白菜型冬油菜β-1,3-葡聚糖酶基因在低温胁迫下的表达

发布时间：2018-11-07 12:24

【摘要】：为探明β-1,3-葡聚糖酶基因(β-1,3-glucanase)对油菜(Brassica campestris)抵御低温胁迫能力的作用,通过蛋白质谱分析得到β-1,3-葡聚糖酶蛋白,采用RT-PCR技术克隆白菜型冬油菜(B.rapa)陇油6号和天油4号β-1,3-葡聚糖酶的c DNA序列;并对该序列进行生物信息学分析;进而采用实时荧光定量PCR及半定量PCR检测β-1,3-葡聚糖酶基因在低温胁迫下的表达模式。结果获得长度为1 032 bp的陇油6号β-1,3-葡聚糖酶基因开放阅读框,编码343个氨基酸,相对分子量为38.102k Da,理论等电点为6.63,其与菜心(B.rapa subsp.chinensis)和甘蓝型油菜(B.napus)的蛋白质氨基酸序列同源性高达93.94%。该基因编码的酶是一个主要由α-螺旋组成的亲水性稳定蛋白,含有1个信号肽,存在2个跨膜结构域。该基因在进化上高度保守,其保守序列属于植物的糖基水解酶家族17特有的保守结构域。β-1,3-葡聚糖酶基因表达模式分析显示,4°C时该基因上调表达,继续低温( 4°C)胁迫处理,该基因上调表达至峰值,至 8°C时其表达下调。研究表明从白菜型冬油菜中克隆的β-1,3-glucanase在冬油菜品种陇油6号抗寒过程中发挥作用。
[Abstract]:In order to find out the effect of 尾 -1n 3-glucanase gene on the ability of rape (Brassica campestris) to resist low temperature stress, 尾 -1N 3-glucanase protein was obtained by protein spectrum analysis. RT-PCR technique was used to clone the c DNA sequences of Longyou 6 and Tianyu 4 尾 -1 glucanase from Brassica campestris (B.rapa). The sequence was analyzed by bioinformatics, and the expression pattern of 尾 -1m3-glucanase gene under low temperature stress was detected by real-time fluorescence quantitative PCR and semi-quantitative PCR. Results A 1 032 bp open reading frame of Longyou 6 尾 -1Glucanase gene was obtained, encoding 343 amino acids, with a relative molecular weight of 38.102 k Da, and a theoretical isoelectric point of 6.63. Its amino acid sequence homology with B.rapa subsp.chinensis and B.napus is 93.94. The enzyme encoded by this gene is a hydrophilic stable protein mainly composed of 伪 -helix, which contains a signal peptide and two transmembrane domains. This gene is highly conserved in evolution and belongs to the conserved domain of the glycosylhydrolase family 17 of plants. The expression pattern of 尾-1 and 3-glucanase gene was up-regulated at 4 掳C. The gene was up-regulated to a peak at 8 掳C and down-regulated at 8 掳C after continued hypothermia (4 掳C) stress. The results showed that 尾 -1N 3-glucanase cloned from winter rape of Chinese cabbage played a role in cold resistance of winter rape variety Longyou 6.
【作者单位】：甘肃农业大学农学院甘肃省油菜工程技术研究中心甘肃省作物遗传改良与种质创新重点实验室;河西学院农业与生物技术学院;
【基金】：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) 甘肃省教育厅科研专项(No.2015A-077)
【分类号】：S565.4

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