甘蔗MOC1基因 (ScMOC1)的克隆与表达分析
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国家自然科学基金(31360359,31601362);云南省基础研究计划青年项目(2015FD063);云南省中青年学术技术带头人后备人才(2014HB038)第一作者主要从事甘蔗分子遗传学研究工作。E-mail:lixujuan2011@163.com。通信作者:刘新龙,主要从事甘蔗分子遗传学研究。E-mail:lxlgood868@163.com


Cloning and Expression Analysis of the MOC1 Gene (ScMOC1) in Sugarcane
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    摘要:

    MONOCULM 1(MOC1) 基因在植物腋分生组织和腋芽的形成中发挥重要作用,是植物分蘖关键调控基因。本研究利用同源基因克隆法结合RT-PCR、RACE技术从甘蔗品种ROC22中克隆到MOC1的同源基因,命名为ScMOC1。该基因的cDNA序列包含一个长度为1299 bp的开放阅读框,编码432个氨基酸残基组成的含有GRAS保守结构域的非分泌蛋白,其分子量为45.43 kD、理化等电点pI为6.98。序列比对分析显示ScMOC1与高粱(Sorghum bicolor)、赖草(Leymus secalinus)、水稻(Oryza sativa)等禾本科植物同源蛋白氨基酸序列一致性较高;系统进化树分析显示其与高粱、小米草(Setaria italica)、玉米(Zea mays)等禾本科植物MOC1同源蛋白亲缘关系最近;ScMOC1在甘蔗品种ROC22中的序列变异分析发现, 30个克隆得到的序列中共有46个SNP位点和29处InDel位点,其中1个位点发生的单个碱基缺失和另一个位点的4个碱基插入是造成基因编码蛋白序列变化的主要原因,中性检测表明,ScMOC1在ROC22中遵循中性进化模型。实时荧光定量PCR(Real time fluorescent quantitative-PCR, q-PCR)表达分析显示ScMOC1基因在分蘖期的ROC22中的表达具有组织特异性,在生命活跃的茎尖生长点处表达量最高;在腋芽形成发育过程中该基因表达总体呈现出“升-降-升-降”的趋势,说明ScMOC1基因可能在甘蔗腋芽形成发育阶段中发挥作用。以上研究可推测ScMOC1在甘蔗的分蘖性状调控上扮演重要的角色。本研究为ScMOC1的功能研究及其在甘蔗产量分子辅助育种中的利用奠定基础。

    Abstract:

    MONOCULM 1 (MOC1), a gene that is important in the regulation of plant tillering, which plays a significant role in the formation of plant axillary meristem and axillary bud. In this study, the homolog of MOC1 was cloned from sugarcane variety, which was named ScMOC1. The cDNA of ScMOC1 contains a 1299 bp ORF which encodes a secreted protein containing a GRAS conservative structure domain. The protein was predicted to encode 432 amino acid residues with a molecular mass of 45.43 kD and bioelectric point value of 6.98. Sequence alignment analysis showed ScMOC1 keep a high sequence identity with some poaceae family plants homologous proteins such as Sorghum bicolor, Leymus secalinus and Oryza sativa. In the phylogenetic tree, ScMOC1 has a closest evolutionary relationship with the homologous protein from Sorghum bicolor, Setaria italica and Zea mays. Sequenced and analysis of ScMOC1 in sugarcane varity ROC22 , totally 46 SNP and 29 InDels were identified in 30 clones. Among these variation sites, one deletion and the other 4 insertion of bases are the main causes of the changes for the gene encoding protein sequences. Neutrality tests showed that no purifying selection occurred of ScMOC1 in ROC22. Real time fluorescent quantitative-PCR (qPCR) shows that ScMOC1 was constitutively expressed in ROC22 and highest expressed in stem apex at the tillering stage; while present a expression tendency of “up-down-up-down” at the axillary buds formation and development period, so we predicted that ScMOC1 plays a role in the formation of sugarcane axillary buds. According to these researches, we speculated that ScMOC1 might play a key role in the regulation of tillering trait of sugarcane. The results obtained above will provide a theoretical foundation for function analysis of ScMOC1 and molecular assisted selection of sugarcane yield in the future.

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李旭娟,李纯佳,徐超华,等.甘蔗MOC1基因 (ScMOC1)的克隆与表达分析[J].植物遗传资源学报,2017,18(4):734-746.

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  • 收稿日期:2016-11-30
  • 最后修改日期:2017-03-03
  • 录用日期:2017-03-23
  • 在线发布日期: 2017-07-14
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