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苦荞转录因子FtNAC11的克隆及其功能分析
杨迪1, 张凯旋2, 赵辉2, 伍小方2, 杨克理2, 周美亮2, 张金林1
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(1.兰州大学草地农业科技学院/草地农业生态系统国家重点实验室;2.中国农业科学院作物科学研究所)
摘要:
NAC(NAM, ATAF1/2, CUC2)转录因子在植物响应非生物胁中发挥着重要的作用。本研究以苦荞品苦1号为材料,从中克隆得到FtNAC11基因。其cDNA全长为774 bp,编码257个氨基酸,分子量为 28954.58 Da,等电点为5.91。氨基酸序列比对和系统进化树分析表明,FtNAC11与拟南芥AtNAC61有较高的同源性。qRT-PCR显示FtNAC11在苦荞中的表达受NaCl和PEG的诱导。转录激活活性分析表明,FtNAC11的C末端具有转录激活活性。亚细胞定位显示,该基因定位于细胞核中。利用根癌农杆菌介导法将FtNAC11基因转入拟南芥,过表达FtNAC11基因显著提高了拟南芥植株的耐盐性和抗旱性。以上结果表明,FtNAC11参与苦荞非生物胁迫适应过程,为进一步研究FtNAC11调控苦荞抗旱和耐盐的分子机制奠定了前期工作基础。
关键词:  苦荞  NAC11  克隆  功能分析  抗旱  耐盐
DOI:10.13430/j.cnki.jpgr.20210218004
投稿时间:2021-02-18修订日期:2021-04-16
基金项目:国家重点研发计划(2019YFD1001300,2019Y FD1001302)
Molecular Cloning and Functional Analysis of FtNAC11 from Tartary Buckwheat (Fagopyrum tataricum)
YANG Di1, ZHANG Kai-xuan2, ZHAO Hui2, WU Xiao-fang2, YANG Ke-li2, ZHOU Mei-liang2, ZHANG Jin-lin1
(1.College of Pastoral Agriculture Science and Technology, Lanzhou University/State Key Laboratory of Grassland Agro-ecosystems;2.Institute of Crop Sciences, Chinese Academy of Agricultural Sciences)
Abstract:
NAC (NAM, ATAF1/2, CUC2) gene plays an important role in responses to plant abiotic stresses. In this study, we cloned FtNAC11 from tartary buckwheat cultivar ‘Pinku No.1’. Sequence analysis showed that the full-length coding sequence of FtNAC11 was 774bp encoding for 257 amino acids residues, with a molecular weight of 8954.58 Da and an isoelectric point of 5.91.The sequence alignment and phylogenetic tree analysis showed that FtNAC11 had higher homology with Arabidopsis thaliana AtNAC61. The expression of FtNAC11 gene was induced under PEG or NaCl treatments. Sub-localization by transient expression and yeast transactivation assay indicated that FtNAC11 encoded for a nuclear transcription activator with an activation domain at the region of C-terminal. Moreover, the coding sequence of FtNAC11 was transformed into Arabidopsis thaliana by Agrobacterium-mediated transformation, and the transgenic plants over-expressing this gene resulted in significant enhancement under drought and salt tolerance treatments. Collectively, these results indicated that FtNAC11 is likely involved in abiotic stress adaptation, providing a basis for further investigation of its molecular mechanism responding to drought and salt tolerance in tartary buckwheat.
Key words:  tartary buckwheat  NAC11  cloning  functional analysis  drought tolerance  salt tolerance

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