Genetic characterization and expression analysis of wheat (Triticum aestivum) line 07OR1074 exhibiting very low polyphenol oxidase (PPO) activity.

Abstract

Characterized novel mutations present at Ppo loci account for the substantial reduction of the total kernel PPO activity present in a putative null Ppo - A1 genetic background. Wheat (Triticum aestivum) polyphenol oxidase (PPO) contributes to the time-dependent discoloration of Asian noodles. Wheat contains multiple paralogous and orthologous Ppo genes, Ppo-A1, Ppo-D1, Ppo-A2, Ppo-D2, and Ppo-B2, expressed in wheat kernels. To date, wheat noodle color improvement efforts have focused on breeding cultivars containing Ppo-D1 and Ppo-A1 alleles conferring reduced PPO activity. A major impediment to wheat quality improvement is a lack of additional Ppo alleles conferring reduced kernel PPO. In this study, a previously reported very low PPO line, 07OR1074, was found to contain a novel allele at Ppo-A2 and null alleles at the Ppo-A1 and Ppo-D1 loci. To examine the impact of each mutation upon kernel PPO, populations were generated from crosses between 07OR1074 and the hard white spring wheat cultivars Choteau and Vida. Expression analysis using RNA-seq demonstrated no detectable Ppo-A1 transcripts in 07OR1074 while Ppo-D1 transcripts were present at less than 10% of that seen in Choteau and Vida. Novel markers specific for the Ppo-D1 and Ppo-A2 mutations discovered in 07OR1074, along with the Ppo-A1 STS marker, were used to screen segregating populations. Evaluation of lines indicated a substantial genotypic effect on PPO with Ppo-A1 and Ppo-D1 alleles contributing significantly to total PPO in both populations. These results show that the novel mutations in Ppo-A1 and Ppo-D1 present in 07OR1074 are both important to lowering overall wheat seed PPO activity and may be useful to produce more desirable and marketable wheat-based products.