Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (20): 4272-4283.doi: 10.3864/j.issn.0578-1752.2013.20.010

• PLANT PROTECTION • Previous Articles     Next Articles

Cloning and Functional Analysis of pdp1 in Ostrinia furnacalis (Lepidoptera: Crambidae)

 CHENG  Xiao-Juan-12, YAN  Shan-Chun-1, HUANG  Yong-Ping-2, TAN  An-Jiang-2   

  1. 1.State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040
    2.Research Center for Insect Sciences, Institute of Plant Physiology and Ecology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences/Key Laboratory of Insect Developmental and Evolutionary Biology, Chinese Academy of Sciences, Shanghai 200032
  • Received:2013-04-07 Online:2013-10-15 Published:2013-05-17

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Abstract: 【Objective】The objectives of this study are to clone pdp1 (protease-activated receptor-domain protein1 gene) which named Ofpdp1 (GenBank accession number: KC857457) from the Asian corn borer, Ostrinia furnacalis (Guenée), and to identify its typical hallmarks, then to analyze the mRNA expression characteristics and physiological function of Ofpdp1. It will provide potential target gene for genetic control of O. furnacalis.【Method】 The cDNA sequence of Ofpdp1 was isolated using RT-PCR and PCR methods. Based on the sequencing results, the bioinformatics analysis of nucleic acid and putative amino acid was conducted. The developmental expression pattern of Ofpdp1 was determined by qPCR. RNAi experiment was performed at proembryo stage to explore the physiological function. 【Result】The cDNA sequence (960 bp) of OfPDP1 was obtained, including a complete open reading frame (ORF) of 804 bp, which encoded 267 amino acids. OfPDP1 was a transcription factor of PAR bZIP (basic leucine zipper) subfamily which included several conserved regions in the carboxy terminal region of the protein. The result of phylogenetic analysis was consistent with the classification of the species in biology. The qPCR results showed a high level expression at the 76 h of eggs’ development process. It could be declined by 71% in the transcript levels of Ofpdp1 in the eggs injected with Ofpdp1-specific dsRNA in 48 h compared to those in the control eggs. 【Conclusion】 A complete ORF of Ofpdp1 was sequenced with clone strategy from Asian corn borer for the first time. Ofpdp1 was highly conserved in the process of evolution, and it belonged to PAR bZIP subfamily. The high levels expression of Ofpdp1 at the time of 72 h suggested that Ofpdp1could take an important part in the changing process of embryo physiological structure during the larva’s forming period.

Key words: Ostrinia furnacalis , ofpdp1 , gene cloning , phylogenetic analysis , mRNA expression , RNA interference

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