作物学报2016，Vol.42Issue(3)：389-398,10.DOI:10.3724/SP.J.1006.2016.00389
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
Cloning of 6-SFT Gene from Leymus racemosus and Analysis of Tolerance to Drought and Cold Stresses in Transgenic Tobacco
摘要
Abstract
The fructan biosynthesis enzyme (6-SFT) plays an important role in plant in response to abiotic stresses. In this study, a full-length cDNA encoding sucrose:fructan-6-fructosyltransferase, designated as Lr-6-SFT (GenBank accession No. KT387273), was cloned from Leymus racemosus (2n=4x=28, NsNsXmXm) using reverse transcriptase PCR (RT-PCR) and rapid-amplifica-tion of cDNA ends (RACE) techniques. The full-length open reading frame comprises 1863 bp and encodes 620 amino acids. The predicted protein structure of the gene contains a conserved fructosyltransferase domain. Multiple sequence alignment and phy-logenetic analysis showed that the Lr-6-SFT protein shared high similarity with 6-SFT proteins from Psathyrostachys huashanica, Triticum aestivum, Aegilops searsii, and Hordeum vulgare subsp. vulgare. The Lr-6-SFT gene was transferred into tobacco (Nico-tiana tabacum L. cv. W38) via Agrobacterium-mediated transformation. The screened plants were tested by PCR and RT-PCR, and the transgenic tobacco plants exhibited much higher tolerance to drought and cold compared with the non-transgenic plants. Under drought and cold stresses, the Lr-6-SFT expressions were associated with the increased accumulation of stored carbohy-drate and proline and the decreased malondialdehyde storage. These results suggest that Lr-6-SFT is a typical member of the gly-coside hydrolase 32 (GH32) family and may be linked to enhanced tolerance to drought and cold stresses.关键词
6-SFT基因/干旱/寒冷/大赖草/生理指标/转基因烟草Key words
6-SFT gene/Drought/Freezing/Leymus racemosus/Physiological indices/Transgenic tobacco引用本文复制引用
贺晓岚,王建伟,李文旭,陈真真,赵继新,武军,王中华,陈新宏..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[J].作物学报,2016,42(3):389-398,10.基金项目
本研究由国家自然科学基金项目(31571650)和西北农林科技大学唐仲英育种基金资助。This study was supported by the National Natural Science Foundation of China (31571650) and the Tang Zhong-Ying Breeding Fund of the Northwest A&F University （31571650）
