蔗茅EfNAC44基因克隆与表达分析

Cloning and Expression Analysis of EfNAC44 Gene in Erianthus fulvus

为了克隆蔗茅EfNAC44基因,并对其进行生物信息学和基因表达模式分析。本研究通过转录组测序和RT-PCR方法相结合首次从蔗茅野生种无性系‘蔗茅99-3’中克隆到一个受低温胁迫响应的NAC家族基因,命名为EfNAC44 (GenBank登录号:MH709118.1),该基因包含一个591个碱基的开放阅读框,编码196个氨基酸。利用ClustalX软件将EfNAC44编码蛋白与其他物种NAC蛋白进行同源性比较分析,发现其与高粱(XP002465328.1)同源性最高达80%。荧光定量PCR检测分析表明,该基因在蔗茅苗期能够对低温胁迫响应,低温胁迫时的相对表达量上调达到20倍。以上研究结果可为进一步挖掘利用蔗茅耐寒相关基因以及研究甘蔗抗逆性品种改良提供理论依据和技术支撑。

In order to clone the EfNAC44 gene from Erianthus fulvus, bioinformatics and gene expression pattern analysis were carried out. The study used transcriptome sequencing and RT-PCR to clone a NAC family gene,named EfNAC44(GenBank No.: MH709118.1), which is responsive to low temperature stress, from a wild Erianthus fulvus clone ’Zhemao 99-3’. The gene contains a 591-base open reading frame encoding 196 amino acids.ClustalX software was used to compare the homology between EfNAC44 and NAC proteins of other species, and it was found that EfNAC44 had the highest homology with sorghum(XP002465328.1), up to 80%. Fluorescence quantitative PCR analysis showed that the gene could respond to low temperature stress at the seedling stage, and its relative expression level was up-regulated by 20 times. The above research results could provide the theoretical basis and technical support for the further exploitation of the related genes of Erianthus fulvus and the improvement of sugarcane resistance varieties.