Acta Entomologica Sinica ›› 2016, Vol. 59 ›› Issue (9): 938-947.doi: 10.16380/j.kcxb.2016.09.003

• RESEARCH PAPERS • Previous Articles     Next Articles

Changes in the expression of trehalose-6-phosphate synthase gene in Antheraea pernyi (Lepidoptera: Saturniidae) during pupal diapause termination

HUANG Ling1, SUN Liang-Zhen1, WANG Yong 1,*, RU Yu-Tao1, Muhammad IRFAN1, JIANG Yi-Ren1, SHI Sheng-Lin1, YANG Rui-Sheng1, LI Xi-Sheng2, QIN Li1,*   

  1. (1. Liaoning Engineering and Technology Resource Center for Insect Resource, College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang 110866, China; 2. The Sericultural Research Institute of Liaoning, Fengcheng, Liaoning 118100, China)
  • Online:2016-09-20 Published:2016-09-20

Abstract: 【Aim】 This study aims to clone the trehalose-6-phosphate synthase (TPS) gene in Antheraea pernyi, to analyze its tissue distribution, and to profile the expression patterns of the gene during diapause termination so as to provide data support for clarifying the relationship between carbohydrate metabolism and diapause termination in diapause pupae of A. pernyi. 【Methods】 The TPS gene was cloned from the fat body of A. pernyi larvae using PCR and 3′ RACE technology, and the putative amino acid sequence was analyzed by bioinformatics methods. The expression pattern of the gene in different tissues of  A. pernyi  larvae was analyzed by RT-PCR. The relative expression levels of the gene in haemolymph and fat body in the process of diapause termination of A. pernyi larvae were measured using real-time PCR technology. 【Results】 The TPS gene of A. pernyi was cloned and named ApTPS. It has an open reading frame (ORF) of 2 487 bp, encoding 828 amino acid residues. The encoded protein is estimated to be 93.19 kD with an isoelectric point (pI) about 4.61, and has no signal peptide and transmembrane region. Subcellular localization analysis indicated that ApTPS targets to cytoplasm. Homology search showed that ApTPS has two conserved domains: TPS (22-497 amino acids) and TPP (532-772 amino acids). Tissue specific mRNA expression profiling showed that ApTPS was mainly expressed in the fat body of A. pernyi larvae. In the process of diapause termination, the expression levels of ApTPS in both haemolymph and fat body were up-regulated and significantly higher than those in the control groups (P<0.05). The elevated expression of the gene in the haemolymph, however, occurred later than that in the fat body. 【Conclusion】 The results suggest that ApTPS participates in the regulation of carbohydrate metabolism and plays a significant role in diapause process of  A. pernyi pupae. There is a close relationship between ApTPS expression and the diapause termination of A. pernyi pupae.

Key words: Antheraea pernyi, diapause, trehalose-6-phosphate synthase, gene expression, diapause termination