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目的从白木香Aquilaria sinensis中克隆sHSP1和sHSP2基因,并对它们的生物信息学及表达模式进行分析。方法根据本课题组白木香转录组数据库进行分析获得2条具有小分子热激蛋白(sHSPs)保守结构域的sHSPs基因序列,利用RT-PCR技术克隆sHSP1和sHSP2基因的全长c DNA序列,并对其进行生物信息学分析;利用实时荧光定量PCR检测sHSP1和sHSP2基因在不同组织和高盐及外源ABA、SA、MJ处理下不同时间的表达差异。结果克隆得到的白木香sHSP1和sHSP2基因开放阅读框都为474 bp,编码157个氨基酸。组织表达分析的结果显示,sHSP1和sHSP2基因主要在根中表达,在茎和叶中的表达量较少;白木香愈伤组织在高盐处理下,sHSP1和sHSP2基因分别在36 h和24 h达到最高表达水平,而愈伤组织在外源ABA、MJ处理下,sHSP1和sHSP2基因都在12 h达到最高表达水平,在SA处理下,sHSP1和sHSP2基因分别在12、24 h达到最高表达水平。结论获得了sHSP1和sHSP2 2基因全长c DNA序列,且2个基因在不同组织根、茎、叶中的表达存在差异性,在受到高盐和外源ABA、SA、MJ处理时,在不同时间的愈伤组织中的表达及积累情况也不同,该研究为后续深入研究白木香防御反应奠定了基础。
Objective To clone sHSP1 and sHSP2 genes from Aquilaria sinensis and analyze their bioinformatics and expression patterns. Methods Two sHSPs gene sequences with conserved domains of small molecular heat-shock proteins (sHSPs) were obtained from the transcriptome database of this group, and the full-length c DNA sequences of sHSP1 and sHSP2 genes were cloned by RT-PCR. Bioinformatics analysis was carried out. The expression differences of sHSP1 and sHSP2 genes in different tissues, high salt and exogenous ABA, SA and MJ were detected by real-time fluorescence quantitative PCR. Results The cloned sHSP1 and sHSP2 genes were 474 bp in open reading frame and encoded 157 amino acids. The results of tissue expression analysis showed that sHSP1 and sHSP2 genes were mainly expressed in roots and less in stems and leaves. Under high salt treatment, sHSP1 and sHSP2 genes were expressed at 36 and 24 h Reached the highest expression levels. However, the expression of sHSP1 and sHSP2 reached the highest levels at 12 h after treatment with exogenous ABA and MJ, respectively. Under SA treatment, the expression levels of sHSP1 and sHSP2 reached their highest levels at 12 and 24 h, respectively. Conclusion The full length c DNA sequences of sHSP1 and sHSP2 2 genes were obtained, and the expression of the two genes in different tissues of root, stem and leaf was different. When treated with high salt and exogenous ABA, SA and MJ, The time of callus expression and accumulation are also different, the study for the follow-up study of the white wood defensive response laid the foundation.