Rice biotechnology
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Rice biotechnology

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  • 06/19/13
  • 06/19/13

Rice biotechnology Rice biotechnology Presentation Transcript

  • 11Women in rice biotechnology:success that will have animpact in the times aheadPresented by:Shafqat FarooqPAEC, Islamabad, Pakistan
  • 22IntroductionRice is the most important crop at global level asit is used as staple food in the most countries ofthe worldSecond major staple cereal being grown on 2.5million hectares in Pakistan, are lowland cultivarsand require a lot of water to grow. The supply ofwater is diminishing with every passing day, Agrain reduction of >30 % occurred during therecent past,Identifying stress-resistance genotypes isessential for genetic improvement of riceespecially for water deficiency tolerance (WD).The use of suitable methodology for measuringstress resistance in large breeding population is
  • 33Some genotypes are more resistant thanothers and out-yield those exposed to thesame degree of water stressThe performance of rice varieties underwater stress conditions at different growthstages varies. It is observed that yieldlosses resulting from water deficit areparticularly sever when drought strikes atbooting stage.Water stress at or before, panicle initiationreduces the most potential spikeletnumber and stress during grain fillingdecreases translocation of assimilates tothe grains which decreases the grainIntroduction cont.
  • 44n order to find WD , we testedRice genotypesconsisting ofi. five local basmati (fine grainaromatic) rice cultivars, IR-6,CP-1 (Chinese variety),ii. 19 lines/land races fromWARDA,iii. and 7 varieties/lines fromCUBA
  • 55ObjectivesObjectivesi)i) To reduce waterTo reduce waterconsumption andconsumption andii) to induce temperatureii) to induce temperaturetolerance in lowlandtolerance in lowlandricericevarietiesvarietiesthrough transferring genethrough transferring gene(s) from upland rice(s) from upland ricevarieties/ land racesvarieties/ land races
  • 66Material was directly sownin the field blocksmaintainedi. at normal flooded conditions(Control) andii. at 75%, 50% and 25% lesswaterthan control,iii. Fertilizer was applied at thenormal recommended doze,iv. Data was collected withreference to plant height,Methodology
  • 77Methodology cont.i. Morphologicalperformanceunder water deficiency,ii. Appearance of stressproteiniii. Efficiency of plantsunderwater deficientconditions,
  • 88Selection from screening forSelection from screening forwater deficiency tolerancewater deficiency toleranceexperimentsexperimentsWe tested all the genotypes under differentwater levels used for irrigation i.e. 100%which is control, and 75%, 50% and 25% lesswater than that required for normal floodedirrigation. Selections were made for theplants that can tolerate at-least 50% lesswater than control and showi) Reduced plant height,ii) Increased number of leaves,iii) Tillers,iv) Plant in which grain yield remainedunaffectedand/or.v) Stay green
  • 99Significant variations were observed for allthe parameters and in all the material.•In 19 entries (IR-6, CP-1, material from WARDAand Cuba), number of tillers and leaves were thehighest at 50% reduced water than control,• In 10 and 8 entries, plant height increased by31.7% and 61.3 %, respectively at 25% and 50%less water than control,• Only 5 out 33 entries performed the best withrespect to all the three parameters at 25% ofcontrol water,• Grain yield in most of the entries (17 out of 33)also increased under 50% water with 9 entriesResults
  • 1010Height reducedHeight reducedYield increasedYield increasedMaterial from WARDA1. WAB-56-502. WAB-450-I-B-P-135-HB3. WAB-450-5-1-BL-1-DV64. WAB56-1045. WAB-450-16-2-BL1-DR26. WAB450-IBP-157-2-17. WAB272-B-B-5+148. WAB-450-15-2-BL1-DR69. WAB 100-B-B-B-B-21-H210. WAB 56-5011. 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-HB17. OS-6 (O.glaberrima)18. CG-14 Oryza sativa19. Oryza glaberrima 103929Material from CUBA20. J104-G3-291-4-221. J104-N2-C53-222. LP-923J104-N2-59-524. LP-825.LP-726. IA-CUBALocal selections27. IR-6 (IRRI)28.NR-1 (Mutant)29.Basmati-Supper30. Basmati-38531. Basmati-37032.LS-1 (Mutant)33. CP-1 (China27.Generally, plant height increased afterwater stress. Maximum increase wasobserved in genotypes collected fromWARDA especially genotypes no. 6where >200, <150 and >100% increasewas observed in plant height under 50,75 and 25% of irrigation water,respectively. Plant height alsoincreased under water stress ingenotypes collected from CUBA and insome of the local plants but themagnitude of increase was less thanthat observed in WARDA genotypes.-500501001502002501 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 10No. of genotypesPlantheight(%ofcontrol)75% 50% 25%
