油菜根内生菌分离、鉴定及其植物益生作用筛选

    Isolation and Identification of Endophytic Bacteria from the Roots of Brassica Napus and the Screening for the Probiotic Effects of Plants

    • 摘要: 从油菜中分离鉴定根内生细菌菌株,检测评估其促生、抑菌潜力和机制,为开发具有植物促生和病害防控的微生物肥料提供依据。以油菜为研究对象,通过分离纯化和16S rRNA基因扩增分离和鉴定油菜根内生细菌,并进一步通过平板法、酶活法以及抑菌试验等分析分离其木制纤维素降解能力、促生作用以及抑菌活性等益生特性及其机理。结果表明:根据细菌菌落形态、颜色和大小,挑取纯化得到14株油菜根内生细菌菌株。经16S rRNA基因测序和鉴定分析,分别为假单胞菌、肠杆菌、芽孢杆菌、葡萄球菌和莱略特菌。在分离纯化的14株内生菌株中,除菌株Y-7、Y-8外都具有明显的木聚糖酶活性,除菌株Y-1、Y-7、Y-8、Y-9、Y-10外都具有纤维素分解能力,进一步对14株内生菌株的植物益生作用进行初步筛选和评价,得到产铁载体菌株Y-2、Y-4、Y-6、Y-10、Y-11、Y-13、Y-14,具有稳定固氮能力的固氮菌株Y-1、Y-2、Y-10、Y-13、Y-14。在拮抗内生菌筛选中,得到拮抗大肠杆菌菌株Y-1、Y-6、Y-7、Y-8、Y-12,拮抗金黄色葡萄球菌菌株Y-3、Y-4,拮抗枯草芽孢杆菌菌株Y-1以及拮抗黑曲霉菌株Y-3、Y-4、Y-6、Y-9、Y-10、Y-14。大多数分离的油菜根内生菌具有一定的木质纤维素降解能力。分离的油菜根内生菌具有广泛而多样性的植物益生特性,如促生长和抑菌活性,这些益生特性可能与其具有的产铁载体、产IAA以及固氮能力有关。结果表明油菜根内生菌可以作为筛选和开发具有植物促生和病害防控作用的微生物菌剂的重要来源。

       

      Abstract: The endophytic bacterial strains were isolated and identified from the roots of Brassica napus, and their growth promotion and antibacterial potential and mechanism were detected and evaluated, thus to provide the basis for the development of microbial fertilizers with the abilities of plant growth promotion and disease prevention. By taking Brassica napus as the research object, the endophytic bacteria in the roots of Brassica napus were isolated and identified by the isolation and purification and 16S rRNA gene amplification. The probiotic characteristics of lignocellulose, such as the degradation ability, growth-promoting effect and antibacterial activity, and its mechanism were further analyzed by using the plate method, enzyme activity method and antibacterial test. The results showed that 14 strains of endophytic bacteria were selected and purified from the roots of Brassica napus according to the morphology, color and size of bacterial colonies. By 16S rRNA gene sequencing and identification & analysis, they were Pseudomonas, Escherichia coli, Bacillus, Staphylococcus and Lelliottia. Among the 14 isolated and purified endophytic strains, all the strains except Y-7 and Y-8 had significant xylanase activity, and all the strains except Y-1, Y-7, Y-8, Y-9 and Y-10 had the cellulose decomposition ability. The preliminary screening and evaluation on the plant probiotic effects of the 14 endophytic strains were further carried out, and the iron carrier-producing strains including strain Y-2, Y-4, Y-6, Y-10, Y-11, Y-13 and Y-14 were obtained, and the azotobacters including strain Y-1, Y-2, Y-10, Y-13 and Y-14 with stable nitrogen fixation ability were also obtained. In the screening of antagonistic endophytic bacteria, the strains Y-1, Y-6, Y-7, Y-8 and Y-12 against Escherichia coli, the strains Y-3 and Y-4 against Staphylococcus aureus, the strain Y-1 against Bacillus subtilis and the strains Y-3, Y-4, Y-6, Y-9, Y-10 and Y-14 against Aspergillus niger were obtained. Most of the endophytic bacteria isolated from the roots of Brassica napus had certain ability of lignocellulose degradation. The endophytic bacteria isolated from the roots of Brassica napus exhibited a wide range of probiotic properties such as growth promotion and bacteriostatic activity, which might be related to their abilities to produce iron carriers & IAA and the nitrogen fixation ability. The results showed that the endophytic bacteria from the roots of Brassica napus could be used as the important source for screening and developing the microbial agents with the abilities of plant growth promotion and disease prevention and control.

       

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