梨小食心虫滞育与非滞育幼虫抑制性消减文库的构建与分析

Construction and analysis of suppression subtractive hybridization cDNA library about the diapause and non-diapause larvae of Grapholita molesta(Busck)

【目的】分离和鉴定梨小食心虫(Grapholita molesta(Busck))滞育相关基因。【方法】分别以梨小食心虫滞育和非滞育幼虫为材料,通过抑制性消减杂交技术(SSH),构建梨小食心虫滞育与非滞育正、反向差减cDNA文库,从中筛选梨小食心虫滞育相关基因,并对其进行测序和分析。【结果】在获得的128个滞育特异和132个非滞育特异的EST中,有滞育差异表达EST 42条(17条功能未知)、非滞育差异表达EST 46条(22条功能未知)。对差异表达EST同源检索后推测,其功能大部分与滞育或非滞育特性相关。在滞育个体中,代谢酶和滞育关联蛋白基因表达量较高;在非滞育个体中,贮存蛋白和信号传递基因表达量较高。【结论】这些EST基本反映了梨小食心虫在滞育与非滞育期间的基因表达谱,为今后深入研究梨小食心虫滞育的分子机理奠定了基础。

【Objective】 This research was to obtain the diapause-related genes from Grapholita molesta（Busck）,and then to get the genes sequenced and analyzed.【Method】 Two suppression subtractive hybridization cDNA libraries were constructed with diapause and non-diapause G.molesta as positive and reverse library.【Result】 128 fragments from the positive library and 132 fragments from the reverse library were respectively selected for sequencing.42 ESTs with diapause difference and 46 ESTs with non-diapause difference were obtained finally,after cluster analysis and ordering repitition sequences.【Conclusion】 Most of the genes screened from two libraries are related to diapause and basically reflect gene expression profiles of G.molesta in duration of diapause and non-diapause by BLASTN.The study establishes a foundation for further research of the molecular mechanism of diapauses of G.molesta.