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中国农学通报 ›› 2014, Vol. 30 ›› Issue (13): 29-33.doi: 10.11924/j.issn.1000-6850.2013-2791

所属专题: 生物技术 园艺

• 林学 园艺 园林 • 上一篇    下一篇

巴西橡胶树 ‘热研 7-33-97’ 单染色体显微分离与扩增

陈学俊 王英 庄南生 高和琼   

  • 收稿日期:2013-10-28 修回日期:2013-12-24 出版日期:2014-05-05 发布日期:2014-05-05
  • 基金资助:
    国家自然科学基金项目“巴西橡胶树单染色体微克隆文库构建的研究”(31260185);海南省高等学校科学研究项目“巴西橡胶树单染色体微克隆文库构建”(Hjkj2011-10);海南省重大科技项目“热带作物抗逆机理与热带植物资源利用研究”(ZDZX2013023);“中央财政支持中西部高校提升综合实力”专项资金(科研平台和创新团队建设专项规划)“植物分子遗传学和遗传资源利用”。

Isolation and Amplification of Single Chromosome in Hevea brasiliensis‘RY7-33-97’

  • Received:2013-10-28 Revised:2013-12-24 Online:2014-05-05 Published:2014-05-05

摘要: 巴西橡胶树是天然橡胶的重要来源。而染色体显微分离技术对基因组研究有重要意义。本研究以巴西橡胶树‘热研7-33-97’幼叶为材料,采用酶解去壁低渗法制片,建立了微细玻璃针法分离巴西橡胶树单条染色体技术体系。成功分离出8条染色体;随机选取其中2条进行了2轮LA-PCR扩增,得到400~1800 bp之间的DNA片段,通过Southern blot杂交对LA-PCR产物进行验证,结果表明扩增片段与巴西橡胶树基因组DNA之间有同源性。从而说明单染色体DNA已被成功的扩增。这就证明利用染色体微分离、微克隆技术对巴西橡胶树基因组进行研究是可行的。为构建巴西橡胶树单染色体文库奠定了基础。

关键词: 蜥蜴, 蜥蜴, 体色, 两性异形, 性选择

Abstract: Hevea brasiliensis was one of the most important sources of natural rubber. The technique of single chromosome microdissection had important significance in genomic study. This experiment took tender leaves of Hevea brasiliensis‘RY-7-33-97’as material. The enzyme and low osmotic method was used to make the chromosom preparation. A technique for single chromosome microdissection by slender glass needles in Hevea brasiliensis was constructed. Eight chromosomes were successfully isolated, and two of them were amplified by linker adaptormediated PCR (LA-PCR) for two rounds, a smear of DNA fragments ranging from 400-1800 bp were generated. Southern blot hybridization results showed that the LA-PCR products were homogeneous with the Hevea brasiliensis genomic DNA, indicating that DNAs from the chromosome have been successfully amplified. This study demonstrated that chromosome microdissection and cloning can be applied in genomic study of Hevea brasiliensis. The results will facilitate for the construction of single chromosomal DNA library of Hevea brasiliensis.