Introgression of shoot fly resistance QTLs into elite postrainy season sorghum varieties using Marker Assisted Backcrossing (MAB)

Transcription

1 Introgression of shoot fly resistance QTLs into elite postrainy season sorghum varieties using Marker Assisted Backcrossing (MAB) Sunita Gorthy Research Scholar Sorghum breeding International Crops Research Institute for Semi-Arid Tropics, Patancheru, Hyderabad Supervisor: A Ashok Kumar Senior Scientist, Sorghum breeding International Crops Research Institute for Semi-Arid Tropics, Patancheru, Hyderabad Co Supervisor: Dr M Lakshmi Narsu Professor, Department of Biotechnology Jawaharlal Nehru Technological University, Hyderabad

2 INTRODUCTION Sorghum [sorghum bicolor (L.) Moench ] is the fifth most important cereal crop in terms of production and utilization. Sorghum is produced in the rainy (hot) season in most parts of the world for various uses: food, feed, fodder, industrial starch, biofuels, etc. In India it is grown in both rainy and postrainy (cold) seasons Rainy Season Sorghum 3.0 m ha and Postrainy Season Sorghum 4.5 m ha The postrainy sorghum grain is preferred for food use owing to its bold globular lustrous nature Sorghum is affected by various biotic factors, shoot fly, aphids and charcoal rot play havoc with postrainy season production

3 SHOOT FLY, ATHERIGONA SOCCATA LIFE CYCLE Shoot fly, Atherigona soccata

4 COMPONENT TRAITS FOR SHOOTFLY RESISTANCE Glossiness Leaf Trichomes Deadhearts Percent Ovipositional nonpreference Seedling vigor

5 PREVIOUS RESEARCH ON SHOOTFLY RESISTANCE Satish et al., 2012, Mol. Breeding

6 PREVIOUS RESEARCH ON SHOOTFLY RESISTANCE Satish et al., 2012, Mol. Breeding

7 Candidate genes underlying these QTLs and their association with shootfly resistance Chromosomes Genes Traits XnhsbmSFC95 (auxin-responsive Ovipositional Non-preference SBI 01 protein IAA30) (Sb01g014130) XnhsbmSFCILP81 (Calcium Seedling vigor dependent protein kinase)(sb01g014730) SBI 05 XnhsbmSFC61 (BZIP transcription factor) (Sb05g003240) Glossiness, Ovipositional Nonpreference and deadhearts XnhsbmSFC110 (caffeic acid-omethyl Lignin Biosynthesis (Glossiness) SBI 07 transferase) (Sb07g003860) LRK(leucine-rich repeat receptor Oviposition Non-preference protein kinase) XnhsbmSFCILP93 (Sb07g003636) Cysteine protease Mir1 Insect resistance (Sb10g028000) Glossy15 (SB10g025053) Glossiness SBI 10 XnhsbmSFCILP2 (NBS LRR protein) (Sb10g010680) Oviposition Non-preference and Deadhearts XnhsbmSFC4 (beta-1, 3-glucanse Glossiness and deadhearts gene) (Sb10g021130) XnhsbmSFCILP24 (tubulin beta- 2/beta-3 chain) (Sb10g027010) Leaf Trichomes Aruna et al., 2011, TAG, Satish et al., 2012 Mol. Breeding

8 OBJECTIVES 1. To develop backcross populations (BC 2 F 2 generation) from crosses involving recurrent recipient and nonrecurrent donor parent 2. Foreground selection for presence of QTL using reported SSR markers 3. To identify genotypes with maximum recovery of recurrent parent genome through background selection 4. Phenotyping of the developed genotypes for both shoot fly resistance and agronomic performance

9 METHODOLOGY Reported chromosomal region selected for Introgression

10 Breeding Scheme for the production of shootfly resistant lines QTLs were previously transferred from IS18551 (Shootfly resistant germplasm accession) to BTx623 Crosses developed 1. ICSB29004 J2658 (SBI01) 2. ICSB29004 J2714 (SBI07) 3. SPV1411 J2614 (SBI10)

