
  • 33. Traits FSL FRL FSW FRW DSW DRW R/S E % 0.2782 0.4332** 0.4555** 0.5009** 0.4479** 0.3204** -0.0508 FSL 0.4688** 0.7518** 0.5672** 0.5909** 0.5542** 0.0606 FRL 0.5536** 0.4769** 0.5793** 0.5411** 0.0506 FSW 0.7682** 0.8105** 0.6978** -0.0060 FRW 0.5633** 0.8800** 0.4457** DSW 0.5176** -0.3404** DRW 0.6158** Table 10:- Simple Correlation coefficients Between seedling traits (conferring Drought Tolerance ) among 63 pearl millet genotypes. Arulselvi and Selvi (2009)Coimbatore (T.N.) ** Significant at 0.01 probability level E %=Emergence %, EI= Emergence index, ERI= Emergence rate, FSL=Fresh Shoot Length, FRL= Fresh Root Length, FSW=Fresh Shoot Weight, FSL= Fresh Root Weight, DSW= Dry ShootWeight, DRW= Dry Root Weight. 33
  • 34. Table 11:-Means and F ratios of genotypes for growth and yield components measured in the irrigated control (c) and drought stress (s) treatments. Characters 1988 (34 Genotypes) 1989 (34 Genotypes) 1990 (32 Genotype) Treatment Means F ratio Means F ratio Means F ratio Time to flowering (days) Control 64 5.80** 66 5.92** 67 11.90** Stress 62 5.40** 65 5.99** 68 15.60** Biomass (g m-2) Control 621 3.02** 858 1.79** 662 2.49** Stress 437 2.49** 585 3.28** 598 2.76** Stover (g m-2) Control 398 3.29** 482 1.91** 431 6.50** Stress 300 4.38** 359 4.57** 423 3.91** Panicle (g m-2) Control 223 2.54** 378 1.77** 230 0.97 Stress 137 2.05** 228 2.19** 175 1.41 Grain yield (g m-2) Control 156 2.30** 271 1.66** 158 1.07 Stress 83 2.48** 140 2.64** 121 0.78 Panicle No. m m-2 Control 9.8 1.58* 11.5 3.61** 10.3 1.50* Stress 8.2 1.47 10.0 2.33** 8.4 1.51* Panicle yield (g) Control 16.2 4.02** 23.7 3.81** 15.0 2.46** Stress 9.8 3.00** 13.7 3.23** 14.2 0.72 No. Grains panicle-1 Control 2440 4.47** 3090 3.75** 2260 1.96** Stress 1840 1.90** 2300 3.16** 2330 0.72 Grain mass (g 100-1) Control 0.67 10.39** 0.77 3.96** 0.66 5.90** Stress 0.53 7.30** 0.53 2.86** 0.61 2.55** No. Grains m-2 (*103) Control 2.306 3.34** 35.3 1.93** 23.5 1.16 Stress 15.4 1.88** 23.3 2.43** 19.6 0.81 Harvest index Control 25.4 4.04** 32.0 3.20** 23.0 2.01** Stress 18.6 4.38** 24.0 3.81** 20.0 0.65 Threshing Percentage Control 70 3.61** 71 1.90** 67 2.30** Stress 58 3.44** 60 3.76** 69 0.61 Sadore Peter (1992) 34
  • 35. Characters Correlation Coefficients Flowering DRI 1988 1989 1990 1988 1989 1990 Time to flowering (days) --- ----- --- -0.07 0.07 -0.02 Biomass (G m-2) 0.53** 0.47** 0.56** 0.56** 0.59** 0.20 Stover (G m-2) 0.76** 0.63** 0.77** 0.26. 0.37** 0.06 Panicle (G m-2) -0.31 -0.21 -0.35* 0.73** 0.74** 0.31 Grain yield (G m-2) -0.46** -0.35* -0.28 0.69** 0.71** 083** Panicle No. M-2 -0.55** -0.47** -066** 050** 0.11 0.10 Panicle yield (g) -0.29 -0.16 0.06 0.62** 0.78** 0.88** No. Grains per panicle 0.13 0.22 0.04 0.55** 0.63** 0.92** Grain mass (G) -0.47** -0.57** 0.04 0.28 0.36* -0.01 No. Grains m-2 (x103) -0.22 -0.08 -0.28 0.65** 0.62** 0.85** Harvest index -0.74** -069** -044* 0.44* 0.25 0.82** Threshing % -0.60** -057** -0.09 0.48** 0.44* 0.44* 0.91** Table 12 :- Correlations of yield parameters in the drought stress treatment to time to flowering under drought stress and Drought Response Index. ** P<0.01 * P<0.05 Peter (1992)Sadore 35
