Development of multiplex-PCR systems for genes related to flour colour in Chinese autumn-sown wheat cultivars uri icon

abstract

  • In wheat kernel, multiple gene loci, Psy-A1, Psy-B1, TaZds-A1, TaZds-D1, TaLox-B1, Ppo-A1 and Ppo-D1 are implicated in the expression of flour colour traits. In this study, eight functional markers, covering seven gene loci, were used to develop three multiplexed polymerase chain reaction (PCR) systems for marker-assisted selection (MAS) of desirable flour colour, i.e. PCR-I (YP7B-1/YP7B-2/YP2A-1), PCR-II (YP2D-1/PPO18) and PCR-III (YP7A/LOX18/STS01). Genotyping of 20 control wheat cultivars by these multiplex-PCR systems completely matched the results from the eight single PCR amplifications. Multiplex-PCR typing was successfully established for a total of 155 Chinese autumn-sown wheat cultivars. In multiplexed PCR-I, the occurring frequencies for three alleles (Psy-B1a, Psy-B1b and Psy-B1c) at the Psy-B1 locus were 33.5, 58.7, and 7.7%; while two alleles (TaZds-A1a and TaZds-A1b) at TaZds-A1 were 40.0 and 60.0%, respectively. In multiplexed PCR-II, the occurring frequencies, TaZds-D1a and TaZds-D1b (98.7 and 1.3%) at the TaZds-D1 locus, Ppo-A1a and Ppo-A1b (39.4 and 60.6%) at the Ppo-A1 locus were obtained, respectively. In multiplexed PCR-III, the occurring frequencies for two predominant alleles at each locus was obtained, Psy-A1a and Psy-A1b (55.5 and 44.5%) at Psy-A1 locus, TaLox-B1a and TaLox-B1b (29.7 and 70.3%) at TaLox-B1 locus, Ppo-D1a and Ppo-D1b (45.8 and 54.2%) at Ppo-D1 locus, respectively. The multiplexed PCRs developed in this study are useful for MAS for wheat flour colour improvement. The genotyping results obtained in this study provide valuable information for DNA-based identification of wheat cultivars.

publication date

  • 2016
  • 2016