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大青杨PuZFP103基因的序列特征及逆境胁迫的表达分析
张馨, 马苗苗, 吕婉秋, LEE Joobin, 杨静莉
南京林业大学学报（自然科学版） ›› 2021, Vol. 45 ›› Issue (1) : 36-44.
PDF(2355 KB)
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大青杨PuZFP103基因的序列特征及逆境胁迫的表达分析
Sequence characteristics and expression pattern analyses of PuZFP103 gene under abiotic stress in Populus ussuriensis
【目的】通过研究大青杨(Populus ussuriensis)PuZFP103基因的序列特征及其在不同组织部位、激素处理与胁迫应答中的表达特性,为揭示PuZFP103在大青杨生长发育中的功能提供依据。【方法】利用各种生物信息学方法对PuZFP103基因进行分析,采用实时荧光定量 PCR(real time quantitative polymerase chain reaction, RT-qpCR)技术分析PuZFP103基因在不同组织、激素处理与胁迫应答中的表达模式,并用基因枪轰击法进行亚细胞定位分析。【结果】生物信息学分析发现,PuZFP103 CDS序列全长504 bp,编码168个氨基酸。分析得出PuZFP103蛋白含有ZnF-C2H2结构域,其二级结构中α螺旋、无规则卷曲和β-折叠分别占24%、84%和1%。进化树构建图显示,其与同属毛果杨、胡杨、毛白杨及番木瓜聚类到一起,亲缘关系最近。RT-qPCR分析结果显示,NaCl、脱落酸(abscisic acid, ABA)和PEG 6000胁迫处理后PuZFP103基因在大青杨叶和根中的表达量在大多数时间内相对于未处理都有所提高。另外,PuZFP103基因定位于细胞核中。【结论】PuZFP103基因在NaCl、ABA和PEG 6000胁迫处理后呈正调控的应答模式,可能参与了大青杨对渗透胁迫的代谢途径或信号传导,属于逆境胁迫下的调控因子。本研究初步证明大青杨PuZFP103基因与胁迫响应相关,但是关于其信号传导与调控机制需进一步研究。
【Objective】 This study aimed to investigate the function of the PuZFP103 gene in the growth and development process of Populus ussuriensis Kom. We analyzed the sequence characteristics, expression patterns in different tissues, and the PuZFP103 gene response to hormones and abiotic stresses. 【Method】 The sequence characteristics of the PuZFP103 gene were analyzed using bioinformatics methods. The expression characteristics of the PuZFP103 gene in different tissues, as well as its response to hormones and abiotic stresses, were analyzed by real-time quantitative PCR (RT-qPCR). In addition, the subcellular localization of the PuZFP103 gene was detected using particle bombardment. 【Result】 The bioinformatic analysis showed that the PuZFP103 gene had 505 bp coding sequences, which encoded a polypeptide of 168 amino acids. The conserved domain analysis revealed that there was a conserved ZnF-C2H2 domain in the PuZFP103 protein. The secondary structure analysis of PuZFP103 showed that the percentage of amino acids of α-helix, random coil, and β-sheet were 24%, 84% and 1%, respectively. The evolutionary tree map showed that it was closely related to the common P. ussuriensis, P. euphratica, P. tomentosa, and Carica papaya. The results of RT-qPCR analysis demonstrated that the expression level of PuZFP103 was increased after NaCl, ABA and PEG 6000 treatment. The subcellular localization assay showed that PuZFP103 was a nuclear protein. 【Conclusion】 The results indicate that the gene is positively regulated by NaCl, ABA and PEG 6000. The PuZFP103 gene may be involved in the metabolic and signaling pathways of osmotic and salt stress and, thus, is a regulatory factor under stress. This study provides preliminarily evidence that the PuZFP103 gene is associated with stress response, but further studies on its signal transduction and regulation mechanism are needed. This is the first study to perform bioinformatics analysis of the PuZFP103 gene in P. ussuriensis.
大青杨 / PuZFP103基因 / 逆境胁迫 / 生物信息学 / RT-qPCR
Populus ussuriensis / PuZFP103 gene / abiotic stress / bioinformation / RT-qPCR
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