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园艺学报 ›› 2013, Vol. 40 ›› Issue (2): 237-246.

• 果树 • 上一篇    下一篇

香蕉茉莉酸合成关键酶基因MaOPR 的克隆和表达分析

 许 奕, 徐碧玉, 宋 顺, 刘菊华, 张建斌, 贾彩红, 金志强   

  1. (1 中国热带农业科学院海口实验站,海口 570102;2 海南省香蕉遗传改良重点实验室,海口 570102;3 中国热带农
    业科学院热带生物技术研究所,农业部热带作物生物学与遗传资源利用重点实验室,海口 571101)
  • 出版日期:2013-02-25 发布日期:2013-02-25

Molecular Cloning and Expression Analysis of MaOPR Gene from Banana

 XU   Yi, XU  Bi-Yu, SONG   Shun, LIU  Ju-Hua, ZHANG  Jian-Bin, JIA  Cai-Hong, JIN  Zhi-Qiang   

  1. (1Haikou Experimental Station,Chinese Academy of Tropical Agricultural Sciences,Haikou 570102,China;2Hainan Key
    Laboratory of Banana Genetic Improvement,Haikou 570102,China;3Key Laboratory of Biology and Genetic Resources of
    Tropical Crops,Ministry of Agriculture;Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical
    Agricultural Sciences,Haikou 571101,China)
  • Online:2013-02-25 Published:2013-02-25

摘要: 从‘巴西’香蕉(Musa acuminata L. AAA group‘Brazilian’)根的cDNA 文库中获得了一段
12–氧–植物二烯酸还原酶OPR(12-oxo-phytodienoic acid reductase)基因的片段,RACE 扩增获得全长,
命名为MaOPR。该基因全长1 512 bp,存在一个完整的开放阅读框1 287 bp,编码429 个氨基酸。生物
信息学分析表明,该蛋白属稳定蛋白,等电点为8.11,具有一个活性位点,一个基质结合位点,一个黄素
单核苷酸结合位点。序列预测分析该蛋白亚细胞定位为细胞质,其不属于跨膜蛋白且不存在信号肽。通
过和已知植物的12–氧–植物二烯酸还原酶基因相比,同源性达到66%以上。其中与苜蓿、谷子、粳稻、
紫云英的OPR 编码的氨基酸序列的同源性均为71%。器官特异性分析表明,MaOPR 在香蕉的根、茎、
叶片、花和果实中均有所表达,其中在茎和果实中表达量较高。通过对其在ABA 抑制剂、乙烯、枯萎病
胁迫下的表达结果分析显示,该基因响应以上3 种胁迫。

关键词: 香蕉, 12–氧–植物二烯酸还原酶基因, 序列分析, 荧光定量PCR

Abstract: Through RACE approaches and bioinformatics analysis,the full-length cDNA of the
cysteine synthase gene,named as MaOPR(12-oxo-phytodienoic acid reductase),was obtained from root
of banana(Musa acuminata L. AAA group‘Brazilian’)cDNA library,and its sequence characters were
also analyzed. The full length of this gene was 1 512 bp,it had an 1 287 bp open reading frame in length
encoding 429 amino acid. The result of bioinformatics showed that this protein was a stable protein with
the active sites,the matrix attachment sites and the flavin mononucleotide binding sites,which pI is 8.11.
The sequence prediction showed that it located in the cytoplasm,it was not a transmembrane protein and
had no signal peptide. Compared with other known plants,the homology of MaOPR was more than 66%.
Amino acids homology analysis indicated that MaOPR had 71% similarity compared with Medicago
sativa,Setaria italic,Oryza sativa,Astragalus sinicus. RT-PCR analysis showed that MaOPR was
constitutively expressed in roots,stems,leaves,flowers and fruits. The expression level was the highest in
fruits. Analysis showed that MaOPR responsed to the stress including the ABA inhibitor,ethylene and wilt
disease stress.

Key words: banana, 12-oxo-phytodienoic acid reductase, sequence character, real-time PCR

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