摩氏假单胞菌环脂肽合成基因簇xtlABC的功能研究

doi:10.16409/j.cnki.2095-039x.2024.02.023

摘要/Abstract

摘要： 摩氏假单胞菌Pseudomonas mosselii PtA1是对多株青枯菌均有良好抑制作用的生防菌，基因组中含有完整的环脂肽合成基因簇xtlABC。为了探明摩氏假单胞菌PtA1中xtlABC基因簇的功能，本研究利用同源重组技术获得缺失突变体ΔxtlA、ΔxtlBC和ΔxtlABC，通过Southern blot验证突变体正确后，分析其对菌株生长速率、游动性、生物膜及生防相关性状等表型的影响。研究结果显示，与野生型菌株PtA1相比，突变体ΔxtlA、ΔxtlBC和ΔxtlABC在LB培养基中的生长速度变慢，在游动培养基上的迁移直径分别减少了12.6、12.5和13.5 mm，生物膜形成量分别降低了30.41%、29.16%和30.70%，对4种青枯菌的抑菌圈直径显著减小。盆栽试验表明，接种青枯菌GMI1000后4 d，处理组PtA1+GMI1000、ΔxtlA+GMI1000、ΔxtlBC+GMI1000和ΔxtlABC+GMI1000对番茄青枯病的防效分别为44.93%、9.49%、10.46%和7.66%，第10 d的防效分别为42.43%、19.56%、31.40%和24.79%，处理组PtA1+GMI1000防治效果最好，与xtlABC突变体处理组差异显著。综合分析表明xtlABC基因簇影响菌株自身的生长及生防相关性状，该基因及其合成的次生代谢产物在抑制青枯菌中起重要作用。

关键词: 摩氏假单胞菌, 环脂肽, 基因簇xtlABC, 青枯菌, 抑菌作用

Abstract: Previous studies have demonstrated that Pseudomonas mosseliiPtA1 displays high levels antibacterial activities against various strains of Ralstonia solanacearum. Its genome encompasses a complete xtlABC gene cluster, which is responsible for the production of cyclic lipopeptide. In order to explore the function of xtlABC gene cluster in P. mosseliiPtA1, we employed homologous recombination to create deletion mutants ΔxtlA, ΔxtlBC and ΔxtlABC, verified through Southern blot analysis, and assessed the phenotypic changes in growth rate, swimming motility, biofilm formation and antagonistic activity. The results revealed that compared with the wild-type strain PtA1, mutants ΔxtlA,ΔxtlBC and ΔxtlABCdisplayed a significant reduction in growth rate. The diameters of swimming rings were diminished by 12.6 mm, 12.5 mm and 13.5 mm, respectively, the amount of biofilm formation decreased by 30.41%, 29.16% and 30.70%, respectively, and the diameters of the antibacterial zone against R. solanacearum were also notably reduced. Pot experiments demonstrated that the control efficacy of treatment groups PtA1+GMI1000, ΔxtlA+GMI1000,ΔxtlBC+GMI1000 and ΔxtlABC+GMI1000 on tomato bacterial wilt were 44.93%, 9.49%, 10.46% and 7.66% 4 days post-inoculation with R. solanacearum GMI1000, respectively. The control efficacies were 42.43%, 19.56%, 31.40% and 24.79% 10 days post-inoculation, respectively. The treatment group PtA1+GMI1000 had the best control effect on tomato bacterial wilt, significantly different compared to the xtlABC mutant treatment groups. These findings underscore the impact of the xtlABC gene cluster on the growth and biocontrol-related characteristics of P. mosselii PtA1, and its secondary metabolite played an important role in inhibiting R. solanacearum.

Key words: Pseudomonas mosselii, cyclic lipopeptide, <xtlABC, Ralstonia solanacearum, antibacterial activity

中图分类号:

S476

郑汉维, 谢文松, 牛世霖, 张旭, 卓涛, 邱思鑫, 范晓静. 摩氏假单胞菌环脂肽合成基因簇xtlABC的功能研究[J]. 中国生物防治学报, 2024, 40(3): 620-629.

ZHENG Hanwei, XIE Wensong, NIU Shilin, ZHANG Xu, ZHUO Tao, QIU Sixin, FAN Xiaojing. Functional Study of the Gene Cluster xtlABC Producing Cyclic Lipopeptide in Pseudomonas mosselii PtA1[J]. Chinese Journal of Biological Control, 2024, 40(3): 620-629.

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