灰葡萄孢菌a说ur1基因对细胞生长和繁殖的影响.docVIP

灰葡萄孢菌AUR1基因对细胞生长和繁殖的影响 赵红霞,孙九丽, 苟萍* (新疆大学生命科学与技术学院, 新疆 乌鲁木齐 830046) 摘 要：目的: 明确灰葡萄孢菌AUR1基因（BcAUR1）对灰葡萄孢菌细胞生长、发育和繁殖的作用。方法: 采用PCR扩增BcAUR1的 P 和T 片段，依次克隆于pTFCM质粒载体的潮霉素基因两侧，构建灰葡萄孢菌AUR1断裂基因表达载体，通过农杆菌介导转化灰葡萄孢菌，在潮霉素抗性培养基上筛选转化子。AbA处理灰葡萄孢菌，观察细胞形态。结果: 断裂基因转化子比空载转化子显著减少，说明BcAUR1断裂基因同源重组替代了宿主AUR1基因，导致转化子不能存活。AbA显著影响野生型灰葡萄孢菌的生长发育，导致孢子萌发延迟、菌丝形态异常，不能形成分生孢子头；但AbA不影响抗AbA突变体BcAUR1a的生长发育。结论： AUR1基因对灰葡萄孢菌生长、发育和繁殖起重要作用 关键词：灰葡萄孢菌, IPC合成酶, 短梗霉素A, 同源重组，转化子 中图分类号：S188+． 2 文献标识码：A Effects of Botrytis cinerea AUR1 gene on cell growth and reproduction ZHAO Hong-xia, GOU Ping* (College of Life and Technology ,Xinjiang University,Urumqi 830046,China) Abstract:Objective: To investigate the influence of AUR1 gene on Botrytis cinerea cell growth and reproduction. Method: BcAUR1 P fragment and T fragment were amplifided by PCR, cloned into both sides of hygromycin gene of plasmid vector pTFCM by sequence. The vector was transformed into B. cinerea by agrobacterium-mediated genetic transformation method and the transformants were screened by hygromycin resistance. B. cinerea was treated by AbA and cell morphology was observed. Result: The interrupted gene transformants were obviously less than empty plasmid transformants, indicating that BcAUR1 interrupted gene had been homologous recombinantion into B. cinerea’s genome, causing B. cinerea not survive. Besides, AbA could inhibit the growth of wild B. cinerea obviously, delay spore germination, induce abnormal of mycelial morphology, block the formation of conidium head. However, AbA did not affect the growth of AbA-resistant mutant BcAUR1a. Conclusion: AUR1 gene plays a key role in B. cinerea cell growth and reproduction. Keywords: Botrytis cinerea, IPC synthase enzyme, aureobasidin A, HYPERLINK javascript:void(0);homologous HYPERLINK javascript:void(0);recombination, transformantsa 基金项目：国家自然科学基金项目 作者简介：赵红霞（1987—），女，甘肃定西市人，硕士研究生，专业方向：生物化学与分子生物学. (E-mail) zhaohongxiazyn@ 通讯作者：苟萍(1955—)，女，新疆乌鲁木齐人，教授，硕士生导师，研究方向：生物化学与分子生物学．(E-mail) HYP