水稻Wx复等位基因基于PCR的功能标记开发与利用

发布时间：2018-04-18 19:30

本文选题：水稻 + 蜡质基因　；参考：《作物学报》2017年11期

【摘要】：直链淀粉含量是影响稻米食味品质的重要因素,主要由蜡质基因(Wx)调控,Wx基因座存在Wxa、Wxb、Wxin、Wxmw等多个复等位基因,通过分子标记辅助选择(marker-assisted selection,MAS)培育中低直链淀粉含量的水稻品种是提高稻米适口性的重要途径。Wx复等位基因的功能由单碱基核苷酸多态性(single nucleotide polymorphisms,SNP)决定,其检测依赖于测序或酶切,存在耗时多、费用昂贵等缺点。为提高选择效率,本文利用Wx复等位基因的SNP差异和四引物扩增受阻突变(tetra-primer ARMS-PCR)原理,进行PCR功能标记的开发。引物设计过程中,针对扩增效率低和非匹配的延伸2个技术难题,提出了调整错配位点的解决方案,成功开发了基于PCR技术的两对功能性标记Waxygt-ARMS2和Waxyac-ARMS2,可以准确区分上述复等位基因型,且具有操作简单、耗费较低的特点。利用新开发标记对辽宁省育成的部分水稻品种进行Wx基因型检测的结果表明,40份辽宁育成品种均携带Wxb基因,遗传基础单一。上述结果为利用Wx基因位点的分子标记培育中低直链淀粉含量的水稻品种,改良辽宁省水稻品种的食味品质提供了技术支撑。
[Abstract]:Amylose content is an important factor that affects the quality of rice taste. The WX locus is regulated by waxy gene (Wxxx), and there are many multiple alleles such as WxaAU WxbCU Wxinwxmw and so on.Breeding rice varieties with low amylose content by molecular marker-assisted selection was an important way to improve palatability of rice. The function of Wx complex allele was determined by single nucleotide polymorphismsNPs, and its detection depended on sequencing or enzyme digestion.There are many time-consuming, expensive and other shortcomings.In order to improve the selection efficiency, PCR functional markers were developed based on the SNP difference of Wx complex allele and the principle of four-primer ARMS-PCR.In the process of primer design, aiming at the two technical problems of low amplification efficiency and unmatched extension, a solution to adjust mismatch sites was proposed.Two pairs of functional markers Waxygt-ARMS2 and Waxyac-ARMS2 based on PCR technology were successfully developed which can accurately distinguish these complex alleles and have the advantages of simple operation and low cost.The WX genotypes of some rice varieties bred in Liaoning Province were detected by using the newly developed markers. The results showed that all 40 cultivated rice varieties in Liaoning province carried Wxb gene and had a single genetic basis.These results provide technical support for the cultivation of rice varieties with low amylose content and the improvement of the edible quality of rice varieties in Liaoning Province by using the molecular markers of WX loci.
【作者单位】：沈阳农业大学水稻研究所;辽宁省盐碱地利用研究所;
【基金】：辽宁省农业领域青年科技创新人才培养计划(2015035,2015036) 国家自然科学基金项目(31371587,31430062)资助~~
【分类号】：S511

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