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Jauhar ali. vol 3. screening for abiotic and biotic stress tolerances

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Jauhar ali. vol 3. screening for abiotic and biotic stress tolerances

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Jauhar ali. vol 3. screening for abiotic and biotic stress tolerances

  1. 1. Jauhar Ali* Xiuqin Zhao# *International Rice Research Institute #Institute of Crop Science, CAAS
  2. 2. Jauhar Ali Plant Breeder, Senior Scientist IRRI-GSR Project Leader & Regional Coordinator (Asia) PBGB, IRRI (J.Ali@irri.org)
  3. 3. Submergence Tolerance Screening in Screen house 1.Pre-germinate healthy seeds by soaking them in a petri dish containing distilled water placed in an incubator (30oC) for 48 hours. 2.Prepare the planting medium by mixing 5 g of ammonium sulphate in 2.5 liter bucket of soil. 3.Put the treated-soil in a seedbox (15x21inches or 38x53cms). 4.Make 12 rows in the seedbox. 5.Seed the pre-germinated seeds in the seedbox with spacing of ~1 cm (20-30 seeds/row). 6.Count the total number of seedlings and measure the average of the plant height of Modified from : Xu K, Mackill DJ (1996) A major locus for submergence tolerance mapped on rice chromosome 9. Mol Breeding 2:219-224. 6.Count the total number of seedlings and measure the average of the plant height of each line before submergence (14 days after emergence). 7.Place the seedbox in submergence tank & fill the tank with fresh water (<1m depth). 8.Monitor the floodwater conditions daily (temperature, dissolved O2, light penetration, and pH). 9.IR42 is used as susceptible check, and FR13A or other donors is used as tolerant check. Check IR42 seedlings 10-14 days after submergence. If they are already 70- 80% chlorotic and very soft, you may remove the seedbox from the tank. 10.Measure the average of plant height of each line after removing the seedbox from the water. 11.Count the percent survival rate at 10 and 21 days after de-submergence.
  4. 4. 1.Sow the seeds of each line in black trays using three seeds per hole (each hill measures 1.5 (W) x 1.5 (L) x2.5 (H) cu cm). If black trays are not available, seed them in seedboxes at the spacing of 4 cm between rows and 1cm between seeds. 2.Prepare the land in field tanks. Apply molluscicide after the first and second harrowing. 3.After the final harrowing, apply 30:30:30:: N: P: K through Submergence Screening in Field Tanks Modified from: Xu K, Mackill DJ (1996) A major locus for submergence tolerance mapped on rice chromosome 9. Mol Breeding 2:219-224. 3.After the final harrowing, apply 30:30:30:: N: P: K through complete fertilizer as basal along with full dose of Zn as Zinc Sulphate (20 kg/ha). 4.Apply the remaining N (60 kg/ha) in to two splits through Urea, first at maximum tillering and the second one at panicle initiation. 5.Transplant the seedling (14 d) in the field using two seedlings per hill at 20X 20 cm2 distance. 6.Transplant extra IR42 seedling at one side of field to monitor the submergence stress.
  5. 5. 7. Submerge seedlings completely two weeks after the transplanting time. Plants will be completely submerged with a water head of 120-125 cm at noon to give plants time to photosynthesize in the morning. 8. Monitor the floodwater conditions (temperature, light penetration, dissolved O2, and pH) daily. 9. After 10d of submergence, uproot 5 plants daily from the extra rows of IR42 to observe their condition. In case of severe submergence plants will be 70-80% chlorotic and stems will be very soft. This condition is Submergence field tanks-(continued) be 70-80% chlorotic and stems will be very soft. This condition is expected to come any day starting from 10 to 14 days depending upon flood water quality and environmental conditions. 10.Just after desubmergence allow field to remain without water for 3-4 days. Afterward fill it with not more than 1-2 cm water until another 15 to 20 days; then increase water level to normal 5-7cm. 11.Measure plant height of the seedlings before and after submergence. 12.Percent survival will be taken 21 days after de-submergence.
