首页|烟草青枯病拮抗菌的高通量筛选及其复配菌群防效评估

烟草青枯病拮抗菌的高通量筛选及其复配菌群防效评估

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  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
构建烟草青枯病的高效生防菌群对生物防控病害至关重要。本研究从高抗烟草青枯病的品种岩烟97根际高通量分离细菌，通过平板喷雾对峙法和趋化特异性检测筛选出趋化性的青枯菌拮抗细菌，采用启发式算法选取拮抗单菌株并组合成生防菌群。通过盆栽试验评估复配菌群对感病烤烟品种红花大金元的抗病效果，利用荧光定量PCR技术检测其对根际土壤中的青枯菌的消减，通过Biolog碳源芯片技术分析该菌群对烟草根际微生态的影响。结果表明，共筛选到青枯菌拮抗菌109株，其中抑菌圈直径超过20mm的菌株25株，从中优选了 3株相互之间具备相容性、具有趋化基因CheA与生物膜形成能力的拮抗菌，组合成复配菌群(Y364+Y832+Y878)。16sDNA分子鉴定表明，Y364、Y832和Y878分别为烟磺假单胞菌(Pseudomonas nicosulfuronedens)、德里假单胞菌(Pseudomonas delhiensis)和高原芽胞杆菌(Bacillus altitudinis)。最终盆栽试验表明，复配菌群防效为79。75％，比单一菌株平均防效高36。22个百分点，发病率较Y364、Y832、Y878处理分别降低33。33、45。68和35。80个百分点，根际土壤青枯菌的数量显著低于CK，对根际土壤微生物功能多样性增加最大。该复配菌群具有潜在的协同增效和根际定殖能力，可为烟草青枯病的生物防治提供更多的生防资源。
High-throughput Screening of Antagonistic Bacteria against Tobacco Bacterial Wilt and Biocontrol Effects of Compound Microbial Agents
It is crucial to construct high-efficiency compound microbial agents for the biological control of tobacco bacterial wilt.Using the tobacco variety Yanyan-97 with high resistance to tobacco bacterial wilt pathogen,Ralstonia.solanacearum,a high-throughput screening of tobacco rhizo-bacteria was performed.R.solanacearum chemotactic antagonistic bacteria to rhizo-bacteria were screened by plate confrontation assay and chemotaxis gene PCR detection.The antagonistic single strains were selected by heuristic algorithms and combined to form biocontrol microbial consortia.The efficacy of consortia on disease resistance with susceptible variety of Honghuadajinyuan was validated by pot experiments.Fluorescence quantitative PCR was utilized to detect R.solanacearum quantities in the rhizosphere soil.Biolog carbon chip technology were used to analyze the impact of the assembled microbial consortia on tobacco rhizosphere microbiome.In total,109 antagonistic bacteria were screened.The inhibition zone diameter of 25 strains was over 20 mm.Three mutually compatible antagonistic strains possessing chemotaxis genes(CheA)and biofilm formation ability were selected and combined to synthesize the compound microbial consortia(Y364+Y832+Y878).Y364,Y832 and Y878 were identified as Pseudomonas nicosulfuronedens,Pseudomonas delhiensis,and Bacillus altitudinis,respectively by 16SrDNA gene sequencing.Ultimately,pot experiments showed that the disease control efficacy of compound microbial agent treatment was 79.75％and 36.22 percentage points higher than that of individual strain treatment.The disease incidence of compound microbial agent treatment was 33.33,45.68 and 35.80 percentage points lower than that of single Y364,Y832 and Y878 treatments,respectively.It also decreased the population of R.solanacearum and increased rhizosphere microbial functional diversity in rhizosphere soil.In conclusion,the composite consortium possesses potential synergistic effects and rhizosphere colonization ability,providing more biocontrol resources for the biological control of tobacco bacterial wilt.

tobacco bacterial wilthigh-throughput screeningcompound microbial agentsBiolog carbon chip technologybiocontrol

陶正朋、刘艳霞、李想、彭宇、李寒、陈丽萍、徐健、王克敏、穆东升、王致尧、蒋天平、宗仪

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贵州大学烟草学院,贵阳 550000

贵州省烟草科学研究院,贵阳 550000

中国烟草总公司贵州省公司,贵阳 550000

江苏中烟工业有限责任公司,南京 210000

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烟草青枯病 高通量筛选 生防菌复配 Biolog碳源芯片技术 生物防控

国家自然科学基金中国烟草总公司科技项目院士工作站资助项目中国烟草总公司贵州省公司科技项目中国烟草总公司贵州省公司科技项目中国烟草总公司贵州省公司科技项目中国烟草总公司贵州省公司科技项目

31860597110202202015黔科合平台人才[2020]40042019052021XM182022XM032023XM41

2024

10.13496/j.issn.1007-5119.2024.02.005

中国烟草科学
中国农业科学院烟草研究所 中国烟草总公司青州烟草研究所

中国烟草科学

CSTPCD北大核心
影响因子：1.318
ISSN：1007-5119
年,卷(期)：2024.45(2)
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