论文部分内容阅读
结合生物信息学的方法,对水稻白叶枯病(Xanthomonas oryzae pv.oryzae,Xoo)抗性基因Xa23的标记C189进行了改良和优化,获得了测序品种日本晴中C189电子克隆的序列,设计改良的分子标记,命名为P23。用P23扩增36个水稻亲本和200份中国栽培稻微核心种质材料,研究了该分子标记的稳定性和特异性。标记多态性检测结果表明,在阳性亲本CBB23中扩增出大约474 bp的条带,其他35个常用水稻亲本中都是扩增出约600 bp的条带;中国栽培稻微核心种质中150个品种扩增出约600 bp的单一条带,50个品种扩增出约474 bp的单一条带。利用P23对含Xa23基因的114个F6代育种家系进行了分子标记辅助选择,获得了43个纯合的抗性株系。这些结果证明标记P23具有特异性强,共显性,PCR扩增产物差异大,利用1%的琼脂糖凝胶电泳即可快速分清等位差异的特点,能广泛用于抗病基因Xa23的辅助选择;该研究中采用的生物信息学的方法对其他基因分子标记的设计或改良也具有一定的借鉴和指导意义。
Combined with bioinformatics method, X183, a marker gene of Xoo resistance to Xanthomonas oryzae pv. Oryzae (Xoo), was modified and optimized. The sequence of C189 was cloned and sequenced. Molecular marker, named P23. Thirty-six rice cultivars and 200 microspore germplasm cultivars from China were amplified by P23, and the stability and specificity of the molecular markers were studied. The results of the marker polymorphism showed that a 474 bp band was amplified in the positive parent CBB23 and a band of about 600 bp was amplified in 35 other common rice parents. A single band of about 600 bp was amplified from 150 cultivars, and a single band of about 474 bp was amplified from 50 cultivars. A total of 114 F6 generation lines with Xa23 gene were selected by molecular marker assisted selection using P23, and 43 homozygous resistant lines were obtained. These results prove that the marker P23 has the characteristics of strong specificity, co-dominant, large difference in PCR amplification products, rapid differentiation of allelic differences using 1% agarose gel electrophoresis, and can be widely used for assisting the resistance gene Xa23 The bioinformatics methods used in this study also have certain reference and guidance significance for the design or improvement of other molecular markers.