  • 36. Character Days to 50 per cent Flowering Days to Maturity Ear head length (cm) Ear head girth (cm) No. of produc- tive tillers per plant Ear head weight per plant (g) Total biomass accumulati on per plant (g) Grain yield per plant (g) Harvest index Panicle harvest index Test weight (g) Root length per plant (cm0 Proline content (µ g g-1 f. w.) Drought response index (DRI) Days to 50 % Flowering 1.00 Days to Maturity 0.14** 1.00 Ear head length -0.22** -0.07 1.00 Ear head Girth -0.06 0.07 0.07 1.00 No of Productive tillers per plant -0.06 0.04 0.03 -0.18** 1.00 Ear head weight per plant -0.08 -0.08 -0.01 -0.16** 0.13* 1.00 Total biomass accumulation per plant -0.05 -0.08 -0.10 -0.06 0.20** 0.24** 1.00 Grain Yield per plant -0.06 -0.12 0.13* -0.20** 0.44** 0.48** 0.19** 1.00 Harvest index -0.18** -0.15** 0.27** -0.08 0.22** 0.11 -0.26** 0.65** 1.00 Panicle harvest index -0.14** -0.08 0.16** -0.08 0.39** -0.21** 0.03 0.74** 0.66** 1.00 Test weight -0.14** -0.15** 0.14** -0.12 0.23** 0.13* 0.09 0.59** 0.42** 0.55** 1.00 Root length per plant -0.00 0.03 0.17** 0.11 0.21** 0.04 0.09 0.35** 0.26** 0.36** 0.33** 1.00 Proline Content 0.04 -0.00 0.04 -0.09 0.36** 0.10 0.13* 0.49** 0.31** 0.45** 0.50** 0.37** 1.00 Drought response index -0.01 0.00 -0.11 -0.30** 0.35** 0.53** 0.01 0.74** 0.28** 0.41** 0.30** 0.17** 0.35** 1.00 Table 13:-Phenotypic correlation coefficient for different characters under terminal drought condition in pearl millet. *, ** Significant at 5 and 1 percent level of probability, respectively AAU (Anand) Patil and Jadeja (2005) 36
  • 38. Fig 4:-Water Conserving mechanisms with the terminal drought tolerance of pearl millet Kholova et al. ( 2010)ICRISAT (A.P.) Drought Tolerant PRTL 2/89-33 863B-P2 Drought Sensitive H 77/833-2 ICMB-841-P3 38
  • 39. ABA content in Leaf 39
  • 40. Fig. 5:- ABA content in pearl millet test cross hybrids (Drought Tolerant Drought Sensitive) in well-water (WW) and water stress (WS) condition Vegetative Stage Reproductive Stage NIL-QTLs Drought Tolerant PRTL 2/89-33 863B-P2 Drought Sensitive H 77/833-2 ICMB-841-P3 Kholova et al. ( 2010) ICRISAT (A.P.) 40
  • 41. Drought Tolerance QTL under salt stress 41
  • 42. 0 2 4 6 8 10 12 14 16 843A X H 77/833- 2 (Drought sensitiveparent) 843A X 01029 (QTL-NIL) 843A X PRTL 2/89-33 (Drought tolerant parent) pH 8.5 pH 9.0 pH 9.4 LeafNa(mg/gdrywt.) FIG. 6:- Na+ accumulation in the leaves of drought- sensitive and drought-tolerant parents, at three Alkalinity levels. Sharma et al. (2010)CSSRI (Karnal) 42
  • 43. LeafNa(mg/gdrywt.) 0 5 10 15 20 25 843A X H77/833-2 (Drought Sensitive parent) 843A X 01029 (QTL-NIL) 834A X PRTL 2/89-33 (Drought tolerant parent) EC 2 ds/m EC 9 ds/m EC 12 ds/m FIG. 7:- Na+ accumulation in the leaves of drought- sensitive and drought-tolerant parents, at three salinity levels. CSSRI (Karnal) Sharma et al. (2010) 43
  • 44. Table 14:- Correlation analysis between enzymatic activities and pigments contents and ratios under well water and water stress condition. Water stress Chlorophyll a Chlorophyll b Carotenoids Chl a/Chl b Chl/Car APX (Ascorbic peroxidase) APX 2 ns ns ns ns 0.9344* APX 4 ns ns ns ns 0.9037*/0.9881* APX 8 ns ns -0.9104* ns ns APX 9 ns ns ns ns 0.8929* SOD (Superoxide dismutase) Mn-SOD2 ns ns ns ns 0.969** Mn-SOD3 ns ns ns ns 0.9516* CAT (Catalse) CAT1 ns ns ns ns 0.9439* CAT2 ns ns ns ns 0.8911* Well-watered Chlorophyll a Chlorophyll b Carotenoids Chl a/Chl b Chl/Car APX (Ascorbic peroxidase) APX5 ns ns ns ns -0.8821* APX 6 ns ns ns ns -0.8972* APX Sum ns ns ns ns -0.9417* Kholova et al. ( 2011)ICRISAT (A.P) 44
  • 45. Fig. 8:- Gene networks involved in drought stress response and tolerance Function in stress tolerance Signal transduction and gene expression Shinozaki and Shinozaki (2007)Japan 45
