小菜蛾V-ATP酶A亚基表达丰度差异

doi:10.3969/j.issn.1004-1524.20240285

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (4): 839-846.DOI: 10.3969/j.issn.1004-1524.20240285

小菜蛾V-ATP酶A亚基表达丰度差异

卢莉娜¹^,²(), 朱庆¹^,², 高美静¹^,², 谢雅晶¹^,², 刘贤金¹^,², 张志勇¹^,²^,^*()

1.省部共建国家重点实验室培育基地—江苏省食品质量安全重点实验室,江苏南京 210014
2.江苏省农业科学院农产品质量安全与营养研究所,江苏南京 210014

收稿日期:2024-03-28 出版日期:2025-04-25 发布日期:2025-05-09
作者简介:卢莉娜(1986—),女,甘肃定西人,博士,助理研究员,主要从事农药毒理学研究。E-mail: linlusailboat@163.com
通讯作者: *张志勇,E-mail:zhangzhiyong@jaas.ac.cn
基金资助:
国家自然科学基金(31901919);国家自然科学基金(31630061)

Expression abundance analysis of the V-ATPase subunit A gene in Plutella xylostella

LU Lina¹^,²(), ZHU Qing¹^,², GAO Meijing¹^,², XIE Yajing¹^,², LIU Xianjin¹^,², ZHANG Zhiyong¹^,²^,^*()

1. Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology, Nanjing 210014, China
2. Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China

Received:2024-03-28 Online:2025-04-25 Published:2025-05-09

摘要/Abstract

摘要： V型ATP酶(V-ATPase)在昆虫体内和Cry毒素作用机制中发挥重要作用。本研究采用分子克隆和实时荧光定量PCR(qRT-PCR)技术探究小菜蛾(Plutella xylostella)幼虫体内V-ATP酶A亚基基因在不同生长发育阶段,以及取食Cry1Ac毒素后的表达量丰度差异和变化情况。结果表明,小菜蛾V-ATP酶A亚基基因CDS区为1 848 bp,编码615个氨基酸(GenBank登录号:ON108624),与亚洲玉米螟(Ostrinia furnacalis)、二化螟(Chilo suppressalis)和棉铃虫(Helicoverpa armigera)的同源性约为94%。V-ATP酶A亚基基因在小菜蛾卵、幼虫、蛹和成虫期均有表达,在卵和蛹期表达量较低,在幼虫2龄期时表达量达到最高值,为卵期的4.16倍,3~4龄期表达量下降。小菜蛾幼虫取食含有Cry1Ac毒素的饲料后,体内V-ATP酶A亚基基因表达量整体呈显著下降趋势,取食48 h后表达量恢复至正常水平。在小菜蛾幼虫体内,V-ATP酶A亚基基因的表达量与生长发育和代谢变化密切相关,并且与Cry1Ac毒素的作用有一定关系。

关键词: 小菜蛾, V-ATP酶A亚基, Cry毒素, 表达丰度

Abstract:

V-ATPase plays important role in insects and mechanism of Cry toxins. In this study, the molecular cloning and real time fluorogenic quantitative PCR(qRT-PCR) was conducted to explore the expression of Plutella xylostella V-ATPase subunit A gene in different growth and developmental stages as well as before/after feeding Cry1Ac toxin. The results showed that the CDS of V-ATPase subunit A gene was 1 848 bp, which encoded 615 amino acids (GenBank No. ON108624) and had approximately 94% homology with V-ATPase subunit A of Ostrinia furnacalis, Chilo suppressalis and Helicoverpa armigera. The qRT-PCR results indicated that V-ATPase subunit A gene was expressed in the whole larval stages, eggs, pupa and adults. In eggs and pupa, the expression level of V-ATPase subunit A gene was low, while in the 2nd instar larvae the expression reached the highest to be 4.16 times as in eggs, and decreased in the 3rd and 4th instar larvae. After feeding the artificial diet with Cry1Ac toxins, the express level of V-ATPase subunit A gene declined and then reached to the normal after feeding 48 h. These results indicated that the express level of V-ATPase subunit A gene in P. xylostella was closely related with development and metabolic changes, and might be involved in the mode action of Cry1Ac toxin.

Key words: diamondback moth (Plutella xylostella), V-ATPase subunit A, Cry toxin, expression level

中图分类号:

S433.4

卢莉娜, 朱庆, 高美静, 谢雅晶, 刘贤金, 张志勇. 小菜蛾V-ATP酶A亚基表达丰度差异[J]. 浙江农业学报, 2025, 37(4): 839-846.

LU Lina, ZHU Qing, GAO Meijing, XIE Yajing, LIU Xianjin, ZHANG Zhiyong. Expression abundance analysis of the V-ATPase subunit A gene in Plutella xylostella[J]. Acta Agriculturae Zhejiangensis, 2025, 37(4): 839-846.

图/表 5

图1 V-ATP酶A亚基的氨基酸序列比对

Fig.1 Amino acid sequence alignment of V-ATPase subunit A

图2 不同昆虫V-ATP酶A亚基同源性分析

Fig.2 Homology analysis of V-ATPase subunit A in different insects

图3 小菜蛾不同生长发育时期V-ATP酶A亚基基因的表达量柱上无相同小写字母表示差异显著(P<0.05)。下同。

Fig.3 The expression level of V-ATPase subunit A gene at different developmental stages of Plutella xylostella Data marked without the same lowercase letters indicate significant difference(P<0.05). The same as below.

图4 不同质量浓度Cry1Ac毒素处理下小菜蛾幼虫的死亡率

Fig.4 Mortality of Plutella xylostella larvae treated with different mass concentrations of Cry1Ac toxin

图5 小菜蛾幼虫取食Cry1Ac毒素后V-ATP酶A亚基基因的表达量

Fig.5 The expression level of V-ATPase subunit A gene in Plutella xylostella larvae after feeding Cry1Ac toxin

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小菜蛾V-ATP酶A亚基表达丰度差异

Expression abundance analysis of the V-ATPase subunit A gene in Plutella xylostella

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