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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2017, Vol. 43 Issue (3): 289-297    DOI: 10.3785/j.issn.1008-9209.2016.10.111
生物科学与技术     
红掌2个SOC1基因的克隆、序列与表达分析
马广莹1，朱开元1，史小华1，邹清成1，刘慧春1，詹菁2，田丹青1*
1.浙江省农业科学院花卉研究开发中心，杭州 311202；2.浙江省杭州萧山技师学院，杭州 311202
Cloning, sequence and expression analysis of two SOC1 genes from Anthurium
MA Guangying1, ZHU Kaiyuan1, SHI Xiaohua1, ZOU Qingcheng1, LIU Huichun1, ZHAN Jing2, TIAN Danqing1*
(1. Flower Research and Development Center, Zhejiang Academy of Agricultural Sciences, Hangzhou 311202, China; 2. Hangzhou Xiaoshan Technician College of Zhejiang Province, Hangzhou 311202, China)
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摘要： 为了明确SOC1转录因子在红掌中的成员及作用机制，采用5＇/3＇末端快速扩增技术获得这2个SOC1 基因的完整编码区，通过生物信息学软件分析这2个基因的相关信息，并结合荧光定量反转录-聚合酶链式反应验证这2个基因的表达模式。结果显示：获得的2个基因核苷酸长度分别为651 bp和642 bp，命名为AaSOC1-1 和AaSOC1-2；2个转录因子在二级结构上均有螺旋、折叠和转角，未发现其他结构。系统进化分析显示，二者与单子叶植物同类蛋白距离较近，与红掌的植物学分类地位一致。表达分析结果显示，2个基因均在营养器官和生殖器官中表达，但表达水平略有差异。研究认为，从红掌中克隆的2个SOC1转录因子可能定位于线粒体而非细胞核中，二者在序列组成和表达模式上既有保守性也存在一定的差异性，预示它们在功能上可能也存在一定的区别。
Abstract: SUPPRESSOR OF OVEREXPRESSION OF CO1 (SOC1) is a flowering integration factor that widely existed in flowering plants, which is necessary in multiple flowering induction pathways, including vernalization, photoperiod, gibberellin, and other endogenous or exogenous environmental signals. Therefore, research of the effect of SOC1 on plant growth and  development can reveal the plant flowering mechanism and direct molecular breeding. Anthurium is a kind of high-grade potted flower in the worldwide. However, molecular biology research on this plant had just started. As a flowering integration factor, SOC1 in Anthurium may be a key to deeply understand the flower conversion process.
In this study, rapid amplification of cDNA ends (RACE) technology was used to obtain the full length of two SOC1 genes of Anthurium. Several biological softwares were used to analyze the biological information, including NCBI Blast, NCBI Conserved Domain Search, ProtParam, TMHMM Server version 2.0, SignalP 4.1 Server, iPSORT, PSIPRED, DNAman, and MEGA 6.0. Gene expression levels of the two SOC1 genes were detected by real-time reverse transcription?polymerase chain reaction (RT-PCR).
The main results were as follows: AaSOC1-1 and AaSOC1-2 were obtained from two independent SOC1 full coding sequence (CDS) regions, which shared 75.6% similarity at the nucleotide level. Their encoded proteins were composed of 216 and 213 amino acid residues, respectively. Both of the putative proteins had secondary structures such as helix, sheet, and turn, but no other special structures were found. Based on the online software analysis results, it was concluded that AaSOC1-1 and AaSOC1- 2 were located in the mitochondria but not in the nucleus. Phylogenetic tree analysis showed that the two SOC1 transcription factors from Anthurium were tightly clustered with each other, and also with other monocotyledons. Fluorescence quantitative RT?PCR analysis revealed that both genes from Anthurium were ubiquitously expressed in the vegetative and reproductive organs, but different in the expression patterns. Particularly, AaSOC1-1 was mainly expressed in the bract and tepal, whereas AaSOC1-2 was mainly expressed in the stem and pedicel.
In sum, this study reported the nucleotide and deduced amino acid composition of the two SOC1 transcription factors. Preliminary analysis of their characteristics, structure, and potential function were performed through bioinformatics software, combined with multiple sequence alignment and phylogenetic tree construction. Based on the results, the two genes found in Anthurium were identified as members of SOC1 gene family.Gene expression experiments confirmed the similarities and  differences of their expression patterns, which were relatively conservative compared with other species. This study lays the foundation for understanding the functional properties of these two transcription factors, which also requires in?depth research to verify the above speculation and views.
收稿日期: 2016-10-11 出版日期: 2017-04-28
CLC:  S 682.14  
基金资助: 国家自然科学基金（31200527）；浙江省农业新品种选育重大专项（2016C02056-13）
通讯作者: 田丹青(http://orcid.org/0000-0002-8646-4206)     E-mail: tdq0123@163.com
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引用本文:

马广莹，朱开元，史小华，邹清成，刘慧春，詹菁，田丹青. 红掌2个SOC1基因的克隆、序列与表达分析[J]. 浙江大学学报(农业与生命科学版), 2017, 43(3): 289-297.

MA Guangying, ZHU Kaiyuan, SHI Xiaohua, ZOU Qingcheng, LIU Huichun, ZHAN Jing, TIAN Danqing. Cloning, sequence and expression analysis of two SOC1 genes from Anthurium. Journal of Zhejiang University (Agriculture and Life Sciences), 2017, 43(3): 289-297.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2016.10.111    或    http://www.zjujournals.com/agr/CN/Y2017/V43/I3/289

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