小菜蛾芳基酰胺酶表达及其与呋喃虫酰肼抗性的关系  

The distribution of aryl-acylamidase in the diamondback moth,Plutella xylostella and its relationship to fufenozide resistance

作　　者：孙璟琰[1] 梁沛[1] 高希武[1] 

机构地区：[1]中国农业大学昆虫学系,北京100193

出　　处：《应用昆虫学报》2012年第2期396-402,共7页Chinese Journal of Applied Entomology

基　　金：国家重点基础研究发展规划项目(973计划)(2012CB114103);国家自然科学基金项目(30671389;30971941;31171873);公益性行(农业)科研专项(201203038)

摘　　要：以对硝基乙酰苯胺为底物,采用紫外分光光度计比色法,研究了小菜蛾Plutella xylostella(L.)体内芳基酰胺酶酶促反应的最适反应条件、各发育阶段和不同组织中芳基酰胺酶的活性及亚细胞分布及其与小菜蛾对呋喃虫酰肼抗性的关系。结果表明,小菜蛾芳基酰胺酶最适反应条件为35℃,pH7.5,底物浓度为1.2×10-3mol/L,反应时间30min。利用非变性聚丙烯凝胶电泳活性染色及SDS-PAGE法测得小菜蛾芳基酰胺酶的分子量为61.1ku。小菜蛾4龄幼虫的芳基酰胺酶活性最高。不同亚细胞层中,芳基酰胺酶活力大小依次为线粒体层>微粒体层>细胞核和细胞碎片层>可溶性部分。芳基酰胺酶主要分布在小菜蛾4龄幼虫的腹部,其余依次为表皮、头部和胸部。小菜蛾呋喃虫酰肼抗性品系芳基酰胺酶的活性是敏感品系的3.89倍,表明该酶参与了小菜蛾对呋喃虫酰肼的抗性形成。The optimum reaction conditions of aryl-acylamidase in Plutella xylostella（L.） were determined using 4-nitroanilide as a substrate.The results show that the optimum pH,temperature,substrate concentration and reaction time were 7.5 and 35℃,1.2×10-3 mol/L and 30 min,respectively.The molecular weight of the enzyme is 61.1 ku,as determined by native-and SDS-PAGE.The hydrolyzing activity of aryl-acylamidase was highest in 4th instar larvae and the highest specific activity was in mitochondria.Specific activity was highest in abdomen,followed by the cuticle,head and thorax.Aryl-acylamidase activity of the fufenozide resistant strain was 3.89 times higher than in the susceptible strain,which suggests that aryl-acylamidase is involved in fufenozide resistance in the P.xylostella.

关 键 词：小菜蛾 芳基酰胺酶 反应体系 发育阶段 组织分布 呋喃虫酰肼 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]