| Nowadays the white rot has become one of the worst diseases of Red Globe in the major grape-producing areas in China, and its incidence is increasing year by year, which directly affect the quality and yield of Red Globe.Aimed at exploring a green and efficient, and sustainable control metheods towards white rot of Red Globe, the present study was undertaken in broad-spectrum antagonistic strain PT2's screening, identification,antifungal inhibitory action, and mixed microbial fermentation research, optimization of fermentation medium process and field experiment.Based on the basic properties observation including cell shape, Gram'stain, sporulation, production of catalase and others chemic characteristics, PT2 was identified as Bacillus spp., and further identified as Bacillus subtilis by the sequences analysis of 16SrRNA. Affirm the strain's antagonistic range initially by doing flat plate inhibition tests of PT2 on Coniella diplodiella (Speq.) Petrak & Sydow..The Bacillus subtilis strain PT2, which has an antagonistic ability on Coniella diplodiella (Speg.) Petrak & Sydow.was irradiated respectively by UV, He-Ne laser and the mutant strains UV10 and HN14 was obtained, both of them to increase the antagonistic ability by 15.0% and 21.7% compared with PT2. The results indicated that the coordinative effect of UV and He-Ne laser was better than using them individually. The cells were treated firstly by UV for 90 seconds and then by He-Ne laser for 10 minutes in 15mW. The best positive mutation strain PT21 was obtained, whose antagonistic ability is 30.9% higher than PT2's. By cultivating it from 1st to 25th generation, its antagonistic ability was proved stable in heredity.Make PT21 mix culture with another broad-spectrum antagonistic Bacillus subtilis QM34. Try to cultivate PT2 and QM34 together through plate crossing streaking, fluid shake flask cultivation, observing the results of A660 and the variation of antagonistic ability after the mixed fermentation. The results show that, PT21 and QM34 can coexist in the same growth condition, and there are complementary effects of increasing their antagonistic abilities at the same time. By improving the ratio of components of corn flour fluid culture to fumble out a industrial fermentation culture fit for the mixed strains PT21 and QM34. Considering the results of A66o and inhibition zoneΦtwo indicators, the orthogonal experimental of L18(3)6 was designed. According to results of range analysis and variance analysis, the optimal fermentation conditions are A2B2C2D2E1F2.The advanced multilevel fermentation process obtained through the patent "Marc fermented biochemical fulvic acid compound microbial fertilizer technology", which was conducted to produce high quality biofertilizer containing biotechnology fulvic acid(BFA) through fermenting apple pomace. Make biofertilizer containing BFA combine reasonably with compound antagonistic agents to produce white rot disease-specific biochemical fulvic acid compound microbial fertilizer. Field experimengt results show that the compound microbial fertilizer can effectively control white rot of Red Globe, and it has a satisfactory prevention effect as well as its manorial effect. Compared with the control group, the average chlorophyll content of leaves of the experiment group increased 21.2%, the incidence of disease decreased 67%, and the sensory evaluation of fruits, residues of chemical pesticides and heavy metals were in all accord with national requeirements of food. |
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