月季<em>RhMYB61</em>基因的克隆及表达特性分析

doi:10.7668/hbnxb.2017.05.010

摘要/Abstract

摘要： 为了研究月季MYB类转录因子在月季花朵开放中的分子特征和表达特性，利用转录组测序获得的序列信息，结合RACE技术，从切花月季萨蔓莎中分离获得一个MYB类型的转录因子基因，命名为RhMYB61（登录号KY921844）。该基因ORF区包含1 323 bp，编码441个aa，分子量为49.1 kDa，等电点为7.66，公式C₂₁₂₃H₃₂₈₀N₆₂₀O₆₈₈S₁₈。系统进化树分析表明，RhMYB61与桃树中的PpMYB86和枇杷EjMYB8聚为一类，属于R2R3-MYB类型转录因子，进一步的蛋白序列多重比较发现，RhMYB61在N端具有保守的R2-MYB和R3-MYB区域，包含有7个保守的基序，在R3-MYB区域包含有核定位序列。利用实时定量PCR技术分析了RhMYB61的时空表达模式，结果显示，随着开花级数的增加，RhMYB61的表达特性呈逐渐增加的趋势，在开花级数5级时表达倍数最高；失水12 h处理提高了RhMYB61的表达倍数；与对照相比较，乙烯处理6，12，24 h显著提高了RhMYB61的表达特性，1-MCP则显著抑制了其表达。上述结果为月季RhMYB61生物学功能的研究奠定了基础，同时也为今后月季的分子育种研究提供了理论参考。

关键词: 切花月季, RhMYB61, qRT-PCR, 表达特性

Abstract: In order to determine the molecular characterization and expression patterns of MYB transcription factors involved in flower opening in Rose. According to the expressed sequence tags from transcriptome assembly and RACE PCR methods,we obtained a new MYB transcription factor gene,and named RhMYB61 (GenBank accession number is KY921844),in Rosa hybrida Samantha petals. The ORF of RhMYB61 was 1 323 bp,which encoded 441 amino acids. Bioinformatics analysis showed that the molecular weight and theoretical isoelectric point of RhMYB61 was 49.1 kDa and 7.66,respectively,and the formula of RhMYB61 was C₂₁₂₃H₃₂₈₀N₆₂₀O₆₈₈S₁₈. Phylogenic tree analysis showed that RhMYB61 was clustered with PpMYB86 and EjMYB8,and belonged to the R2R3-MYB type transcription factor. Multi-alignment of RhMYB61 of several plants found that RhMYB61 had the seven characteristic conservative motifs.Two conserved domains,including R2-MYB domain and R3-MYB domain,were found in its N-terminal. The nuclear localization sites was found in R3-MYB domain. Qualitative PCR analysis showed that RhMYB61 expression was induced as the rose flower opening. RhMYB61 expression was accumulated at 12 h dehydration treatment,compared with control. In addition, RhMYB61 transcript was rapidly and dramatically induced by 6,12,24 h ethylene treatment compared with control,and was inhibited following 1-methylcyclopropene (1-MCP) treatment.These results above gave a clear data for understanding the biological function of RhMYB61, and provided a foundation for further research of molecular breeding of rose plant.

Key words: Cut roses, RhMYB61, qRT-PCR, Expression characteristics

中图分类号:

Q78
S685.03

李绍翠, 姜新强, 丁爱琴, 刘庆超, 王奎玲, 李伟, 刘庆华. 月季RhMYB61基因的克隆及表达特性分析[J]. 华北农学报, 2017, 32(5): 61-68. doi: 10.7668/hbnxb.2017.05.010.

LI Shaocui, JIANG Xinqiang, DING Aiqin, LIU Qingchao, WANG Kuiling, LI Wei, LIU Qinghua. Cloning and Expression Characteristics of RhMYB61 Gene from Rosa hybrida[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(5): 61-68. doi: 10.7668/hbnxb.2017.05.010.

http://www.hbnxb.net/CN/Y2017/V32/I5/61

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月季RhMYB61基因的克隆及表达特性分析

Cloning and Expression Characteristics of RhMYB61 Gene from Rosa hybrida

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Cloning and Expression Characteristics of RhMYB61 Gene from Rosa hybrida

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