• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

木薯MebZIP34基因克隆及功能分析

Cloning and Functional of MebZIP34 in Cassava

  • 摘要:
    目的 解析木薯MebZIP34基因功能,探讨其在抗氧化调控中的作用机制,为木薯采后保鲜与抗逆育种提供理论基础。
    方法 以SC8品种木薯块根为材料,克隆MebZIP34编码序列,并开展生物信息学分析,包括蛋白结构、保守结构域、系统进化关系和启动子顺式元件预测。构建植物过表达载体并瞬时转化木薯叶片,检测其对下游抗氧化基因MeMDAR1表达的调控能力;同时获得MebZIP34转基因拟南芥植株,测定其抗氧化相关生理指标。
    结果 成功克隆MebZIP34全长CDS序列(975 bp),编码324个氨基酸,含典型的bZIP保守结构域,蛋白以α螺旋和无规则卷曲结构为主。系统进化分析显示其与橡胶树(Hevea brasiliensis)具有较近亲缘关系。启动子分析揭示其受光照、激素及胁迫等多种顺式元件调控。瞬时表达结果表明MebZIP34显著上调MeMDAR1表达,增强SOD活性,降低MDA含量。转基因拟南芥中也观察到相同的表达与生理变化,表明MebZIP34具有调节植物抗氧化能力。MebZIP34 在有序胚性结构(OES)和茎尖分生组织(SAM)中表达水平较高,而在须根、储藏根和茎等营养器官中表现为中等表达水平。
    结论 MebZIP34是一个具有保守结构和功能特征的bZIP转录因子,能通过调控下游基因表达提升植物抗氧化能力,在木薯采后保鲜及木薯贮藏性育种中具有潜在应用价值。

     

    Abstract:
    Objective Functions in responses to abiotic stress and antioxidant regulation of bZIP34 in Manihot esculenta, an important tropical food crop prone to postharvest shelf life-shortening discoloration, were studied.
    Methods Coding sequence of MebZIP34 was cloned from the storage roots of M. esculenta SC8. Bioinformatics analysis was performed to determine the protein structure, conserved motif, phylogenetic tree, and promoter cis-elements of the gene. An overexpression vector was constructed and transiently expressed in cassava leaves to examine its regulatory effect on the downstream antioxidant gene MeMDAR1. Transgenic Arabidopsis lines with overexpressed MebZIP34 were generated to assess the physiological indicators of antioxidant capacity.
    Results The full-length CDS of MebZIP34 was 975 bp encoded a 324-amino-acid protein containing a typical bZIP domain and the structure consisting primarily of α-helices and random coils. Phylogenetically, it closely related to Hevea brasiliensis. Its promoter expression was regulated by multiple cis-acting elements associated with light, hormonal signaling, and stress responses. Transient expression of the gene significantly upregulated MeMDAR1, enhanced SOD, and reduced MDA. Similar phenomena were observed in the MebZIP34-overexpressed Arabidopsis as well, which confirmed the role of the gene in regulating antioxidation of the plant. It also exhibited relatively high levels of expression in organized embryogenic structures and shoot apical meristem, but moderate in vegetative organs, such as fibrous roots, storage roots, and stems.
    Conclusion A bZIP transcription factor, MebZIP34 had a conserved structure and functions that enhanced the plant antioxidant capacity by regulating the downstream gene expression. It displayed significant potential for applications in cassava post-harvest preservation and breeding stress-resistant varieties with prolonged shelf life.

     

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