Development of hull-less barley with ultra-low gluten content via target genes combination. I. Isolation of triple mutants and black grained genotypes

Abstract

Aim. The purpose of the research, presented in this paper, is to develop (as the first one in Ukraine) hull-less barley breeding material with ultra-low gluten, combining three hordein-deficient mutations, derived from Risø56, Risø1508, and R118, with black grain color as an indication of bioactive pigments content, enhancing the functional status of grain as a food product. Methods. We used electrophoretic analysis of proteins in polyacrylamide gel, DNA isolation by the CTAB method, polymerase chain reaction with DNA-markers, restriction analysis and electrophoretic separa tion of DNA in agarose gel. Results. The article presents the results of the first stage of combining target hordeindefi cient mutations by binary crosses. The F2/3 and F3/4 populations were derived from binary crosses of Risø56 × Risø1508 (reciprocal Risø1508 × Risø56) and R118 × hull-less barley Achilles. The new PCR protocol was optimized. Markers B1hor, 3a5F and Dhor-m were selected to identify mutations in B, C and D hordeins. Conclusions. Several barley genotypes with one or two target hordein-deficient mutations and black grain color were isolated. The genotypes obtained will be used in a further backcrosses program, aimed at developing of a black hull-less barley variety with ultra-low gluten content.