title: RNA interference of a putative S-adenosyl-L-homocysteine hydrolase gene affects larval performance in Leptinotarsa decemlineata (Say).
abstract: In Leptinotarsa decemlineata, juvenile hormones (JHs) play primary roles in the regulation of metamorphosis, reproduction and diapause. In JH biosynthetic pathway in insect corpora allata, methylation of farnesoic acid or JH acid using S-adenosyl-L-methionine generates a potent feedback inhibitor S-adenosyl-L-homocysteine (AdoHcy). Rapid removal of AdoHcy is hypothesized to be essential for JH synthesis. AdoHcy hydrolase (SAHase) is the only eukaryotic enzyme catalyzing the removal. In the present paper, we firstly cloned a putative LdSAHase gene from L. decemlineata. The cDNA consists of 1806 bp and encodes a 525 amino acid protein. LdSAHase was expressed in all developmental stages. The gene had the highest and the lowest level of transcription respectively in the 3rd- and 4th-instars' heads that contain corpora allata, which was positively correlated with JH titer in the haemolymph and the mRNA level of a JH early-inducible gene, the Krüppel homolog 1 gene (Kr-h1). Secondly, dietary ingestion of bacterially-expressed LdSAHase-dsRNA significantly decreased LdSAHase and LdKr-h1 mRNA levels, reduced JH titer, and caused the death of the larvae, and the failure of pupation and adult emergence. After continuous exposure for 12 days, 42% of the larvae died, 65% of the prepupae failed to pupate and 100% of the pupae failed to emerge. Moreover, RNAi-mediated LdSAHase knockdown also reduced larval developing time, and decreased larval weight. Lastly, application of JH analogue pyriproxyfen to LdSAHase-dsRNA-exposed larvae did not greatly increase LdSAHase expression level and JH content, but up-regulated LdKr-h1 mRNA level. Expectedly, pyriproxyfen application could partially rescue the negative effects on the survival and the development. Thus, our results support the hypothesis that SAHase plays a critical role in JH biosynthesis in insects.
authors: Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
authors: Li-Tao
authors: LT
authors: Zhou
authors: Shuang
authors: S
authors: Jia
authors: Pin-Jun
authors: PJ
authors: Wan
authors: Ye
authors: Y
authors: Kong
authors: Wen-Chao
authors: WC
authors: Guo
authors: Tursun
authors: T
authors: Ahmat
authors: Guo-Qing
authors: GQ
authors: Li
mesh_terms: D043383:Adenosylhomocysteinase / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001517:Coleoptera / Q000201:enzymology* / Q000235:genetics; D005260:Female; D017344:Genes, Insect; D019476:Insect Proteins / Q000378:metabolism; D007605:Juvenile Hormones / Q000096:biosynthesis; D007814:Larva; D008297:Male; D008675:Metamorphosis, Biological; D011725:Pyridines; D034622:RNA Interference
chemical_list: D019476:Insect Proteins; D007605:Juvenile Hormones; D011725:Pyridines; C055613:pyriproxyfen; D043383:Adenosylhomocysteinase
keywords: Development; Double-stranded RNA; Leptinotarsa decemlineata; Pyriproxyfen; S-adenosyl-L-homocysteine hydrolase; Survival