优质新疆拉面品种品质评价的多重PCR体系构建与应用

新疆拉面是深受人们喜爱的特色面食之一,建立优质新疆拉面品质评价体系对其专用品种选育和评价具有重要意义。本研究构建了3套检测新疆拉面优质基因的多重PCR体系,并对46份新疆冬小麦(Triticum aestivum L.)进行了检测。体系Ⅰ可同时直接检测Ax2、Pinb-D1b和ω-secalin(1B·1R易位)3种基因,体系Ⅱ可同时直接检测两个位点Psy-A1(Psy-A1a、Psy-A1b、Psy-A1c)和Ppo-D1(Ppo-D1a、Ppo-D1b)的5种基因,体系Ⅲ可同时直接检测Glu-B3a、Wx-B1a和Wx-B1b3种基因,3套体系检测对照品种(系)的结果均与已知基因型完全一致。对46份新疆冬小麦的检测结果表明,Ax2、Pinb-D1b和不含ω-secalin(非1B·1R易位)基因出现的频率依次为30.4%、45.7%和78.3%,Psy-A1a、Psy-A1b、Psy-A1c、Ppo-D1a和Ppo-D1b基因的频率分别为94.5%、4.3%、2.2%、91.3%和8.7%,Glu-B3a、Wx-B1a和Wx-B1b基因的频率依次为21.7%、91.3%和8.7%;其中,与新疆拉面优质性状相关的5个基因Ppo-D1a、不含ω-secalin、Pinb-D1b、Glu-B3a和Ax2的频率明显高于全国平均水平;共检测到19种优质拉面基因的组合类型,其中含1、2、3、4和5个优质基因的组合类型依次占参试品种(系)的13.0%、32.6%、26.1%、23.9%和4.3%,没有发现同时含6或7个优质基因的品种(系),出现频率最高的优质基因组合类型为Ppo-D1a/不含ω-secalin(非1B·1R易位)、Ppo-D1a/Glu-B3a/不含ω-secalin和Ppo-D1a/Ax2/Pinb-D1b/不含ω-secalin。研究结果表明,本研究建立的3套多重PCR体系,结果稳定可靠,可作为新疆拉面品种品质性状快速评价的方法;利用本研究构建的多重PCR体系和筛选出的含优质基因(组合)的品种(系),开展小麦品质评价和分子聚合育种,将会提高新疆拉面品种评价和选育的效率。

Establishment and Application of Multiplex PCR Systems for Cultivar Evaluation of High Quality Xinjiang Hand Stretched Noodle

Xinjiang hand stretched noodle(XHSN) is one of the most popular noodles in China,and evaluation for high quality XHSN is essential to success of breeding program.Based on past research achievement of the XHSN and relevant molecular markers for the genes coding high quality XHSN,three types of multiplex PCR systems were developed and validated with 13 Xinjiang winter wheat cultivars and advanced lines with known genes,and used to evaluate the genes associated with XHSN quality in 46 Xinjiang winter wheat(Triticum aestivum L.) cultivars and advanced lines.The first multiplex PCR was used to simultaneously detect genes Ax2*,Pinb-D1b and ω-secalin,the second one was to detect the genes Psy-A1a,Psy-A1b,Psy-A1c,Ppo-D1a and Ppo-D1b,and the third one was to detect Glu-B3a,Wx-B1a and Wx-B1b.The test results showed that the frequencies of genes Ax2*,Pinb-D1b,without ω-secalin,Psy-A1a,Psy-A1b,Psy-A1c,Ppo-D1a,Ppo-D1b,Glu-B3a,Wx-B1a and Wx-B1b in the 46 winter wheat cultivars and advanced lines were 30.4%,45.7%,78.3%,94.5%,4.3%,2.2%,91.3%,8.7%,21.7%,91.3% and 8.7%,respectively.Five(Ppo-D1a,without ω-secalin,Pinb-D1b,Glu-B3a and Ax2*) of those genes related with the high quality for the XHSN in Xinjiang wheats had higher frequencies than those in wheats from other Chinese regions,but the high quality genes Psy-A1b and Wx-B1b had lower frequencies.Meanwhile,19 combinations of the high quality genes appeared in the tested cultivars and advanced lines,the combinations with one,two,three,four or five high quality genes accounted for 13.0%,32.6%,26.1%,23.9% and 4.3%,respectively.The cultivar with six or seven high quality genes was not found.In all combinations of the high quality genes,Ppo-D1a/without ω-secalin,Ppo-D1a/Glu-B3a/without ω-secalin and Ppo-D1a/Ax2*/Pinb-D1b/without ω-secalin had the highest frequencies.Those results showed that the three systems developed in this study were effective and stable to amplify target bands for these genes and could be used to evaluate the XHSN quality quickly.In order to improve the quality of the XHSN in the future,those high quality genes and their combinations should be maintained for the XHSN.Developing cultivars with lower in both yellow pigment(Psy-A1b) and amylose(Wx-B1b) contents will be important objectives in the XHSN wheat breeding programs in the future.Developed three multiplex PCR systems and identified wheat genetic resources with the high quality genes in this study can be provide valuable information for estimation and breeding of the XHSN cultivars in the future.