title: The putative Halloween gene phantom involved in ecdysteroidogenesis in the white-backed planthopper Sogatella furcifera.
abstract: Postembryonic development of insects is highly dependent on ecdysteroid hormones ecdysone (E) and 20-hydroxyecdysone (20E). A cytochrome P450 monooxygenase CYP306A1, the product of the Halloween gene phantom (phm), is involved in the ecdysteroidogenesis in representative insects in Diptera, Lepidoptera and Orthoptera. In the present paper, Sfphm was cloned from a hemipteran insect species, the white-backed planthopper Sogatella furcifera. SfPHM has five insect conserved P450 motifs, i.e., Helix-C, Helix-I, Helix-K, PERF and heme-binding motifs. Temporal and spatial expression patterns of Sfphm were evaluated by q-PCR. Sfphm showed three expression peaks in late second-, third- and fourth-instar stages. In contrast, the expression levels were lower and formed three troughs in the newly-molted second-, third- and fourth-instar nymphs. The relative 20E levels exhibited similar temporal patterns to Sfphm expression levels. On day 3 of the fourth-instar nymphs, Sfphm clearly had a high transcript level in the thorax where PGs were located. Dietary introduction of double-stranded RNA of Sfphm into the second instars successfully knocked down the target gene, and greatly reduced 20E level and ecdysone receptor (EcR) expression level. Moreover, knockdown of Sfphm caused lethality and slowed down ecdysis during nymphal stages. Furthermore, ingestion of 20-hydroxyecdysone did not alter Sfphm expression level, but almost completely rescued SfEcR expression level, and relieved the negative effects on nymphal survival and ecdysis in Sfphm-dsRNA-exposed planthoppers. Thus, our results suggest that Sfphm plays a critical role in ecdysteroidogenesis in S. furcifera.
authors.affiliation: Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
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.
authors.forename: Shuang
authors.initials: S
authors.lastname: Jia
authors.affiliation: Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Na
authors.initials: N
authors.lastname: Li
authors.affiliation: Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Jin-Mei
authors.initials: JM
authors.lastname: Fan
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: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins / Q000378:metabolism; D026461:Ecdysteroids / Q000378:metabolism*; D004441:Ecdysterone / Q000378:metabolism / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D006430:Hemiptera / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism*; D007814:Larva / Q000235:genetics; D008675:Metamorphosis, Biological / Q000235:genetics; D006899:Mixed Function Oxygenases / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D012330:RNA, Double-Stranded / Q000494:pharmacology; D011987:Receptors, Steroid / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D026461:Ecdysteroids; D019476:Insect Proteins; D012330:RNA, Double-Stranded; D011987:Receptors, Steroid; C055469:ecdysone receptor; D004441:Ecdysterone; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C496261:Phm protein, Drosophila
keywords: Double-stranded RNA; Ecdysteroidogenesis; Performance; Phantom; Sogatella furcifera