  • 47. Lines of pearl millet used in crosses to developed genetic maps Lines Characteristics 1 H 77/833-2 Elite male parent of grain hybrids in north-western India, Susceptible to downy mildew but with seedling thermotolerance, high tillering capacity and earliness 2 PRLT 2/89-33 Inbred 33 in ICRISAT potential R- line Trail conducted in 1989; derived by selfing in the ICRISAT bold seeded Early Composite, low tillering, large seeds, drought tolerant which is largely based on lniadi landrace germplasm from West Africa. 47
  • 48. Fig. 9:- Strategy for development of Genetic linkage map to identify QTLs linked to traits Yadav et al. 2010ICRISAT 48
  • 49. Fig:-10 Genetic map Genetic map of a pearl millet population that segregates for drought tolerance showing the distribution of molecular marker on the different linkage group. The highlighted regions indicate parts of the genome controlling grain yield, and its components, during drought stress. 49
  • 50. Linkage Group (QTL on) Drought Response for Grain yield Genetic Background References LG2 Up to 32 % H 77/833-2 x PRTL 2/89-33 Yadav et al., 1999,2002 LG 3 & 4 11.6-17.3 % ICMB 841 x 863 B Bidinger et al., 2007 LG 5 14.8 % ICMB 841 x 863 B Yadav et al., 2004 LG 6 & 1 QTL has Pleiotropic to decreased panicle number H 77/833-2 x PRTL 2/833 Yadav et al., 2010 Table:- 15 QTLs associated with drought tolerance of grain yield 50
  • 51. Fig:-11 Marker Assisted Backcross Breeding for Drought Tolerance 51
  • 52. Fig. 12:- Fine mapping population or High Resolution Cross Yadav et al. 2010 Objective of HRC 1. To fine map the DT-QTL interval on LG 2 2. To Pyramid this DT-QTL with the Downy-mildew resistance QTLs on LG 1 & 4 52ICRISAT
  • 53. Table 16:-Comparison of the DT-QTL based and Field performance- based Hybrids. Moisture Environment Crop Trait QTL topcross Hybrids Field topcross hybrids LSD (P=0.05) Non- Stress Flowering (d) 39.1 41.3 0.19 Biomass (g /m2) 777 845 15.6 Harvest Index (%) 49.6 45.9 0.55 Grain Yield (g/m2) 381 393 7.7 Terminal Stress Flowering (d) 41.1 43.5 0.15 Biomass (g /m2) 581 619 11.4 Harvest Index (%) 41.7 38.3 0.60 Grain Yield (g/m2) 245 239 5.6 Line Source Flowering (d) 35.1 38.1 0.12 Biomass (g /m2) 537 562 7.9 Harvest Index (%) 49.5 43.8 0.5 Grain Yield (g/m2) 268 255 5.1 Serraj et al.(2005)ICRISAT 53
  • 54. Fig. 13:-Marker Assisted Selection 1. Marker-assisted selection. Genetic composition at the drought tolerance QTL ( to constitute a MAS TCP) 2. Phenotypic selection. Field performance (best 16) in the drought trials used to identify QTLs (to constitute a phenotype TCP) 3. A Random control. A random sample from within the mapping population (to constitute a random TCP). Bidinger et al. (2005)ICRISAT 54
  • 55. 55
  • 56. The mapped progeny were phenotyped as testcross hybrids rather than as the skeleton-mapped F2 plants. • To restore heterotic vigour to partially inbred mapping progeny that might otherwise be too weak for effective screening under stress conditions (Inbreeding depression) • To reduced variation in flowering time among the test units, in order to focus the mapping on specific drought tolerance traits rather than traits or responses associated with variation on capacity for drought escape • To have test units that approximate the genetic structure of the F1 hybrids grown by farmers rather than partially inbred F3 or F4 lines. 56
  • 57. Framework of an integrated strategy for genetic enhancement of crop grain yield (GY) and its components under water-limited conditions at ICRISAT. TR=total plant water transpired; TE=transpiration efficiency; HI= harvest index. 57
  • 58. Breeding Approaches  Breeding under optimum (water-stress free) condition  Breeding under actual drought condition  Breeding under artificially created environment  Incorporation of drought tolerance 58
