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核农学报  2011 

玉米海藻糖含量测定及其合成酶(TPS)基因序列分析

, PP. 1107-1116

Keywords: 玉米,海藻糖,高效液相色谱,海藻糖-6-磷酸合成酶,序列分析

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Abstract:

自然界生物体内广泛存在着海藻糖。海藻糖不仅能在正常情况下为生物体新陈代谢提供能量贮备,更能在干旱、高热、低温等逆境胁迫下通过大量积累来帮助生物体抵御逆境伤害。因此,对海藻糖积累及其合成途径关键酶类的研究已越来越受到人们关注。本研究以垫状卷柏(Selaginellapulvinata(Hook.etGrev)Maxim.)为对照,利用高效液相色谱法(HPLC)检测了玉米自交系在不同干旱条件下的叶片海藻糖含量,结果显示,在重度、中度和轻度干旱胁迫下,玉米中均未检测到海藻糖,而正常状态下的垫状卷柏海藻糖含量达10.31mg/g干重。另外,本研究结合电子克隆对玉米自交系18-599白的海藻糖-6-磷酸合成酶(trehalose-phosphatesynthase,TPS)基因进行了序列扩增,并用RT-PCR进行验证,得到1条全长3055bp的cDNA全序列,命名为ZmTPS,GenBank登录号为EU659122。通过对不同物种TPS氨基酸序列进行比对分析,发现在玉米中存在许多重要的保守位点,但同其他TPS活性低的植物一样,缺失了部分保守区,推测这可能是玉米海藻糖含量太低以至于HPLC未能检测出的原因。

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