花生果糖-1,6-二磷酸醛缩酶基因AhFBA1的克隆与表达

Cloning and Expression of Fructose-1,6-Bisphosphate Aldolase Gene AhFBA1 in Peanut(Arachis hypogaea L.)

以花生品种花育33为试材,根据cDNA文库中已知的果糖-1,6-二磷酸醛缩酶(fructose-1,6-bisphosphate aldolase,FBA)基因全长序列设计引物,通过RT-PCR克隆到该基因,命名为AhFBA1。AhFBA1全长为1489 bp,开放阅读框为1200 bp,编码400个氨基酸。预测该基因编码的蛋白含有Glycolytic保守结构域,可能定位于叶绿体中。蛋白序列比对和进化树分析表明,花生与大豆(Glycine max)、苜蓿(Medicago truncatula)、鹰嘴豆(Cicer arietinum)和菜豆(Phaseolus vulgaris)等豆科植物中的FBA序列相似性最高,亲缘关系最近。荧光定量PCR结果显示,在高盐和干旱胁迫下,AhFBA1在花生叶和根中的表达均受明显诱导,说明该基因可能参与花生对高盐和干旱胁迫的适应性调控;AhFBA1在花生根和叶中均受ABA的明显诱导,说明该基因对花生非生物胁迫的调控可能是依赖ABA的。

In this article, a fructose-1,6-bisphosphate aldolase （FBA） gene was cloned from the leaf of peanut （Arachis hypogaea L. cultivar Huayu 33） using RT-PCR, and was designated as AhFBA1. The whole sequence of AhFBA1 is 1489 bp and its open reading frame is 1200 bp, encoding a polypeptide of 400 amino acids. Its protein was predicted to be located in chloroplast, con- taining the conserved glycolytic domain. Multiple sequence alignments and phylogenetic analysis of FBA proteins indicated Ah- FBA1 was most similar with FBA from Glycine max, Medicago truncatula, Cicer arietinum, and Phaseolus vulgaris. The results of Real-time RT-PCR showed that the expression of AhFBA1 was induced distinctly in both peanut root and leaf under salt and drought conditions, suggesting that AhFBA1 may participate in the salt and drought stress regulation of peanut. The expression of AhFBA1 was also induced by exogenous ABA in both peanut leaf and root, which indicated that AhFBA1 may regulate peanut abiotic stresses resistance through ABA-dependent pathway.