Cloning and mRNA expression analysis of Vacuolar H+-ATPase d subunit gene in the brown planthopper, Nilaparvata lugens (St?l)

Author of the article:Zhu You-Li Wang Jian Xie Zhi-Juan Liu Ya-Ping

Author's Workplace：College of Horticulture and Plant Protection, Yangzhou University

Key Words：Nilaparvata lugens; V-ATPase gene; qRT-PCR; subleathal dose; triazophos

Abstract：The vacuolar H+-ATPase (V-ATPase) plays an important role in growth and development of insects. In this study, a cDNA sequence of V-ATPase d subunit gene was obtained from the brown planthopper, Nilaparvata lugens (Stål), by RT-PCR and named as NlVHA-d, and its expression was analyzed by qRT-PCR. The results showed that the open reading frame of NlVHA-d is 1047bp in length, encoding 349 amino acid residues, and the V-ATPase d genes are highly conserved among different insects. The qRT-PCR analysis revealed that the mRNA expression level of NlVHA-d was the highest in second instar nymphs, and the female adults showed higher levels of expression than the male adults. Furthermore, at third day after eclosion, the relative expressions of NlVHA-d in brachypterous male adults treated with LD10 and LD30 of triazophos were 2.15 and 2.46 times higher than that in control treatment with acetone, respectively. The over-expression of NlVHA-d gene in brachypterous male adults treated with subleathal dose of triazophos might contribute to the resurgence of brown planthopper.