11 QTLs SSRs tested for polymorphism Polymorphic SSRs SBI-01/ LG A SBI-07/ LG E SSR MARKERS USED FOR UNDERTAKING FOREGROUND SELECTION 6 (Xtxp329, Xtxp149, Xtxp088, Xisep1035, Xisep1028, Xtxp075) 18 (Xtxp417, Xtxp413, msfcilp85, Xtxp481, Xtxp093, msfc107, msfcilp88, SFC105, msfc106, SFCILP93,mSFC110, msfcilp94, msfc111, SFC112,mSFC113, msfc114, Xtxp159, Xtxp278) 5 (Xtxp329, Xtxp149, Xisep1035, Xisep1028 and Xtxp075) 6 ( msfc107, msfc106, msfcilp94, msfc112, Xtxp159 and Xtxp278) SBI-10/ LG G SBI-05/ LG J 12 (Xisep0634, Xgap001, Xnhsbm1008, Xsbarslbk10.06, Xnhsbm1011, Xisep0643, Xtxp320, Xisep0639, msbcir227, Xcup16, Xtxp141, Xcup07) 22 (msfc43, msfc44, Dsenhsbm057, msfc45, msfc51, msfcilp43, msfc48, msfc50, Dsenhsbm092, Xiabtp420, Xiabtp454, msfc6,msfc62, msfc65, Xtxp112, msfc67, msfc69, msfc71, msfcilp58, Xisep1029, msfc47, Xisp10258) 11 (Xisep0634, Xgap001, Xnhsbm1008, Xnhsbm1011, Xisep0643, Xtxp320, Xisep0639, msbcir227, Xcup16, Xtxp141 and Xcup07) 11 (msfc44, Dsenhsbm057, msfcilp43, Xiabtp420, Xiabtp454, msfc61, msfc62, msfc67, msfc71, Xisep1029, Xisp10258)

12 MARKERS FOR BACKGROUND SCREENING

13 FIELD EVALUATION OF SHOOT FLY RESISTANCE Field evaluations were conducted under high stress condition using Interlard fish meal technique Shoot fly Evaluation was done in two seasons 2014 post rainy and 2015 rainy season

14 PHENOTYPING FOR SHOOTFLY RESISTANCE Glossiness Leaf trichomes Deadhearts Percent Ovipositional nonpreference Seedling vigor

15 AGRONOMIC PERFORMANCE Days to 50% flowering 100 seed weight Grain yield per plot Plant aspect score

16 RESULTS SELECTION OF MOLECULAR MARKERS Graphical representation (chromatogram) of parental lines screening with SSR markers, analysed through ABI prism 3700xl.

17 Results contd. MARKER ASSISTED FOREGROUND SELECTION Heterozygotes with three QTLs on three different chromosomes SBI-01, SBI-07 and SBI-10

18 Results contd. MARKER ASSISTED BACKGROUND SELECTION Background screening using 4% agarose gel. M: 100bp ladder, RP: Recurrent Parent, DP: Donor Parent

19 Results contd. Graphical genotypes of selected lines using Polymorphic SSR markers

20 Results contd. RECOVERY OF RECURRENT GENOME IN INTROGRESSED LINES CROSS ICSB J 2658 (SBI 01) CROSS ICSB29004 J2714 (SBI 07) CROSS SPV1411 J2614 (SBI-10) Genotypes RGC% ICSB J Genotypes RGC% ICSB J Genotypes RGC% SPV J In all the three crosses we have identified the plants with more than 80% recurrent genome content

