大麦气孔发育转录因子基因HvMUTE的克隆及生物学功能初步鉴定

Gene Cloning and Preliminary Function Identification of HvMUTE Transcription Factor for Barley Stomatal Development

为了对大麦气孔发育转录因子基因HvMUTE的功能进行初步鉴定,并对其生物学特性及表达模式进行分析,通过生物信息学分析、PCR扩增及qRT-PCR等方法来对大麦气孔发育转录因子基因HvMUTE进行研究。由于大麦G1614为大麦MOREX品种的姊妹系,且大麦MOREX品种公布了参考基因组数据,所以通过将短穗二柄草BdMUTE基因同大麦G1614基因组进行序列同源比对,得到大麦HvMUTE基因序列。从大麦材料G1614幼芽中的第1片叶片中取样,克隆得到大麦HvMUTE基因651 bp的编码区。生物信息学分析结果表明,HvMUTE蛋白是位于细胞核中的无明显亲水疏水性的不稳定蛋白质,并且其蛋白质三维结构含有螺旋-环-螺旋(HLH)结构。系统进化树分析结果表明,HvMUTE蛋白同小麦和山羊草的MUTE-Like蛋白亲缘关系较近。qRT-PCR结果显示,在干旱条件下,HvMUTE基因表达量无明显变化,这一结果同前人研究有所不同,猜测与大麦为单子叶植物,气孔结构中存在副卫细胞同双子叶植物气孔结构形态不同有关。

In order to preliminarily identify the function of HvMUTE, a transcription factor gene for stomatal development in barley, and by analyzing its biological characteristics and expression patterns, HvMUTE, a transcription factor gene for stomatal development in barley, was studied by bioinformatics analysis, PCR amplification and qRT-PCR. Because barley G1614 is the sister line of barley MOREX, and the reference genome data of barley MOREX has been published. So HvMUTE gene sequence was obtained by homology Blast of BdMUTE against between the referencre genome of barely G1614. The coding region of the HvMUTE gene with 651 bp in length was isolated from the first leaf emerging from the bud of barley G1614. Bioinformatics analysis indicated that HvMUTE protein was an unstable protein with no obvious hydrophilicity and hydrophobicity, located in the nucleus, and hold a Helix-Loop-Helix(HLH) structure in three demision level. Phylogenetic tree analysis showed that HvMUTE was closely related to MUTEs of wheat and Aegilops. qRT-PCR results showed that there was no significant change in the expression of HvMUTE gene under drought conditions. This result was different from previous studies, and it was speculated that barley was a monocotyledonous plant, and the existence of parathyroid cells in the stomatal structure was different from that of dicotyledonous plants.