Application of streptomyces diastatochromogenes metabolite in prevention and treatment of black spot and anthracnose of Zhejiang ophiopogon japonicus
A technology for producing Streptomyces chromogenes and metabolites, applied in the field of microorganisms, can solve problems such as insecurity, and achieve the effect of good environmental compatibility
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Embodiment 1
[0017] Embodiment 1: (the liquid fermentation of amylase Streptomyces chromogenicum tz3-1)
[0018] Streptomyces diastatochromogenes tz3-1 was obtained from Streptomyces diastatochromogenes D through ribosome engineering. For details, please refer to the applied invention patent (application number: 2019114179541).
[0019] Fermentation broth contains 20g of soluble starch per 1000mL, CaCO 3 5g, KH 2 PO4 3g, NH 4 Cl 3g, FeCl 2 1g, 43g of soybean powder, 4g of yeast extract, 6g of malt extract, and the balance is water; 60mL of fermentation culture broth is packed in each 300mL triangular bottle, sterilized at 121°C for 20min after preparation, cooled to 50°C, and picked under aseptic conditions 1. Platinum cycloamylase Streptomyces chromogenes D or corresponding mutant strain inoculation, 28℃±1℃, fermented for 5 days; the fermentation broth was centrifuged at 5000r / min for 10min, and the supernatant was filtered through a 0.22μm membrane for antibacterial activity Determ...
Embodiment 2
[0020] Embodiment 2: (Amylase Streptomyces chromogenes tz3-1 metabolite antibacterial activity assay)
[0021] Take 1mL of the fermentation supernatant filtered through a 0.22μm membrane in a sterile petri dish, and quickly mix it with 9mL of PDA medium cooled to 50°C. After cooling, place a test bacteria with a diameter of 5mm on the medium plane The fungus cake, the fungus cake is connected to the center of the petri dish (the diameter of the petri dish is 9cm), placed in an incubator for 36 hours at 28°C, and treated with sterile water as a control, repeated 3 times; the diameter of the colony is measured by the cross method, and the following formula is used: Calculation of inhibition rate: Mycelia growth inhibition rate (%)=[(control colony diameter-5)-(treatment colony diameter-5)] / (control colony diameter-5)×100%.
[0022] The results showed that the metabolites of amylase Streptomyces diastatochromogenes tz3-1 had good inhibitory effects on the mycelium growth of the t...
Embodiment 3
[0026] Example 3: (Field Control Effect Measurement of Amylase Streptomyces Chromogenes tz3-1 Metabolites)
[0027] Select 100%, 50% amylase Streptomyces chromogenes tz3-1 metabolites and 50% carbendazim as the test agents for field control efficacy determination, clean water is the negative control, and each treatment field is treated with clean water The negative control was used as an interval field. On the 7th day, the incidence was investigated, the disease index was counted, and the corrected control effect was calculated. According to the severity of the disease, it is divided into 4 grades: grade 0, healthy seedlings; grade 1, at least 25% of the leaves are dead; grade 2, 26% to 50% of the leaves are dead; grade 3, 51% to 75% of the leaves are dead; level, 76% to 100% of the leaves are dead. Disease index=[∑(number of diseased plants at all levels×representative value corresponding to each level) / (total number of plants under investigation×representative value of the...
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