黄花棘豆脱水素OoY2K4的鉴定及表达分析
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农业部“十二五”公益性行业(农业)重大科研专项(201203062);


Identification and Expression Analysis of Dehydrated Gene OoY2K4 from Oxytropis ochrocephala Bunge
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    摘要:

    该研究以黄花棘豆cDNA为模板,采用同源克隆法,从黄花棘豆转录组数据库中克隆获得1个响应逆境胁迫的胚胎发育晚期丰富蛋白基因,命名为OoY2K4OoY2K4基因ORF为786 bp,编码261个氨基酸,含有2个保守的Y片段和4个K片段,为典型的Y2K4类脱水蛋白亚家族成员;OoY2K4蛋白不具有跨膜结构域,不存在信号肽,亲水性极强,含有1个糖基化位点和17个磷酸化位点;亚细胞定位显示,OoY2K4蛋白定位于细胞质中。多序列比对发现,OoY2K4蛋白与其他物种第二组LEA蛋白(脱水素)序列高度保守;进化树分析显示,该序列与三叶草、蒺藜苜蓿和紫花苜蓿相似度最高,亲缘关系最近。采用qRTPCR对OoY2K4基因在干旱、高盐、低温以及脱落酸、乙烯、赤霉素处理下的表达分析显示,干旱和高盐胁迫可显著诱导OoY2K4基因表达,而低温胁迫下基本无变化;激素处理均可诱导OoY2K4基因高效表达,其中脱落酸诱导下OoY2K4基因表达最显著。研究推测,OoY2K4基因可能通过依赖ABA的信号途径参与黄花棘豆对干旱和高盐逆境胁迫的应答反应。

    Abstract:

    The late embryogenesis abundant protein gene OoY2K4 that participated in different stress response was isolated from the transcriptome database of Oxytropis ochrocephala Bunge by homologous cloning in this study. OoY2K4 contained an open reading frame(ORF)of 786 bp and encoded a protein of 261 amino acid residues, and was considered to be a member of Y2K4 protein families for containing a conservative 2 Y fragments and 4 K pieces. OoY2K4 protein did not have the transmembrane domains, signal peptide, and had strong hydrophilic, and contained one glycosylation sites and seventeen phosphorylation sites. Subcellular localization prediction and experiment showed that OoY2K4 protein was localized to cytoplasm. Multiple sequence alignment found that OoY2K4 protein had highly conservative LEAⅡ sequence with other species. A homology analysis based on the deduced amino sequence indicated that OoY2K4 protein had a closer relationship with LEAⅡ from Trifolium repens, Medicago truncutula and Medicago sativa than that from other plants. The expression patterns of OoY2K4 under abiotic stress and exogenous hormone treatments were investigated by qRTPCR and the results showed that drought and salt stresses could significantly affect the expression of OoY2K4, while cold stress could not; Hormone treatments could make OoY2K4 express efficiently, in which OoY2K4 gene expression induced by abscisic acid is of most significance. All results above speculated that OoY2K4 gene may be involved in the response to drought and high salt stress by relying on ABA signaling pathway.

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管慧锐,傅艳萍,刘 新,等.黄花棘豆脱水素OoY2K4的鉴定及表达分析[J].西北植物学报,2017,37(10):1910-1918

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  • 在线发布日期: 2017-12-01
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