Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (22): 4469-4482.doi: 10.3864/j.issn.0578-1752.2015.22.008

• PLANT PROTECTION • Previous Articles     Next Articles

Identification of Ecdysone Synthesis Pathway Genes and Analysis on the Impact of Food Deprivation on Larvae Development of Bactrocera dorsalis Hendel

CONG Lin1, 2, JIANG Xuan-zhao1, YANG Wen-jia1, 3, XU Kang-kang1, 3, DOU Wei1, RAN Chun2, WANG Jin-jun1   

  1. 1College of Plant Protection, Southwest University, Chongqing 400715
    2Citrus Research Institute, Southwest University, Chongqing 400712
    3College of Biology and Environmental Engineering, Guiyang University, Guiyang 550005
  • Received:2015-06-18 Online:2015-11-16 Published:2015-11-16

Abstract: 【Objective】 As one of the most important endohormones, ecdysone plays an important role in regulating the development of insects and responding to environmental factors. The objective of this study is to investigate the expression profiles of ecdysone synthesis pathway genes in Bactrocera dorsalis Hendel in various tissues and different conditions, providing theoretical knowledge to understand the mechanisms of metamorphosis and the stress response. 【Method】RT-PCR and RACE technologies were applied to clone the ecdysone synthesis pathway genes BdCyp302a1, BdCyp315a1 and BdCyp314a1. And quantitative PCR (qPCR) was carried to evaluate the mRNA expression patterns of ecdysone synthesis pathway genes BdNvd, BdCyp306a1, BdCyp302a1, BdCyp315a1 and BdCyp314a1 at different larval developmental stages (1- to 8-day-old larvae), tissues (prothoracic gland mixture, fat body, midgut, Malpighian tubes, integument and trachea), and starvation condition. Further, the impacts of food deprivation on larvae development were also examined in this research.【Result】The open reading frames (ORF) of BdCyp302a1 (GenBank accession number: JQ027284), BdCyp315a1 (GenBank accession number: KC515377) and BdCyp314a1 (GenBank accession number: JQ229645) were finally obtained. The analysis indicated that the protein sequences were highly conserved, which harbored typical and P450 motifs, such as Helix-C/I/K, PERF motifs, heme-binding domains and prolin/glycine rich domain. The results of qPCR showed that the expression levels of BdNvd, BdCyp306a1 and BdCyp314a1 were stable in the first four days at the larval stage, but significantly elevated in the last two days, which were up-regulated for 7.33, 10.89 and 7.82 folds comparing to the lowest level, respectively. And the expression levels BdCyp302a1 and BdCyp315a1 were stable at the larval stage. The expression profiles showed that all of the ecdysone synthesis pathway genes were traceable in the selected tissues. The highest relative expression levels of BdNvd, BdCyp306a1 and BdCyp315a1 were observed in the prothoracic glands, and no significant differences among other tissues. The expression levels of BdCyp302a1 followed the precedence order: fat body > prothoracic glands > integument/Malpighian tubes > trachea/midgut. The expression level of BdCyp302a1 in the fat body was 30 folds greater than that in midgut. And the expression abundances of BdCyp314a1 were extremely higher in the midgut, Malpighian tubes and fat body than in other tissues. Starvation could shorten the larval duration by prepupation. The larvae began to pupate in 6 h after food deprivation and the pupation rate increased with time extended. And the measurements indicated that the length and width of pupa were affected by starvation, which were significantly decreased in the treated group. However, no significant difference was detected in the survival rate. The expression levels of BdNvd, BdCyp302a1 and BdCyp314a1 were significantly elevated at 6 h post food deprivation, and down-regulated at 48 h. However, the expression levels of BdCyp315a1 were not relevant to nutrition condition.【Conclusion】Ecdysone synthesis pathway genes were transcribed differently in larval tissues, and played a vital role in mediating larva-pupa metamorphosis and nutrition stress.

Key words: Bactrocera dorsalis, ecdysone synthesis pathway genes, food deprivation, larvae development, gene expression

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