  • 1111Height reducedHeight reducedYield increasedYield increasedMaterial from WARDA1. WAB-56-502. WAB-450-I-B-P-135-HB3. WAB-450-5-1-BL-1-DV64. WAB56-1045. WAB-450-16-2-BL1-DR26. WAB450-IBP-157-2-17. WAB272-B-B-5+148. WAB-450-15-2-BL1-DR69. WAB 100-B-B-B-B-21-H210. WAB 56-5011. 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-HB17. OS-6 (O.glaberrima)18. CG-14 Oryza sativa19. Oryza glaberrima 103929Material from CUBA20. J104-G3-291-4-221. J104-N2-C53-222. LP-923J104-N2-59-524. LP-825.LP-726. IA-CUBALocal selections27. IR-6 (IRRI)28.NR-1 (Mutant)29.Basmati-Supper30. Basmati-38531. Basmati-37032.LS-1 (Mutant)33. CP-1 (China27.Contrary to plant height, no. ofleaves decreased in generalexcept genotype no. 18 ofWARDA, 7 of CUBA and 4, 5 and8 of local selection where no.of leaves increasedsignificantly after waterstress.-150-100-50050100150135791113151357246810No of genotypesNoofleaves(%ofcontrol)75% 50% 25%
  • 1212Height reducedHeight reducedYield increasedYield increasedMaterial from WARDA1. WAB-56-502. WAB-450-I-B-P-135-HB3. WAB-450-5-1-BL-1-DV64. WAB56-1045. WAB-450-16-2-BL1-DR26. WAB450-IBP-157-2-17. WAB272-B-B-5+148. WAB-450-15-2-BL1-DR69. WAB 100-B-B-B-B-21-H210. WAB 56-5011. 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-HB17. OS-6 (O.glaberrima)18. CG-14 Oryza sativa19. Oryza glaberrima 103929Material from CUBA20. J104-G3-291-4-221. J104-N2-C53-222. LP-923J104-N2-59-524. LP-825.LP-726. IA-CUBALocal selections27. IR-6 (IRRI)28.NR-1 (Mutant)29.Basmati-Supper30. Basmati-38531. Basmati-37032.LS-1 (Mutant)33. CP-1 (China27.Number of tillers remains unaffectedin most of the genotypes irrespectiveof it origin, except in some of WARDAselections growing at 25% ofirrigation levels especially thegenotypes no.13 and 14 whereincrease in tillers was 7000 % morecompared to control.-1000010002000300040005000600070008000135791113151357246810No. of genotypesNo.oftillers(%ofcontrol)75% 50% 25%
  • 1313Height reducedHeight reducedYield increasedYield increasedMaterial from WARDA1. WAB-56-502. WAB-450-I-B-P-135-HB3. WAB-450-5-1-BL-1-DV64. WAB56-1045. WAB-450-16-2-BL1-DR26. WAB450-IBP-157-2-17. WAB272-B-B-5+148. WAB-450-15-2-BL1-DR69. WAB 100-B-B-B-B-21-H210. WAB 56-5011. 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-HB17. OS-6 (O.glaberrima)18. CG-14 Oryza sativa19. Oryza glaberrima 103929Material from CUBA20. J104-G3-291-4-221. J104-N2-C53-222. LP-923J104-N2-59-524. LP-825.LP-726. IA-CUBALocal selections27. IR-6 (IRRI)28.NR-1 (Mutant)29.Basmati-Supper30. Basmati-38531. Basmati-37032.LS-1 (Mutant)33. CP-1 (China27.Grain yield either remain unaffectedor reduced compared to control. Insome of the WARDA and CUBAcollections, it increased significantlyespecially in genotypes nos. 20, 22and 24 of CUBA collections whereincrease in yield was between 100 to>200% of the control.-400-300-200-10001002003001 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 10No of genotypesGrainyield(%ofcontrol)75% 50% 25%
  • 1414•The plants with the ability to produce highernumber of tillers and grain yield were further testedfor heat tolerance by giving heat shocks of 40, 45and 50 oC . We observed several heat shock proteinsespecially the induction of 20kDa fraction in OS-2after heat shock of 50 oC•The were then subjected to efficiency analysis. Theresults indicated that EF-11, O. glebrrima (CG-14)and WAB-56-104 maintained optimum Fv/Fmvalues (0.80) at 30% light intensity given for longerperiods.
  • 1515OS-64 3 2 1 M11697846655453629****O. glaberrima CG-14M 1 2 3 4***116978466554536292016****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 proteinbands expressed after heat treatment. “M” represents protein molecular weight marker.**
  • 1616Local selection (LS)1 2 3 4 M****1169784665545362920*M 4 3 2 111697846655453629WAB-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.
