2024年12月2日 星期一
薄荷McZFP1基因克隆及表达分析
Gene cloning and expression analysis on McZFP1 in Mentha canadensis
2024年 第33卷 第1期 页码[35-46]    下载全文[9.1MB]  
摘要

根据前期茉莉酸甲酯处理的薄荷(Mentha canadensis Linn.)转录组数据分析结果,从薄荷中克隆到McZFP1,并进行了基因和蛋白质特性、组织表达、非生物胁迫处理下的表达、亚细胞定位以及转录自激活活性分析。结果显示:薄荷McZFP1的开放阅读框长度为558 bp,编码185个氨基酸。McZFP1具亲水性,无跨膜结构域,属于不稳定蛋白,包含C2H2保守结构域,含有13个丝氨酸、3个酪氨酸和3个苏氨酸的磷酸化位点。进化树分析结果表明McZFP1与一串红(Salvia splendens KerGawler)SsZFP7的亲缘关系最近。实时荧光定量PCR结果表明:McZFP1在薄荷不定根、茎、根状茎、幼叶、成熟叶和花中均有表达,在根状茎中的相对表达量最高,且受150 mmol·L-1 NaCl、300 mmol·L-1甘露醇、200 μmol·L-1脱落酸和200 μmol·L-1茉莉酸甲酯处理诱导表达。McZFP1定位于细胞核,无转录自激活活性。综上所述,薄荷McZFP1在不同组织中存在表达差异,参与调节激素和非生物胁迫响应过程。

Abstract

Based on the previous analysis result of transcriptome data of Mentha canadensis Linn. treated by methyl jasmonate, McZFP1 was cloned from M. canadensis, and analyses on gene and protein characteristics, tissue expression, expression under abiotic stress treatments, subcellular localization, and transcriptional self-activating activity were performed. The results show that the open reading frame. of McZFP1 in M. canadensis is 558 bp in length, encoding 185 amino acids. McZFP1 is hydrophilic, has no transmembrane domain, is of unstable protein, contains a C2H2 conserved domain, and possesses phosphorylation sites of 13 serine, 3 tyrosine, and 3 threonine. The phylogenetic tree analysis result shows that McZFP1 is most closely related to SsZFP7 in Salvia splendens Ker-Gawler. The result of real-time fluorescent quantitative PCR shows that McZFP1 is expressed in adventitious root, stem, rhizome, young leaf, mature leaf, and flower of M. canadensis, its relative expression is the highest in rhizome, and its expression is induced by 150 mmol·L-1 NaCl, 300 mmol·L-1 mannitol, 200 μmol·L-1 abscisic acid, and 200 μmol·L-1 methyl jasmonate treatments. McZFP1 is localized in cell nucleus, and has no transcriptional self-activating activity. In conclusion, there are differences in expression of McZFP1 among different tissues of M. canadensis, and McZFP1 is involved in regulation of hormone and abiotic stress response processes.

关键词薄荷; 锌指蛋白(ZFP)转录因子; 生物信息学; 亚细胞定位; 表达
Key wordsMentha canadensis Linn.; zinc finger protein (ZFP) transcription factor; bioinformatics; subcellular localization; expression
作者郑晓薇, 柏杨, 亓希武, 于盱, 房海灵, 李莉, 刘冬梅, 梁呈元
所在单位江苏省中国科学院植物研究所(南京中山植物园) 江苏省植物资源研究与利用重点实验室, 江苏 南京 210014
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基金项目国家自然科学基金项目(31970353; 32100313); 江苏省自然科学基金项目(BK20210164); 江苏省植物资源研究与利用重点实验室开放基金项目(JSPKLB202029)