摘要
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(Selaginella pulvinata(Hook.et Grev)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-phosphate synthase,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。
The trehalose exists abroad in natural organisms.It is not only providing energy reservior for metabolism in general condition,but also resisting hurt from adversity stress such as drought,heat,cold and so on by accumulation excessively.So,studying on the trehalose accumulating mechanism and key enzyme in its synthesis approach has been giving more and more attentions.The leaf trehalose contents of maize inbred lines under different drought treatments were tested by high performance liquid chromatography（HPLC） in this study,Selaginella pulvinata（Hook.et Grev） Maxim with in better water.The results showed that trehalose was not tested in all samples of maize,and 10.31mg/g drought weight in Selaginella pulvinata（Hook.et Grev） Maxim was measured.All 3055bp cDNA sequence of trehalose-phosphate synthase gene（TPS） was cloned from maize inbred line ＂18-599 Bai＂ by combining electronic cloning method and RT-PCR.This sequence named as ZmTPS and logging in GenBank as EU659122.Comparing and analysing with other species＇ amino acid sequences of TPS,some of conservative sequences was discoverred in maize TPS,but existed partial conservative sequences,which was same as other plant species with low activity of TPS enzyme.It is estimated that the differences between TPS of maize and other species may result in too low content trehalose in maize to be tested by HPLC.
出处
《核农学报》
CAS
CSCD
北大核心
2011年第6期1107-1116,共10页
Journal of Nuclear Agricultural Sciences
基金
国家自然科学基金(31071433)(30971795)
抗逆转基因专项(2008ZX08003-004)
