title: Response of the vacuolar ATPase subunit E to RNA interference and four chemical pesticides in Leptinotarsa decemlineata (Say).
abstract: Vacuolar-type H(+)-ATPases (vATPases) are localized in the apical membranes of nearly all epithelial tissues of insects, energize the membranes to absorb and/or secrete ions and fluids, and play essential roles in many physiological functions. Here we cloned and characterized a 1041-bp full-length vATPase subunit E cDNA (named as LdATPaseE) that encoded a 226-amino acid protein in Leptinotarsa decemlineata. LdATPaseE mRNA levels were constantly increased from egg to the third- and fourth-instar stages, dropped in wandering and pupal stages and were elevated again in the adult stage. It was highly expressed in ileum and rectum, moderately expressed in Malpighian tubules, midgut and foregut, and lowly expressed in fat body, ventral ganglion, epidermis and haemocytes in the fourth instars. After continuously ingested double-stranded RNAs originated from two LdATPaseE fragments LdATPaseE1 and LdATPaseE2, the target mRNA levels in the larvae were reduced by 85% and 55%, the larval growth and survival were significantly affected. Furthermore, topical application of fipronil, butane-fipronil, endosulfan and cypermethrin significantly upregulated LdATPaseE expression up to 8.3, 4.2, 2.8 and 6.2-fold 1 day after experiment, and up to 15.8, 3.4, 3.6 and 4.5-fold 2 days after treatment. It seems that depletion of vATPase subunit E is lethal, indicating that targeting vATPases by dsRNA appears a promising means of combating L. decemlineata. Moreover, vATPase subunit E is a pesticide inducible gene and may play a role in pesticide toxicity.
authors.affiliation: Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: fukaiyun000@foxmail.com.
authors.forename: Kai-Yun
authors.initials: KY
authors.lastname: Fu
authors.affiliation: Department of Plant Protection, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China. Electronic address: gwc1966@163.com.
authors.forename: Wen-Chao
authors.initials: WC
authors.lastname: Guo
authors.affiliation: Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: 287148308@qq.com.
authors.forename: Feng-gong
authors.initials: FG
authors.lastname: Lü
authors.affiliation: Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: 853422201@qq.com.
authors.forename: Xin-ping
authors.initials: XP
authors.lastname: Liu
authors.affiliation: Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: ligq@njau.edu.cn.
authors.forename: Guo-qing
authors.initials: GQ
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001517:Coleoptera / Q000201:enzymology* / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D004726:Endosulfan / Q000633:toxicity; D006846:Hydrocarbons, Halogenated / Q000633:toxicity; D019476:Insect Proteins / Q000235:genetics*; D007306:Insecticides / Q000633:toxicity*; D007814:Larva / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D021122:Protein Subunits / Q000235:genetics; D011720:Pyrazoles / Q000633:toxicity; D011722:Pyrethrins / Q000633:toxicity; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics*
chemical_list: C561658:1-(2,6-dichloro-alpha,alpha,alpha-trifluoro-p-tolyl)-5-(2-methylamino)-4-(trifluoromethylsulfinyl)pyrazole-3-carbonitrile; D018076:DNA, Complementary; D006846:Hydrocarbons, Halogenated; D019476:Insect Proteins; D007306:Insecticides; D021122:Protein Subunits; D011720:Pyrazoles; D011722:Pyrethrins; D012333:RNA, Messenger; C017160:cypermethrin; D025262:Vacuolar Proton-Translocating ATPases; D004726:Endosulfan; C082360:fipronil
keywords: Double-stranded RNA; Growth; Insecticide; Leptinotarsa decemlineata; Lethal; Vacuolar-type H(+)-ATPase