This document summarizes research on screening rice genotypes for water deficiency tolerance. 33 rice genotypes from Pakistan, WARDA, and Cuba were tested under normal water conditions and 3 levels of reduced water (75%, 50%, 25% less than normal). Some genotypes from WARDA showed increased plant height and tillers but reduced leaves under reduced water. Grain yield increased in some genotypes from WARDA and Cuba with reduced water. Further tests on heat tolerance identified heat shock proteins induced in some genotypes. Efficiency tests found 3 genotypes maintained photosynthesis under low light conditions, including WAB-56-104 which reduced plant height but increased leaves and unaffected yield with reduced water.

1. 11
Women in rice biotechnology:
success that will have an
impact in the times ahead
Presented by:
Shafqat Farooq
PAEC, Islamabad, Pakistan

2. 22
Introduction
Rice is the most important crop at global level as
it is used as staple food in the most countries of
the world
Second major staple cereal being grown on 2.5
million hectares in Pakistan, are lowland cultivars
and require a lot of water to grow. The supply of
water is diminishing with every passing day, A
grain reduction of >30 % occurred during the
recent past,
Identifying stress-resistance genotypes is
essential for genetic improvement of rice
especially for water deficiency tolerance (WD).
The use of suitable methodology for measuring
stress resistance in large breeding population is

3. 33
Some genotypes are more resistant than
others and out-yield those exposed to the
same degree of water stress
The performance of rice varieties under
water stress conditions at different growth
stages varies. It is observed that yield
losses resulting from water deficit are
particularly sever when drought strikes at
booting stage.
Water stress at or before, panicle initiation
reduces the most potential spikelet
number and stress during grain filling
decreases translocation of assimilates to
the grains which decreases the grain
Introduction cont.

4. 44
n order to find WD , we tested
Rice genotypes
consisting of
i. five local basmati (fine grain
aromatic) rice cultivars, IR-6,
CP-1 (Chinese variety),
ii. 19 lines/land races from
WARDA,
iii. and 7 varieties/lines from
CUBA

5. 55
ObjectivesObjectives
i)i) To reduce waterTo reduce water
consumption andconsumption and
ii) to induce temperatureii) to induce temperature
tolerance in lowlandtolerance in lowland
ricerice
varietiesvarieties
through transferring genethrough transferring gene
(s) from upland rice(s) from upland rice
varieties/ land racesvarieties/ land races

6. 66
Material was directly sown
in the field blocks
maintained
i. at normal flooded conditions
(Control) and
ii. at 75%, 50% and 25% less
water
than control,
iii. Fertilizer was applied at the
normal recommended doze,
iv. Data was collected with
reference to plant height,
Methodology

7. 77
Methodology cont.
i. Morphological
performance
under water deficiency,
ii. Appearance of stress
protein
iii. Efficiency of plants
under
water deficient
conditions,

8. 88
Selection from screening forSelection from screening for
water deficiency tolerancewater deficiency tolerance
experimentsexperimentsWe tested all the genotypes under different
water levels used for irrigation i.e. 100%
which is control, and 75%, 50% and 25% less
water than that required for normal flooded
irrigation. Selections were made for the
plants that can tolerate at-least 50% less
water than control and show
i) Reduced plant height,
ii) Increased number of leaves,
iii) Tillers,
iv) Plant in which grain yield remained
unaffected
and/or.
v) Stay green

9. 99
Significant variations were observed for all
the parameters and in all the material.
•In 19 entries (IR-6, CP-1, material from WARDA
and Cuba), number of tillers and leaves were the
highest at 50% reduced water than control,
• In 10 and 8 entries, plant height increased by
31.7% and 61.3 %, respectively at 25% and 50%
less water than control,
• Only 5 out 33 entries performed the best with
respect to all the three parameters at 25% of
control water,
• Grain yield in most of the entries (17 out of 33)
also increased under 50% water with 9 entries
Results

