利用mRNA差异显示技术筛选线果芥抗旱相关基因

doi:10.6048/j.issn.1001-4330.2017.10.002

摘要/Abstract

摘要： 【目的】十字花科短命植物线果芥在新疆干旱环境下生长，具有较强的抗旱性，研究十字花科短命植物线果芥的抗旱机制。为进一步了解线果芥的抗旱机理和从野生植物资源中发掘一些优异的抗逆基因奠定基础。【方法】用20% PEG6000处理3~4周的线果芥幼苗，分别提取0、3、6、9、12、24及48 h的线果芥叶片总RNA。用mRNA差异显示技术筛选差异表达的基因。【结果】使用80对引物组合共筛选获得差异片段18个，分别命名为DF1-DF18，其中表达上调的有12个，下调的有6个。按照基因的功能可以划分为6大类:基础代谢、转录因子、抗病相关、假想蛋白、未知蛋白和光周期蛋白。其中以基础代谢相关基因最多。对其中的DF-2、DF-6及DF-14进行高盐和干旱胁迫下的表达模式分析，这三个基因均受干旱和盐调控表达。【结论】从短命植物线果芥中筛选了18个抗旱相关的基因，并对其中的3个基因进行表达模式分析，的确受干旱和盐胁迫的诱导表达。

关键词: 线果芥; 抗旱; mRNA差异显示

Abstract: 【Objective】 The ephemeral species Conringia planisiliqua belongs to Brassicaceae which grows in Xinjiang arid environment, which has strong drought resistance, but its drought resistance mechanism is not clear. In order to understand the mechanism of drought resistance and discover some excellent resistance genes from wild plant resources, it has laid an important foundation for further understanding the mechanism of drought resistance in conringia planisiliqua.【Method】Using the plant that has grown for three to four weeks treated by 20% PEG6000, total RNA of the leaves at 0 h, 3 h, 6 h, 9 h, 12 h, 24 h and 48 h was extracted. Then the differentially expressed genes were screened by mRNA differential display techniques.【Result】Eighty primer combinations were used to screen the differentially expressed cDNAs, and a total of 18 differentially expressed cDNAs were found. They were named of DF1-DF18, 12 of the cDNAs were the up-regulated and 6 of them were down-regulated gene fragments. According to the gene function they were divided into 6 categories: basal metabolism, transcription factor, disease resistance protein, hypothetical protein, unknown protein and photoperiod protein. Most of them were the basal metabolism related genes. We analyzed the expression pattern of the DF-2,DF-6 and DF-14 under drought and salt stress, and the finding showed that the three genes were regulated by drought and salt.【Conclusion】Eighteen of the drought-resistant genes were screened from ephemeral plant Conringia planisiliqua. Three of the genes expression patterns were detected, they were induced by drought and salt stress.

Key words: Conringia planisiliqua; drought stress; mRNA different display

中图分类号:

S188

朱燕飞, 陈全家, 曲延英. 利用mRNA差异显示技术筛选线果芥抗旱相关基因[J]. 新疆农业科学, 2017, 54(10): 1775-1784.

ZHU Yan-fei, CHEN Quan-jia, QU Yan-ying. Application of mRNA Differential Display Technique in Screening Drought Resistance Related Genes in C. planisiliqua[J]. Xinjiang Agricultural Sciences, 2017, 54(10): 1775-1784.

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