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园艺学报 ›› 2019, Vol. 46 ›› Issue (11): 2188-2200.doi: 10.16420/j.issn.0513-353x.2019-0032

• 研究论文 • 上一篇    下一篇

‘金叶’弯刺蔷薇叶绿体结构、叶绿素合成物质含量及相关基因表达分析

闫 菲1,杨树华1,*,卫晶晶2,龙 瑜3,贾瑞冬1,赵 鑫1,葛 红1,*   

  1. 1中国农业科学院蔬菜花卉研究所,国家花卉改良中心,北京 100081;2北京市植物园,北京 100093;3黔西南州金农生态产业发展有限公司,贵州兴义 562400
  • 出版日期:2019-11-25 发布日期:2019-11-25
  • 基金资助:
    国家自然科学基金项目(31672183);“十三五”重点研发项目(2018YFD1000401);北京市科委科技计划课题(D161100001916004);中央级公益性科研院所基本科研业务费专项(Y2017PT52);农业部园艺作物生物学与种质创制重点试验室项目

Chloroplast Ultrastructure,Metabolite Contents and Gene Expression Involved in the Pathway of Chlorophyll Biosynthesis of Rosa beggeriana ‘Aurea’

YAN Fei1,YANG Shuhua1,*,WEI Jingjing2,LONG Yu3,JIA Ruidong1,ZHAO Xin1,and GE Hong1,*   

  1. 1National Flowers Improvement Center of China,Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China;2Beijing Botanical Garden,Beijing 100093,China;3Qianxinan Jinnong Ecological Industry Development Co.,Ltd,Xingyi,Guizhou 562400,China
  • Online:2019-11-25 Published:2019-11-25

摘要: ‘金叶’弯刺蔷薇(Rosa beggeriana‘Aurea’)是弯刺蔷薇(R. beggeriana)经60Co–γ辐射处理获得的金色叶突变体。以弯刺蔷薇为对照,对‘金叶’弯刺蔷薇叶绿体超微结构和光合色素含量进行分析,结果显示,‘金叶’弯刺蔷薇叶绿体内基粒片层较少且垛叠稀疏不齐,叶绿体发育受到了一定程度的影响;叶绿素a和b、类胡萝卜素含量均显著低于弯刺蔷薇,但叶绿素a/b比值显著提高,属于叶绿素b缺乏型突变体。对叶绿素生物合成前体物质含量及催化其合成的关键酶基因表达分析发现,胆色素原(porphobilinogen)合成后,‘金叶’弯刺蔷薇叶片中尿卟啉原Ⅲ(uroporphyrinogen Ⅲ)、粪卟啉原Ⅲ(coproporphyrinogen Ⅲ)、原卟啉Ⅸ(protoporphyrin Ⅸ)和镁原卟啉Ⅸ(Mg-Protoporphyrin Ⅸ)含量及其合成关键酶基因RbHEMC、RbHEME1/RbHEME和RbCHLI表达水平都显著低于弯刺蔷薇,表明其叶绿素生物合成可能在由胆色素原合成尿卟啉原Ⅲ的步骤受到了阻碍。进一步对下调基因进行克隆分析,发现4个基因编码区的碱基序列在野生型和突变体之间没有差别,表明‘金叶’弯刺蔷薇叶绿素生物合成受阻不是由上述关键酶基因编码区的碱基突变造成,其叶色变异的调控机制还需要进一步探寻。

关键词: 蔷薇, ‘金叶’弯刺蔷薇, 叶色突变, 叶绿素生物合成, 基因克隆, 表达分析

Abstract: Rosa beggeriana‘Aurea’is a yellow-leaf mutant of R. beggeriana which is obtained by 60Co-γ irradiation treatment. The chloroplast ultrastructure and photosynthetic pigment content were analyzed in R. beggeriana‘Aurea’and R. beggeriana. The results showed that fewer grana lamellae,with sparsely and irregularly stacking,were observed in the chloroplasts of R. beggeriana‘Aurea’,indicating the chloroplast development of R. beggeriana‘Aurea’was impaired. The content of chlorophyll a,chlorophyll b and carotenoid in leaves of R. beggeriana‘Aurea’was significantly lower,while the chlorophyll a/b ratio was significantly increased in R. beggeriana‘Aurea’,indicating R. beggeriana ‘Aurea’was a chlorophyll b-deficient mutant. The quantification of intermediates and the gene expression of key enzymes in chlorophyll biosynthesis pathway revealed that the content of uroporphyrinogen Ⅲ,coproporphyrinogen Ⅲ,protoporphyrin Ⅸ and Mg-protoporphyrin Ⅸ,as well as the gene expression level of RbHEMC,RbHEME1/RbHEME and RbCHLI,was significantly lower in leaves of R. beggeriana‘Aurea’,indicating the chlorophyll biosynthesis of R. beggeriana ‘Aurea’may be hinderd at the process of synthesizing uroporphyrinogen Ⅲ from porphobilinogen. The coding region of 4 down-regulated genes were cloned and showed the exactly same DNA sequences between the mutant and wild type,which suggested that the blockage in the chlorophyll biosynthesis of R. beggeriana ‘Aurea’was not due to the mutation in the coding region of genes mentioned above,reflecting that the regulation mechanism of leaf color variation remains further exploration.

Key words: rose, Rosa beggeriana‘Aurea’, leaf-color mutant, chlorophyll biosynthesis, gene cloning, gene expression level

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