植物学报2017，Vol.52Issue(5)：568-578,11.DOI:10.11983/CBB16225
白菜型冬油菜β-1,3-葡聚糖酶基因在低温胁迫下的表达
Expression Analysis of β-1,3-Glucanase Gene from Winter Brassica rapa Under Low Temperature Stress
摘要
Abstract
This study examined the function of β-1,3-glucanase in Brassica rapa with low temperature stress,whose encoding protein was identified by protein mass spectrometry analysis.The cDNA sequence of β-1,3-glucanase was cloned by RT-PCR of winter rapeseed from Longyou6 and Tianyou4 cultivars.Semi-quantitative PCR and real-time fluorescence quantitative PCR were used to study the expression pattern of β-1,3-glucanase in response to low temperature stress.The open reading frame of the β-1,3-glucanase gene was obtained at a length of 1 032 bp,and encoded 343 amino acid.The molecular weight was 38.102 kDa and the isoelectric point 6.63,with 93.94％ amino sequence similarity to B.rapa subsp.chinensis and B.napus.The protein encoded by this gene is a hydrophilic protein with one signal peptide and two transmembrane domains.The prediction of the secondary structures indicated that β-1,3-glucanase is a protein with more g-helices.The predicted β-1,3-glucanase contains a conserved amino acid sequence corresponding to the plant Glycol-Hydro 17 superfamily.RT-PCR and semi-quantitative RT-PCR results showed that β-1,3-cglucanase was upregulated in response to 4℃,and the gene was upregulated to peak in expression at-4℃;However,the expression was inhibited at lower temperature (-8℃).The β-1,3-glucanase gene cloned from the winter B.rapa cv.‘Longyou6’ might play a role in cold tolerance.关键词
β-1,3-葡聚糖酶基因/低温/分子克隆/表达分析Key words
β-1,3-glucanase gene/low temperature/molecular cloning/expression analysis引用本文复制引用
马骊,董小云,何辉立,孙万仓,袁金海,刘自刚,武军艳,方彦,许耀照,蒲媛媛,白静..白菜型冬油菜β-1,3-葡聚糖酶基因在低温胁迫下的表达[J].植物学报,2017,52(5):568-578,11.基金项目
国家现代农业产业技术体系(No.CARS-13)、国家自然科学基金(No.31460356,No.31560397,No.3166040,No.31660404)、国家重点基础研究发展计划(No.2015CB150206)、科技部油菜杂种优势利用技术与强优势杂交种创制(No.2016YFD0101300)和甘肃省教育厅科研专项(No.2015A-077) （No.CARS-13）
