与黄麻炭疽病抗性相关的SSR分子标记筛选及新型SNP标记开发

摘要

为筛选出与黄麻炭疽病抗性相关的新型分子标记,本研究在前期黄麻炭疽病抗性QTL定位结果的基础上,开发出7对可能与黄麻炭疽病抗性基因连锁的新型SNP标记.同时,在前期对重组自交系群体进行田间炭疽病抗性鉴定的基础上,分别以6个抗病株系和6个感病株系的基因组DNA构建了抗池和感池.以抗、感池的DNA为模板,对36对SSR引物和7对SNP引物进行了多态性筛选,从中初步筛选出具有多态性的SSR引物15对,SNP引物2对.继而,分别以12份抗病和感病株系的基因组DNA为模板,对上述具有多态性的SSR和SNP引物进行进一步验证,最终获得1对与黄麻炭疽病抗性相关的SNP标记,其产生的多态性片段大小约为600 bp.%This study aims to identify new molecular markers associated with anthracnose resistance of white jute ( Corchorus capsularis L.), as well as to provide a convenient and reliable molecular biology method for anthracnose resistance identification. Firstly, based on quantitative traits loci ( QTL) identification for anthracnose resistance of white jute, 7 pairs of new single nucleotide polymorphisms (SNP) markers which might link to anthracnose resistance of jute were identified. Secondly, by examining the anthracnose resistance of the recombining inbred lines of white jute in our previous study, the resistant and susceptible pools were generated using the genomic DNA of 6 resistant and 6 susceptible lines, respectively. Thirdly, 36 pairs of simple sequence repeat ( SSR) and 7 pairs of SNP primers were screened using genomic DNA from the two pools as templates, and 17 polymorphic primers including 15 pairs of SSR primers and 2 pairs of SNP primers were identified. Moreover, those polymorphic SSR and SNP primers were further verified using 12 copies of the genomic DNA of resistant and susceptible white jute lines as templates. Finally, 1 pair of SNP primer was identified with polymorphism significantly associated with anthracnose resistance of white jute, which produced a polymorphic DNA fragment with the size of around 600 bp.