title: A putative Δ1-pyrroline-5-carboxylate synthetase involved in the biosynthesis of proline and arginine in Leptinotarsa decemlineata.
abstract: Delta 1-pyrroline-5-carboxylate synthetase (P5CS) catalyzes the conversion of glutamate (Glu) to Glu semialdehyde (GSA). GSA spontaneously cyclizes to form P5C. P5C is then reduced to proline (Pro) or is converted to ornithine, the intermediate for arginine (Arg) biosynthesis. In the present study, a full-length Ldp5cs complementary DNA was cloned from the Colorado potato beetle Leptinotarsa decemlineata, a notorious insect defoliator of potato in most potato-growing regions of the world. Ldp5cs encodes a 792-amino-acid protein which shares high identity to homologues from other insect species. Quantitative reverse transcription polymerase chain reaction revealed that Ldp5cs was ubiquitously expressed in the eggs, first to fourth-instar larvae, wandering larvae, pupae and sexually mature adults. In the adults, Ldp5cs mRNA levels were higher in the fat body, foregut, midgut and hindgut, moderate in the ventral ganglion, lower in the thorax muscles, epidermis and Malpighian tubules. Two double-stranded RNAs (dsRNAs) (dsLdp5cs1 and dsLdp5cs2) targeting Ldp5cs were constructed and bacterially expressed. Ingestion during 3 consecutive days of dsLdp5cs1 or dsLdp5cs2 successfully silenced Ldp5cs, significantly reduced the contents of Pro and Arg in the hemolymph, decreased flight speed and shortened flight distance of the resulting adults. Furthermore, knocking down Ldp5cs significantly increased adult mortality. Thus, our results suggest that identified Ldp5cs encodes a functional P5CS enzyme that is involved in the biosynthesis of Pro and Arg 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: 176061610@qq.com.
authors.forename: Kai-Yun
authors.initials: KY
authors.lastname: Fu
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: 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: ligq@njau.edu.cn.
authors.forename: Guo-Qing
authors.initials: GQ
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000097:blood; D000818:Animals; D003001:Cloning, Molecular; D001517:Coleoptera / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D005260:Female; D006458:Hemolymph / Q000737:chemistry; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009953:Ornithine-Oxo-Acid Transaminase / Q000235:genetics* / Q000378:metabolism; D010063:Ovum / Q000201:enzymology / Q000378:metabolism; D010802:Phylogeny; D011679:Pupa / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D016415:Sequence Alignment
chemical_list: D000596:Amino Acids; D019476:Insect Proteins; D009953:Ornithine-Oxo-Acid Transaminase
keywords: Amino acid; Flight; Leptinotarsa decemlineata; Pyrroline-5-carboxylate synthetase; Ribonucleic acid interference