10. 1010
Height reducedHeight reduced
Yield increasedYield increased
Material from WARDA
1. WAB-56-50
2. WAB-450-I-B-P-135-HB
3. WAB-450-5-1-BL-1-DV6
4. WAB56-104
5. WAB-450-16-2-BL1-DR2
6. WAB450-IBP-157-2-1
7. WAB272-B-B-5+14
8. WAB-450-15-2-BL1-DR6
9. WAB 100-B-B-B-B-21-H2
10. WAB 56-50
11. WAB 450-5-1-BL1-DR5
12. WAB 450-11-2-BL1-DR2
13. WAB 272-B-B-5H3
14. WAB 272-B-B-2H3
15. WAB-450-IBP-163-2
16. WAB 450-IBP-20-HB
17. OS-6 (O.glaberrima)
18. CG-14 Oryza sativa
19. Oryza glaberrima 103929
Material from CUBA
20. J104-G3-291-4-2
21. J104-N2-C53-2
22. LP-9
23J104-N2-59-5
24. LP-8
25.LP-7
26. IA-CUBA
Local selections
27. IR-6 (IRRI)
28.NR-1 (Mutant)
29.Basmati-Supper
30. Basmati-385
31. Basmati-370
32.LS-1 (Mutant)
33. CP-1 (China
27.
Generally, plant height increased after
water stress. Maximum increase was
observed in genotypes collected from
WARDA especially genotypes no. 6
where >200, <150 and >100% increase
was observed in plant height under 50,
75 and 25% of irrigation water,
respectively. Plant height also
increased under water stress in
genotypes collected from CUBA and in
some of the local plants but the
magnitude of increase was less than
that observed in WARDA genotypes.
-50
0
50
100
150
200
250
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1 2 3 4 5 6 7 1 2 3 4 5 6 7 8 9 10
No. of genotypes
Plantheight(%ofcontrol)
75% 50% 25%

11. 1111
Height reducedHeight reduced
Yield increasedYield increased
Material from WARDA
1. WAB-56-50
2. WAB-450-I-B-P-135-HB
3. WAB-450-5-1-BL-1-DV6
4. WAB56-104
5. WAB-450-16-2-BL1-DR2
6. WAB450-IBP-157-2-1
7. WAB272-B-B-5+14
8. WAB-450-15-2-BL1-DR6
9. WAB 100-B-B-B-B-21-H2
10. WAB 56-50
11. WAB 450-5-1-BL1-DR5
12. WAB 450-11-2-BL1-DR2
13. WAB 272-B-B-5H3
14. WAB 272-B-B-2H3
15. WAB-450-IBP-163-2
16. WAB 450-IBP-20-HB
17. OS-6 (O.glaberrima)
18. CG-14 Oryza sativa
19. Oryza glaberrima 103929
Material from CUBA
20. J104-G3-291-4-2
21. J104-N2-C53-2
22. LP-9
23J104-N2-59-5
24. LP-8
25.LP-7
26. IA-CUBA
Local selections
27. IR-6 (IRRI)
28.NR-1 (Mutant)
29.Basmati-Supper
30. Basmati-385
31. Basmati-370
32.LS-1 (Mutant)
33. CP-1 (China
27.
Contrary to plant height, no. of
leaves decreased in general
except genotype no. 18 of
WARDA, 7 of CUBA and 4, 5 and
8 of local selection where no.
of leaves increased
significantly after water
stress.
-150
-100
-50
0
50
100
150
1
3
5
7
9
11
13
15
1
3
5
7
2
4
6
8
10
No of genotypes
Noofleaves(%ofcontrol)
75% 50% 25%

