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Applying genotyping (TILLING) and phenotyping analyses to elucidate gene function in a chemically induced sorghum mutant population

DOI: 10.1186/1471-2229-8-103

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A sorghum mutant population consisting of 1,600 lines was generated from the inbred line BTx623 by treatment with the chemical agent ethyl methanesulfonate (EMS). Numerous phenotypes with altered morphological and agronomic traits were observed from M2 and M3 lines in the field. A subset of 768 mutant lines was analyzed by TILLING using four target genes. A total of five mutations were identified resulting in a calculated mutation density of 1/526 kb. Two of the mutations identified by TILLING and verified by sequencing were detected in the gene encoding caffeic acid O-methyltransferase (COMT) in two independent mutant lines. The two mutant lines segregated for the expected brown midrib (bmr) phenotype, a trait associated with altered lignin content and increased digestibility.TILLING as a reverse genetic approach has been successfully applied to sorghum. The diversity of the mutant phenotypes observed in the field, and the density of induced mutations calculated from TILLING indicate that this mutant population represents a useful resource for members of the sorghum research community. Moreover, TILLING has been demonstrated to be applicable for sorghum functional genomics by evaluating a small subset of the EMS-induced mutant lines.Sorghum (2n = 2x = 20, 7.35 × 108 bp for 1C nucleus) is a C4 crop that displays excellent tolerance to both drought and high temperature stresses [1]. Sorghum has the highest water use efficiency among major crop plants and is unusually tolerant to low soil fertility, traits essential for survival and productivity in arid and semi-arid areas with limited irrigation capability. Worldwide, sorghum is the 5th most important grain crop, providing food and fodder for the inhabitants of drought-susceptible regions webcite. Recently, sorghum has been demonstrated as a viable bioenergy feedstock [2]. Compared with other bioenergy grain crops, sorghum is particularly advantageous because it can be grown profitably on marginal


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