›› 2014, Vol. 57 ›› Issue (8): 889-896.

• 研究论文 • 上一篇    下一篇

拟黑多刺蚁成虫头部USP基因的定位与定量分析及其与EcR基因相关性

党亮, 奚耕思*, 樊瑾瑛, 杨栋梁, 王彦男   

  1. (陕西师范大学生命科学学院, 西安 710062)
  • 出版日期:2014-08-20 发布日期:2014-08-20

Positioning and quantitative analysis of ultraspiracle protein gene in the head and its correlation with ecdysone receptor gene in adult Ployrhachis vicina (Hymenoptera: Formicidae)

DANG Liang, XI Geng-Si*, FAN Jin-Ying, YANG Dong-Liang, WANG Yan-Nan   

  1. (College of Life Sciences, Shaanxi Normal University, Xi’an 710062, China)
  • Online:2014-08-20 Published:2014-08-20

摘要: 【目的】超气门蛋白(ultraspiracle protein, USP)是蜕皮激素作用靶标的重要组成部分。本研究拟通过分析在拟黑多刺蚁Polyrhachis vicina Roger USP基因PvUSP在不同品级成虫头部mRNA表达的差异,推测PvUSP对其脑神经功能或行为的影响;拟通过饲喂PvUSP dsRNA对拟黑多刺蚁不同品级成虫PvUSP进行RNA干扰(RNAi),推测PvUSP对虫体生理功能的影响及其与蜕皮激素受体(ecdysone receptor, EcR)基因PvEcR之间功能的相关性。【方法】利用荧光原位杂交及荧光实时定量PCR技术对拟黑多刺蚁不同品级成虫头部PvUSP mRNA组织分布与表达水平进行检测;通过饲喂PvUSP dsRNA对拟黑多刺蚁不同品级成虫PvUSP进行RNAi,采用荧光实时定量PCR检测拟黑多刺蚁PvUSPPvEcR表达水平的变化。【结果】PvUSP mRNA广泛表达于拟黑多刺蚁不同品级成虫头部(主要分布于蕈形体),不同品级成虫头部PvUSP mRNA的表达量不同,工蚁头部表达量最高,雄蚁头部次之,雌蚁头部的表达量最低;RNAi沉默PvUSP后,与对照组相比,PvUSP mRNA在不同品级拟黑多刺蚁成虫体内表达量均明显降低,并存在显著差异(P<0.01);与对照组相比,PvEcR mRNA在各品级表达量均有增加,工蚁和雄蚁表达量变化不显著(P>0.05),但雌蚁体内PvEcR mRNA表达量显著增加(P<0.05)。【结论】PvUSP基因对拟黑多刺蚁不同品级头部神经系统的构建、功能作用的发挥有关;拟黑多刺蚁PvUSP基因与PvEcR基因可能在异源二聚体形成过程中存在关联性或功能的互补性;PvUSP可能对雌蚁的生殖功能产生重要影响。

关键词: 拟黑多刺蚁, 超气门蛋白(USP), RNA干扰, 蜕皮激素受体(EcR);发育调节

Abstract: 【Aim】 Ultraspiracle protein (USP) belongs to an essential component of a heterodimeric receptor complex with the ecdysone receptor (EcR) which binds ecdysteroids. This study aimed to explore the distribution of USP mRNA in heads of different castes of Polyrhachis vicina Roger adults for inferring the influence of PvUSP on the function or behavior of cranial nerve of P. vicina, and to assess the influence of PvUSP on physiological functions in the body and the relationship with EcR gene by feeding PvUSP dsRNA as RNAi to different castes of P. vicina adults. 【Methods】 We used real-time fluorescent quantitative PCR technique and fluorescence in situ hybridization to test the distribution of PvUSP mRNA expression level in the head of different castes of P. vicina adults, conducted RNAi on different castes of P. vicina adults by feeding PvUSP dsRNA, and used the real-time fluorescent quantitative PCR technique to detect the changes in PvUSP and PvEcR mRNA levels. 【Results】 PvUSP was widely expressed in the head of different castes of P. vicina adults, and especially concentrated in the mushroom body. However, its expression level was quite different in the three castes. The highest expression level was found in the head of workers, the moderate expression level was in the head of male adults, and the lowest was in the head of female adults. After using RNAi technique to silence PvUSP, the expression level of PvUSP mRNA decreased obviously in bodies of different castes of P. vicina adults as compared to the control, and an extremely significant difference was observed (P<0.01), while the expression levels of PvEcR mRNA all increased in different castes. The changes in PvEcR mRNA level in workers and male adults were not distinct (P>0.05), while those in the female body showed a dramatic increase (P<0.05). 【Conclusion】 PvUSP may be related to the structure and functions of nervous system of P. vicina. PvUSP and PvEcR may have relevance in the formation of heterogeneous dimers or function complementarily. PvUSP may influence the reproductive functions of female adults of P. vicina.

Key words: Polyrhachis vicina, ultraspiracle protein (USP), RNAi, ecdysone receptor (EcR), developmental regulation