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中国农学通报

中国农学通报 ›› 2025, Vol. 41 ›› Issue (33): 131-138.doi: 10.11924/j.issn.1000-6850.casb2025-0016

• 生物科学 • 上一篇    下一篇

常温常压等离子体诱变选育荧光假单胞菌CLW17高效促生长突变株

陈艳彬(), 支梦涛, 白变霞, 贾瑞, 任嘉红()   

  1. 长治学院生命科学系，山西长治 046011
  • 收稿日期:2025-01-10 修回日期:2025-03-15 出版日期:2025-11-25 发布日期:2025-12-01
  • 通讯作者:
    任嘉红，女，1976年生，云南文山人，教授，博士，研究方向：森林微生物。通信地址：046011 山西省长治市潞州区太行东街街道长治学院北校区 长治学院，Tel：15235535898，E-mail：。
  • 作者简介:

    陈艳彬，女，1981年生，山西沁县人，高级实验师，硕士，研究方向：根际微生物及食品分析检测。通信地址：046011 山西省长治市潞州区太行东街街道长治学院北校区 长治学院，Tel：15235591986，E-mail：。

  • 基金资助:
    国家自然科学基金“群体感应(QS)介导内生Burkholderia pyrrocinia的生防分子机制”(32071770); 山西省“1331工程”“太行山药用植物生物肥料工程研究中心”(1331KSC); 山西省回国留学人员科研资助项目“款冬褐斑病拮抗合成菌群构建及其活性因子研究”(2023-172); 长治学院大学生创新创业训练计划项目“款冬褐斑病的生物防治研究”(2023DC08)

Breeding of Highly Efficient Growth-promoting Mutant Strain of Pseudomonas fluorescens CLW17 by Atmospheric and Room Temperature Plasma

CHEN Yanbin(), ZHI Mengtao, BAI Bianxia, JIA Rui, REN Jiahong()   

  1. Department of Biological Sciences, Changzhi College, Changzhi, Shanxi 046011
  • Received:2025-01-10 Revised:2025-03-15 Published:2025-11-25 Online:2025-12-01

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摘要：

为定向改良荧光假单胞菌(Pseudomonads fluorescens) CLW17高效优良促生长特性，为微生物菌肥生产提供优质菌种。本研究以具有解磷、产嗜铁素能力的CLW17野生株为出发菌株，采用常温常压等离子(ARTP)诱变技术进行菌株改良，通过“定性初筛-定量复筛-分子鉴定”三级筛选体系，获得高效促生长突变株并优化诱变条件。结果表明：CLW17菌株的最佳ARTP诱变条件为输出功率100 W，照射距离2 mm，处理时间240 s，此条件下菌株致死率达91.67%，正突变率为11.27%。经NBRIP-BPB平板解磷圈定性筛选（SI值）、钼锑抗比色法解磷定量测定及CAS平板/液体法产嗜铁素能力测定，最终筛选出2株高效优良促生长突变株CLW17-240-2-1和CLW17-240-3-2，其解磷量分别为553.08 mg/L、584.47 mg/L，较野生株(453.77 mg/L)提升21.89%、30.40%；产嗜铁素能力分别为62.52%、72.33%，较野生株(51.55%)提升21.27%、40.29%。16S rDNA分子鉴定证实，2株突变株均为CLW17的遗传突变体，无外源污染。本研究通过ARTP诱变成功实现CLW17菌株“解磷-产嗜铁素”双促生性状的协同提升，为植物根际促生菌(PGPR)的定向育种提供了技术参考，也为南方红豆杉等林木专用微生物菌肥的研发奠定了菌种基础。

关键词: 荧光假单胞菌CLW17, 常温常压等离子体(ARTP)诱变, 解磷, 产嗜铁素, 促生长

Abstract:

The purpose of this study is to select and breed highly efficient and excellent growth-promoting mutant strains of Pseudomonas fluorescent CLW17, to provide high-quality bacterial strain resources for the production of microbial fertilizer. The wild strain CLW17 with the phosphate-solubilizing and siderophore-producing abilities was used as the starting strain, and the strain was improved by ARTP mutagenesis technology. Through the three-stage screening system of 'qualitative primary screening-quantitative re-screening-molecular identification', the high-efficiency growth-promoting mutant strain was obtained and the mutagenesis conditions were optimized. The results showed that the optimal mutagenesis for CLW17 was as follows: output power of 100 W, irradiation distance of 2 mm, and treatment time of 240 s. Under these conditions, the mortality rate of the strain was 91.67%, and the positive mutation rate was 11.27%. After ARTP mutagenesis, two mutants with high efficiency and excellent growth-promoting characteristics (CLW17-240-2-1 and CLW17-240-3-2) were finally screened out. Compared to the original strain, the phosphate-solubilizing capacity of CLW17-240-2-1 and CLW17-240-3-2 was enhanced by 21.89% and 30.40% respectively, and their siderophore-producing ability was enhanced by 21.27% and 40.29%. 16S rDNA molecular identification confirmed that the two mutants were genetic mutants of CLW17 without exogenous pollution. In this study, the synergistic improvement of the dual growth-promoting traits of CLW17 strain 'phosphate-solubilizing and siderophore-producing' was successfully achieved through ARTP mutagenesis, which provided a technical reference for the directional breeding of plant growth-promoting rhizobacteria (PGPR), and also laid a strain foundation for the research and development of microbial fertilizer for trees such as Taxus chinensis var.mairei.

Key words: Pseudomonas fluorescens CLW17, atmospheric and room temperature plasma mutagenesis, phosphorus-solubilizing, siderophore-producing, growth-promoting