Abstract
Chitin synthase is responsible for chitin synthesis in the cuticles and cuticular linings of other tissues in insects. We cloned two alternative splicing variants of the chitin synthase 1 gene (SfCHS1) from the white-backed planthopper, Sogatella furcifera. The full-length cDNA of the two variants (SfCHS1a and SfCHS1b) consists of 6408 bp, contains a 4719-bp open reading frame encoding 1572 amino acids, and has 5′ and 3′ non-coding regions of 283 and 1406 bp, respectively. The two splicing variants occur at the same position in the cDNA sequence between base pairs 4115 and 4291, and consist of 177 nucleotides that encode 59 amino acids but show 74.6% identity at the amino acid level. Analysis in different developmental stages showed that expression of SfCHS1 and SfCHS1a were highest just after molting, whereas SfCHS1b reached its highest expression level 2 days after molting. Further, SfCHS1 and SfCHS1a were mainly expressed in the integument, whereas SfCHS1b was predominately expressed in the gut and fat body. RNAi-based gene silencing inhibited transcript levels of the corresponding mRNAs in S. furcifera nymphs injected with double-stranded RNA of SfCHS1, SfCHS1a, and SfCHS1b, resulted in malformed phenotypes, and killed most of the treated nymphs. Our results indicate that SfCHS1 may be a potential target gene for RNAi-based S. furcifera control.
Highlights
Chitin synthase is responsible for chitin synthesis in the cuticles and cuticular linings of other tissues in insects
The full-length cDNA sequence of SfCHS1 was obtained by multiple PCR amplifications and RACE
Our results indicated that injecting double-stranded RNA (dsRNA) of CHS1 into S. furcifera nymphs could lead to a significant mortality, suggesting that SfCHS1 may be a candidate gene for use in S. furcifera control
Summary
Chitin synthase is responsible for chitin synthesis in the cuticles and cuticular linings of other tissues in insects. The functions of the CHS genes have been extensively investigated using RNA interference (RNAi) in both holometabolous and hemimetabolous insects such as Tribolium castaneum[29,30], Anthonomus grandis[31], Spodoptera exigua[32], Bactrocera dorsalis[23], Drosophila melanogaster[33,34], Locusta migratoria[16,17], Laodelphax striatellus and N. lugens[19] These results have indicated that CHS genes are essential for survival, ecdysis, fecundity, and egg hatching. We cloned and characterized a full-length cDNA encoding chitin synthase 1 (SfCHS1) from S. furcifera, identified two alternative splicing variants (SfCHS1a and SfCHS1b) of SfCHS1, and analyzed the expression patterns of SfCHS1 and the two alternative variants at different developmental stages and in different tissues. We describe lethal phenotypes of S. furcifera induced by target gene silencing
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