昆虫学报 ›› 2016, Vol. 59 ›› Issue (2): 172-184.doi: 10.16380/j.kcxb.2016.02.007

• 研究论文 • 上一篇    下一篇

基于转录组的葱蝇表皮蛋白基因家族的鉴定及在非滞育和冬滞育条件下的表达分析

付丹影, 司风玲, 陈斌, 徐燕玲, 郝友进*   

  1. (重庆师范大学生命科学学院, 昆虫与分子生物学研究所, 重庆 401331)
  • 出版日期:2016-02-20 发布日期:2016-02-20
  • 作者简介:付丹影, 女, 1990年3月生, 吉林省公主岭人, 硕士研究生, 研究方向为昆虫分子生物学, E-mail: a1249990416@163.com

Transcriptome-guided identification of the cuticular protein gene family of Delia antiqua (Diptera: Anthomyiidae) and expression analysis under non- and winter-diapause conditions

FU Dan-Ying, SI Feng-Ling, CHEN Bin, XU Yan-Ling, HAO You-Jin*   

  1. (Institute of Entomology and Molecular Biology, College of Life Science, Chongqing Normal University, Chongqing 401331, China)
  • Online:2016-02-20 Published:2016-02-20

摘要: 【目的】昆虫表皮蛋白在昆虫的生长发育、抑制水分蒸发和抵御其他外界不良环境等方面起到重要的作用。通过转录组水平上葱蝇 Delia antiqua 表皮蛋白基因家族的鉴定及部分基因在非滞育和冬滞育条件下的表达规律分析,为进一步研究葱蝇表皮蛋白在滞育期的抗逆功能奠定基础。【方法】基于转录组数据,通过生物信息学方法对葱蝇表皮蛋白基因进行鉴定分析,系统进化和保守结构域分析并采用实时荧光定量PCR法比较分析了12个基因在葱蝇非滞育和冬滞育不同发育时期的表达变化。【结果】 在葱蝇中鉴定出81个表皮蛋白基因,分别属于RR (71个基因)、CPAP (5个基因)和CPLC (5个基因)亚家族,其编码的蛋白质含有92~495个氨基酸, 分子量约为 8.692~52.2 kD,等电点为3.90~9.93。亚细胞定位预测结果显示67个全长表皮蛋白均为外分泌型。12个葱蝇表皮基因在非滞育和冬滞育过程中有明显不同的表达规律。【结论】葱蝇表皮蛋白基因RR-2型的保守基序中发生了基因突变,引起关键氨基酸的替换,这可能暗示它们在非滞育和滞育不同条件下,不同发育阶段中蛹皮结构重建和某些器官的发生方面发挥不同作用。该研究结果为进一步阐明葱蝇滞育蛹皮结构及抗逆功能的研究提供了重要理论基础。

关键词: 葱蝇, 表皮蛋白, 基因家族, 滞育, 生物信息学, 表达分析

Abstract: 【Aim】 Insect cuticular proteins (CPs) play important roles in development, evaporation resistance and defense against other adverse environmental factors. In this study, the CPs gene family in Delia antiqua was identified based on transcriptome data and their expression in non-diapausing and winter-diapausing puape were investigated, so as to provide a basis for further investigation of the role of CPs in stress tolerance during diapausing period. 【Methods】 The CPs gene family of D. antiqua was identified based on transcriptome data using bioinformatics approaches, and analyses of subcellular localization, phylogeny and conserved domains were also performed. The expression of 12 genes in non-diapausing and winter-diapausing pupae was evaluated by RT-qPCR. 【Results】 Eighty-one CP genes were identified and categorized into CPR (71 genes), CPAP (5 genes) and CPLC (5 genes) subfamily, respectively. They encode proteins with 92-495 amino acids and pI of 3.90-9.93. Bioinformatic prediction showed that the molecular weight of CPs ranges from 8.692 to 52.2 kD. Subcellular localizationprediction revealed that 67 CPs are excreted protein. Among them, 12 genes had significantly different expression patterns between non-diapausing and winter-diapausing pupae, suggesting that they have stage-specific response to low temperature or short-day condition. 【Conclusion】 Functional diversity of some CPs genes from D. antiqua may contribute to the reconstruction of pupal cuticle. Our results would provide a basis for further studies on pupal cuticle structure and their potential functions in stress resistance during the diapausing period.

Key words: Dalia antiqua, cuticular proteins (CPs), gene family, diapause, bioinformatics, expression analysis