›› 2015, Vol. 58 ›› Issue (2): 103-114.

• 研究论文 •    下一篇

禾谷缢管蚜鞣化激素基因克隆及功能分析

刘孝明#, 燕赛英#, 孙成贤, 高杜军, 杜孟芳, 安世恒*, 尹新明*   

  1. (河南农业大学植物保护学院, 郑州 450002)
  • 出版日期:2015-02-20 发布日期:2015-02-20
  • 作者简介:刘孝明, 男, 1989年生, 河南商丘人, 硕士研究生, 研究方向为昆虫分子生物学, E-mail: liuxiaoming0318@126.com; 燕赛英, 女, 1989年生, 湖南常德人, 硕士研究生, 研究方向为昆虫生理生化和分子生物学, E-mail: yansaiying200707@163.com

Molecular cloning and functional analysis of bursicon genes in Rhopalosiphum padi (Hemiptera: Aphididae)

LIU Xiao-Ming#, YAN Sai-Ying#, SUN Cheng-Xian, GAO Du-Jun, DU Meng-Fang, AN Shi-Heng* , YIN Xin-Ming*   

  1. (College of Plant Protection, Henan Agricultural University, Zhengzhou 450002, China)
  • Online:2015-02-20 Published:2015-02-20

摘要: 【目的】探讨禾谷缢管蚜 Rhopalosiphum padi (Linnaeus)鞣化激素基因的发育表达模式及功能。【方法】采用转录组测序得到禾谷缢管蚜鞣化激素基因bursicon-α bursicon-β cDNA序列。通过实时荧光定量PCR方法,分析该基因的发育表达模式。利用RNA干扰(RNA interference, RNAi)介导 bursicon-αbursicon-β 沉默,分析鞣化激素的功能。【结果】序列分析结果显示,禾谷缢管蚜鞣化激素α亚基基因(bursicon-α)cDNA序列开放阅读框为480 bp,编码159个氨基酸残基;β亚基基因(bursicon-β)cDNA序列开放阅读框为417 bp,编码138个氨基酸残基。时序表达分析表明,鞣化激素两个亚基基因在禾谷缢管蚜整个发育期均有表达,以1龄若蚜期表达量最高;成蚜有翅个体中表达量显著高于无翅个体。RNAi介导的 bursicon-α bursicon-β 沉默均能显著抑制禾谷缢管蚜成蚜表皮的黑化。【结论】研究结果表明,鞣化激素在禾谷缢管蚜体壁黑化中发挥着重要作用。该结果为进一步研究鞣化激素在蚜虫生长发育过程中的生理功能提供了基础资料。

关键词: 禾谷缢管蚜, 鞣化激素, 表皮鞣化, 发育表达模式, 荧光定量PCR, RNA干扰

Abstract: 【Aim】 This study aims to explore the developmental expression pattern and functional analysis of bursicon genes  bursicon-α and  bursicon-β in Rhopalosiphum padi (Linnaeus). 【Methods】 In this study, the cDNA sequences encoding bursicon-α and bursicon-β were obtained from RNA-seq transcriptome sequencing of  R. padi . The expression patterns of  bursicon-α  and  bursicon-β are analyzed by real-time quantitative PCR and the functional analyses of bursicon-α and bursicon-β were carried out through RNA interference technology. 【Results】 The sequence analysis showed that the open reading frame (ORF) of bursicon-α from R. padi  is 480 bp in length, encoding a protein with 159 amino acid residues. The ORF of bursicon-β  is 417 bp in length, encoding a protein with 138 amino acid residues. The developmental expression patterns showed that bursicon-α and bursicon-β were ordinarily expressed during the whole developmental stage of  R. padi, and arrived at the highest expression level in the 1st instar nymph. The expression levels of these two genes in winged adult aphids were significantly higher than those in wingless adult aphids. RNA interference-mediated knockdown of bursicon-α and bursicon-β significantly inhibited the cuticular melanization of adult aphids. 【Conclusion】 These results revealed that bursicon plays an important role in the process of the cuticular melanization in R. padi, providing a foundation for further study for the physiological functions of bursicon in aphids.

Key words: Rhopalosiphum padi , bursicon, cuticular tanning, developmental expression pattern, real-time quantitative PCR, RNA interference