  • 59. 1. Introduction (PRLT 2/89-33, lniadi landrace germplasm from West Africa.) 2. Interspecific and intergeneric hybridization Single backcross Three way cross (Gene pyramiding) 3. Pedigree selection 4. Back cross breeding 5. Mutation breeding 6. Ideotype breeding : Breeding activity aimed at producing new genotypes with novel morpho-physiological features that fit a pre-defined architecture thought to be advantageous based on experimental physiology and/or modelling. Breeding Methods 59
  • 60. 7. Marker assisted breeding 8. Marker-assisted backcrossing (MABC): Repeated backcrossing of the F1’s to reconstitute the recipient genome without losing the desirable gene. 9. Tissue culture (in vitro Screening by using PEG 6000) 10.Development of Transgenic 60
  • 62. Table:-17 Recommended drought-tolerant/drought-avoiding hybrids/varieties of pearl millet in India State Hybrids Varieties Maharashtra Nandi 35,Saburi, PAC 903 PPC 6, HC 20, JBV 2, ICTP 8203, ICMV 221, AIMP 92901 Tamil Nadu GHB 558, CoH (Cu)8, X7 Co7, ICMV 221, ICMV 155 Andhra Pradesh PB 106, GHB 558 AIMP 92901, ASP-1, ICTP 8203 Karnataka PB 106, GHB 558 ICMV 221, ICTP 8203 Rajasthan HHB 67, RBH 121, GHB 538, PB 180 CZP 9802, Raj 171 Gujarat GHB 577, GHB 526, PB 172, PB 112, ICMB 356 JBV 2, HC 20 Haryana HHB 67, GHB 538, HHB 117, ICMB 356 CZP 9802 ICRISAT REPORT (2007) 62
  • 63. 63
  • 64. Limitations • There is no single major gene, which has a remarkable effect on the drought tolerance • Drought tolerance is an environmental and developmental stage specific character • Drought reduces nutrients uptake, and is associated with temperature stress and at higher elevation with cold. This associations make the breeding programme more complicated. • Most of the physiological and metabolic processes are affected by water deficits: cell growth, stomatal regulation, photosynthesis, translocation, etc. • Large number of genes regulated up- or down- by drought • Large genetic populations and replicates are required • Even drought component traits are often complex and difficult to screen 64
  • 65. Conclusion • Tillering stage (30-45 DAS) is most susceptible to drought; wherein 23-25 % of yield reductions occurres, followed by drought at grain filling and maturity stage. • The osmotic (PEG 6000) stress at the seedling stage is the most suitable method for drought tolerance screening owing to their significant relationship with declining the germination percentage, root and shoot length. • Selection for seedling traits conferring drought tolerance such as root length, root weight and root shoot ratio will be useful for identifying genotypes with drought tolerance capacity. • Post-flowering drought stress is one of the most important environmental factors reducing the grain yield and yield stability of pearl millet and increasing the incidence of crop failure in dryland production environments • Marker Assisted Selection is the most appropriate method to improve drought tolerance genotype. 65
  • 66. Future thrust • Consolidation of yield grains through multiple resistance to various abiotic and biotic stresses. • To develop a plant ideotype by restructuring the morphological attributes these can withstand drought conditions. • Need to saturate QTL areas to increase the efficiency • Pyramiding with other QTLs • In pearl millet, it need to be tested whether high leaf ABA content and the lower Tr are linked or not. • Need to develop Transgenic for Drought Tolerance, though difficult because of polygenic inheritance. 66