Rice biotechnology

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Women in rice biotechnology: success that will have an impact in the times ahead

Presented by:Shafqat Farooq

PAEC, Islamabad, Pakistan

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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 also very important.

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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 weight and induces sterility

Introduction cont.

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In 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 for tolerance to water deficiency

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ObjectivesObjectivesi)i) To reduce waterTo reduce water consumption and consumption and ii) to induce temperatureii) to induce temperature tolerance in lowland ricetolerance in lowland rice varieties varieties through transferring gene (s) through transferring gene (s)

from upland rice varieties/ from upland rice varieties/ land racesland races

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Material was directly sown in the field blocks maintained

i. at normal flooded conditions (Control) andii. 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, number of leaves and productive tillers, and grain yield.

Methodology

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Methodology cont.

i. Morphological performance under water deficiency,ii. Appearance of stress

protein iii. Efficiency of plants under water deficient conditions, andiv. Stay Green were considered as criteria to measure water deficiency

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Selection from screening for Selection from screening for water deficiency tolerance water 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

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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 simply out yielded the rest under 25% of control water

Results

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Height reducedHeight reducedYield 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-DR69. WAB 100-B-B-B-B-21-H2 10. WAB 56-50 11. WAB 450-5-1-BL1-DR512. WAB 450-11-2-BL1-DR213. WAB 272-B-B-5H314. WAB 272-B-B-2H315. WAB-450-IBP-163-216. WAB 450-IBP-20-HB 17. OS-6 (O.glaberrima)18. CG-14 Oryza sativa 19. Oryza glaberrima 103929 Material from CUBA 20. J104-G3-291-4-221. J104-N2-C53-222. LP-923J104-N2-59-524. LP-825.LP-726. IA-CUBA Local selections 27. IR-6 (IRRI)28.NR-1 (Mutant)29.Basmati-Supper30. Basmati-38531. Basmati-37032.LS-1 (Mutant)33. CP-1 (China

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.

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

Plan

t hei

ght (

% o

f con

trol

)

75% 50% 25%

1111


Material from WARDA


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.

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

No of genotypes

No

of

leav

es (

% o

f co

ntr

ol)

75% 50% 25%

1212


Material from WARDA


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

No. of genotypes

No. o

f till

ers

(% o

f con

trol)

75% 50% 25%

1313


Material from WARDA


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.

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

Gra

in y

ield

(% o

f con

trol

)

75% 50% 25%

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 oC . We observed several heat shock proteins especially the induction of 20kDa fraction in OS-2 after heat shock of 50 oC

•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.

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OS-6

4 3 2 1 M

1169784

6655

45

36

29

**

**

O. glaberrima CG-14

M 1 2 3 4

***

116978466

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.

*

*

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Local selection (LS)

1 2 3 4 M

**

*

*

116978466

55

45

36

2920

*

M 4 3 2 1

116978466

55

45

36

29

WAB-56-104

* *

Figure showing protein profiles of Local selection (left) and WAB-56-104 (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.

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0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

385 LS CG-14

EF-11 IR-56 IR-5908

OS-6 O. gl 56-104

FV

/Fm

20 30 40 50 60

Efficiency of PSII as affected by 50% light intensity given for various timings to

different rice genotypes

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WAB-56-104: WAB-56-104: Plant height reduced up Plant height reduced up

to 40% of control,to 40% of control,

• OS-6OS-6 No. of leaves increased, No. of leaves increased, Grain yield unaffectedGrain yield unaffected

• CG-14CG-14 No. of tillers increasedNo. of tillers increased

• NR-1 & NR-1 & Grain yield increased by Grain yield increased by

• Local line (LS)Local line (LS) 100 to 200 % of control 100 to 200 % of control

Based on these criteria, selected (under 50% less water)

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• Since number of leaves is an important Since number of leaves is an important criteria to get biomass,criteria to get biomass,

• higher productive tillers and grain yield higher productive tillers and grain yield

is necessary to get increased grain yield is necessary to get increased grain yield under stress conditions and,under stress conditions and,

• stay green prolong the grain filling by stay green prolong the grain filling by

continuously providing photosynthate continuously 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 grain 56-104 in crosses with EF-11 (fine grain local selection) and IRRI varieties in local selection) and IRRI varieties in order to combine water deficiency order to combine water deficiency tolerance with high grain yield in tolerance with high grain yield in lowland local varieties.lowland local varieties.

