【作者】 鲁琰； 邹潇潇； 黄惠琴； 刘敏； 鲍时翔；

【Author】 Lu Yan;Zou Xiaoxiao;Huang Huiqin;Liu Min;Bao Shixiang;Institute of Tropical Bioscience and Biotechnology,CATAS;

【机构】 中国热带农业科学院热带生物技术研究所；

【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。笔者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。然后以穿刺巴斯德芽菌孢子体外吸附的南方根结线虫为实验材料,在显微镜下观察供试线虫的存活情况,每24h计数一次死亡线虫的条数,总共观察12d。并在被穿刺巴斯德芽菌吸附的第三天收集根结线虫提取线虫总RNA。采用实时荧光定量PCR方法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。通过分子鉴定笔者们确认纯化的根结线虫是南方根结线虫;且由吸附试验结果得知,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存活率降低,抑制其生殖能力,该作用机理可能与cyp-33、lin-45和mek-2的调控有关。更多还原

【Abstract】 In order to investigate the influence of Pasteuria penetrans on the changes of survival of Meloidogyne incognita and the expression of life-related gene and reproductive-related gene at the beginning of adsorption,authors purified Meloidogyne incognita collected from wild with single egg and identified it with molecular biology methods firstly.Then Meloidogyne incognita was adsorpted by Pasteuria penetrans in vitro.Within twelve days researchers observed Meloidogyne incognita with microscope and counted the number of death Meloidogyne incognita every 24 hours.The total RNA of Meloidogyne incognita was extracted at the third day after Meloidogyne incognita wasadsorpted.Real time quantitative PCR was used to detected the expression of cyp-33、lin-45 and mek-2.Authors confirmed that the purified nematode was Meloidogyne incognita and learned from adsorption test that the survival of Meloidogyne incognita significantly increased when infected by Pasteuria Penetrans.At the same time the expression of life-related gene cyp-33 and reproductive-related genes lin-45 and mek-2 reduced at the beginning of adsorption compared with the control group(P<0.05).Pasteuria penetrans could reduce the lifespan and reproduction of Meloidogyne incognita.The mechanism may be related to the regulation of cyp-33、lin-45 and mek-2.更多还原