南京林业大学学报(自然科学版) ›› 2018, Vol. 61 ›› Issue (06): 27-34.doi: 10.3969/j.issn.1000-2006.201803002

• 研究论文 • 上一篇    下一篇

3个白桦细胞色素P450基因生物信息学及表达分析

杨 杰1,孙 璐1,王思瑶1,李 影1,翟 睿1,林香雨1,詹亚光1,2,尹 静1*   

  1. (1.东北林业大学生命科学学院,黑龙江 哈尔滨 150040; 2.林木遗传育种与生物技术国家重点实验室,黑龙江 哈尔滨 150040)
  • 出版日期:2018-11-30 发布日期:2018-11-30
  • 基金资助:
    收稿日期:2018-03-01 修回日期:2018-09-11
    基金项目:国家自然科学基金项目(31570589); 中央高校基本科研业务费专项资金项目(257-2017DY02); 东北林业大学创新创业训练项目(201710225109)
    第一作者:杨杰(yangjiesolo@163.com)。*通信作者:尹静(yinjing20135@163.com),副教授。

Tissue specificity and hormone induced expression of three cytochrome P450 genes from Betula platyphylla Suk

YANG Jie1, SUN Lu1, WANG Siyao1, LI Ying1, ZHAI Rui1, LIN Xiangyu1, ZHAN Yaguang1,2, YIN Jing1*   

  1. (1. School of Life Science, Northeast Forestry University, Harbin 150040, China; 2. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040, China)
  • Online:2018-11-30 Published:2018-11-30

摘要: 【目的】研究白桦细胞色素P450基因表达的组织特异性,以及在茉莉酸甲酯(MeJA)、水杨酸(SA)、赤霉素(GA3)、脱落酸(ABA)、乙烯利和伤害处理下的基因表达模式。【方法】筛选白桦转录组,获得3个BpCYP450 基因,分别命名为BpCYP4、BpCYP5、 BpCYP14,利用生物信息学分析BpCYP450蛋白的分子结构特征及其与其他物种CYP450蛋白的亲缘关系。采用QRT-PCR技术,对白桦BpCYP450组织特异性、激素信号及伤害诱导下的表达特征进行分析。【结果】生物信息学结果表明BpCYP4、BpCYP5、BpCYP14 cDNA序列长度分别为1 569、1 584和1 530 bp,具有完整的开放阅读框(ORF),分别编码522、527和509个氨基酸,均为亲水性跨膜蛋白,主要定位于叶绿体。白桦BpCYP450与百脉根、扁豆及豌豆等豆科植物CYP450蛋白亲缘性较高。组织特异性分析结果显示,3个BpCYP450 基因在叶和根中表达较高,茎中较低。激素信号及伤害诱导结果表明,3个BpCYP450基因不同程度地响应MeJA、SA、GA3、ABA、乙烯利激素信号及伤害诱导。【结论】3个BpCYP450基因均为CYP450基因家族的新成员,具有组织特异性及激素诱导表达特性,可能在白桦生长发育、抵御胁迫及代谢物合成中发挥重要作用。

Abstract: 【Objective】Explore the tissue specificity of the cytochrome P450 genes in the triterpenoids from birch(Betula platyphylla Suk.)and expression pattern under treatments of methyl jasmonate(MeJA), salicylic acid(SA), gibberellin(GA3), abscisic acid(ABA), ethephon and leaf injury.【Method】Three BpCYP450 genes were screened from the transcriptome of B. platyphylla Suk. and named BpCYP4, BpCYP5 and BpCYP14, respectively. The molecular structure of BpCYP450 proteins and relationship with CYP450 proteins of other species of plants were described by bioinformatics analysis. The tissue specificity of BpCYP450 and the expression characteristics under hormone and injury were analyzed by QRT-PCR.【Result】Bioinformatics results showed that the size of BpCYP4, BpCYP5 and BpCYP14 were 1 569, 1 584 and 1 530 bp, respectively, and contained the complete ORF, encoding 522, 527 and 509 amino acids, which were hydrophilic transmembrane proteins and mainly located in the chloroplast. Three BpCYP450 were closely related to leguminous plant CYP450 proteins such as Lotus japonicus, Lens culinaris and Pisum sativum. The ana-lysis of tissue-specific showed that BpCYP450s expression were higher in leaves and in roots,and lower in stems. The results of hormone and injury induction showed that BpCYP450 genes was induced by MeJA, SA, GA3 and ABA, ethephon and injury. 【Conclusion】Three BpCYP450 are new members of the CYP450 genes family, with expression of tissue-specific and hormone induction. These results suggested that three CYP450 genes may play an important role in the growth and development the defense stress and the metabolites synthesis of birch.

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