21 Results contd. PHENOTYPING OF INTROGRESSED LINES FOR SHOOTFLY RESISTANCE CROSS ICSB J 2658 (SBI-01)

22 Results contd. PHENOTYPING OF INTROGRESSED LINES FOR SHOOTFLY RESISTANCE CROSS ICSB29004 J2714 (SBI-07)

23 Results contd. PHENOTYPING OF INTROGRESSED LINES FOR SHOOTFLY RESISTANCE CROSS SPV 1411 J2614 (SBI-10)

24 Results contd. Genotypes Days to 50% flowering 2014 PR 100 Seed weight (grams) 2014 PR AGRONOMIC DATA FOR 2014 POST RAINY AND 2015 RAINY SEASON Panicle weight 2014 PR Grain Weight 2014 PR Plant Aspect score 2015 R ICSB J ICSB J SPV J Genotypes Days to 50% flowering 2015 R 100 Seed weight (grams) 2015 R Panicle weight (grams) 2015 R Grain Weight 2015 R Plant Aspect score 2015 R ICSB J ICSB J SPV J

25 Results contd. AGRONOMIC PERFORMANCE OF INTROGRESSED LINES CROSS ICSB J 2658 (SBI 01)

26 Results contd. AGRONOMIC PERFORMANCE OF INTROGRESSED LINES ICSB29004 RP J-2714 DP CROSS ICSB J 2714 (SBI 07)

27 Results contd. AGRONOMIC PERFORMANCE OF INTROGRESSED LINES SPV 1411 RP J-2614 DP SPV1411 J 2614 (SBI 10)

28 Images from Shootfly screening block Recurrent Parent ICSB29004 Introgression lines with QTL on SBI-01 Donor Parent

29 Images from Shootfly screening block Recurrent Parent ICSB29004 Introgression lines with QTL on SBI-07 Donor Parent

30 Images from Shootfly screening block Recurrent Parent SPV1411 Introgression lines with QTL on SBI-10 Donor Parent

31 CONCLUSION This is the first report on the successful introgression of shootfly resistant QTLs into the elite sorghum cultivar SPV1411 and ICSB29004 DNA markers for shootfly resistance QTLs are reliable for markerassisted selection of shootfly resistant sorghum lines The recovery of the recurrent parent along with the introgression of QTLs with marker-assisted backcross breeding was much faster than that with conventional breeding Six improved shootfly resistance lines were produced from three backcrosses between the recurrent parents SPV1411, ICSB and donor parents J2658, J2714, J2614. These improved shootfly resistant lines have a practical breeding value without yield penalty by providing shootfly resistance to the elite sorghum lines

32 ACKNOWLEDGEMENT Department of Science & Technology (DST), New Delhi Dr A Ashok Kumar, Sorghum Breeding, ICRISAT Dr Santosh P Deshpande Dr Kotla Anuradha P L Ranga Reddy Mrs. Pochamma Sorghum Breeding Staff Dr H C Sharma, Dept of Entomology, ICRISAT Dr Rajeev Varshney, CEG, ICRISAT Prof. M Lakshmi Narsu JNTU Hyderabad Dr Stefania Grando & Management Team ICRISAT, Patancheru, Hyderabad

33 THANK YOU

34 QTL G Heterozygote Recurrent Parent Allele Recurrent Parent Allele Marker Used Xhnsbm1011 Donor Parent Allele Donor Parent Allele

35 QTL E Heterozygote Recurrent Parent Allele Donor Parent Allele Marker Used Xtxp159

36 QTL G Heterozygote Recurrent Parent Allele Donor Parent Allele Marker Used Xhnsbm1011

37 QTL G Heterozygote Recurrent Parent Allele Donor Parent Allele Marker Used Xhnsbm1011

38 QTL J Heterozygote Recurrent Parent Allele Donor Parent Allele Marker Used Xtxp065

39 QTL J Heterozygote Recurrent Parent Allele Donor Parent Allele Marker Used Dsenhsbm057

40 QTL A Heterozygote Recurrent Parent Allele Donor Parent Allele Marker Used Xisep 1035

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