与Pi-2（t）基因连锁的栽培稻BAC克隆在药用野生稻和栽培稻中的比较物理定位

与Pi-2（t）基因连锁的栽培稻BAC克隆在药用野生稻和栽培稻中的比较物理定位 Comparative Physical Map of Linked Markers of Pi-2（t） Gene in Oryza officinalis and Oryza sativa with BAC-FISH 李霞 LI Xia 1 2 first-author 李宗芸 LI Zong-yun 1 2 覃瑞 QIN Rui 1 2 何光存 HE Guang-cun 1 2 宋运淳 SONG Yun-chun 1 2 武汉大学植物发育生物学教育部重点实验室 武汉大学植物发育生物学教育部重点实验室 Key Laboratory of MOE for Plant Developmental Biology, Wuhan University Key Laboratory of MOE for Plant Developmental Biology, Wuhan University 武汉大学植物发育生物学教育部重点实验室,湖北,武汉,430072 武汉大学植物发育生物学教育部重点实验室,湖北,武汉,430072 用栽培稻遗传图第 6连锁群中与 Pi- 2 （ t）紧密连锁的 RFL P标记 RG64及其筛选出来的 BAC克隆3 8D17作为探针 ,对药用野生稻和栽培稻进行荧光原位杂交 （ FISH） ,供试探针 RG64及 BAC克隆 3 8D17均被定位于药用野生稻和栽培稻第 6染色体 .药用野生稻杂交信号的百分距离分别为 4 4 .87± 5 .3 3和 4 6.5 0± 4 .5 7,而栽培稻则为 3 5 .85± 3 .0 6和 3 6.0 5± 2 .4 4 ,信号检出率相应地为 7.14 %、4 2 .5 3 %、8.0 9%和 4 0 .78% .BAC克隆和RFL P探针杂交位置几乎一致 ,由此推知 ,与抗性基因 Pi- 2 （ t）连锁的 BAC克隆在药用野生稻中的同源顺序就在第6染色体杂交信号出现的相应位置 .在未封阻的情况下 ,药用野生稻多个染色体上具有信号 ,这表明它和栽培稻的Cot- DNA重复序列在一定的程度上具有同源性 .文中讨论了栽培稻 BAC克隆对药用野生稻原位杂交物理作图的可行性和 Cot- DNA的制备所涉及的一些技术问题 . A FISH procedure was adopted to physically localize rice BAC clone 38D17 linked with Pi-2（t） gene in O.sativa and O.officinalis. The FISH results showed that both the RG64 and BAC clone 38D17 were localized on the chromosome 6 in O.officinalis and O.sativa, the percentage distances from the centromere to the hybridization sites were 44.87±5.33, 46.50±4.67 （O.officinalis）, and 35.85±3.06, 36.05±2.44（O.sativa）, the detection rates were 7.14%, 42.53% and 8.09%, 40.78% respectively. The results suggested that the RG64 of cultivated rice and its homologous sequence of O.officinalis were in the same BAC clone. We observed many background signals when probe was not blocked with Cot-Ⅰ DNA. It demonstrated that highly repetitive sequences were possibly homologous in a certain degree between cultivated rice and O.officinalis. The identification of chromosome 6 of O.officinalis was based on the comparative genetics map of Jena et al. （1994） and our research results. In our study, the feasibility of physical mapping in O.officinalis with rice BAC clones and preparation of Cot-Ⅰ DNA were discussed. 药用野生稻 细菌人工染色体-荧光原位杂交 比较物理定位 Pi-2（t） O.officinalis BAC-FISH comparative physical map Pi-2（t） S511.01 国家自然科学基金 （ 39870 42 3）;高等学校博士学科点专项研究基金 （ 2 0 7980 112 ）资助项目 2001-04-01 2021-04-01 4