  • 1717Efficiency of PSII as affected by 50%light intensity given for various timingsto different rice genotypes
  • 1818WAB-56-104:WAB-56-104: Plant height reduced upPlant height reduced upto 40% of control,to 40% of control,• OS-6OS-6 No. of leaves increased,No. of leaves increased,Grain yieldGrain yieldunaffectedunaffected• 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 controlBased on these criteria, selected(under 50% less water)
  • 1919• Since number of leaves is an importantSince number of leaves is an importantcriteria to get biomass,criteria to get biomass,• higher productive tillers and grain yieldhigher productive tillers and grain yieldis necessary to get increased grain yieldis necessary to get increased grain yieldunder stress conditions and,under stress conditions and,• stay green prolong the grain filling bystay green prolong the grain filling bycontinuously 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 grainlocal selection) and IRRI varieties inlocal selection) and IRRI varieties inorder to combine water deficiencyorder to combine water deficiencytolerance with high grain yield in lowlandtolerance with high grain yield in lowlandlocal varieties.local varieties.
  • 2020Crosses attempted in 2004Crosses attempted in 2004• These crosses that were madeThese crosses that were madebetweenbetween• 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 segregationpopulation is growing on 50% ofpopulation is growing on 50% ofthe water required for normalthe water required for normalirrigation of rice.irrigation of rice.• The data recorded so far is beingThe data recorded so far is beingpresented below.presented below.
  • 2121Table 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 segregationpopulation of various crosses attempted during 2004population of various crosses attempted during 2004CrossCrossCombinationCombinationssF1F1 Range of grain yield in F2 segregatingRange of grain yield in F2 segregatingpopulationpopulation0-100-10 10-2010-20 21-3021-30 31-4031-40 41-50 51-41-50 51-6060 >60>60IR56/CG-14IR56/CG-14(79)(79)8888 1616 8181 2626 1818 88 11 22IR56/OS-6IR56/OS-6(128)(128)6969 2323 6262 4040 3030 1515 22 22IR56/WAB56-IR56/WAB56-104 (139)104 (139)9393 2424 6565 2525 3030 2323 1515 77EF11/OS-6EF11/OS-6(148)(148)6868 1919 88 5353 1717 11 NilNil NilNilFigures within brackets are total number of plants in F2 population. Grainyield of parents was 61 (IR56), 53.5 (CG-14), 25.5 (OS-6), 29 (WAB 56-104)and 38.5 (EF11) gram plant-1
  • 2222Stay Green
  • 232375%75% 50%50% 25%25%How OS-6 behaved after reducing waterHow OS-6 behaved after reducing water100100%%
  • 2424100%100% 75%75% 50%50% 25%25%disease appeardisease appear Yield un-affectedYield un-affected
  • 2525WAB-56-104WAB-56-10475%75% 50%50%DiseasedDiseased
  • 2626EF-11 Height affectedEF-11 Height affectedDisease appearedDisease appeared100%100% 75%75% 50%50% 25%25%
  • 2727EF-11/OS-6EF-11/OS-675%75% 50%50%Height reduced,Height reduced,yield increasedyield increased
  • 2828EF-11/OS-6EF-11/OS-650%50% 25%25%Disease spot appearedDisease spot appeared
  • 2929Variation in F2 segregating populationVariation in F2 segregating populationof EF-11 (LS)/CG-14of EF-11 (LS)/CG-14Height reducedHeight reducedYieldYieldunaffectedunaffected75%75% 50%50% 25%25%100%100%
  • 3030Variation in F2 segregation populationVariation in F2 segregation populationIR56/CG-14IR56/CG-14100%100% 75%75% 50%50% 25%25%
  • 3131No significant difference in yield, no sterilityNo significant difference in yield, no sterility
  • 3232Efficiency of PS-II in one of the F2selections made from the cross of LS-1x OS-6.
  • 3333Variation in F2 segregation populationVariation in F2 segregation populationIR56/CG-14IR56/CG-14Hybridization induced sterilityHybridization induced sterility
  • 3434Sterility andSterility anddiseasediseaseIR-56/CG-14 controlIR-56/CG-14 control
  • 3535FF22 IR-56/WAB-56-104IR-56/WAB-56-104Variation in plant height, grainVariation in plant height, grainyield not affectedyield not affected
  • 3636100% 75% 50% 25%100% 75% 50% 25%IR-56/OS-6IR-56/OS-6Height and yield comparable to controlHeight and yield comparable to control
  • 3737F2 IR-56/WAB-56-104F2 IR-56/WAB-56-104Disease appearedDisease appeared50%50% 25%25%
  • 3838Disease appearedDisease appeared
  • 3939
  • 4040
  • 4141Future PlanFuture PlanDuring 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
  • 4242Future Plan (cont.)Future Plan (cont.) All the plants in this experiment would also beAll the plants in this experiment would also besubjected 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 begreenness of the selected plants would also bedetermined to know the actual cause of reduction ( ifdetermined to know the actual cause of reduction ( ifany) in the ratio of Fv/Fm: the normal of which shouldany) in the ratio of Fv/Fm: the normal of which shouldbe 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 beselected irrespective of grain yieldselected irrespective of grain yield
  • 4343•We believe that together wewould be able to select someof the plants that would growon only 25-50% of waterbeing used now a days togrow rice,•This would make an impactin era when water shortagewill enter into real waterscarcity: the time for whichConclusion:
  • 4444Thank you for giving usthe opportunityAnd for wonderful hospitality