大赖草6-SFT基因的克隆及其转基因烟草抗旱和抗寒性分析

发布时间：2018-06-05 21:15

本文选题：-SFT基因 + 干旱　；参考：《作物学报》2016年03期

【摘要】：果聚糖合成酶(6-SFT)在植物抵御逆境胁迫中起重要作用。利用RACE结合RT-PCR技术从大赖草(Leymus racemosus,2n=4x=28,NsNsXmXm)中克隆到6-SFT基因,其完整开放阅读框为1863 bp。该基因被命名为Lr-6-SFT(GenBank登录号KT387273),编码620个氨基酸,其推导氨基酸序列含有保守的果糖基转移酶结构域。氨基酸序列比对及进化树分析表明,大赖草6-SFT与华山新麦草、普通小麦、西尔斯山羊草和大麦6-SFT具有高度相似性。采用基因重组技术构建p1300-35SN-Lr-6-SFT表达载体,利用农杆菌介导法将该载体转入烟草品种W38中。对经过抗性筛选、PCR和RT-PCR验证的转基因植株进行抗旱和抗寒性鉴定,发现转基因植株与对照相比,抗旱和抗寒性明显增强;在逆境胁迫条件下,转基因植株的果聚糖、可溶性糖、脯氨酸含量都显著高于对照,而丙二醛的含量显著低于对照。本研究表明,Lr-6-SFT基因是典型的GH32家族成员,其表达能够提高烟草对干旱和寒冷胁迫的抗性。
[Abstract]:Fructan synthase 6-SFT plays an important role in plant resistance to stress. The 6-SFT gene was cloned from Leymus racemosus 2nC4xC28NsNsXmXm by RACE and RT-PCR technique. The complete open reading frame was 1863 BP. The gene was named Lr-6-SFT(GenBank accession number KT387273, encoding 620 amino acids. The deduced amino acid sequence contains conserved fructosyltransferase domain. Amino acid sequence alignment and phylogenetic tree analysis showed that Leymus chinensis 6-SFT had a high similarity with wheat, Sears and barley 6-SFT. P1300-35SN-Lr-6-SFT expression vector was constructed by gene recombination technique and transferred into tobacco variety W38 by Agrobacterium tumefaciens. The drought resistance and cold resistance of transgenic plants verified by RT-PCR and resistance screening showed that the drought resistance and cold resistance of transgenic plants were significantly higher than those of control, and the fructan and soluble sugar of transgenic plants were increased under stress. The content of proline was significantly higher than that of the control, but the content of malondialdehyde was significantly lower than that of the control. This study suggests that Lr-6-SFT gene is a typical member of GH32 family, and its expression can enhance the resistance of tobacco to drought and cold stress.
【作者单位】：西北农林科技大学农学院/陕西省植物遗传工程育种重点实验室;凯里学院环境与生命科学学院;河南省农业科学院小麦研究所;
【基金】：国家自然科学基金项目(31571650) 西北农林科技大学唐仲英育种基金资助~~
【分类号】：Q943.2

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