甘肃贝母生防菌分离及其对腐烂病防治的研究

Isolation of Biocontrol Microbes from Fritillaria przewalskii and Its Prevention and Treatment of Rot Disease

目的:从甘肃贝母中分离生防菌,并进行腐烂病防治研究,为甘肃贝母及其他作物腐烂病的防治提供一定的参考。方法:通过常规方法分离鉴定内生菌,无氮培养基培养定性检测微生物固氮活性,平板拮抗试验检测抗病性,LC-MS检测蒂莫内马赛菌Massilia timonae激素水平,分光光度法检测酶活,盆栽试验检测不同微生物对贝母腐烂病防治的影响。结果:从甘肃贝母中分离得到解淀粉芽孢杆菌Bacillus amyloliquefaciens、荧光假单胞菌Pseudomonas fluorescens、嗜根寡养单胞菌Rhizophobic oligomonas、粉红螺旋聚孢霉Clonostachys rosea及互生细菌蒂莫内马赛菌;粉红螺旋聚孢霉具有固氮和拮抗镰刀菌的作用,蒂莫内马赛菌单位浊度菌液生长素类物质含量为8.89 ng/mL,细胞分裂素类物质含量为0.55 ng/mL,赤霉素类物质含量为13.51 ng/mL,显著促进粉红螺旋聚孢霉菌球的生长;蒂莫内马赛菌单位浊度菌液具有很高的几丁质酶、木质素酶、纤维素酶活性,其活性分别达到了0.28 mg/mg·h prot、7.33 U/mg prot和27.41μg/mg·min prot,增强了粉红螺旋聚孢霉在植物中的定殖能力。结论:蒂莫内马赛菌与粉红螺旋聚孢霉的混合物对甘肃贝母腐烂病的相对防治效果达到92.64%,效果优于解淀粉芽孢杆菌。

Objective:To isolate biocontrol microbes from Fritillaria przewalskii and study the prevention and treatment of rot disease,so as to provide some reference for the prevention and treatment of rot disease in Fritillaria przewalskii and other crops.Methods:Conventional methods were used to isolate and identify endophytes,nitrogen free culture medium was used to qualitatively detect the nitrogen-fixing activity of microorganisms,the disease resistance was detected by plate antagonism test.LC-MS was used to detect the hormone levels of Massilia timonae,spectrophotometry was used to detect the enzyme activity,pot experiment was used to detect the influence of different microorganisms on the prevention and treatment of Fritillaria przewalskii rot disease.Results:Bacillus amyloliquefaciens,Pseudomonas fluorescens,Rhizophobic oligomonas,Clonostachys rosea and alternant bacteria Massilia timonae were isolated from Fritillaria przewalskii,Clonostachys rosea could fix nitrogen and antagonize fusarium,The contents of auxin substance in bacterial fluid per unit turbidity,cytokinin,gibberellin in Massilia timonae were 8.89,0.55,13.51 ng/mL,respectively,which could significantly promote the growth of Clonostachys rosea.The unit turbidity of Massilia timonae showed high activities of chitinase,ligninase and cellulose,reaching 0.28 mg/mg·h prot,7.33 U/mg prot and 27.41μg/mg·min prot,respectively,which enhanced the colonization ability of Clonostachys rosea in plants.Conclusion:The relative control effect of the mixture of Massilia timonae and Clonostachys rosea on rot disease in Fritillaria przewalskii is 92.64%,which is better than that of Bacillus amyloliquefaciens.