背景:在普通小麦中(TriticumaestivumL.),高分子量谷蛋白亚基Glu-B1基因座的等位基因变异对谷物最终用途质量具有重要影响。Glu-B1基因座由编码表现出高度可变频率的x型和y型亚基的两个紧密连锁的基因组成。然而,对已报道的等位基因区分标记的研究有限。这里,我们开发了11个基于琼脂糖凝胶的PCR标记,用于检测Glu-1Bx和Glu-1By等位基因。
结果:通过将新开发的标记与先前发表的PCR标记整合,九个Glu-1Bx基因座等位基因(Glu-1Bx6,Glu-1Bx7,Glu-1Bx7*,Glu-1Bx7OE,Glu-1Bx13,Glu-1Bx14(-),Glu-1Bx14(+)/Bx20和Glu-1Bx17)和七个Glu-1By基因座等位基因(Glu-1By8,Glu-1By8*,Glu-1By9,Glu-1By15/By20,Glu-1By16和Glu-1By18)在25个小麦品种中得到了区分。使用新开发的PCR标记区分Glu-1Bx6、Glu-1Bx13、Glu-1Bx14(+)/Bx20、Glu-1By16和Glu-1By18。此外,Glu-1Bx13和Glu-1Bx14(+)/Bx20通过其启动子区的插入和缺失来区分。Glu-1Bx6,Glu-1Bx7,Glu-1By9,Glu-1Bx14(-),通过使用基因编码区的插入和缺失来区分Glu-1By15/By20等位基因。在基因编码区中主要鉴定了Glu-1By13,Glu-1By16和Glu-1By18。我们还开发了区分两个Glu-1Bx14等位基因的标记。然而,使用PCR标记无法区分Glu-1Bx14(+)+Glu-1By15和Glu-1Bx20+Glu-1By20等位基因组合。采用超高效液相色谱和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对小麦品种的高分子量谷蛋白亚基进行了分析,并将结果与PCR分析结果进行比较。
结论:开发了七个Glu-1Bx和四个Glu-1By等位基因检测标记,以检测九个Glu-1Bx和七个Glu-1By基因座等位基因,分别。整合先前报道的标记和11个新开发的PCR标记可以改善Glu-B1基因座的等位基因鉴定,并有助于更有效地分析Glu-B1等位基因分子变异,这可能会提高小麦的最终用途品质。
BACKGROUND: In common wheat (Triticum aestivum L.), allelic variations in the high-molecular-weight glutenin subunits Glu-B1 locus have important effects on grain end-use quality. The Glu-B1 locus consists of two tightly linked genes encoding x- and y-type subunits that exhibit highly variable frequencies. However, studies on the discriminating markers of the alleles that have been reported are limited. Here, we developed 11 agarose gel-based PCR markers for detecting Glu-1Bx and Glu-1By alleles.
RESULTS: By integrating the newly developed markers with previously published PCR markers, nine Glu-1Bx locus alleles (Glu-1Bx6, Glu-1Bx7, Glu-1Bx7*, Glu-1Bx7 OE, Glu-1Bx13, Glu-1Bx14 (-) , Glu-1Bx14 (+)/Bx20, and Glu-1Bx17) and seven Glu-1By locus alleles (Glu-1By8, Glu-1By8*, Glu-1By9, Glu-1By15/By20, Glu-1By16, and Glu-1By18) were distinguished in 25 wheat cultivars. Glu-1Bx6, Glu-1Bx13, Glu-1Bx14 (+)/Bx20, Glu-1By16, and Glu-1By18 were distinguished using the newly developed PCR markers. Additionally, the Glu-1Bx13 and Glu-1Bx14 (+)/Bx20 were distinguished by insertions and deletions in their promoter regions. The Glu-1Bx6, Glu-1Bx7, Glu-1By9, Glu-1Bx14 (-), and Glu-1By15/By20 alleles were distinguished by using insertions and deletions in the gene-coding region. Glu-1By13, Glu-1By16, and Glu-1By18 were dominantly identified in the gene-coding region. We also developed a marker to distinguish between the two Glu-1Bx14 alleles. However, the Glu-1Bx14 (+) + Glu-1By15 and Glu-1Bx20 + Glu-1By20 allele combinations could not be distinguished using PCR markers. The high-molecular-weight glutenin subunits of wheat varieties were analyzed by ultra-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the findings were compared with the results of PCR analysis.
CONCLUSIONS: Seven Glu-1Bx and four Glu-1By allele detection markers were developed to detect nine Glu-1Bx and seven Glu-1By locus alleles, respectively. Integrating previously reported markers and 11 newly developed PCR markers improves allelic identification of the Glu-B1 locus and facilitates more effective analysis of Glu-B1 alleles molecular variations, which may improve the end-use quality of wheat.