12. 1212
Height reducedHeight reduced
Yield increasedYield increased
Material from WARDA
1. WAB-56-50
2. WAB-450-I-B-P-135-HB
3. WAB-450-5-1-BL-1-DV6
4. WAB56-104
5. WAB-450-16-2-BL1-DR2
6. WAB450-IBP-157-2-1
7. WAB272-B-B-5+14
8. WAB-450-15-2-BL1-DR6
9. WAB 100-B-B-B-B-21-H2
10. WAB 56-50
11. WAB 450-5-1-BL1-DR5
12. WAB 450-11-2-BL1-DR2
13. WAB 272-B-B-5H3
14. WAB 272-B-B-2H3
15. WAB-450-IBP-163-2
16. WAB 450-IBP-20-HB
17. OS-6 (O.glaberrima)
18. CG-14 Oryza sativa
19. Oryza glaberrima 103929
Material from CUBA
20. J104-G3-291-4-2
21. J104-N2-C53-2
22. LP-9
23J104-N2-59-5
24. LP-8
25.LP-7
26. IA-CUBA
Local selections
27. IR-6 (IRRI)
28.NR-1 (Mutant)
29.Basmati-Supper
30. Basmati-385
31. Basmati-370
32.LS-1 (Mutant)
33. CP-1 (China
27.
Number of tillers remains unaffected
in most of the genotypes irrespective
of it origin, except in some of WARDA
selections growing at 25% of
irrigation levels especially the
genotypes no.13 and 14 where
increase in tillers was 7000 % more
compared to control.
-1000
0
1000
2000
3000
4000
5000
6000
7000
8000
1
3
5
7
9
11
13
15
1
3
5
7
2
4
6
8
10
No. of genotypes
No.oftillers(%ofcontrol)
75% 50% 25%

13. 1313
Height reducedHeight reduced
Yield increasedYield increased
Material from WARDA
1. WAB-56-50
2. WAB-450-I-B-P-135-HB
3. WAB-450-5-1-BL-1-DV6
4. WAB56-104
5. WAB-450-16-2-BL1-DR2
6. WAB450-IBP-157-2-1
7. WAB272-B-B-5+14
8. WAB-450-15-2-BL1-DR6
9. WAB 100-B-B-B-B-21-H2
10. WAB 56-50
11. WAB 450-5-1-BL1-DR5
12. WAB 450-11-2-BL1-DR2
13. WAB 272-B-B-5H3
14. WAB 272-B-B-2H3
15. WAB-450-IBP-163-2
16. WAB 450-IBP-20-HB
17. OS-6 (O.glaberrima)
18. CG-14 Oryza sativa
19. Oryza glaberrima 103929
Material from CUBA
20. J104-G3-291-4-2
21. J104-N2-C53-2
22. LP-9
23J104-N2-59-5
24. LP-8
25.LP-7
26. IA-CUBA
Local selections
27. IR-6 (IRRI)
28.NR-1 (Mutant)
29.Basmati-Supper
30. Basmati-385
31. Basmati-370
32.LS-1 (Mutant)
33. CP-1 (China
27.
Grain yield either remain unaffected
or reduced compared to control. In
some of the WARDA and CUBA
collections, it increased significantly
especially in genotypes nos. 20, 22
and 24 of CUBA collections where
increase in yield was between 100 to
>200% of the control.
-400
-300
-200
-100
0
100
200
300
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1 2 3 4 5 6 7 1 2 3 4 5 6 7 8 9 10
No of genotypes
Grainyield(%ofcontrol)
75% 50% 25%

14. 1414
•The plants with the ability to produce higher
number of tillers and grain yield were further tested
for heat tolerance by giving heat shocks of 40, 45
and 50 o
C . We observed several heat shock proteins
especially the induction of 20kDa fraction in OS-2
after heat shock of 50 o
C
•The were then subjected to efficiency analysis. The
results indicated that EF-11, O. glebrrima (CG-14)
and WAB-56-104 maintained optimum Fv/Fm
values (0.80) at 30% light intensity given for longer
periods.

15. 1515
OS-6
4 3 2 1 M
116
97
84
66
55
45
36
29
*
*
*
*
O. glaberrima CG-14
M 1 2 3 4
*
*
*
116
97
84
66
55
45
36
29
20
16
*
*
*
*
Figure showing protein profiles of OS-6 (left) and O.glaberrima CG-14(right) for control
(lane-1), 40oC (lane 2) 45oC (lane 3) and 50oC (lane 4). Symbol * indicate the protein
bands expressed after heat treatment. “M” represents protein molecular weight marker.
*
*

16. 1616
Local selection (LS)
1 2 3 4 M
*
*
*
*
116
97
84
66
55
45
36
29
20
*
M 4 3 2 1
116
97
84
66
55
45
36
29
WAB-56-104
*
*
Figure showing protein profiles of Local selection (left) and WAB-56-104
(right) for control (lane-1), 40o
C (lane 2) 45o
C (lane 3) and 50o
C (lane 4).
Symbol * indicate the protein bands expressed after heat treatment.
“M” represents protein molecular weight marker.

