棉铃虫JNKs基因的克隆及表达分析  

Cloning and expression analysis of JNKs gene in cotton bollworm Helicoverpa armigera

作　　者：于思琪 汤金荣 张彩虹 EI THINZAR SOE 梁革梅[1,2] YU Siqi;TANG Jinrong;ZHANG Caihong;EI THINZAR SOE;LIANG Gemei(State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China;West Agricultural Research Center,Chinese Academy of Agricultural Sciences,Changji 831100,China)

机构地区：[1]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 [2]中国农业科学院西部农业研究中心,昌吉831100

出　　处：《植物保护》2023年第1期139-146,共8页Plant Protection

基　　金：国家棉花产业技术体系(CARS-15-20)。

摘　　要：c-Jun氨基末端激酶(c-Jun N-terminal kinase, JNK)是MAPK家族(mitogen-activated protein kinases,丝裂原活化蛋白激酶)的重要成员,具有参与昆虫抗逆反应等多种功能。为明确JNK在棉铃虫Helicoverpa armigera中的表达特性及其对Bt杀虫蛋白的应激与免疫反应,本研究通过PCR克隆得到2个棉铃虫JNK基因HaJNK1和HaJNK2;生物信息学分析结果显示:HaJNK1和HaJNK2基因开放阅读框分别为1 191、1 143 bp,分别编码397、381个氨基酸。系统进化树分析结果表明棉铃虫HaJNK1与黏虫Mythimna separata聚为一支,亲缘关系较近,HaJNK2与家蚕Bombyx mori聚为一支,同源性较高。利用实时荧光定量PCR技术分析HaJNK1与HaJNK2在棉铃虫不同发育时期、不同组织中的表达量,发现HaJNK1与HaJNK2在卵中表达量最高,其次是雌成虫;HaJNK1在性腺中表达量最高,其次是唾液腺;HaJNK2在头部表达量最高,其次是性腺。取食Cry1Ac的4龄棉铃虫幼虫的中肠组织中,HaJNK1与HaJNK2的表达量均显著升高。推测HaJNKs基因可能参与棉铃虫抵御Bt杀虫蛋白伤害的应激和抗逆反应。c-Jun N-terminal kinase(JNK) is an important member of the MAPK family(mitogen-activated protein kinases), which has various functions such as participating in insect anti-stress response. In order to clarify the expression characteristics of HaJNK and its role in defense against Bt insecticidal protein in cotton bollworm Helicoverpa armigera, we cloned two HaJNK gene coding sequences HaJNK1 and HaJNK2 by PCR amplification. Bioinformatics analysis showed that the open reading frames of HaJNK1 and HaJNK2 genes were 1 191 bp and 1 143 bp, encoding 397 and 381 amino acids, respectively. Phylogenetic tree analysis showed that HaJNK1 of H.armigera had high homology with that of Mythimna separata and HaJNK2 of H.armigera is clustered with that of Bombyx mori. Quantitative real-time PCR was used to analyze the expression levels of HaJNK1 and HaJNK2 in different developmental stages and tissues. It was found that the expression levels of HaJNK1 and HaJNK2 were the highest in the stage of egg, following by female adults. The highest expression of HaJNK1 was observed in gonads followed by salivary glands, and the highest expression of HaJNK2 was in head followed by gonads. For the fourth instar larvae, after cotton bollworm fed on Cry1Ac, the expression levels of HaJNK1 and HaJNK2 in midgut were significantly increased. It was speculated that the HaJNKs of H.armigera might be involved in emergency response and antistress response against Bt insecticidal protein.

关 键 词：棉铃虫 Cry1Ac蛋白 JNK基因 表达水平 

分 类 号：S435.622.3[农业科学—农业昆虫与害虫防治]