Molecular characterization of the ryanodine receptor from Adoxophyes orana and its response to lethal and sublethal doses of chlorantraniliprole

doi:10.1016/S2095-3119(20)63356-1

摘要/Abstract

摘要：

昆虫鱼尼丁受体（RyRs）是邻甲酰氨基苯甲酰胺类和邻苯二甲酰胺类等二酰胺类杀虫剂的靶标。目前，已经有几种二酰胺类杀虫剂在我国果园害虫防治中应用。为了加强二酰胺类杀虫剂作用机制的了解，本文克隆了危害仁果和核果类较为严重的害虫苹小卷叶蛾的RyR（AoRyR）基因cDNA全长。此外，还研究了二酰胺杀虫剂对AoRyR mRNA表达调控的影响。通过本研究获得AoRyR mRNA的开放阅读框(ORF)为15402 bp，可编码5113个氨基酸。氨基酸序列比对结果显示，AoRyR与其他昆虫和哺乳动物RyRs分别具有77-92%和45-47%的一致性。在AoRyR中发现了一个可变剪接位点为互斥外显子(a/b)，其在卵和幼虫中的使用频率高于蛹和成虫。实时荧光定量qPCR显示，AoRyR mRNA在各发育阶段均有表达，特别是在卵、蛹和雄成虫中高表达。3龄幼虫经LC₁₀、LC₂₀和LC₅₀剂量氯虫苯甲酰胺处理后，AoRyR mRNA表达水平明显上调。研究结果为进一步研究苹小卷叶蛾鱼尼丁受体的功能和开发具有抗虫选择性活性的新化合物提供了依据。

Abstract:

The insect ryanodine receptor (RyR) is a novel target of the anthranilic and phthalic insecticides, which have high activity against lepidopteran insects. Several diamide insecticides have been used to control pests in orchards in China. To enhance our understanding of the effects of diamides on RyRs, full-length cDNAs were isolated and characterized from the summer fruit tortrix moth, Adoxophyes orana, which is the most severe pest of stone and pome trees worldwide. In addition, the modulation of AoRyR mRNA expression by diamide insecticides was investigated. The AoRyR mRNA obtained had an open reading frame (ORF) of 15 402 bp nucleotides encoding 5 113 amino acids, and shared high and low identity with its orthologs in other insects and mammals of 77–92 and 45–47% identity, respectively. One alternative splice site with two exclusive exons was revealed in AoRyR (a/b). The usage of exon was more frequent in eggs and larvae than in pupae and adults. Quantitative real-time reverse transcription PCR (qRT-PCR) showed that AoRyR mRNA was expressed at all developmental stages, especially in eggs, male pupae and male adults. The expression levels of AoRyR mRNA in the whole body were up-regulated markedly after 3rd instar larvae were treated with chlorantraniliprole at LC₁₀, LC₂₀ and LC₅₀ dosages. The results could provide the basis for further functional studies of AoRyR and for the development of new chemicals with selective activity against insects.

Key words: ryanodine receptor , Adoxophyes orana , chlorantraniliprole , mRNA expression

. [J]. Journal of Integrative Agriculture, 2021, 20(6): 1585-1595.

SUN Li-na, LIU Yan-di, ZHANG Huai-jiang, YAN Wen-tao, YUE Qiang, QIU Gui-sheng. Molecular characterization of the ryanodine receptor from Adoxophyes orana and its response to lethal and sublethal doses of chlorantraniliprole[J]. Journal of Integrative Agriculture, 2021, 20(6): 1585-1595.

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