木薯海藻糖合成酶基因MeTPS6克隆及其在非生物胁迫下的表达分析

发布时间：2018-09-08 18:46

【摘要】：【目的】克隆木薯海藻糖合成酶基因MeTPS6,并分析其在干旱、低温、遮荫等非生物胁迫应答中的表达情况,为研究TPS基因功能及抗逆分子机理提供参考。【方法】采用RT-PCR从木薯中克隆MeTPS6基因,并进行生物信息学分析及构建系统发育进化树。采用荧光定量PCR(q PCR)分析MeTPS6基因在干旱、低温和遮荫胁迫下不同木薯品种不同组织中的表达特性及模式。【结果】克隆获得的MeTPS6基因具有一个2565 bp的开放阅读框,编码854个氨基酸,含有TPS家族保守结构域(Glyco_transf_20)。MeTPS6蛋白与杨树(Potri.010G104500)和杞柳(Sapur V1A.0015s0610)同源蛋白氨基酸序列的相似性分别达92.6%和90.0%,表明木薯与杨树和杞柳的亲缘关系较近。MeTPS6基因的启动子序列中存在大量非生物胁迫相关元件(低温相关元件LTR、干旱相关元件MBS等)、激素相关元件(脱落酸相关的元件ABRE和CE3、水杨酸相关元件TCA-element等)及光响应相关元件(ACE、Box I等)。MeTPS6基因在木薯储藏根中的表达量最高,须根次之,二者均显著高于茎、叶柄和叶片中的表达量(P0.05,下同),且在干旱、低温和遮荫胁迫下,不同组织中的表达模式存在明显差异。对于不同木薯品种,干旱、低温和遮荫胁迫处理均显著诱导其MeTPS6基因表达。在高置信度(Confidence=0.8)的情况下,共找到13个共表达基因,其中有10个为TPS同源基因,表明这些同源基因可能相互协调,共同参与植物生物学过程;另外3个基因(CAT、CAT1和F5M15.5)均编码过氧化氢酶,主要参与保护细胞免受过氧化氢的毒性作用。【结论】MeTPS6基因主要在木薯的根(特别是储藏根)中表达,并从转录水平参与干旱、低温和遮荫胁迫响应,可作为候选基因进一步研究其在木薯抗逆中的功能。
[Abstract]:[objective] to clone cassava trehalose synthase gene MeTPS6, and analyze its expression in abiotic stress responses such as drought, low temperature and shade. [methods] RT-PCR was used to clone MeTPS6 gene from cassava, and bioinformatics analysis was carried out and phylogenetic tree was constructed. Fluorescence quantitative PCR (q PCR) was used to analyze the expression characteristics and patterns of MeTPS6 gene in different tissues of cassava varieties under drought, low temperature and shade stress. [results] the cloned MeTPS6 gene had an open reading frame of 2565 bp. Encoding 854 amino acids, The amino acid sequence of conserved domain (Glyco_transf_20). MeTPS6 of TPS family and poplar (Potri.010G104500) and willow (Sapur V1A.0015s0610) homologous protein were 92.6% and 90.0%, respectively, indicating that the promoter sequence of .MeTPS6 gene of cassava and poplar and willow were close to each other. A large number of abiotic stress related elements (low temperature related element LTR, drought related element MBS et al.), hormone related element (abscisic acid related element ABRE and CE3, salicylic acid related element TCA-element etc.) and photoresponse related element (ACE,Box I et al.). MeTPS6 gene The highest expression level was found in cassava storage roots. The expression of fibrous roots was significantly higher than that of stems, petioles and leaves (P0.05, the same below), and there were significant differences in expression patterns in different tissues under drought, low temperature and shading stress. For different cassava varieties, drought, low temperature and shading stress significantly induced the expression of MeTPS6 gene. Under the condition of high confidence (Confidence=0.8), a total of 13 co-expressed genes were found, 10 of which were TPS homologous genes, indicating that these homologous genes may coordinate with each other and participate in the plant biological process. The other three genes (CAT,CAT1 and F5M15.5) all encode catalase and are mainly involved in protecting cells from the toxicity of hydrogen peroxide. [conclusion] the MeTPS6 gene is mainly expressed in cassava roots, especially in storage roots, and is involved in drought at the transcriptional level. The response to low temperature and shade stress can be used as candidate genes to further study its function in cassava stress resistance.
【作者单位】：中国热带农业科学院热带生物技术研究所;
【基金】：国家自然科学基金项目(31600198) 海南省自然科学基金项目(20163120)
【分类号】：S533

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