柱花草WRKY转录因子在低磷胁迫下的克隆与分析
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国家自然科学基金(31560665,31160481)


Cloning and Analysis of WRKY Transcription Factors of Stylosanthes guianensis under Low Phosphorus Stress
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    摘要:

    该研究依据生物信息学分析,采用RTPCR方法从格拉姆柱花草[Stylosanthes guianensis (Aubl.)]中克隆出1个WRKY转录因子基因,命名为StWRKY45。该基因最大开放阅读框(ORF)为924 bp,编码307个氨基酸,分子量为35.62 kD,等电点为9.81。系统进化树分析表明,StWRKY45属于WRKY转录因子第Ⅱ类WRKY基因,与拟南芥AtWRKY45、AtWRKY57亲缘关系最近。实时荧光定量PCR结果表明,在非磷胁迫下(对照),StWRKY45基因在柱花草幼苗的根、茎、叶中都有表达,但表达量均相对较低;在低磷胁迫下,StWRKY45基因在格拉姆柱花草根、茎、叶中的表达基本随胁迫时间的延长逐渐升高,且均在叶中的表达量最高;在低磷胁迫96 h时叶、茎中的相对表达量均达到最高,分别是对照的20.47倍、9.38倍,但在低磷胁迫72 h时根中的相对表达量达到最高,为对照的9.29倍。研究表明,StWRKY45基因受低磷胁迫诱导高表达,推测StWRKY45基因可能参与柱花草对低磷胁迫的响应。

    Abstract:

    In this study, based on bioinformatics analysis, a WRKY transcription factor gene was cloned from Stylosanthes guianensis (Aubl.) by RTPCR and named as StWRKY45. The largest open reading frame (ORF) of this gene is 924 bp, encoding 307 amino acids ,with a molecular mass of 35.62 kD and an isoelectric point of 9.81. Phylogenetic tree analysis indicated that StWRKY45 belongs to the WRKY gene of WRKY transcription factor class Ⅱ, and has the closest relationship with Arabidopsis AtWRKY45 and AtWRKY57. The results of Realtime PCR showed that under the nonphosphorus stress (control), StWRKY45 gene was expressed in roots, stems and leaves of S. guianensis seedlings, but the expression level was relatively low; under low phosphorus stress, the expression of StWRKY45 gene in roots, stems and leaves increased gradually with the prolongation of stress time, and the expression level was the highest in leaves. The relative expression of leaves and stems reached the highest at 96 h after low phosphorus stress. They were 20.47 times and 9.38 times of the control, respectively, but the relative expression in roots reached the highest at 72 h after low phosphorus stress, which was 9.29 times of the control. Studies have shown that StWRKY45 gene is induced by lowphosphorus stress, suggesting that StWRKY45 gene may be involved in the response of S. guianensis to low phosphorus stress.

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文亦芾,单贵莲,姜 飞,等.柱花草WRKY转录因子在低磷胁迫下的克隆与分析[J].西北植物学报,2019,39(2):226-233

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  • 在线发布日期: 2019-03-22
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