1. Marker validation for high grain protein content Genes/QTLs in some elite wheat ( T. aestivum L. em. Thell) varieties of Indo-Gangetic plains Manish K. Vishwakarma a * , B. Arun a , V. K. Mishra a , Punam S. Yadav a , H. Kumar a , P.K. Gupta b and Arun K. Joshi a,c a Department of Genetics and Plant Breeding, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi, India; b C.C.S. University, Meerut; c CIMMYT, South Asia Regional Office, P.O. Box 5186, Kathmandu, Nepal; *manishvis@gmail.com Grain protein content (GPC) constitutes the most important economic component of bread wheat determining the end use quality. Selection for high GPC is expensive and time-consuming. Therefore, an attempt was made to validate seven putative SSR/STS/CAPS/STSM markers for desirable GPC genes/QTLs in ten genetic background for effective introgression of grain protein gene into the background of indigenous wheat varieties of Indo-Gangetic plains. Validated markers were utilized in foreground selection of BC1F1 generation . For background selection , 74 Xgmw, Xwmc and Xgdm series markers were used to survey the parental polymorphism , of which 31 markers were found to be polymorphic in the cross HUW234/Glu pro, 27 in HUW468/Glu Pro, 28 in HUW510/Glu Pro. M 2 3 1 4 5 6 7 9 10 8 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 27 28 26 29 30 P2 P1 M-Ladder 100bp, P1 – Doner parent, P2- Recipient parent, Selected Progeny- 4,10,17,18,19,20,21,23,24,25,26,27,28 Ten wheat varieties consisted four Glu Pro derived lines, three CIMMYT deriverd lines viz., Waxwing/Kiritati, Waxwing/Kukuna, Kauz/Pasture and EGPZ popular lines i e. HUW234, HUW 468 and HUW510 were validated with seven putative markers linked to Grain protein content (Table 1). Popular varieties of Indo-Gangetic plains viz., HUW 234, HUW468 and HUW 510 were chosen for developing isogenic lines carrying genes/QTLs for Grain Protein Content (GPC B1), using marker assisted foreground and background selection. These genotypes were crossed with donor Glu pro derived lines and F1s and BC1F1s were screened to identify plants validating for the GPC B1 gene along with maximum recovery of recipient genome. Table 1- Gene/QTLs linked primers and there chromosomal location to be used for foreground selection Wheat Grain Wheat Introduction Plant Materials and Methods Result and Discussion M-Ladder 100bp, Parent 1-HUW234, 2-HUW468, 3-HUW510, 4- Glu p 1, 5-Glu p2, 6-Glu p3, 7-Glu p4, 8-WAX/KIRITATI, 9-WAX/KUKUNA, 10-KAUZ /PASTURE M 1 3 5 6 4 8 7 9 2 10 200 bp Results revealed that except UHW89/6BS and QGpc.ccsu-2D.1/ 2DL the remaining molecular markers associated with high GPC viz. QGpc.ccsu-2D.1 / 2DL, CAPS/ XNor-B2, XNor-B2, WMC415/5A, Xucw108 and Xucw109 were present in all four Glu Pro derived lines and waxwing/Kiritati but not in Waxwing/Kukuna, Kauz/Pasture, HUW234, HUW468 and HUW510. From all above validated molecular markers Xucw 108 ( 217 bp ) appeared to be more useful in molecular assisted breeding program because of its gene specificity. 200 bp Conclusion Validation of markers linked to grain protein content in Glu pro derived lines shall facilitate marker assisted breeding for developing indigenous lines of Indo-Gangetic plain. Development of near isogenic lines for Grain protein content gene GPC B1 in the background of HUW 234, HUW 468, HUW 510 will lead to development of genetically enhanced wheat germplasm for utilization in wheat breeding program. Development of Isogenic Lines Foreground Selection HUW-234/Glu Pro (Marker - Xucw108 ) Validation of Xucw 108 marker in different lines of glu pro and some CIMMYT lines and three recurrent parents Department of Genetics and Plant Breeding, Inst. of Agri. Sciences, Banaras Hindu University, Varanasi, India gwm120 gwm501 S urvey of p arental polymorphism f or background selection Five set of Primers, Lane 1 - Ladder 100 bp , Lane 1-5, 6-10, 11-15, 16-20,21-25 (Glu Pro, waxwing kiritati, HUW234, HUW468, HUW510), Two polymorphic marker Xgwm 120 and Xgwm 501 M 3 25 4 5 6 7 9 10 8 11 12 13 14 15 16 17 18 19 20 21 22 23 24 2 1 S no. Genes/QTLs For GPC B1 Marker types Marker location References 1 Xuhw89 SSR UHW89 6BS Assaf Distelfeld, 2005 2 QGpc.ccsu-2D.1 SSR Xgwm1264 2DL M Prasad,1999 3 XNor-B2 CAPS XNor-B2 6Bs I A KHAN, 2000 4 XNor-B21 ASA XNor-B2 6Bs I A KHAN, 2000 5 QGpc.ccsu-5A1 STMS WMC415 5A H SINGH, 2001 6 Xucw108 SSR Xucw108 6B C Uauy, 2006 7 Xucw109 SSR Xucw109 6B C Uauy, 2006