与大白菜抗霜霉病基因连锁的分子标记研究.pdfVIP

中国农学通报 2010,26(15):66-70 Chinese Agricultural Science Bulletin 与大白菜抗霜霉病基因连锁的分子标记研究 虞慧芳，钟新民，李必元，顾宏辉 （浙江省农业科学院蔬菜研究所，杭州310021） 摘 要：【研究目的】研究与大白菜抗霜霉病基因紧密连锁的DNA 分子标记。【方法】利用高抗自交系 ‘660 ’和高感自交系‘654B ’及其杂交F 群体162 个单株为材料，采用构建抗、感病池，利用BSA 法筛选 2 了87 对SSR 引物，35 对拟南芥抗霜霉病相关基因的特异引物，其中特异引物RPP 13P2 和RPP 131-2R 在 抗病亲本和抗病池中扩增出一条多态性片段RPP 13MK ，利用这对引物对F 代单株构建的分离群体进 2 行扩增，验证标记RPP 13MK 与目的基因的连锁关系，Mapmaker 3.0 软件计算遗传距离。【结果】通过F2 单株验证后证明RPP 13MK 与抗霜霉病基因紧密连锁，其遗传距离为5.6 cM。【结论】获得了一个与大白 菜抗霜霉病基因紧密连锁的分子标记RPP 13MK 。 关键词：大白菜；霜霉病；DNA 分子标记；遗传距离 中图分类号：S3 文献标志码：A 论文编号：2010-0500 A Molecular Marker Linked to Downy Mildew Resistant Gene in Heading Chinese Cabbage (Brassica rapa L.ssp .pekinensis (Lour) Olsson) Yu Huifang, Zhong Xinming, Li Biyuan, Gu Honghui (Institute of Vegetables, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021) Abstract: 【OBJECTIVE】The objective of this study was to identify molecular markers for downy mildew resistance in heading Chinese cabbage. 【METHOD】SSR-PCR and the PCR using specific primers from downy mildew resistance genes in Arabidopsis were conducted using BSA (Bulked Segregant Analysis) of a segregating F2 population which has 162 individuals from a cross between 660 (resistant) and 654B (susceptible). After performing PCR analysis with 87 pairs of SSR primers and 35 pairs of specific primers from downy mildew resistance genes in Arabidopsis and agarose-gel electrophoresis, only one molecular marker RPP13MK which is PCR product using primers RPP13P2 and RPP131-2R was obtained. Then linkage was analyzed