蠋蝽热激蛋白基因AcHsp83a和AcHsp83b的克隆、表达谱及对高低温和UV-B胁迫的响应

doi:10.16380/j.kcxb.2023.11.001

摘要/Abstract

摘要： 【目的】探索天敌昆虫蠋蝽Arma chinensis响应高低温和UV-B胁迫的分子机制。【方法】利用RT-PCR克隆蠋蝽热激蛋白基因Hsp83a和Hsp83b，并利用生物信息学分析其序列特征；利用RT-qPCR检测Hsp83a和Hsp83b在蠋蝽不同发育阶段(卵、1-5龄若虫、雌成虫和雄成虫)、成虫不同组织(头、胸、腹、翅、触角、脂肪体、足、马氏管、口器、中肠、卵巢和精巢)及38 ℃高温、4 ℃低温0(CK), 6和24 h时和UV-B胁迫0(CK), 6和12 h时雌成虫和雄成虫中的表达量。【结果】克隆获得蠋蝽2个Hsp90基因，分别命名为AcHsp83a(GenBank登录号: OP791883)和AcHsp83b(GenBank登录号: OP791884)，开放阅读框(ORF)分别长2 172和2 163 bp，分别编码723和720个氨基酸，编码蛋白相对分子量分别为83.12和82.90 kD，等电点(pI)分别为4.94和4.97，C末端序列都含保守基序EEVD，均为胞质型热激蛋白。AcHsp83a和AcHsp83b高度保守。AcHsp83a在卵中表达量最高，AcHsp83b在成虫中表达量最高；AcHsp83a在雄成虫精巢中表达量最高，AcHsp83b在雌成虫中肠中表达量最高。对于雌成虫，随着38 ℃, 4 ℃和UV-B处理时间的延长，AcHsp83a和AcHsp83b的表达量均先上升后下降，在6 h时达到最高。对于雄成虫，随着38 ℃和UV-B处理时间的延长，AcHsp83a的表达量均先上升后下降，在6 h时达到最高；随着4 ℃处理时间的延长，AcHsp83a的表达量在雄成虫中先下降后上升，在24 h时达到最高；在3种胁迫条件下，雄成虫体内AcHsp83b的表达量均较对照显著降低。【结论】蠋蝽AcHsp83a和AcHsp83b的差异表达说明其在蠋蝽的生长发育及适应极端温度和UV-B胁迫过程中发挥重要作用。

关键词: 蠋蝽, Hsp90, 基因克隆, 表达谱分析, 环境胁迫

Abstract: 【Aim】 To explore the molecular mechanism of the natural enemy insect Arma chinensis in response to high or low temperature and UV-B stresses.【Methods】 The heat shock protein genes AcHsp83a and AcHsp83b of A. chinensis were cloned by RT-PCR, and their sequence characteristics were analyzed by bioinformatics methods. RT-qPCR was used to detect the expression levels of AcHsp83a and AcHsp83b in different developmental stages (egg, 1st-5th instar nymphs, female adult and male adult), different adult tissues (head, thorax, abdomen, wing, antenna, fat body, leg, Malpighian tubules, mouthparts, midgut, ovary and testis), and female and male adults at 0 (CK), 6 and 24 h after exposure to high temperature of 38 ℃, and low temperature of 4 ℃, and at 0 (CK), 6 and 12 h after UV-B stress.【Results】 Two Hsp90 genes were cloned from A. chinensis, and namely AcHsp83a (GenBank accession no.: OP791883) and AcHsp83b (GenBank accession no.: OP791884). Their open reading frames (ORFs) were 2 172 and 2 163 bp in length, encoding 723 and 720 amino acids with the relative molecular weight of 83.12 and 82.90 kD and the isoelectric point (pI) of 4.94 and 4.97, respectively, and the C-terminal sequences both contain the conserved motif EEVD, showing that they belong to the cytoplasmic Hsps. AcHsp83a and AcHsp83b are highly conserved. The expression level of AcHsp83a was the highest in eggs, whereas that of AcHsp83b was the highest in adults. The expression level of AcHsp83a was the highest in the testis of male adults, whereas that of AcHsp83b was the highest in the midgut of female adults. When the female adults were exposed to 38 ℃, 4 ℃ or UV-B, the expression levels of AcHsp83a and AcHsp83b increased firstly and then decreased with treatment time, reaching the peak at 6 h. For the male adults, the expression level of AcHsp83a increased firstly and then decreased with treatment time, and reached the peak at 6 h after exposure to 38 ℃ or UV-B. The expression level of AcHsp83a in male adults decreased firstly and then increased with treatment time, and reached the peak at 24 h after exposure to 4 ℃. Compared with the control group, 38 ℃, 4 ℃ or UV-B exposure significantly decreased the expression levels of AcHsp83b in male adults. 【Conclusion】 The differential expression of AcHsp83a and AcHsp83b in A. chinensis suggests that the two genes play important roles in the growth and development of A. chinensis and contribute to its tolerance to extreme temperatures and UV-B stress.

Key words: Arma chinensis, Hsp90, gene cloning, expression profiling, environmental stress

孟建玉, 金鑫, 何龙春, 张雪霞, 杨昌利, 张长禹. 蠋蝽热激蛋白基因AcHsp83a和AcHsp83b的克隆、表达谱及对高低温和UV-B胁迫的响应[J]. 昆虫学报, 2023, 66(11): 1425-1434.

MENG Jian-Yu, JIN Xin, HE Long-Chun, ZHANG Xue-Xia, YANG Chang-Li, ZHANG Chang-Yu. Cloning and expression profiling of heat shock protein genes AcHsp83a and AcHsp83b from Arma chinensis (Hemiptera: Pentatomidae) and their response to high or low temperature and UV-B stresses[J]. Acta Entomologica Sinica, 2023, 66(11): 1425-1434.

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蠋蝽热激蛋白基因AcHsp83a和AcHsp83b的克隆、表达谱及对高低温和UV-B胁迫的响应

Cloning and expression profiling of heat shock protein genes AcHsp83a and AcHsp83b from Arma chinensis (Hemiptera: Pentatomidae) and their response to high or low temperature and UV-B stresses

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