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我们用辣椒(Capsicum annuum)栽培种(已导入了灯笼椒Capsicum chinenseL3基因)的种内F2代群体(2016株)和种间F2代群体(3391株)(由灯笼椒与Capsicum frutescence杂交产生)对灯笼椒抗烟草花叶病毒属病毒的L3基因进行定位。通过L3基因抗性紧密相关的AFLP分子标记的BAC文库的分析,揭示出番茄抗病同源基因I2的存在。通过简并PCR技术,对来自35株不同辣椒的同源基因I2的部分或全部编码序列进行克隆,且在种间组合中产生了17个遗传标记。图谱显示:L3基因位于I2同源基因标记IH1-04和BAC-end标记189D23M中间,L3基因定位于包含两个不同BAC重叠群的区内,这两个不同的BAC重叠群分别由4个和1个无性系组成。DNA纤维荧光原位杂交揭示这两个重叠群被约30kb隔开。DNA纤维荧光原位杂交结果和BAC无性系的Southern杂交表明在高度重复序列中富集包含L3基因位点的区。Southern杂交表明两个BAC重叠群包含多于十个的I2同源基因拷贝体。相反,对于种间F2代群体,,重组后代没有结合位点,在种内F2代群体中,该结合位点存在于两个不同的BAC重叠群内,这两个不同的BAC重叠群分别由7个和2个无性系组成。而且,两个群体间结合位点分配的不同表明在含有L基因位点的区域连锁不平衡。
We used the F2 population (2016 strain) and the F2 population (3391 strain) of Capsicum annuum cultivar (Capsicum chinenseL3 gene introduced) (produced by the cross between Capsicum frutescence) The bell pepper locates the anti-tobacco mosaic virus L3 gene. Analysis of AFLP-tagged BAC libraries closely related to L3 gene resistance revealed the presence of the resistance gene 12 in tomato. Some or all of the coding sequences for the homologous gene 12 from 35 different capsicums were cloned by degenerate PCR technology and 17 genetic markers were generated in interspecies combinations. The map shows that the L3 gene is located between the I2 homologous gene marker IH1-04 and the BAC-end marker 189D23M, and the L3 gene is located in a region containing two different BAC contigs, the two different BAC contigs respectively consisting of 4 and 1 clones. DNA fiber fluorescence in situ hybridization revealed that these two contigs were separated by about 30 kb. Southern hybridization of DNA fiber fluorescence in situ hybridization results to BAC clones revealed enrichment of regions containing L3 loci in highly repetitive sequences. Southern blotting showed that two BAC contigs contained more than ten copies of I2 homologous genes. In contrast, there was no binding site for recombined offspring for interspecific F2 population, and in the F2 population within the species, the binding site was present in two different BAC contigs, the two different BAC contigs, respectively 7 and 2 clones. Moreover, differences in the binding site assignments between the two populations indicate a linkage disequilibrium in the region containing the L gene locus.