  6. 6. Submergence screening: submergence tankSubmergence screening: submergence tankSubmergence screening: submergence tankSubmergence screening: submergence tank Populations (BC1F2) Single plant selections (BC1F3-5) Day 0: Seed soaking (pre-germinate at 30OC for 48 hours) Planting medium – seed boxes Day 17: Submergence (count total no. of seedlings) (Place in submergence tank with fresh water at <1m depth) Day 31: De-submergence (Remove seed boxes from tank)Planting medium – seed boxes mix 5 grams Ammonium sulphate in 2.5 kg soil and put in seed box (15in x 21in); make 12 rows in the seed box Day 2: Sowing (Sow pre-germinated seeds in seedbox with spacing ~1cm; 20-30 seeds/row ) (Remove seed boxes from tank) (count no. of surviving seedling at 10 & 21 days after de-submergence) Day 52: Transplanting (transplant surviving plants) (Single seedling per hill) Maturity: Harvesting (Single plant harvesting)
  7. 7. SUBMERGEN CE screeningCE screening in screen house
  8. 8. Submergence mass screening: field tankSubmergence mass screening: field tankSubmergence mass screening: field tankSubmergence mass screening: field tank Populations (BC1F2, 20g/pop’n.) Single plant selections (BC1F3-5, 2g/line) Seed beds: From prepared land in field tanks, prepare wet beds, <1m wide, make rows at ~7cm distance. Day 31:De-submergence (Drain water from the tank) Day 52: Scoring (count for percent survival 21 days after de-submergence)rows at ~7cm distance. Day 0: Sowing (sow seeds at 10rows/population or 1row/line, cover with thin layer of soil) Day 17: Submergence (score for germination/emergence) (measure average plant height) (14 DAE, fill the tank with ~1m depth of fresh water for 14 days) 21 days after de-submergence) Transplanting (transplant surviving plants) (Single seedling per hill) Maturity: Harvesting (Single plant harvesting)
  9. 9. Screening of BC2F2 populations for submergence tolerance in a deep-water pond Thirty-five-day old seedlings were submerged under deep water for two weeks, then allowed to recover
  10. 10. Anaerobic germination screeningAnaerobic germination screeningAnaerobic germination screeningAnaerobic germination screening Seeds are direct seeded and immediately submerged in waterSeeds are direct seeded and immediately submerged in waterSeeds are direct seeded and immediately submerged in waterSeeds are direct seeded and immediately submerged in water with 10 cm depth for 21 days. Lines showing high germinationwith 10 cm depth for 21 days. Lines showing high germinationwith 10 cm depth for 21 days. Lines showing high germinationwith 10 cm depth for 21 days. Lines showing high germination score under low oxygen condition are identified.score under low oxygen condition are identified.score under low oxygen condition are identified.score under low oxygen condition are identified.
  11. 11. Jauhar Ali Plant Breeder, Senior Scientist IRRI-GSR Project Leader & Regional Coordinator (Asia) PBGB, IRRI (J.Ali@irri.org)
  12. 12. •Sieve soil and transfer it in a plastic tray (plastic tray has 17 holes/row) •Prepare 17-34 dry seeds/line (clean, fully filled and not discolored) •Sow the dry seeds (dry seeding) with one seed one hill (total of 17-34 seeds/line), which each hill measures 1.5 (W) X 1.5 (L) X 2.5 (H) cu cm. •Place the seed about 1 cm (not more) below the soil surface. When all the rows in the plastic tray have seeds, cover the seeds entirely using the sieved soil, filling up the hill. Anaerobic Germination Screening Protocol Modified from protocol developed by Dr. A. Ismail’s group, unpublished, CESD, IRRI soil, filling up the hill. •Submergence is done in the concrete table. Maintain the water depth of 5-7 cm. Observe daily, remove weeds and algae. • Daily measure the water conditions (light, pH, O2, and temp level of water) •Score for survival 21 days after seeding. [The percent survival (seeds that germinated and seedlings emerged out of the submerged condition) was recorded for each BC population and the surviving plants were transferred to the field for seed production. Seeds from the surviving plants were harvested and the progeny was tested under the same conditions in the following season to confirm the tolerance of the selected AGT lines.]
