Generation of Mutant Plants by Gamma Ray Exposure and Development of Low-Cost TILLING Population in Chickpea (Cicer arietinum L.)

Abstract

Induced mutations have been used to facilitate plant breeding for more than 80 years. Success requires the development of a mutant population and methods to evaluate that population. In this protocol we provide methods for the development of a chickpea mutant population using gamma irradiation, and low-cost methods for the molecular characterization of the mutant population. Specifically, this chapter provides detailed methods for (1) mutation induction by gamma rays and determination of LD₅₀ and RD₅₀, (2) phenotypic assessment of the M₂ generation, (3) low-cost extraction of genomic DNA, and (4) identification of induced mutations using low-cost agarose-gel based TILLING. The methods are low-cost and designed to be applicable in most research settings.

Keywords: Enzymatic mismatch cleavage; Legumes; Low-cost TILLING; M2 generation; Mutation discovery; Mutation induction; Phenotypic screening; Radiosensitivity.