白菜型冬油菜类甜蛋白的筛选、克隆及其在低温胁迫下的表达
Selection and Cloning of Thaumatin-like Protein (TLP) Gene from Winter Brassica rapa and Its Expression under Low Temperature Stress
摘要
Abstract
Thaumatin-like protein (TLP) is an essential protein induced by low temperature stress and enhances plant resistance to adverse circumstances. In our study, we selected differential protein spots from Longyou 7 leaves under low temperature stress and separated TLP closely related to cold resistance by using two-dimensional electrophoresis and mass spectrometry. Using the primers ofTLP cDNA sequences and reverse transcription PCR (RT-PCR), we cloned the complete open reading frame ofTLP gene of winterBrassica rapa cultivar Longyou 7. The sequence length ofTLP fromB. rapa was 732 bp, encoding a protein of 243 amino acid residues with a predicted molecular weight of 26.02 kD and a theoretical pI of 9.02. Bioinformatics analysis showed a similarity of 99.18% in TLP amino acid sequence betweenB. rapa andBrassica napus, the predicted TLP contained a conserved amino acid sequence belonging to the plant GH69-TLP-SF superfamily. The TLP had one signal peptide which belongs to hydrophilic protein. Moreover, the detection of subcellular localization suggested the synthetic process of this protein is in the endoplasmic reticulum. The prediction of the second structures indicated that TLP is a labile protein consisting of random coils and extended strands. The expression analysis of TLP gene by utilizing real time and Semi-quantitative RT-PCR suggested that the TLP expression could be up-regulated in response to lower temperature. In conclusion,TLP gene of the winterB. rapa cultivar Longyou 7 might play a significant role in response to cold stress.关键词
白菜型冬油菜/双向电泳/低温/类甜蛋白Key words
WinterBrassica rapa/Two-dimensional gel electrophoresis/Lower temperature/TLP引用本文复制引用
马骊,许耀照,蒲媛媛,刘海卿,杨刚,刘林波,袁金海,孙万仓,刘自刚,曾秀存,武军艳,方彦,李学才,陈奇..白菜型冬油菜类甜蛋白的筛选、克隆及其在低温胁迫下的表达[J].作物学报,2017,43(4):620-628,9.基金项目
本研究由国家现代农业产业技术体系建设专项(CARS-13),国家自然科学基金项目(31460356, 31560397),国家重点基础研究发展计划项目(973计划) (2015CB150206),国家农业科技成果转化项目(2014G10000317),甘肃省自然科学基金项目(145RJZG050),科技部油菜杂种优势利用技术与强优势杂交种创制项目(2016YFD0101300)和丝绸之路经济带新疆核心区农田周年覆盖高效种植模式研究与示范项目(2016E02009)资助. This study was supported by the China Agriculture Research System (CARS-13), the National Natural Science Foundation of China (31460356, 31560397), the National Basic Research Program of China (973 Program) (2015CB150206), the Natural Science Foundation of Gansu Province (145RJZG050), the part of National Key Research and Development Program (2016YFD0101300), and the Research and Demonstration Project of High Efficient Planting Pattern in the Core Area of the Silk Road Economic Zone in Xinjiang (2016E02009). （CARS-13）