  13. 13. Jauhar Ali Plant Breeder, Senior Scientist IRRI-GSR Project Leader & Regional Coordinator (Asia) PBGB, IRRI (J.Ali@irri.org)
  14. 14. Screening for seedling cold tolerance Twelve-day old seedlings were subjected to cold temperature for 18 days at the mean daily temperature of 11.8 Co, including 3-day of low temperature at 8 Co between April 24-26 (LAAS, 2002).
  15. 15. Selection of 861 C418 plants with seedling cold tolerance from 28 C418 BC2F2 populations 2002 (LAAS) # of populations 28 2 26 BC2F2 CT donors Non-CT donors Seedling Cold Tolerance (from NARES) Range 1.4 – 19.3% # of surviving plants per population 10.3% The mean population size was 310, ranging from 196 – 465, the recipient, C418 (japonica) was killed by the stress. 10 – 16% 0 – 3.0% 0.314% # of surviving plants per BC population 10.3% 10.5%7.6%
  16. 16. NAFREC Lao PDR Luang Prabang Lao PDR FC&RI, Vietnam FC&RI, Vietnam CARDI Cambodia Batalagoda,Sri Lanka Batalagoda,Sri Lanka CARDI Cambodia Phka Khgnei, NIA, TandoJam, Pakistan PT Sang Hyan Seri,Phka Khgnei, Cambodia PT Sang Hyan Seri, Sukamandi, Indonesia NIBGE, Faisalabad, Pakistan RRI, Kala Shah Kaku BRAC, Ghazipur Irrigated ICRR Pusakanegara Irrigated ICRR Sukamandi Irrigated
  17. 17. CAMBODIA GSR Hybrid CNH 9101 IR56383IR56383--3535--33--22--11 OR142-99 CAR9 PRM CAMBODIA GSR Hybrid TRIALS IG 80+5 cks CNH 9107CNH 9111 Partially water stressed under irrigated conditions CNH 9097 CNH 9099
  18. 18. Initial GSR Success stories in Asia: Sri Lanka CNH9050 3½ month hybrid desirable identified 4½ month hybrid undesirable to farmers BG 407 H GSR inbred performing well in Sri Lanka under severely water stressed rainfed lowland conditions against their checks CNI9024 BG358
  19. 19. Line FGW- stress(g) FGW-S.Stress (g) %reduction in yield DM-Stress DM- S.Stress CNI25 222.0 204.0 8.8 116 123 CNI26 145.3 143.8 1.0 109 130 CNI 7 159.3 141.5 12.6 103 130 CNI21 141.9 132.9 6.8 113 130 CNI13 171.8 122.4 40.4 109 124 CNI10 166.8 112.1 48.8 109 130 CNI23 138.1 103.2 33.8 109 124 CNI14 145.6 100.7 44.6 103 130 SRI LANKAN RAINFED GSR INBRED SCREENING CNI14 145.6 100.7 44.6 103 130 At308 150.4 93.4 61.0 103 130 CNI24 158.4 88.2 79.6 94 117 CNI28 135.2 76.4 77.0 90 86 Bg250 140.3 74.6 88.1 109 117 At581 174.4 74.5 134.1 103 130 Bg3R 156.0 58.7 165.8 103 130 At1382 120.1 45.5 164.0 103 124 Bg 304 165.4 40.9 304.4 94 124
  20. 20. SRI LANKASRI LANKA --RRDIRRDI--STRESSSTRESS SRI LANKASRI LANKA --RRDIRRDI--SEVERE STRESSSEVERE STRESS PAKISTANPAKISTAN--KSKKSK--IGIG PAKISTANPAKISTAN--NIANIA--IGIG EntryEntry Yld(t/ha)Yld(t/ha) StressedStressed % over% over checkcheck EntryEntry Yld(t/ha)Yld(t/ha) S.StressedS.Stressed % over% over checkcheck EntryEntry Yld(t/ha)Yld(t/ha) (IG)(IG) %% overover checkcheck EntryEntry Yld(t/ha)Yld(t/ha) (IG)(IG) % over% over checkcheck CNI9003CNI9003 2353.02353.0 34.934.9 CNI9025CNI9025 20402040 118.4118.4 CNI9021CNI9021 34463446 18.6518.65 