作物学报2018，Vol.44Issue(6)：814-823,10.DOI:10.3724/SP.J.1006.2018.00814
小麦近缘属植物1-FFT基因的克隆及功能分析
Molecular Cloning and Functional Analysis of 1-FFT in Wheat Relatives
摘要
Abstract
Fructan is closely related to carbon distribution and the response to adverse stresses in plant, protecting plant cell membrane stabilization in stress conditions. To determine the effects of fructan:fructan-1-fructosyltransferase genes from different species, we cloned three genes of full-length cDNA from Psathyrostachys huashanica (2n = 2x = 14, NsNs), Dasypyrum villosum (2n = 2x = 14, VV) and Leymus racemosus (2n = 4x = 28, NsNsXmXm), designated Ph-1-FFT, Dv-1-FFT, and Lr-1-FFT, respectively, using reverse transcriptase PCR (RT-PCR) and rapid-amplification of cDNA ends (RACE) techniques. The full-length open reading frames (ORF) are 1989, 1950, and 1989 bp in length, encoding 662, 649, and 662 amino acids, respectively. The predicted protein contains a conserved fructosyltransferase domain. Multiple sequences alignment and phylogenetic analysis indicated that the Dv-1-FFT shared a high similarity with 1-FFT from Triticum aestivum, T. turgidum subsp. durum, Aegilops searsii and T. urartu, Ph-1-FFT and Lr-1-FFT were clustered into the same evolutionary branch, which was different from those of Dv-1-FFT. Furthermore, the plant expression vector pCAMBIA1300-35SN-Ph-1-FFT/Dv-1-FFT/Lr-1-FFT was constructed and transferred into tobacco (Nicotiana tabacum) cv. W38 via Agrobacterium-mediated transformation. The validation with PCR and RT-PCR assay showed that drought and cold tolerance was obviously improved in all types of transgenic tobacco compared with the wild type, and no significant difference between transgenic genotypes. Under drought and cold stresses, the contents of carbohydrate and proline were significantly higher in the transgenic tobacco lines than in the wild type, whereas, malondialdehyde content was significantly lower in the transgenic lines, which was no significant difference between the transgenic plants with 1-FFT from different species. These results suggest that Ph-1-FFT, Dv-1-FFT, and Lr-1-FFT are typical members of the gene family coding glycoside hydrolase 32 (GH32) and play a role in drought and cold resistance in tobacco.关键词
1-FFT基因/抗旱性/抗寒性/小麦近缘属植物/转基因烟草Key words
1-FFT gene/drought tolerance/cold tolerance/wheat relatives/transgenic tobacco引用本文复制引用
王建伟,贺晓岚,李文旭,陈新宏..小麦近缘属植物1-FFT基因的克隆及功能分析[J].作物学报,2018,44(6):814-823,10.基金项目
本研究由凯里学院博士专项基金(BS201606)和贵州省教育厅科技拔尖人才支持计划项目(黔教合KY字[2017]094)资助. This study was supported by the Special Fund for the Doctoral Program of Kaili University (BS201606) and the Science and Technology Top Talents Support Program of Guizhou Provincial Education Department (Guizhou Education KY [2017]094). （BS201606）
