白桦BpMYB21基因的克隆及其表达模式分析
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摘要
【目的】研究了白桦(Betula platyphylla Suk.)MYB基因功能,分析其在茉莉酸(Me JA)和水杨酸(SA)诱导下的表达特性。【方法】以白桦为试材,通过RT-PCR等方法克隆得到1条白桦MYB转录因子家族基因序列,运用生物信息学方法,分析其蛋白质性质和亲缘关系,通过实时荧光定量PCR分析Me JA和SA诱导处理下该基因的表达特性及组织特异性。【结果】获得了1条新的白桦MYB转录因子家族基因,命名为BpMYB21,包含完整的开放阅读框,长度为1 014 bp,编码337个氨基酸。BpMYB 21蛋白与其他植物MYB氨基酸同源性为51%~64%,其中与胡桃(Juglans regia)MYB30-Like的亲缘关系最近。BpMYB21在白桦根、茎、叶中均有表达,且在叶中的表达量高于茎和根,但差异不显著;与对照相比,100μmol/L Me JA和50 mg/L SA分别处理白桦苗6 h时,BpMYB21在茎和叶中的表达较高,处理12~48 h逐渐降低。【结论】BpMYB21为MYB转录因子家族新成员,与胡桃MYB基因亲缘关系较近。该基因响应激素诱导,推测其可能在白桦生长发育及在次生代谢过程发挥重要作用。
【Objective】In this study,we explored the biological functions of the MYB gene in triterpenoid metabolism and examine MYB expression patterns induced by Me JA and SA treatment and expression specificity in different birch( Betula platyphylla Suk.) tissues.【Method】A new MYB transcription factor gene from birch was obtained by RT-PCR,and its protein characteristics and phylogenetic relationships were analyzed using bioinformatics methods. The expression pattern induced by Me JA and SA treatment was determined using real-time fluorescence quantitative PCR.【Result】A new gene of the MYB family transcription factor,named BpMYB21,was cloned from birch. The gene contains a complete1 014 bp open reading frame( ORF),encoding a 337 amino acid protein. The BpMYB21 protein has an amino acid homology of 51%-64% with other plants and is most closely related to the walnut( Juglans regia) MYB30-Like protein.BpMYB21 was expressed in the roots,stems and leaves of birch and the expression level in the leaves was higher than those in the stems and roots; however,the difference was not significant. In birch culture plants treated with 100 μmol/L Me JA and 50 mg/L SA for 6 h,the expression of BpMYB21 was the highest in the stems and leaves and expression levels gradually decreased with time from 12 to 48 h. 【Conclusion】BpMYB21 is a new member of the MYB transcription factor family,which is closely related to walnut( Juglans regia). This gene is responsive to hormone induction and may play an important role in the growth and development and secondary metabolism of birch.
【Objective】In this study,we explored the biological functions of the MYB gene in triterpenoid metabolism and examine MYB expression patterns induced by Me JA and SA treatment and expression specificity in different birch( Betula platyphylla Suk.) tissues.【Method】A new MYB transcription factor gene from birch was obtained by RT-PCR,and its protein characteristics and phylogenetic relationships were analyzed using bioinformatics methods. The expression pattern induced by Me JA and SA treatment was determined using real-time fluorescence quantitative PCR.【Result】A new gene of the MYB family transcription factor,named BpMYB21,was cloned from birch. The gene contains a complete1 014 bp open reading frame( ORF),encoding a 337 amino acid protein. The BpMYB21 protein has an amino acid homology of 51%-64% with other plants and is most closely related to the walnut( Juglans regia) MYB30-Like protein.BpMYB21 was expressed in the roots,stems and leaves of birch and the expression level in the leaves was higher than those in the stems and roots; however,the difference was not significant. In birch culture plants treated with 100 μmol/L Me JA and 50 mg/L SA for 6 h,the expression of BpMYB21 was the highest in the stems and leaves and expression levels gradually decreased with time from 12 to 48 h. 【Conclusion】BpMYB21 is a new member of the MYB transcription factor family,which is closely related to walnut( Juglans regia). This gene is responsive to hormone induction and may play an important role in the growth and development and secondary metabolism of birch.
引文
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[9]牛义岭,姜秀明,许向阳.植物转录因子MYB基因家族的研究进展[J].分子植物育种,2016,14(8):2050-2059.DOI:10.13271/j.mpb.014.002050.NIU Y L,JIANG X M,XU X Y.Reaserch advances on transcription factor MYB gene family in plant[J].Molecular Plant Breeding,2016,14(8):2050-2059.
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