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MOLECULAR CLONING AND CHARACTERIZATION OF A SID‐1‐LIKE GENE IN Plutella xylostella
Author(s) -
Wang Huidong,
Gong Liang,
Qi Jiangwei,
Hu Meiying,
Zhong Guohua,
Gong Liang
Publication year - 2014
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.21189
Subject(s) - rna interference , biology , plutella , rna silencing , complementary dna , open reading frame , midgut , hemolymph , microbiology and biotechnology , gene , messenger rna , gene expression , caenorhabditis elegans , rna , bombyx mori , genetics , biochemistry , peptide sequence , botany , lepidoptera genitalia , larva
RNA interference (RNAi) signal can spread from the point where the double‐stranded RNA (dsRNA) was initially applied to other cells or tissues. SID‐related genes in Caenorhabditis elegans help in the spreading of this signal. However, the mechanisms of systemic RNAi are still not unveiled in insects. In this study, we cloned a full‐length cDNA of sid‐1‐like gene, Pxylsid‐1, from Plutella xylostella that contains 1,047 bp opening reading frame encoding a putative protein of 348 amino acids. This transcript is very much similar to the sil‐1 in Bombyx mori (68.8%). The higher expression levels of Pxylsid‐1 were found at the adult and fourth‐instar stages compared to the second‐instar stage with 21.48‐ and 10.36‐fold increase, respectively. Its expression levels in different tissues were confirmed with the highest expression in the hemolymph, which showed 21.09‐fold increase than the midgut; however it was lower in other tissues. The result of RNAi by feeding bacterially expressed dsRNA targeting Pxylace‐1 , which showed that the mRNA level of Pxylace‐1 decreased by 34.52 and 64.04% after 36‐ and 72‐h treatment, respectively. However, the mRNA level of Pxylsid‐1 was not significantly induced when the Pxylace‐1 was downregulated. Furthermore, we found that downregulation of Pxylsid‐1 did not affect the RNAi effect of Pxylace‐1 . Hence, the Pxylsid‐1 may not be involved in absorption of dsRNA from the midgut fluid. A further study is needed to uncover the function of Pxylsid‐1.