[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 Cry1Ac resistance mechanism and constituting reasonable resistance management strategy,the difference of ALP1 expression level between Cry1Ac-resistant and susceptible Helicoverpa armigera,the effect of Cry1Ac toxin on ALP1 expression level in Cry1Ac-resistant larvae and the relationship between ALP 1 and Cry 1 Ac-resistant strains were analyzed.[Method] The expression levels of ALP1 in susceptible and Cry1Ac-resistant H.armigera larvae feeding on artificial diet containing Cry1Ac and no-Cry1Ac were compared using real-time quantitative PCR analysis.[Result] The ALP1 could express in whole development period ofH.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 Cry1Ac.The ALP1 expression of the resistant larvae significantly increased compared with the susceptible strain after they were fed on the artificial diet containing Cry1Ac,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 ALP1 in the 4th instar larval midgut of all resistant strains were obviously higher than that in susceptible strain,and the expression 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 Cry1Ac and metabolism of Cry1Ac.
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