滞育七星瓢虫酰基辅酶AΔ11去饱和酶基因的克隆及表达分析

doi:10.16409/j.cnki.2095-039x.2018.05.005

摘要/Abstract

摘要： 去饱和酶（Fatty acid desaturase，FAD）是生物体不饱和脂肪酸合成中的关键酶，酰基辅酶AΔ11去饱和酶基因是其中重要的一种。本试验基于七星瓢虫Coccinella septempunctata L.转录组数据，利用RT-PCR和RACE技术从滞育七星瓢虫中克隆得到酰基辅酶AΔ11去饱和酶基因全长，并命名为CsFadΔ11（GenBank登录号：MF458996），该基因全长1447 bp，开放阅读框（ORF）1095 bp，编码364个氨基酸，预测蛋白质分子量为42.99 kD，等电点（pI）为8.87，无信号肽。氨基酸序列分析结果表明，CsFadΔ11有3个组氨酸富集区和6个跨膜结构域，与膜翅目的内华达古白蚁Zootermopsis nevadensis同源性较高。利用实时荧光定量PCR技术研究其时间表达模式，发现CsFadΔ11基因在七星瓢虫初羽化、滞育诱导10 d、滞育诱导20 d、滞育初期、滞育后期、滞育解除期以及正常发育状态下均有所表达，且在滞育早期表达量最高，其次是在滞育解除个体中。其整体表达趋势与相应时期脂积累变化情况基本相符，推测CsFadΔ11基因参与七星瓢虫脂积累调控，并进一步影响七星瓢虫滞育。

关键词: 七星瓢虫, 酰基辅酶AΔ11去饱和酶, 基因克隆, 表达分析, 滞育, 脂积累

Abstract: Acyl-CoA Δ11 desaturase is one of the fatty acid desaturases that are key enzymes in the synthesis pathway of unsaturated fatty acids. Based on the transcriptome database of Coccinella septempunctata L., we cloned the full length cDNA of a FadΔ11 gene using RT-PCR and RACE technique, which was named CsFadΔ11 (GenBank accession no. MF458996). CsFadΔ11 is 1447 bp in length and contains an open reading frame (ORF) of 1095 bp, encoding 364 amino acid residues. The putative protein is 42.99 kD with an isoelectric point (pI) of 8.87 and has no signal peptide. Amino acid sequence analysis showed that CsFadΔ11 contains three histidine boxes and six transmembrane domains, which are composite characteristics of a fatty acid desaturase. Meanwhile, CsFadΔ11 showed high homology with Zootermopsis nevadensis. The expression profile of this gene in different treatments was detected by quantitative real-time PCR. The results indicated that CsFadΔ11 was constently expressed in newly emerged adult, day 10 or 20 after diapause induction, early diapause, late diapause, post diapause and normal development adults, with the highest expression levels in early diapause and the second levels in post diapause. The change trend of the gene was consistent with the lipid content in different developmental stages of C. septempunctata. These results suggested that CsFadΔ11 gene may participate in lipid metabolism, and further affect the diapause development of C. septempunctata.

Key words: Coccinella septempunctata, acyl-CoA Delta (11) desaturase, gene cloning, gene expression, diapause, lipid accumulation

中图分类号:

S476.2

韩艳华, 刘梦姚, 高飞, 王曼姿, 张洪志, 孔琳, 陈红印, 张礼生. 滞育七星瓢虫酰基辅酶AΔ11去饱和酶基因的克隆及表达分析[J]. 中国生物防治学报, 2018, 34(5): 676-685.

HAN Yanhua, LIU Mengyao, GAO Fei, WANG Manzi, ZHANG Hongzhi, KONG Lin, CHEN Hongyin, ZHANG Lisheng. Molecular Cloning and Expression Analysis of Acyl-CoA Delta (11) Desaturase in the Diapausing Ladybird, Coccinella septempunctata L. (Coleoptera: Coccinellidae)[J]. journal1, 2018, 34(5): 676-685.

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