新疆核桃早实特性及RAPD分析

Early Fruiting and its RAPD Analysis of Walnut in Xinjiang Uygur Autonomous

用RAPD技术进行新疆核桃早实特性的分子标记研究.用180个10-m er随机引物分别扩增早实和晚实近等基因池DNA,筛选出5个多态性引物,结果只有引物OPG 15(5-′ACT GGG ACT C-3′)在早实近等基因池及其个体中能重复扩增出一条约710 bp的特异片段OPG 15710,而晚实近等基因池及其个体中无此特异片段.将OPG 15710克隆于pUCm-T载体,筛选阳性克隆用限制性内切酶P stⅠ消化,表明克隆片段大小正确.初步分析认为,OPG 15710可能是与核桃早实特性相关的分子标记,该标记仍在研究之中.该研究将为进一步克隆早实基因,实现早实优质,探索早实机制奠定基础.

RAPD was adopted to explore the molecular markers of early fruiting of walnut growing in Xinjiang. 180 10-mer random primers were used to amplify the DNA in early-fruiting and late-fruiting nearisogenic pools ,and five polymorphic primers were screened ;of the five primers ,only OPG 15 （5＇-ACT GGG ACT C-3＇ ） was capable of repeatedly amplifying OPG15710,a specific band of about 710 bp,from the earlyfruiting near-isogenic pool and its individuals but not capable of amplifying this band from the late-fruiting near-isogenic pool. OPG15710 was cloned onto pUCm-T vector and then its positive clones were screened and digested by Pst I ;The digestion proved that the cloned band had the correct size. It was preliminarily shown that OPG15710 is preliminarily proved probably to be the molecular marker relating to the early fruiting of walnut and is still under study. This study has laid down the basis for further cloning early-fruiting genes and making early-fruiting and high-quality come true and exploring early fruiting mechanism.