17. 1717
Efficiency of PSII as affected by 50%
light intensity given for various timings
to different rice genotypes

18. 1818
WAB-56-104:WAB-56-104: Plant height reduced upPlant height reduced up
to 40% of control,to 40% of control,
• OS-6OS-6 No. of leaves increased,No. of leaves increased,
Grain yieldGrain yield
unaffectedunaffected
• CG-14CG-14 No. of tillers increasedNo. of tillers increased
• NR-1 &NR-1 & Grain yield increased byGrain yield increased by
• Local line (LS)Local line (LS) 100 to 200 % of control100 to 200 % of control
Based on these criteria, selected
(under 50% less water)

19. 1919
• Since number of leaves is an importantSince number of leaves is an important
criteria to get biomass,criteria to get biomass,
• higher productive tillers and grain yieldhigher productive tillers and grain yield
is necessary to get increased grain yieldis necessary to get increased grain yield
under stress conditions and,under stress conditions and,
• stay green prolong the grain filling bystay green prolong the grain filling by
continuously providing photosynthatecontinuously providing photosynthate
• therefore, we used OS-6, CG-14, WAB-therefore, we used OS-6, CG-14, WAB-
56-104 in crosses with EF-11 (fine grain56-104 in crosses with EF-11 (fine grain
local selection) and IRRI varieties inlocal selection) and IRRI varieties in
order to combine water deficiencyorder to combine water deficiency
tolerance with high grain yield in lowlandtolerance with high grain yield in lowland
local varieties.local varieties.

20. 2020
Crosses attempted in 2004Crosses attempted in 2004
• These crosses that were madeThese crosses that were made
betweenbetween
• IR56/CG-14,IR56/CG-14,
• IR56/OS-6,IR56/OS-6,
• IR56/WAB56-104,IR56/WAB56-104,
• EF11/OS-6,EF11/OS-6,
• Were raised to F1 during 2005Were raised to F1 during 2005
• During 2006, F2 segregationDuring 2006, F2 segregation
population is growing on 50% ofpopulation is growing on 50% of
the water required for normalthe water required for normal
irrigation of rice.irrigation of rice.
• The data recorded so far is beingThe data recorded so far is being
presented below.presented below.

21. 2121
Table 1. Grain yield (gram plantTable 1. Grain yield (gram plant-1-1
) obtained under 50%) obtained under 50%
less water given to the plants growing in F2 segregationless water given to the plants growing in F2 segregation
population of various crosses attempted during 2004population of various crosses attempted during 2004
CrossCross
CombinationCombination
ss
F1F1 Range of grain yield in F2 segregatingRange of grain yield in F2 segregating
populationpopulation
0-100-10 10-2010-20 21-3021-30 31-4031-40 41-50 51-41-50 51-
6060 >60>60
IR56/CG-14IR56/CG-14
(79)(79)
8888 1616 8181 2626 1818 88 11 22
IR56/OS-6IR56/OS-6
(128)(128)
6969 2323 6262 4040 3030 1515 22 22
IR56/WAB56-IR56/WAB56-
104 (139)104 (139)
9393 2424 6565 2525 3030 2323 1515 77
EF11/OS-6EF11/OS-6
(148)(148)
6868 1919 88 5353 1717 11 NilNil NilNil
Figures within brackets are total number of plants in F2 population. Grain
yield of parents was 61 (IR56), 53.5 (CG-14), 25.5 (OS-6), 29 (WAB 56-104)
and 38.5 (EF11) gram plant-1

22. 2222
Stay Green

23. 2323
75%75% 50%50% 25%25%
How OS-6 behaved after reducing waterHow OS-6 behaved after reducing water
100100%%

