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粉红螺旋聚孢霉与枯草芽胞杆菌共培养发酵滤液生防与促生活性

  


Biocontrol and Growth-promoting Activities of Co-culture Fermentation Filtrate of Clonostachys rosea and Bacillus subtilis


摘要: 粉红螺旋聚孢霉是一种重要的植病生防真菌。本文研究了不同营养条件下粉红螺旋聚孢霉67-1与枯草芽胞杆菌B006共培养发酵滤液的生物活性和生防作用。采用平板培养测定共培养发酵滤液对大豆菌核病菌菌核形成,以及甘蓝枯萎病菌和西瓜枯萎病菌菌丝生长的影响;采用6孔组织培养板测定对根结线虫2龄幼虫的抑杀活性;通过平皿测定和温室盆栽试验研究发酵滤液对黄瓜种子萌发及幼苗生长的影响。结果表明,不同营养条件下粉红螺旋聚孢霉与枯草芽胞杆菌共培养发酵滤液能够显著降低立枯丝核菌菌核数量和尖孢镰孢菌生物量,其中对甘蓝枯萎病菌生长的抑制率提高40%以上,葡萄糖/硫酸铵培养基中发酵滤液抑菌率比粉红螺旋聚孢霉67-1和枯草芽胞杆菌B006分别提高了86.6%和62.7%。共培养发酵滤液杀线活性显著提高,玉米粉/豆粕粉和葡萄糖/硫酸铵培养基中真菌-细菌共培养无菌滤液处理后线虫死亡率达到65%以上,分别比菌株67-1发酵滤液提高了37.0%和50.8%,比菌株B006提高79.1%和94.7%。黄瓜种子浸种后活力指数、幼苗株高及地上部鲜重增加7.4%~30.4%。本研究为提高粉红螺旋聚孢霉防病效果、研发高效生防新产品提供了新思路。

关键词:

共培养, 粉红螺旋聚孢霉, 枯草芽胞杆菌, 拮抗作用, 促生, 杀线活性

Abstract: Clonostachys rosea is an important biocontrol fungus against many kinds of plant diseases. In this paper, the biological characters and biocontrol activities of the fermentation filtrates of C. rosea 67-1 and Bacillus subtilis B006 co-cultured in different nutritional media were investigated. The effects of the filtrates on sclerotial formation of Rhizoctonia solani and hyphal growth of Fusarium oxysporum f. sp. conglutinans and F. oxysporum f. sp. niveum were determined on agar plates. The nematicidal activities of the filtrates to the juveniles of root-knot nematode were tested by using 6-well tissue culture plates. The effects of the co-culture filtrates on germination of cucumber seeds and the growth of seedlings were determined in lab and in greenhouse. Results showed that the co-cultured fermentation filtrates of C. rosea and B. subtilis significantly reduced the sclerotia formation ofR. solani and the biomass of F. oxysporum, in which the inhibition rates of the growth of F. oxysporum f. sp. conglutinansincreased by over 40%; in the media of glucose/ammonium sulfate the rates decreased by 86.6% and 62.7% compared to the strain 67-1 and B006, respectively. The nematicidal activities of the co-cultured filtrates significantly increased.  After treated with the fermentation filtrates co-cultured in the media of corn/soybean flour and glucose/ammonium sulfate, the mortality rates of the juveniles reached more than 65%, which were 37.0% and 50.8% higher than those in 67-1 filtrates and 79.1% and 94.7% higher than B006, respectively. After soaked in the co-culture filtrates, the vigor index of cucumber seeds, seedling length and root fresh weight increased by 7.4-30.4%. The research provides new insight into the improvement of control efficiency of C. roseaand the development of new highly efficient biocontrol agents.

Key words:

co-culture; Clonostachys rosea, Bacillus subtilis, antagonism, growth-promoting, nematicidal activity