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Establishment of Multiplex-PCR for Quality Traits in Common Wheat

小麦品质性状分子标记多重PCR体系的建立


小麦高分子量谷蛋白亚基组成、1B/1R易位、籽粒硬度、直链淀粉含量和穗发芽抗性等性状与加工品质密切相关,建立相关性状的多重PCR体系在小麦品质分子育种中具有重要意义。本研究利用现有品质性状基因的分子标记,建立了适合不同品质类型品种评价和分子聚合育种的3个多重PCR体系,并用已知基因的品种(系)进行验证。多重PCR体系Ⅰ包括ω-secalin(1B/1R)、Vp1B3和Pinb-D1b基因的分子标记检测,可用于一般的品质检测;体系Ⅱ包括ω-secalin、Ax2*、Bx17和Dx5 基因的检测,可望用于强筋小麦品种的选育;体系Ⅲ包括Wx-A1、Wx-B1和Wx-D1位点的检测,可用于淀粉品质或糯小麦的选育。每个体系内的引物之间不存在相互抑制作用和错配,检测品种(系)的结果可靠、重复性好,成本低。3个多重PCR体系用于小麦品质育种的亲本评价和杂交后代优质基因的聚合,将会提高优质专用小麦品种评价和选育的效率。

Wheat (Triticum aestivum L.) quality properties are strongly affected by the compositions of high-molecular-weight glutenin subunits, kernel hardness, amylase content, pre-harvest sprouting tolerance and presence or absence of 1B/1R translocation. It is very important to develop multiplex PCR for wheat quality improvement in molecular marker assisted breeding to reduce the cost and improve the efficiency. Three types of multiplex PCRs were developed and validated with 13, 30, and 11 Chinese wheat cultivars and advanced lines with known genes, respectively. The first multiplex PCR was used to simultaneously detect genes ω-secalin (1B/1R), Vp1B3, and Pinb-D1b for improving wheat processing quality. The second one was to detect the genes ω-secalin, Ax2*, Bx17, and Dx5 for improving gluten quality and bread making quality. The third multiplex PCR included three markers for Wx-7A, Wx-4A, and Wx-7D to improve starch quality and breed waxy wheat cultivars. The genotypes of all tested wheat cultivars and advanced lines identified by three multiplex PCRs were in agreement with those detected by other methods. The primer-primer interactions in each multiplex PCR were not found. Genomic DNAs extracted from both wheat kernels and leaves were feasible for three multiplex PCR amplifications. The three multiplex PCRs were highly effective in the test of Chinese wheat cultivars, demonstrating good repeatability and low cost for the evaluation of wheat quality properties in wheat breeding program.


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