转几丁质酶和葡聚糖酶双价基因棉花对土壤细菌种群多样性的影响.pdfVIP

Hereditas (Beijing) 2015 年 8 月, 37(8): 821 ―827 研究报告 转几丁质酶和葡聚糖酶双价基因棉花对土壤细菌种群 多样性的影响 李志芳，冯自力，赵丽红，师勇强，冯鸿杰，朱荷琴 中国农业科学院棉花研究所，棉花生物学国家重点实验室，安阳 455000 摘要 : 以转几丁质酶和葡聚糖酶双价基因棉花为研究对象，非转基因受体棉花为对照，通过比较可培养细菌数 量和基于 16S rRNA 克隆文库细菌种群分析，评价外源双价基因的导入在苗期、蕾期、花铃期和吐絮期对棉花 根际细菌群落多样性的影响。结果表明，可培养细菌的数量不受外源双价基因的影响，随着棉花生育期的交替 而变化，以代谢旺盛的花铃期最多。构建的转基因和非转基因不同生育期根际土壤细菌 16S rRNA 文库容量为 2400 个克隆，涵盖了细菌的283 个属。其中，Acidobacterium 是最大优势类群，共包括 624 个克隆，其次为未 知细菌种群和 Flavisolibacter 。比较转基因和非转基因棉花根际土壤细菌的种群结构，结果显示，同一生育期 内前者种群的多样性显著低于后者，二者的共有类群随着生长发育的进行而增多。研究结果说明几丁质酶基因 和葡聚糖酶基因对棉花根际细菌种群多样性有着不同程度的削减作用，但是随着种植时间的延长，该差异呈现 逐渐缩小的趋势。 关键词 : 转基因棉花；可培养细菌；16S rRNA 克隆文库；土壤细菌群落 Effects of transgenic cotton expressing chitinase and glucanase genes on the diversity of soil bacterial community Zhifang Li, Zili Feng, Lihong Zhao, Yongqiang Shi, Hongjie Feng, Heqin Zhu State Key Laboratory of Cotton Biology, Institute of Cotton Research of Chinese Academy of Agricultural Sciences, Anyang 455000, China Abstract: The transgenic cotton expressing chitinase and glucanase genes was studied using nontransgenic cotton as a control. Specifically, the effects of exogenous genes on bacterial community diversity in rhizospheres of cotton at stages of seedling, budding, boll forming and boll opening were evaluated through comparing the number of cultiva- ble bacteria and analyzing 16S rRNA gene clone libraries. The results showed that the number of cultivable bacteria was not affected by exogenous genes but the cotton growth period, and the number peaked at the stage of boll forming with vigorous metabolism. The 16S rRNA gene clone library prepared from soil bacteria in rhizospheres of transgenic and nontransgenic cotton at different stages contained 2400 clones which covered 283 genera. Among them, Acid o- bacterium was the most dominant group which contained 642 clones, followed by unclassified bacterium and 收稿日期: 2014−12−09