Journal of Agricultural Science and Technology ›› 2021, Vol. 23 ›› Issue (2): 81-88.DOI: 10.13304/j.nykjdb.2020.0248

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Impact of PoprL Promoter on Biosynthesis of Coronatine in Pseudomonas Syringae DC3000

HE Yan1, JIANG Feng2, YU Sha1, YU Chunxin1, TAN Weiming1, LI Zhaohu1, ZHANG Jie3, DUAN Liusheng1*   

  1. 1.State Key Laboratory of Plant Physiology and Biochemistry, Engineering Research Center of Plant Growth Regulator, Ministry of Education; College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China;
    2.College of Horticulture, China Agricultural University, Beijing  100193, China; 
    3.State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
  • Received:2020-03-23 Online:2021-02-15 Published:2020-04-30

PoprL启动子对丁香假单胞菌DC3000合成冠菌素的影响

何彦1,姜峰2,于莎1,于春欣1,谭伟明1,李召虎1,张杰3,段留生1*   

  1. 1.中国农业大学农学院, 植物生理生化国家重点实验室, 教育部植物生长调节剂工程研究中心, 北京 100193;
    2.中国农业大学园艺学院, 北京 100193;
    3.中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193
  • 通讯作者: 段留生 E-mail:duanlsh@cau.edu.cn
  • 作者简介:何彦 E-mail:heyan_571187@126.com
  • 基金资助:
    国家重点研发计划重点专项(2017YFD0201300)

Abstract: Coronatine (COR) is a secondary metabolite produced by several pathovars of Pseudomonas syringae, and it is a novel plant growth regulator. In order to improve the production of strains and optimize the culture conditions of fermentation, the strong constitutive promoter PoprL was selected to construct the recombinant strain CO by homologous recombination using the plasmid pUCP24/recTE in Pseudomonas syringae DC3000. The results showed that the productions of COR  increased in CO strain. The yield of CO strain was 1.8 times as much as that of the original strain under 18 ℃ culture and 4.1 times than under 28 ℃ culture. In this study, promoter substitution was used to effectively increase the yield of COR, and the restriction effect of high-temperature fermentation conditions on the yield of COR was improved, which provided strong support and reference for the strain modification and industrial production of COR.

Key words: coronatine, secondary metabolites, promoter, pseudomonas syringae

摘要: 冠菌素(coronatine, COR)是由多种丁香假单胞菌致病变种产生的次生代谢产物,是一种新型植物生长调节剂。为了提高菌株冠菌素产量并优化发酵培养条件,将来源于假单胞菌肽聚糖脂蛋白编码基因oprL高效表达组成型启动子PoprL,通过质粒pUCP24/recTE同源重组到丁香假单胞菌DC3000冠菌素合成基因cfl中,得到启动子重组菌株CO,在18和28 ℃培养条件下发酵培养,结果表明,CO菌株在18 ℃培养条件下冠菌素产量是出发菌株DC3000的1.8倍,在28 ℃培养条件下冠菌素产量是出发菌株DC3000的4.1倍。利用启动子替换的方法有效提高了冠菌素的产量,并且改善了高温发酵条件对菌株冠菌素产量的制约影响,为冠菌素的菌种改造和工业化生产应用提供有力支持与借鉴。

关键词: 冠菌素, 次生代谢产物, 启动子, 丁香假单胞菌