title: RNAi suppression of the ryanodine receptor gene results in decreased susceptibility to chlorantraniliprole in Colorado potato beetle Leptinotarsa decemlineata.
abstract: Leptinotarsadecemlineata is the most important pest in potato and causes serious yield loss each year. Chlorantraniliprole acts on insect ryanodine receptors (RyRs) and is among the most active compounds against L. decemlineata. Here we cloned and characterized a 15,792-bp full-length LdRyR cDNA that encoded a 5128-amino acid protein. LdRyR shares 85-92% amino acid similarities with other insect RyR homologues, and 59-61% similarities with those from Caenorhabditis elegans and Homo sapiens. All hallmarks of the RyR proteins are conserved in LdRyR. LdRyR has a MIR domain, two RIH domains, three SPRY domains, four copies of RyR domain and a RIH-associated domain in the N-terminus, and it possesses two consensus calcium ion-binding EF-hand motifs and six predicted transmembrane helices in the C-terminus. Temporal, spatial and tissue-specific expression patterns of LdRyR were evaluated. LdRyR expression level was increased constantly from egg to wandering stages, dropped in pupal stage and was increased again in the adult stage. It was widely expressed in the head, thorax and abdomen of day 3 fourth-instar larvae. Moreover, it was ubiquitously expressed in all inspected tissues including epidermis, foregut, midgut, ileum, rectum, fat body, ventral ganglia and Malpighian tubules in day 3 fourth-instar larvae. Dietary introduction of double-stranded RNA of LdRyR significantly reduced the mRNA levels of the target gene in the larvae and adults, respectively, and significantly decreased chlorantraniliprole-induced mortalities. Thus, our results suggested that LdRyR encoded a functional ryanodine receptor in L. decemlineata.
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: pinjunwan@gmail.com.
authors.forename: Pin-Jun
authors.initials: PJ
authors.lastname: Wan
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: guo-weiyan2005@hotmail.com.
authors.forename: Wei-Yan
authors.initials: WY
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: 787226557@qq.com.
authors.forename: Yao
authors.initials: Y
authors.lastname: Yang
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: 2008102089@njau.edu.cn.
authors.forename: Wei-Ping
authors.initials: WP
authors.lastname: Lu
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 / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D005786:Gene Expression Regulation; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D007306:Insecticides / Q000633:toxicity*; D007814:Larva / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D011679:Pupa / Q000187:drug effects / Q000378:metabolism; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism; D019837:Ryanodine Receptor Calcium Release Channel / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014018:Tissue Distribution; D062367:ortho-Aminobenzoates / Q000633:toxicity*
chemical_list: D019476:Insect Proteins; D007306:Insecticides; D012330:RNA, Double-Stranded; D019837:Ryanodine Receptor Calcium Release Channel; D062367:ortho-Aminobenzoates; C517733:chlorantranilipole
keywords: Chlorantraniliprole; Double-stranded RNA; Leptinotarsa decemlineata; Mortality; Ryanodine receptor; mRNA expression