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甘蔗TB1基因的克隆与生物信息学分析

2015-10-21李旭娟林秀琴刘洪博李纯佳徐超华刘新龙

热带作物学报 2015年11期

李旭娟 林秀琴 刘洪博 李纯佳 徐超华 刘新龙

摘 要 以负调控禾本科植物分蘖的关键基因TB1为研究对象,根据水稻、玉米、高粱TB1同源基因保守序列设计引物,使用RT-PCR方法和RACE技术从甘蔗茎尖处克隆到该基因的同源基因ScTB1。ScTB1 cDNA全长1 668 bp,包含274 bp的5′ UTR,1 149 bp的CDS和245 bp的3′ UTR。生物信息学分析表明该基因可编码382个氨基酸残基,编码产物含有保守的SP区、TCP区和R区,属于TCP家族转录因子。其分子量为40.7 ku,理论等电点为8.55,蛋白亚细胞定位预测其主要定位于细胞质。系统进化分析表明ScTB1属于CYC/TB1亚族蛋白,与高粱TB1和玉米TB1聚为一个亚类。根据序列的保守性和预测结果可推测ScTB1很可能也参与了甘蔗分蘖性状的调控。

关键词 甘蔗;分蘖;TB1基因;克隆;生物信息学分析

中图分类号 S566.1 文献标识码 A

Abstract In this study, TB1 homolog of sugarcane was cloned from the stem apex by using RT-PCR and RACE method referring to the conserved sequence of TB1 homologs in rice, maize and sorghum. The cDNA length of ScTB1 is 1 668 bp, which contains 274 bp 5′ UTR, 1 149 bp CDS and 245 bp 3′ UTR. Bioinformatics analysis showed that ScTB1 belonged to the TCP family of transcription factors, which is composed of 382 amino acid residues and contains three conservative domains(SP, TCP and R domains). The molecular weight and theoretical isoelectric point of ScTB1 was 40.7 ku and 8.55, respectively, and the gene was likely located in cytoplasm. Phylogenetic tree analysis indicates that ScTB1 belonged to CYC/TB1 subfamily and had a very close relationship with TB1 homologs of maize and sorghum. Based on the sequence conservation, it is suggested that ScTB1 may be required for the regulation of tillering trait. This study provides important information for further research on the functional analysis and regulating mechanisms of ScTB1 in future.

Key words Sugarcane; Tillering; TB1 gene; Cloning; Bioinformatics analysis

doi 10.3969/j.issn.1000-2561.2015.11.010

甘蔗(Saccharum spp.)是主產于热带和亚热带区域的重要糖料经济作物,全世界近80%以上的食糖来自于甘蔗。此外,甘蔗还因蕴含较高的生物量而成为世界上最重要的能源作物之一[1-2]。甘蔗属无性繁殖作物,蔗茎为其主要收获产品,主要由单一的主茎和有效分蘖茎组成,其中有效分蘖茎数量的多少对于甘蔗最终产量的大小具有重要影响,由此可见,分蘖性状表现对于甘蔗产量具有举足轻重的作用。而从甘蔗分蘖性状着手,研究其分子基础,对于利用分蘖性状改良甘蔗品种产量具有重要意义。

TB1(TEOSINTE BRANCHED 1)基因,是一个作用于独脚金内酯(Strigolactones, SLs)下游的基因,属植物所特有的TCP家族转录因子,编码含有不典型的螺旋-环-螺旋结构(non-canonical basic-Helix-Loop-Helix strueture,bHLH)的DNA结合蛋白,主要在分生组织中起作用,与细胞分化和生长有很大的关系[3]。……

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