CNI9023CNI9023 67736773 46.8146.81 CNI9025CNI9025 2220.02220.0 27.327.3 CNI9031CNI9031 16571657 77.477.4 CNI9031CNI9031 32853285 13.1213.12 CNI9015CNI9015 65336533 41.6141.61 CNI9011CNI9011 2153.02153.0 23.523.5 CNI9026CNI9026 14381438 54.054.0 CNI9001CNI9001 30853085 6.226.22 CNI9016CNI9016 58675867 27.1627.16 CNI9018CNI9018 1913.01913.0 9.79.7 CNI9007CNI9007 14151415 51.551.5 CNI9009CNI9009 30263026 4.194.19 CNI9003CNI9003 56005600 21.3821.38 CNI9029CNI9029 1861.01861.0 6.76.7 CNI9021CNI9021 13291329 42.342.3 CNI9012CNI9012 29362936 1.101.10 CNI9026CNI9026 52805280 14.4414.44 At581At581 1744.01744.0 0.00.0 CNI9013CNI9013 12241224 31.031.0 IR6IR6 29002900 --0.130.13 CNI9018CNI9018 50675067 9.829.82 CNI9013CNI9013 1718.01718.0 --1.51.5 CNI9010CNI9010 11211121 20.020.0 CNI9030CNI9030 28692869 --1.191.19 CNI9019CNI9019 50675067 9.829.82 Adaptability GSR inbred trial (31) in South Asia At 308At 308 1698.01698.0 --2.62.6 CNI9017CNI9017 10831083 16.016.0 CNI9004CNI9004 28592859 --1.551.55 CNI9012CNI9012 46134613 0.000.00 CNI9030CNI9030 1669.01669.0 --4.34.3 CNI9023CNI9023 10321032 10.510.5 CNI9020CNI9020 27312731 --5.955.95 KSKKSK--282282 46134613 0.000.00 CNI9010CNI9010 1668.01668.0 --4.44.4 At308At308 934934 0.00.0 CNI9011CNI9011 27082708 --6.766.76 CNI9001CNI9001 45604560 --1.161.16 Bg 304Bg 304 1654.01654.0 --5.25.2 Bg250Bg250 746746 --20.120.1 CNI9007CNI9007 26732673 --7.967.96 CNI9007CNI9007 43734373 --5.215.21 Bg3RBg3R 1560.01560.0 --10.610.6 At581At581 745745 --20.220.2 KSK 133KSK 133 20622062 --29.0029.00 CNI9014CNI9014 43474347 --5.795.79 At308At308 1504.01504.0 --13.813.8 At 308At 308 637637 --31.831.8 DRDR--8383 17301730 --40.4440.44 IR6IR6 42934293 --6.946.94 Bg250Bg250 1403.01403.0 --19.619.6 Bg3RBg3R 587587 --37.237.2 At1382At1382 1201.01201.0 --31.131.1 At1382At1382 455455 --51.351.3 Bg 304Bg 304 409409 --56.256.2 PlotPlot SizeSize 1.0m1.0m22 1.0m1.0m22 1.25m1.25m22 1.25m1.25m22 No. ofNo. of Lines >Lines > CheckCheck 55 1010 55 88
  21. 21. Adaptability GSR inbred trial (31) in S. E. Asia
  22. 22. Adaptability GSR hybrid trial (122) in South Asia
  23. 23. BG407H GSR-H-0158 Irrigated condition, Batalgoda, Sri Lanka
  24. 24. ThanksThanks
  25. 25. Thanks Reference:Reference: Training manual Fischer K.2003 ”BreedingTraining manual Fischer K.2003 ”Breeding rice for droughtrice for drought--prone environments”prone environments” available in theavailable in the web:http://www.knowledgebank.irri.org/drought/web:http://www.knowledgebank.irri.org/drought/web:http://www.knowledgebank.irri.org/drought/web:http://www.knowledgebank.irri.org/drought/ web:http://www.plantstress.com/web:http://www.plantstress.com/
  26. 26. Jauhar Ali*Jauhar Ali* Yongli Zhou# *International Rice Research Institute #Institute of Crop Science, CAAS
