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澳洲坚果锌指蛋白MiZFP17的亚细胞定位及非生物胁迫下的基因表达分析

Subcellular localization of the Macadamia integrifolia Maiden & Betche zinc finger protein MiZFP17 and gene expression analysis under abiotic stress

  • 摘要: C3H型锌指蛋白基因在植物生长发育及响应非生物胁迫中发挥重要的调控作用。本研究以澳洲坚果(Macadamia integrifolia Maiden & Betche)‘JW’品种为材料,采用RT-PCR技术克隆锌指蛋白基因MiZFP17,对其编码蛋白进行生物信息学和亚细胞定位分析,并应用实时荧光定量PCR检测该基因在不同器官组织中和非生物胁迫下的表达情况。结果显示,MiZFP17含有3个外显子和2个内含子,其启动子含有丰富的与植物激素、转录因子互作、逆境胁迫等相关的作用元件。MiZFP17蛋白属于串联C3H型锌指蛋白,含有卷曲螺旋区域及2个典型的ZnF_C3H结构域。本氏烟草(Nicotiana benthamiana Domin)瞬时表达系统的亚细胞定位显示,MiZFP17蛋白主要定位于细胞核。实时荧光定量PCR分析显示,MiZFP17在澳洲坚果根、茎、叶、小花、小果及果仁中都有表达,其中小果和果仁的表达量最高,显著高于其他器官组织。在低温和干旱胁迫下,MiZFP17在澳洲坚果叶片中的表达受到显著抑制;在高盐、高温和除草剂胁迫下,其表达则受到显著的诱导作用。因此,主要定位于细胞核的MiZFP17在器官组织的生长发育以及应答低温、干旱、高盐、高温、除草剂等非生物胁迫中可能发挥重要的调控作用。

     

    Abstract: C3H-type zinc finger protein genes function as key regulators of plant growth, development, and responses to abiotic stress. In this study, a C3H zinc finger protein gene, MiZFP17, was cloned from macadamia (Macadamia integrifolia Maiden & Betche) cultivar ‘ JW’ using RT-PCR, followed by bioinformatic characterization and subcellular localization. Spatial and stress-responsive expression patterns of MiZFP17 were quantified across macadamia organs and under multiple abiotic stress conditions using quantitative real-time PCR (qRT-PCR). Results showed that MiZFP17 contained three exons and two introns, and promoter analysis identified abundant cis-acting elements associated with phytohormone signaling, transcriptional regulation, and stress responsiveness. Structural features classified MiZFP17 as a tandem C3H zinc finger protein containing a coiled-coil region and two conserved ZnF-C3H domains. Transient expression of tobacco (Nicotiana benthamiana Domin) leaves demonstrated predominant nuclear localization. Expression profiling indicated broad transcription of MiZFP17 in roots, stems, leaves, flowers, small fruits, and kernels, with markedly elevated expression in small fruits and kernels relative to other tissues. Transcript abundance in leaves was significantly inhibited under low-temperature and drought stresses, whereas exposure to high salinity, elevated temperature, and herbicide treatment resulted in pronounced induction. These findings suggest that MiZFP17 encodes a nuclear-localized tandem C3H zinc finger protein with potential regulatory roles in macadamia organ development and adaptive responses to diverse abiotic stresses.

     

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