南方根结线虫被穿刺巴斯德芽菌侵染后寿命和生殖相关基因的表达变化  

Expression Changes of Life-related and Reproduction-related Genes of Meloidogyne Incognita Infected by Pasteuria Penetrans

作　　者：鲁琰[1,2,3] 邹潇潇 黄惠琴[2] 刘敏[2] 鲍时翔[2] 

机构地区：[1]海南大学农学院,海南海口570228 [2]中国热带农业科学院生物技术研究所,海南海口571101 [3]海南医学院,海南海口571199

出　　处：《热带作物学报》2015年第10期1845-1850,共6页Chinese Journal of Tropical Crops

基　　金：973计划(No.2013CB127502);国家自然科学基金项目(No.31170062);948项目(No.2011-G25);公益性行业农业科研专项(No.201403075);海南省科技项目(No.KJHZ2013-11;No.311080;No.sys2014005);海口市重点科技计划项目(No.2011-0099);中央级公益性科研院所基金(No.ITBB11-0302)

摘　　要：为了探讨南方根结线虫在植物体外被穿刺巴斯德芽菌感染后存活率的变化及感染初期线虫寿命和生殖相关基因的表达情况。首先利用单卵块孵化法对从野外采集的根结线虫进行纯化和分子鉴定。以穿刺巴斯德芽菌孢子体外吸附的南方根结线虫为材料,在显微镜下观察供试线虫的存活情况,每24 h计数一次死亡线虫的条数,总共观察12 d。在被穿刺巴斯德芽菌吸附的第3天收集根结线虫,提取线虫总RNA,采用实时荧光定量PCR检测线虫寿命相关基因cyp-33和生殖相关基因lin-45/mek-2的表达情况。结果表明:通过分子鉴定确认纯化的根结线虫为南方根结线虫;感染了穿刺巴斯德芽菌的南方根结线虫死亡率显著升高;同时在穿刺巴斯德芽菌吸附南方根结线虫的初期,线虫寿命相关基因cyp-33和生殖相关基因lin-45/mek-2的表达水平与对照组相比均下调(p<0.05)。这说明穿刺巴斯德芽菌在吸附南方根结线虫的初期能够促使线虫的存活率降低,并抑制其生殖能力,这可能与基因cyp-33、lin-45和mek-2的调控有关。In order to investigate the influence of Pasteuria penetrans on the survival rate of Meloidogyne incognita and the expression of life-related gene and reproduction-related gene at the beginning of adsorption, the authors purified M. incognita collected from wild and identified it with molecular biology methods firstly. Then M.incognita was adsorpted by P penetrans in vitro. Within twelve days the researchers observed M. incognita with microscope and counted the number of dead M. incognita every 24 hours. The total RNA of M. incognita was extracted at the third day after M. incognita was adsorpted. Real time quantitative PCR was used to detect the expression of cyp- 33 、 lin- 45 and mek- 2 gene. The authors confirmed that the purified nematode was M. incognita and the death rate of M. incognita significantly increased when infected by P.penetrans. At the same time the expression of life-related gene cyp-33 and reproduction-related genes lin-45 and mek-2 reduced at the beginning of adsorption comparing with the control group（p 0.05）. P. penetrans could reduce the lifespan and reproduction of M. incognita. The mechanism ma y be related to the regulation of cyp-33 、 lin-45 and mek-2 gene.

关 键 词：南方根结线虫 穿刺巴斯德芽菌 荧光定量PCR 存活率 

分 类 号：R383.1[医药卫生—医学寄生虫学]