24. 2424
100%100% 75%75% 50%50% 25%25%
disease appeardisease appear Yield un-affectedYield un-affected

25. 2525
WAB-56-104WAB-56-104
75%75% 50%50%
DiseasedDiseased

26. 2626
EF-11 Height affectedEF-11 Height affected
Disease appearedDisease appeared
100%100% 75%75% 50%50% 25%25%

27. 2727
EF-11/OS-6EF-11/OS-6
75%75% 50%50%
Height reduced,Height reduced,
yield increasedyield increased

28. 2828
EF-11/OS-6EF-11/OS-6
50%50% 25%25%
Disease spot appearedDisease spot appeared

29. 2929
Variation in F2 segregating populationVariation in F2 segregating population
of EF-11 (LS)/CG-14of EF-11 (LS)/CG-14
Height reducedHeight reduced
YieldYield
unaffectedunaffected
75%75% 50%50% 25%25%100%100%

30. 3030
Variation in F2 segregation populationVariation in F2 segregation population
IR56/CG-14IR56/CG-14
100%100% 75%75% 50%50% 25%25%

31. 3131No significant difference in yield, no sterilityNo significant difference in yield, no sterility

32. 3232
Efficiency of PS-II in one of the F2
selections made from the cross of LS-1
x OS-6.

33. 3333
Variation in F2 segregation populationVariation in F2 segregation population
IR56/CG-14IR56/CG-14
Hybridization induced sterilityHybridization induced sterility

34. 3434
Sterility andSterility and
diseasedisease
IR-56/CG-14 controlIR-56/CG-14 control

35. 3535
FF22 IR-56/WAB-56-104IR-56/WAB-56-104
Variation in plant height, grainVariation in plant height, grain
yield not affectedyield not affected

36. 3636
100% 75% 50% 25%100% 75% 50% 25%
IR-56/OS-6IR-56/OS-6
Height and yield comparable to controlHeight and yield comparable to control

37. 3737
F2 IR-56/WAB-56-104F2 IR-56/WAB-56-104
Disease appearedDisease appeared
50%50% 25%25%

38. 3838
Disease appearedDisease appeared

39. 3939

40. 4040

41. 4141
Future PlanFuture Plan
During 2007, plants selected currently inDuring 2007, plants selected currently in
the grain yield range of 30-60 grams plantthe grain yield range of 30-60 grams plant
are being tested in replicated trials under 4are being tested in replicated trials under 4
water levels i.e. full flooded water (100%water levels i.e. full flooded water (100%
control) and 75%, 50% and 25% less thancontrol) and 75%, 50% and 25% less than
that to see the actual potential of thethat to see the actual potential of the
selected plants for water deficiencyselected plants for water deficiency
tolerancetolerance

42. 4242
Future Plan (cont.)Future Plan (cont.)
 All the plants in this experiment would also beAll the plants in this experiment would also be
subjected to Plant Efficiency Analysis to see thesubjected to Plant Efficiency Analysis to see the
status of Fv/Fm and its relationship (if any) with waterstatus of Fv/Fm and its relationship (if any) with water
deficiency tolerancedeficiency tolerance
 Since Plant Efficiency Analysis is based onSince Plant Efficiency Analysis is based on
chlorophyll fluorescence therefore, the actualchlorophyll fluorescence therefore, the actual
greenness of the selected plants would also begreenness of the selected plants would also be
determined to know the actual cause of reduction ( ifdetermined to know the actual cause of reduction ( if
any) in the ratio of Fv/Fm: the normal of which shouldany) in the ratio of Fv/Fm: the normal of which should
be equal to 0.8 or more,be equal to 0.8 or more,
 Plant showing “Stay Green” character would bePlant showing “Stay Green” character would be
selected irrespective of grain yieldselected irrespective of grain yield

43. 4343
•We believe that together we
would be able to select some
of the plants that would grow
on only 25-50% of water
being used now a days to
grow rice,
•This would make an impact
in era when water shortage
will enter into real water
scarcity: the time for which
Conclusion:

44. 4444
Thank you for giving us
the opportunity
And for wonderful hospitality