抗浓核病毒中国株家蚕品系中肠羧酸酯酶活力变化和基因表达差异的研究.docVIP

抗浓核病毒中国株家蚕品系中肠羧酸酯酶活力变化和 基因表达差异的研究 高贵田1,2，陈克平1，姚 勤1，陈慧卿1，王林玲1，徐家萍1，赵 远1，王永杰1 （1江苏大学生命科学研究院，镇江212013；2陕西师范大学食品工程系，西安710062） 摘要：【目的】探讨家蚕中肠羧酸酯酶活力变化及羧酸酯酶基因表达差异与家蚕抗浓核病毒之间的关系，阐明其抗性的分子机理。【方法】以易感浓核病毒中国株家蚕品系华八和完全抗性品系BC8（华八的近等基因系）为材料，用BIO-TEK SYNERGY测定经口接种病毒(以下简称接种)后12 h、36 h、72 h 2个品系中肠的羧酸酯酶活力，并用实时荧光定量PCR研究接种后12 h、36 h、72 h中肠羧酸酯酶基因的表达差异。【结果】接种后12 h中肠羧酸酯酶活力BC8接种组分别是BC8对照组、华八接种组和华八对照组的3.28倍、2.26倍和3.02倍,差异达到极显著水平（P＜0.01）其它时间段的供试组之间无统计学差异；接种后12 h中肠羧酸酯酶基因的相对表达量BC8接种组分别是华八接种组、华八对照组、BC8对照组的17.714倍、21.76倍和15.09倍,差异达到极显著水平（P＜0.01），其它时间段的供试组之间也无统计学差异。【结论】接种后12 h抗性品系BC8中肠羧酸酯酶活力升高及中肠羧酸酯酶基因表达水平升高可能与家蚕品系是否有抗性基因(nsd/nsd)有关,也与浓核病毒中国株的感染刺激有关；抗性品系BC8中肠羧酸酯酶活力升高的分子基础可能是羧酸酯酶基因表达水平的改变。 关键词：家蚕；抗性；浓核病毒中国株；中肠；羧酸酯酶；酶活力；表达差异 Activity and Differential Display of Carboxylesterase in Bombyx mori Midgets of Against BmDNV-Z GAO Gui-tian1,2, CHEN Ke-ping1, YAO Qin1, CHEN Hui-qing1, WANG Lin-ling1, XU Jia-ping1, ZHAO Yuan1,WANG Yong-jie1 (1Institute of Life Sciences, Jiangsu University, Zhenjiang, 212013；2Department of Food Enginneering,Shaanxi Normal University, Xian 710062) Abstract: 【Objective】 The study was used to discuss the relationship between the activity of Bombyx mori carboxylesterase (BmCarE) in the midgets. It was also needed to study the differential display of bmcare against Bombyx mori densonucleosis virus (BmDNV), and to depict the molecular mechanism of resistance to BmDNV. 【Method】 The activity of BmCarE in the midgets was detected by BIO-TEK SYNERGY after 12h, 36h and 72h inoculation with virus between the silkworm strain Huaba. This strain is susceptible to BmDNV-Z, and BC8 (a near isogenic line of Huaba),which is resistant to the same virus. The expression differential of the BmCar was investigated after 12h, 36h and 72h inoculation with virus by real-time fluorescence quantitative PCR. 【Result】 The activity of BmCarE in the midgets of BC8 inoculated with virus after 12h was 3.28, 2.26 and 3.02 times of that in the BC8 control, HUABA inoculated with virus and HUA