Cry1Ac抗、感棉铃虫碱性磷酸酯酶(ALP1)的表达量比较  

The Expression Level of Alkaline Phosphatase(ALP1) in Cry1Ac-Resistant and Susceptible Cotton Bollworm Helicoverpa armigera Hübner

作　　者：张涛[1,2] 张丽丽[2] 魏纪珍[2] 肖玉涛[2] 梁革梅[2] 周瑞阳[1] 

机构地区：[1]广西大学农学院,南宁530004 [2]中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室,北京100193

出　　处：《中国农业科学》2013年第17期3580-3586,共7页Scientia Agricultura Sinica

基　　金：国家转基因生物新品种培育重大专项课题(2013ZX08011-002)

摘　　要：【目的】通过比较Cry1Ac抗、感棉铃虫昆虫中肠碱性磷酸酶(ALP1)表达量的差异及抗性棉铃虫取食Cry1Ac蛋白对ALP1表达量的影响,分析ALP1表达量变化与抗性的关系,为进一步明确Bt抗性机制、制定抗性治理策略提供理论依据。【方法】利用实时荧光定量PCR技术,比较敏感棉铃虫、Cry1Ac抗性棉铃虫取食含Cry1Ac蛋白的饲料和正常饲料后ALP1表达量的差异。【结果】ALP1在棉铃虫整个发育期都表达,幼虫的表达量最高,蛹期表达量最低,在成虫体内也有较高的表达;ALP1在幼虫中肠表达量最高,其次是后肠、前肠、马氏管,表皮中的表达量最低;与敏感品系相比,对Cry1Ac具有中等水平抗性的棉铃虫ALP1表达量明显增加,尤其是取食含Cry1Ac蛋白饲料的抗性棉铃虫幼虫的ALP1的表达量显著升高,但抗性棉铃虫取食正常饲料后,2、3龄幼虫的ALP1的表达量与敏感棉铃虫差异不显著;所有的抗性品系4龄棉铃虫幼虫中肠ALP1的表达量都显著高于敏感品系,而且随着棉铃虫抗性倍数的升高,ALP1的表达量呈逐渐降低的趋势。【结论】抗性棉铃虫中肠ALP1表达量的改变可能与Cry1Ac抗性、Cry1Ac代谢有一定的关系。[Objective] The objective of this study is to provide a theoretical basis for further clarifying CrylAc resistance mechanism and constituting reasonable resistance management strategy, the difference of ALP1 expression level between CrylAc-resistant and susceptible Helicoverpa armigera, the effect of CrylAc toxin on ALP1 expression level in CrylAc-resistant larvae and the relationship between ALP1 and CrylAc-resistant strains were analyzed. [Method] The expression levels of ALP1 in susceptible and CrylAc-resistant H. armigera larvae feeding on artificial diet containing CrylAc and no-CrylAc were compared using real-time quantitative PCR analysis. [Result] The ALP1 could express in whole development period of/-/, armigera, the highest expression level occurred in larvae, while the lowest was in pupae, and there was also relatively high expression level in moth. The highest expression level of ALP1 was in midgut of H. armigera larvae, followed by hindgut, foregut, malpighian tubules and epidermis. Compared with the susceptible H. armigera, the expression of ALP1 obviously increased in resistant larvae which had moderate resistance to CrylAc. The ALP1 expression of the resistant larvae significantly increased compared with the susceptible strain after they were fed on the artificial diet containing CrylAc, while the difference of ALP1 expression in 2nd and 3rd instar larvae was not significant between resistant strain and susceptible strain after they were fed on the normal artificial diet. The expression of ALP 1 in the 4th instar larval midgnt of all resistant strains were obviously higher than that in susceptible strain, and theexpression level of ALP1 appeared downtrend along with the resistance ration increased. [Conclusion] The changes of ALP1 expression in midgut of resistant H. armigera may have a relationship with its resistance to CrylAc and metabolism of CrylAc.

关 键 词：棉铃虫 BT 抗性 碱性磷酸酶 表达量 

分 类 号：S435.622.3[农业科学—农业昆虫与害虫防治]