中国农业科学 ›› 2016, Vol. 49 ›› Issue (19): 3786-3797.doi: 10.3864/j.issn.0578-1752.2016.19.010

• 园艺 • 上一篇    下一篇

葡萄OVATE基因家族生物信息学及表达

袁 月,张亚光,高世敏,陶建敏   

  1. 南京农业大学园艺学院,南京 210095
  • 收稿日期:2016-03-16 出版日期:2016-10-01 发布日期:2016-10-01
  • 通讯作者: 陶建敏,Tel:13905160976,E-mail:tjm266@sina.com
  • 作者简介:袁月,Tel:15150536081;E-mail:2013104040@njau.edu.cn
  • 基金资助:
    国家现代农业葡萄产业技术体系项目(CARS-30)、国家“948”重点项目(2016-X19)、淮安科技局葡萄新品种引进及设施栽培技术研究与应用项目(HAC2014019)、江苏省科技厅设施条件下葡萄高效栽培新技术研究与应用项目(SBE2014030811)

Bioinformatics and Expression of the OVATE Gene Family in Grape

YUAN Yue, ZHANG Ya-guang, GAO Shi-min, TAO Jian-min   

  1. College of Horticulture, Nanjing Agricultural University, Nanjing 210095
  • Received:2016-03-16 Online:2016-10-01 Published:2016-10-01

摘要: 【目的】OVATE是一类调控植物生长发育的转录抑制因子,对葡萄OVATE基因家族(VvOFPs)进行生物信息学和组织特异性表达分析,为该类基因的功能研究奠定基础。【方法】根据OVATE保守域蛋白序列(PF04844)对葡萄OVATE基因家族进行鉴定,利用生物信息学方法对葡萄OVATE基因家族染色体定位、基因结构、保守结构域、亚细胞定位等方面进行预测和分析,并分析葡萄和拟南芥OVATE基因家族的进化关系。采用实时荧光定量PCR技术检测VvOFPs组织表达特性。【结果】葡萄OVATE基因家族包含17个成员,不均匀地分布在11条染色体上,均没有内含子结构,编码115—444个氨基酸,等电点4.55—9.69,均为亲水蛋白;所有蛋白均包含完整的OVATE保守结构域,亚细胞定位主要在细胞核中。根据进化树拓扑结构,将葡萄和拟南芥OVATE蛋白家族聚为六类(I—VI),其中,Ⅱ、Ⅲ、Ⅴ类仅包含两个基因,Ⅰ、Ⅳ、Ⅵ类中VvOFPs和AtOFPs相互交错地聚类在一起;VvOFPs和AtOFPs共包含10个未知基序,保守元件1和2位于OVATE结构域区域,此外,在OVATE结构域外每个类别均包含特有基序。VvOFPs具有组织表达特异性,12个基因在根、茎、叶、花和果实中均可以检测到表达,其余5个基因仅在特定组织中表达。多数VvOFPs在根、嫩茎和花中表达量较高,在嫩叶和果实中仅检测到少数基因表达;VvOFPs在不同发育期表达量存在差异,通常在开花前1周和开花期表达量较高,而花后4周果实中表达量较低。1对旁系同源基因表达模式相似,3对旁系同源基因产生了新的表达模式。【结论】葡萄OVATE结构域序列比较保守,其在不同组织中呈现出多种表达模式,推测其可能参与了葡萄生长发育的调控。

关键词: 葡萄, OVATE基因家族;生物信息学;表达

Abstract: 【Objective】OVATE is a class of transcriptional repressor that regulates plant growth and development. In this study, the authors analyzed the grape OVATE gene family (VvOFPs) from the aspects of bioinformatics and tissue-specific expression in order to lay a foundation for the functional studies in future. 【Method】Based on the conserved OVATE domain (PF04844), OVATE genes were identified from grape genome. Chromosome location, gene structure, conserved domain and subcellular localization were also analyzed by the bioinformatics methods. A phylogenetic analysis was conducted based on the alignment of the OVATE proteins from grape and Arabidopsis genomes. The expression patterns of VvOFPs were tested via quantitative real-time polymerase chain reaction (qRT-PCR). 【Result】Seventeen putative VvOFPs were identified in the grape genome and they unevenly distributed on the eleven chromosomes. No intron was found in the gene structures of VvOFPs. The VvOFPs encoded hydrophilic proteins, containing 115-444 amino acids, and their isoelectric points ranged from 4.55-9.69. In addition, all the VvOFPs contained the entire OVATE domain and were predicted to be located in the nucleus. According to the topology of phylogenetic tree, OVATE proteins from grape and Arabidopsis were divided into six groups (I-VI). Group II, III and V contained only two members, whilst members of groups I, IV and VI were clustered in the species-specific pattern. VvOFPs and AtOFPs contained 10 uncharacterized motifs. Motifs 1 and 2 were located in the OVATE domain region. Besides, group-specific motifs were also found outside the OVATE domain. qRT-PCR analysis showed that VvOFPs exhibited different tissue-specific expression patterns. Interestingly, twelve VvOFPs were found to be expressed in the detected organs of roots, stems, leaves, flowers and fruits while the others were expressed in the specific organ tissues. Most of the VvOFPs were more highly expressed in roots, stems and flowers, whereas a little gene was detected with low level in leaves and fruits. Meanwhile, VvOFPs expressed differently at different developmental stage. Transcripts in flowers a week before or at anthesis were at a high level, while low in fruits four weeks after anthesis. In addition, one pair of paralogues exhibited the same expression patterns, whereas the other three pairs of paralogues exhibited divergent expression patterns. 【Conclusion】Results of this study showed that grape OVATE domain is conserved. VvOFPs exhibited different expression patterns and may be involved in regulation of plant growth and development.

Key words: grape, OVATE gene family, bioinformatics, expression