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为了探讨草莓中单脱氢抗坏血酸还原酶(Monodehydroascorbate reductase,MDHAR)和谷胱甘肽还原酶(Glutathione reductase,GR)基因的功能,采用同源克隆的方法从‘丰香’草莓中克隆得到c DNA全长序列,并采用实时定量PCR方法对其在不同组织和不同发育阶段果实中的表达模式进行分析。结果表明:草莓MDHAR基因c DNA(Fa MDHAR,Gen Bank登录号:KP025946)全长1 305 bp,编码434个氨基酸,分子量约为47 k D,含有保守的FAD结合结构域,定位于细胞质中。GR基因c DNA(Fa GR,Gen Bank登录号:JQ339738)开放阅读框全长1 491 bp,编码496个氨基酸,分子量为53 k D,定位于细胞质中。Fa MDHAR在各组织中均有表达,在成熟果实中表达量最高,叶和花次之,根中最低;在果实发育过程中Fa MDHAR在绿果期有相对较高的表达,随后急剧增加,到白熟期最高,之后下降并维持在相对稳定水平。Fa GR在叶和花中表达较高,在成熟果实中较低;在果实发育过程中,表达量从小绿期呈现增加趋势,至转红果期达最高,随后逐渐下降,在成熟果实中较低。果实发育过程中酶活性变化呈现出与各自基因表达量相似的变化规律。经过4℃低温处理24 h后的草莓叶片中,Fa MDHAR相对表达量较对照显著增加,而Fa GR无显著变化。草莓中Fa MDHAR和Fa GR表达存在时空差异,并对低温逆境响应存在差异。
In order to investigate the function of Monodehydroascorbate reductase (MDHAR) and Glutathione reductase (GR) genes in strawberry, c DNA was cloned from ’Fengxiang’ strawberry by homologous cloning Full-length sequence, and using real-time quantitative PCR method to analyze its expression patterns in different tissues and different developmental stages of fruit. The results showed that the F MDHAR (GenBank accession no. KP025946) cDNA of strawberry MDHAR gene was 1 305 bp in length and encoded 434 amino acids with a molecular mass of 47 kD and contained a conserved FAD binding domain, which located in the cytoplasm. The full-length open reading frame of GR gene cDNA (Fa GR, Gen Bank accession number: JQ339738) was 1 491 bp, encoding 496 amino acids with a molecular mass of 53 kD and localized in the cytoplasm. Fa MDHAR expression in all tissues, the highest expression in mature fruits, leaves and flowers, the lowest root; Fa MDHAR in fruit development during the relatively high expression, then increased sharply, To the highest white maturity, then decline and maintain a relatively stable level. The expression of Fa GR was higher in leaves and flowers and lower in mature fruits. During the fruit development, the expression level of Fa GR increased from small green stage to highest stage of red fruit stage, then decreased gradually, and lower in mature fruit . During the process of fruit development, the changes of enzyme activity showed similar changes with the expression of their respective genes. Compared with the control, the relative expression level of Fa MDHAR in strawberry leaves after 4 ℃ low temperature treatment for 24 h did not change significantly. The expression of Fa MDHAR and Fa GR in strawberry was spatially and temporally different, and its response to low temperature stress differed.