TY - PRO AU - Rajaram,V. AU - Bhasker Raj,A.G. AU - Blummel,M. AU - Choudhary,S. AU - Hash,C.T. AU - Kavi Kishor,P.B. AU - Kholova,J. AU - Kocova,M. AU - Kumar,S. AU - Lakshmi NarasuM. AU - Nepolean,T. AU - Pandurangarao,V. AU - Rai,R.N. AU - Sahrawat,K.L. AU - Senthilvel,S. AU - Singh,G. AU - Supriya AU - Thakur,R.P. AU - Vadez,V. AU - Varshney,R.K. AU - Yadav,R.C. AU - Velu,G. AU - Sharma,R.K. TI - Development of EST resources in pearl millet and their use in development and mapping of EST-SSRs in four RIL populations PY - 2010/// N2 - Elite pearl millet inbreds ICMB 841-P3 and 863B-P2 (RIL population parents differing for drought and salinity tolerance, downy mildew and blast resistance, and stover and grain quality) were drought stressed at boot-leaf stage using a standard dry-down protocol. Total RNA extracted separately from stressed root and leaf tissues of each inbred was used for cDNA synthesis. 454-sequencing of cDNAs yielded raw EST sequences that were cleaned and assembled using the PLANTTA pipeline. A total of 112,864 unigenes were obtained from the four cDNA samples. A total of 5,800 SNPs from 2,146 contigs were identified between the two parents. Similarly, 499 sequences containing class I SSRs were identified from the unigenes and from these 236 non-redundant EST-SSR primer pairs were designed. PCR products from 139 amplifying EST-SSR primer pairs and parental DNA of crosses ICMB 841-P3 x 863B-P2, H 77/833-2 x PRLT 2/89-33, 81B-P6 x ICMP 451-P8 and PT 732B-P2 x P1449-2-P1 were screened for polymorphism. >50 new EST-SSR markers were mapped for each mapping population and a consensus map developed. These four EST-SSR skeleton-mapped RIL populations have been phenotyped for grain Fe and Zn concentration, salinity tolerance, canopy conductance, downy mildew reaction, rust reaction, and/or blast reaction, and QTLs with closely flanking EST-SSR markers were detected for each of these traits. This study increases the number of mapped pearl millet SSRs by >40%, filling gaps in the earlier linkage maps, and will facilitate SSR-based marker-assisted foreground selection in parts of the pearl millet genome where this was not previously possible ER -