山皂荚GjCYP71D1基因表达分析及在三萜合成中的功能鉴定  

Expression analysis of GjCYP71D1 gene from Gleditsia japonica and identification of its function in triterpene synthesis

作　　者：董明 张玉琦 李影[2] 武文博 王硕 孟春雨 刘梦轻 尹静[1,2] DONG Ming;ZHANG Yuqi;LI Ying;WU Wenbo;WANG Shuo;MENG Chunyu;LIU Mengqing;YIN Jing(Key Laboratory of Saline-alkali Vegetation Ecology Restoration,Ministry of Education,Northeast Forestry University,Harbin 150040,China;College of Life Sciences,Northeast Forestry University,Harbin 150040,China)

机构地区：[1]东北林业大学东北盐碱植被恢复与重建教育部重点实验室,哈尔滨150040 [2]东北林业大学生命科学学院,哈尔滨150040

出　　处：《植物生理学报》2023年第11期2085-2094,共10页Plant Physiology Journal

基　　金：黑龙江省自然科学基金(ZD2023C004);东北林业大学本科创新项目(202110225336)。

摘　　要：皂荚树含有重要的五环三萜化合物,因其广泛的药理活性而具有重要价值。本研究以山皂荚组培苗叶片为材料,通过RT-PCR方法克隆了一个CYP450新基因,并对其进行一系列生物信息学分析;采用qRT-PCR方法检测该CYP450基因、GjLus(羽扇豆醇合酶)基因在不同处理下(10μmol·L^(-1)ABA、100mg·L^(-1)GA3、50 mg·L^(-1)SA、100μmol·L^(-1)MeJA、5μmol·L^(-1)乙烯利和机械伤害)及不同组织部位的表达模式。研究结果表明,该CYP450为新基因,基因开放阅读框(ORF)为1533 bp,编码含有510个氨基酸,与决明(Senna tora)CYP71D10的相似度最高,为75.8%,暂命名为GjCYP71D1。GjCYP71D1的表达具有组织特异性,该基因在茎中和叶中表达量分别是根中的87.0%和89.9%。GjCYP71D1和GjLus响应不同生长调节物质的诱导,在MeJA、SA、GA3、ABA、乙烯利和机械伤害诱导下,均表现出相似的表达模式。通过瞬时侵染本氏烟草(Nicotiana benthamiana)发现GjCYP71D1具有催化白桦脂醇合成的功能,为山皂荚CYP450基因功能挖掘及鉴定奠定基础。The Gleditsia tree contains important pentacyclic triterpenoid compounds that are valuable for their broad pharmacological activity.In this study,a new CYP450 gene was cloned from the leaves of Gleditsia japonica tissue culture seedlings by RT-PCR,and a series of bioinformatics analyses were carried out.The expression patterns of GjCYP71D1 and GjLus(lupinol synthase)genes under different treatments(10μmol·L^(-1)ABA,100 mg·L^(-1)GA3,50 mg·L^(-1)SA,100μmol·L^(-1)MeJA,5μmol·L^(-1)ethephon and mechanical damage)and different tissue parts were detected by qRT-PCR.The results showed that the CYP450 was a new gene,the gene open reading frame(ORF)was 1533 bp,contained 510 amino acids,and the similarity with cassia(Senna tora)CYP71D10 was the highest,at 75.8%,tentatively named GjCYP71D1.GjCYP71D1 has tissue specificity,and its expression in stem and leaf was 87.0%and 89.9%of that in root,respectively.GjCYP71D1 and Gj Lus showed similar expression trends under the induction of different treatments of MeJA,SA,GA3,ABA,ethephon and mechanical injury.Through transient infection of Nicotiana benthamiana,GjCYP71D1 has the function of catalyzing betulin synthesis,which lays a foundation for the excavation and identification of CYP450 gene function in G.japonica.

关 键 词：山皂荚 CYP450 三萜化合物 表达特性 基因功能 

分 类 号：S792.99[农业科学—林木遗传育种]