番茄LeGlyⅠ基因的生物信息学特征与表达模式分析

Analysis on Bioinformatics Characteristics and Expression Pattern of LeGlyⅠGene in Tomato

植物在遭受逆境胁迫时会产生过量的甲基乙二醛(MG)等有毒醛类化合物,而乙二醛酶系统能帮助其有效清除过量的MG,其中乙二醛酶Ⅰ(GlyⅠ)是级联反应的第一个酶,在此过程中发挥了重要作用。本研究以野生番茄‘Heinz 1706’的幼苗为试材,克隆了番茄LeGlyⅠ基因,并对其生物信息学特征、表达模式以及亚细胞定位进行了分析。结果表明,LeGlyⅠ的CDS序列全长为489 bp,共编码162个氨基酸;LeGlyⅠ蛋白分子式为C_(798)H_(1226)N_(214)O_(235)S_(4),分子量为17 706.05 Da,等电点为6.51,亲水性平均值为-0.386,不稳定系数为39.32,预测为亲水性稳定蛋白,含有一个VOC-like结构域。LeGlyⅠ的启动子序列中包含多种顺式作用元件,包括无氧诱导必需的顺式作用元件、光响应元件、低温响应元件、干旱响应元件以及茉莉酸甲酯、脱落酸、生长素、赤霉素等激素响应元件。通过qRT-PCR分析发现,LeGlyⅠ基因在番茄的根、茎、叶中均有表达,且在叶中的表达量最高,并且其表达水平受氧化胁迫的影响。亚细胞定位结果显示,LeGlyⅠ蛋白主要在线粒体和细胞核中发挥功能。本研究结果可为深入研究LeGlyⅠ基因在番茄抗逆中的功能及其分子机制提供理论依据。

The plant may produce excessive toxic aldehydes such as methylglyoxal(MG)when facing adversity stress,and glyoxal enzyme system can effectively clear the excessive MG.Glyoxalase I(GlyⅠ)as the first enzyme in the cascade reaction plays an important role.In this study,LeGlyⅠgene was successfully cloned from the seedlings of tomato‘Heinz 1706’,and its bioinformatics characteristics,expression pattern and subcellular localization were analyzed.The results showed that the total length of LeGlyⅠCDS sequence was 489 bp,which encoding 162 amino acids.The LeGlyⅠwas predicted to be a hydrophilic stable protein with molecular formula as C_(798)H_(1226)N_(214)O_(235)S_(4),molecular weight as 17706.05 Da,isoelectric point as 6.51,GRAVY coefficient as-0.386 and instability index as 39.32,and it had a VOC⁃like domain.The promoter se⁃quence of LeGlyⅠcontained a variety of cis⁃acting elements,including cis⁃acting elements necessary for anae⁃robic induction,light response elements,low⁃temperature response elements,drought response elements,and hormone response elements such as methyl jasmonate,abscisic acid,auxin and gibberellin.Through qRT⁃PCR analysis,it was found that LeGlyⅠexpressed in roots,stems and leaves of tomato with the highest expression level in leaves,and the expression could be affected by oxidative stress.The subcellular localization results showed that LeGlyⅠprotein was mainly localized in mitochondria and nucleus.These results could provide theoretical bases for further research on the function and molecular mechanism ofLeGlyⅠ gene in tomato resistantto stresses.