2020

Crosses attempted in 2004Crosses attempted in 2004• These crosses that were made between These crosses that were made between • 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 2005 Were raised to F1 during 2005 • During 2006, F2 segregation population During 2006, F2 segregation population

is growing on 50% of the water required is growing on 50% of the water required for normal irrigation of rice. for normal irrigation of rice.

• The data recorded so far is being The data recorded so far is being presented below.presented below.

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Table 1. Grain yield (gram plantTable 1. Grain yield (gram plant-1-1) obtained under 50% less ) obtained under 50% less water given to the plants growing in F2 segregation population water given to the plants growing in F2 segregation population of various crosses attempted during 2004of various crosses attempted during 2004

Cross Cross

CombinationsCombinations F1F1 Range of grain yield in F2 segregating Range of grain yield in F2 segregating

populationpopulation

0-10 0-10 10-20 10-20 21-30 21-30 31-40 31-40 41-50 41-50 51-6051-60 >60>60

IR56/CG-14 IR56/CG-14

(79)(79) 8888 1616 8181 2626 1818 88 11 22

IR56/OS-6 IR56/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-6 EF11/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

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Stay Green

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75%75% 5050%%

25%25%

How OS-6 behaved after reducing waterHow OS-6 behaved after reducing water

100100%%

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100%100% 75%75% 50%50% 25%25% disease appeardisease appear Yield un-affected Yield un-affected

2525

WAB-56-104WAB-56-10475%75% 50%50%

Diseased Diseased

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EF-11 Height affectedEF-11 Height affectedDisease appearedDisease appeared

100%100% 75%75% 50% 50% 25%25%

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EF-11/OS-6EF-11/OS-6

75%75% 50%50% Height reduced, Height reduced, yield increased yield increased

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EF-11/OS-6EF-11/OS-650%50% 25% 25%

Disease spot appearedDisease spot appeared

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Variation in F2 segregating populationVariation in F2 segregating population of EF-11 (LS)/CG-14of EF-11 (LS)/CG-14

Height reducedHeight reduced

Yield Yield unaffectedunaffected

75%75% 50%50% 25%25%100%100%

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Variation in F2 segregation populationVariation in F2 segregation populationIR56/CG-14IR56/CG-14

100%100% 75% 75% 50% 50% 25%25%

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

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

0.5

0.6

0.7

0.8

0.9

5 10 15 20 25Time of exposure

Fv/

Fm

20 30 40 50 60

F2-1- flag leaf July 10th, 2004

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Variation in F2 segregation Variation in F2 segregation populationpopulation

IR56/CG-14IR56/CG-14Hybridization induced Hybridization induced

sterility sterility

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Sterility and Sterility and diseasedisease

IR-56/CG-14 controlIR-56/CG-14 control

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FF22 IR-56/WAB-56-104 IR-56/WAB-56-104Variation in plant height, grain Variation in plant height, grain

yield not affectedyield not affected

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100% 75% 50% 25%100% 75% 50% 25%

IR-56/OS-6IR-56/OS-6Height and yield comparable to controlHeight and yield comparable to control

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F2 IR-56/WAB-56-104F2 IR-56/WAB-56-104Disease appearedDisease appeared

50%50% 25%25%

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Disease appeared Disease appeared

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Future PlanFuture Plan

During 2007, plants selected currently inDuring 2007, plants selected currently inthe grain yield range of 30-60 grams plantthe grain yield range of 30-60 grams plantare being tested in replicated trials under 4are being tested in replicated trials under 4water 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 thanthat to see the actual potential of thethat to see the actual potential of theselected plants for water deficiencyselected plants for water deficiencytolerancetolerance

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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 thestatus of Fv/Fm and its relationship (if any) with waterstatus of Fv/Fm and its relationship (if any) with waterdeficiency tolerancedeficiency tolerance

Since Plant Efficiency Analysis is based onSince Plant Efficiency Analysis is based onchlorophyll fluorescence therefore, the actualchlorophyll fluorescence therefore, the actualgreenness of the selected plants would also be greenness of the selected plants would also be determined to know the actual cause of reduction ( if determined to know the actual cause of reduction ( if any) in the ratio of Fv/Fm: the normal of which should any) 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 be Plant showing “Stay Green” character would be selected irrespective of grain yieldselected irrespective of grain yield

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•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 is not very far

Conclusion:

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Thank you for giving us the opportunity

And for wonderful hospitality

Rice biotechnology

Technology

Transcript of Rice biotechnology