茶树中2个NAC转录因子基因的克隆及温度胁迫的响应
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国家自然科学基金(31200520);江苏省自然科学基金(BK2012774)


Isolation and Expression Profiles of NAC Transcription Factor Genes under Temperature Stress in Camellia sinensis
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    摘要:

    利用茶树转录组数据库,检索得到2个NAC家族转录因子基因CsNAC1和CsNAC2。通过RT-PCR方法,将其从茶树‘迎霜’中分离克隆,利用荧光定量PCR方法,对CsNAC1和CsNAC2基因在‘迎霜’和‘安吉白茶’2个茶树品种不同组织以及温度胁迫处理下的表达进行分析,以探讨NAC家族转录因子在温度胁迫下的响应特征。结果表明:(1)CsNAC1和CsNAC2基因开放阅读框长度分别为1 044和1 047 bp,分别编码347个和348个氨基酸;蛋白功能域预测和多重对比显示,CsNAC1和CsNAC2蛋白N端均含有典型NAC家族成员所具有的NAM保守结构域。(2)进化分析表明,CsNAC1和CsNAC2分别属于NAC家族的NAP和AtNAC3亚家族。(3)三维分子模型建模显示,CsNAC1和CsNAC2蛋白分别含有3个和2个α-螺旋,6个和7个β-折叠。(4)荧光定量PCR结果显示,CsNAC1在2个茶树品种中具有较相似的组织特异性,均在茶树成熟叶中表达量最高;CsNAC2则分别在‘安吉白茶’的幼叶中,‘迎霜’的根中表达量最高;高温(38 ℃)和低温(4 ℃)处理下,CsNAC1和CsNAC2基因的表达均受不同温度胁迫影响,不同茶树品种、不同时间段的表达存在差异。

    Abstract:

    In this study,the CsNAC1 and CsNAC2 genes,which encode the NAC transcription factors,were cloned from Camellia sinensis cultivar ‘Yingshuang’ by RT-PCR based on the tea transcriptome database.(1)Sequence analysis indicated that the open reading frame of CsNAC1 is 1 044 bp,which encodes 347 amino acids;and the CsNAC2 gene is 1 047 bp,which encodes 348 amino acids.Domain prediction and multiple alignments of CsNAC1 and CsNAC2 showed that N-terminals of the proteins contain typical NAM conservative structure which belongs to NAC family.(2)Phylogenetic analysis showed that the CsNAC1 and CsNAC2 belong to NAP and AtNAC3 subgroups of NAC family,respectively.(3)Molecular modeling showed that the CsNAC1 and CsNAC2 proteins contain 3 and 2 α-helix,6 and 7 β-strands,respectively.(4)Expression profiles of CsNAC1 and CsNAC2 genes in different tissues of two tea cultivars ‘Yingshuang’ and ‘Anjibaicha’ and under different temperature stresses were detected by quantitative real-time PCR.Results showed that the expression profiles of CsNAC1 gene between two tea cultivars exhibit similar tissue specificity and both the highest expression levels were found in mature leaves;while CsNAC2 gene showed the highest expression levels in the tender leaf of ‘Anjibaicha’ and root of ‘Yingshuang’,respectively.Under high (38 ℃) and low (4 ℃) temperature treatments,the expression profiles of CsNAC1 and CsNAC2 genes were different between the two tea cultivars and processing time,and were also influenced by high and low temperature stress treatments.

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王永鑫,刘志薇,吴致君,等.茶树中2个NAC转录因子基因的克隆及温度胁迫的响应[J].西北植物学报,2015,35(11):2148-2156

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  • 在线发布日期: 2015-12-07
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