  27. 27. Selection Strategy Identify a widely adaptable genotype Use several donors 15 to 45 donors BC1F2 is better than higher generations Screening BC1F2 under higher levels of stress Level of stress should be able to kill the recipient parent It should allow us to select a good number of Ils that can be managed based on breeding capacity of a given centermanaged based on breeding capacity of a given center Progeny confirmation is essential for at least two rounds before we can be sure that these trait is stably being inherited without segregating Extreme transgressive segregants must be carefully utilized for further pyramiding across different donors
  28. 28. 15 20 25 30 Hua 564 Hua 565 Wanxian 763 Wanxian 77 Huanghuazhan Hexi 41 Yunjing 23 SAGC-4 SAGC-7 Zhonghua 1 Weed Tolerant Rice 1 Wuyujing 3 Wuyujing 20 Bacterialblightlesionlength(cm) Resistant Susceptible 0 5 10 0 5 10 15 Blast (%DLA) 053A-3 BD007 Cau 1 Cau 2 Yundao 1 Luyin 46 RC8 6527 JH15-1-1-1 PD29 D4098 Resistant Susceptible Bacterialblightlesionlength(cm)
  29. 29. Population Cross Donor Gen. Number of lines (10 DS) HHZ5 Huang-Hua-Zhan*2/OM1723 OM1723 BC1F5 75 HHZ8 Huang-Hua-Zhan*2/Phalguna Phalguna BC1F5 56 HHZ9 Huang-Hua-Zhan*2/IR50 IR50 BC1F5 62 HHZ11 Huang-Hua-Zhan*2/IR64 IR64 BC1F5 56 Materials evaluated for blast and bacterial blight resistance HHZ12 Huang-Hua-Zhan*2/Teqing Teqing BC1F5 66 HHZ15 Huang-Hua-Zhan*2/PSBRC66 PSBRc66 BC1F5 45 HHZ17 Huang-Hua-Zhan*2/CDR22 CDR22 BC1F5 70 HHZ19 Huang-Hua-Zhan*2/PSBRC28 PSBRc28 BC1F5 82 by Dr. Xu in 2010-2011
  30. 30. 10 15 20 25 30 PXO 61 PXO 86 PXO 79 PXO 340 PXO 71 PXO 112 PXO 99 PXO 145 PXO 280 PXO 339 PXO 341 PXO 347 0 5 PXO 349 PXO 363 Interaction among 14 Xoo strains and selected lines in HHZ15 population.
  31. 31. Blast evaluation of virulent strainsBlast evaluation of virulent strains Evaluation of BB resistance of >500 linesEvaluation of BB resistance of >500 lines (HHZ background) against 14 strains of(HHZ background) against 14 strains of 1010 XooXoo races, 2010 WSraces, 2010 WS HHZ PSBRc66 BC1F5 # 329 BC1F5 #350Meirong Xu et al
  32. 32. Zhongzu14-ski-4-1
  33. 33. BPH and Virus Resistance Screening IRRI-ICRR joint project collaborators: Prof.Baehaki/Drs Muhsin,Untung • 30 BC3F2 and BC2F3 population (CS 3) • 39 BC3F3 and BC2F4 population (CS 4; 3rd year)ongoing BC2 F3 HHZ populations screened against virulent BPH strain that caused outbreak in Sukamandi in 2010 Several populations showed ILs with comparable resistance with the checks in second round of screening. ICRR 8.2011
  34. 34. Ragged stunt virus- BPH outbreak in Sukamandi, Indonesia Ragged stunt virus- BPH –Tolerant inbred lines Promising Multiple disease resistant GSR hybrids at Jakenan rainfed conditions DS2010
  35. 35. Jakenan, Indonesia GSR trials: 42 DT hybrids + 3 checks & 31 DT inbreds + 4 checks (4th March 2010) GSR hybrid under rainfed hand dibbled direct seeded Ciherang check variety
  36. 36. Adaptability GSR hybrid trial (122) in South East Asia

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