酿酒葡萄‘赤霞珠’VvbHLH93基因的克隆及分析
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国家葡萄产业技术体系(CARS-29-zp-6);陕西省现代农业葡萄产业技术体系建设(NYKJ-2021[XN]25);陕西省葡萄现代化提质增效栽培技术创新团队(No.2020TD-047)


Cloning and Analysis of VvbHLH93 Gene in Cabernet Sauvignon
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    摘要:

    bHLH93转录因子参与调节植物的生长发育、应对各种胁迫等多种生理过程,该研究以酿酒葡萄‘赤霞珠’为试验材料,采用RTPCR方法克隆葡萄VvbHLH93基因全长,并进行生物信息学分析;用qRTPCR法分析bHLH93在不同组织和果实不同发育时期的表达量,为进一步探索VvbHLH93基因的功能及其机制奠定基础。结果表明:(1)成功克隆获得葡萄VvbHLH93,该基因cDNA全长为1 319 bp,开放阅读框长度为939 bp,编码312个氨基酸,相对分子量为35.18 kD,理论等电点为4.68,无信号肽和跨膜域,属于bHLH转录因子家族。(2)葡萄bHLH93蛋白与荷花的亲缘关系较近;VvbHLH93蛋白的C端为酸性结构区,富含丝氨酸、苏氨酸磷酸化位点;VvbHLH93基因的启动子含有光响应元件和低温、干旱、赤霉素等应答元件。(3)qRTPCR分析表明,VvbHLH93在‘赤霞珠’中的表达具有组织特异性,在叶片中基本不表达,在茎中的表达量最高;随着赤霞珠葡萄果实的发育成熟,VvbHLH93相对表达量不断降低,到幼果期(直径 > 2 mm)后第5周开始相对表达量基本为0,总体呈现降低的变化趋势;与对照相比,在低温胁迫、盐胁迫、高温胁迫及充分灌溉胁迫下VvbHLH93表达量显著降低。研究推测, VvbHLH93基因可能是葡萄抗逆胁迫的负转录因子。

    Abstract:

    bHLH93 transcription factor is involved in regulating the plant growth and development and various physiological processes in plants. The VvbHLH93 gene was cloned from Cabernet Sauvignon by RTPCR and analyzed by bioinformatics. The expression of VvbHLH93 in different tissues and fruits at different developmental stages was analyzed by qRTPCR, which laid a foundation for further exploring the function and mechanism of VvbHLH93 gene. The results showed that: (1) VvbHLH93 was cloned successfully. The full length cDNA of VvbHLH93 was 1 319 bp, containing a 939 bp open reading frame which encoded 312 amino acids. The relative molecular weight of VvbHLH93 was 35.18 kDa, and its isoelectric point was 4.68. There was no signal peptide or transmembrane domain in this protein, belonging to the bHLH family. (2) bHLH93 in Vitis vinifera L. was closely related to that in Nelumbo nucifera. VvbHLH93 possesses an acidic region in C terminus rich in serine and threonine putative phosphorylation sites. The promoter of VvbHLH93 gene contained light response element, low temperature, drought and GA response element. (3) qRTqPCR analysis showed that the expression of VvbHLH93 in Cabernet Sauvignon was tissuespecific, it was not expressed in leaves, and had the highest expression in stems. The expression level of VvbHLH93 was decreased continuously with the development and maturity of fruit, and it was basically 0 in the 5th week after young fruit stage (> 2 mm in diameter), showing a unimodal downward trend during seed development. Compared with the control group, the expression of VvbHLH93 was significantly decreased under low temperature stress, salt stress, high temperature stress and full irrigation stress. The study speculated that VvbHLH93 may be a negative transcription factor of grape stress resistance.

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徐学蕾,鞠延仑,王婉妮,等.酿酒葡萄‘赤霞珠’VvbHLH93基因的克隆及分析[J].西北植物学报,2022,42(11):1844-1850

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  • 在线发布日期: 2022-12-06
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