香蕉MaPFK基因家族鉴定、MaPFK3克隆和表达

Identification of MaPFK gene family and MaPFK3 cloning and expression in bananas

为研究香蕉中MaPFK基因的功能和生物学特性,采用生物信息学分析法对香蕉A基因组的MaPFK基因家族成员进行鉴定、蛋白特性分析、分子进化树分析、FPKM值分析,同时以‘天宝蕉’为材料,通过RT-PCR技术克隆MaPFK3基因,进行生物信息学分析;采用qRT-PCR技术进行低温胁迫下的表达分析.结果表明,MaPFK家族包含12个成员,分子进化树分析可分为3类,FPKM值分析表明MaPFK成员在13、4、0℃不同低温胁迫下有不同程度的上调或下调表达.采用RT-PCR技术克隆了MaPFK3,其基因编码区长1 617 bp,预测编码538个氨基酸,MaPFK3编码蛋白属于PFK超家族,是不稳定的酸性亲水蛋白,无信号肽,亚细胞预测定位于细胞质;MaPFK3启动子顺式作用元件预测分析结果显示,MaPFK3启动子含有光响应、激素响应以及与逆境胁迫相关的作用元件.qRT-PCR分析结果显示,香蕉MaPFK3基因的表达具有组织差异性,且表达量为叶片>假茎>根;低温胁迫引起‘天宝蕉’中MaPFK3基因下调表达,在13、4、0℃不同低温胁迫下,MaPFK3的表达量为13℃<4℃<0℃.本研究表明MaPFK基因家族成员能在香蕉应对低温胁迫的过程中发挥着重要作用.(图11表2参32)

To further study the characteristics and biological function of the MaPFK gene family in bananas, the MaPFK gene family members were identified in the whole A genome of Musa acuminata. The protein characteristics, phylogenetic tree, and FPKM values of the MaPFK gene family were analyzed using bioinformatics methods. RT-PCR was used to clone MaPFK3 from the ‘Tianbaojiao’ banana using RT-PCR, and qRT-PCR was used to detect the expression of MaPFK3 under low temperature stress. The results showed that the MaPFK gene family contained 12 members. The phylogenetic tree indicated that the MaPFK gene family could be divided into three groups. FPKM values showed that MaPFK members are upregulated or downregulated to varying degrees under 13 ℃, 14 ℃, and 0 ℃, under different low temperature stresses. The coding sequence of MaPFK3 was 1617 bp in length and was predicted to encode 538 amino acids. The encoded protein of MaPFK3 belongs to the PFK superfamily. It is an unstable acidic hydrophilic protein without a signal peptide and has transmembrane structures. Subcellular localization prediction results showed that MaPFK3 was located in the cytoplasm. The MaPFK3 promoter cis-acting element prediction analysis results showed that the promoter contains light response, hormone response, and stress-related elements. The results of qRT-PCR analysis showed that the expression of the MaPFK3 gene was tissue-differentiated(leaves > pseudostems > roots), downregulated under low temperature stress, and the expression level of MaPFK3 was 13 ℃ < 4 ℃ < 0 ℃. This study shows that members of the MaPFK gene family can respond to low temperature stress in bananas.