Trichoderma viride fungi and preparation and application of fungicide thereof
A technology of Trichoderma viridans and fungi, applied in the direction of application, fungicides, fungi, etc., to achieve the effect of broad antibacterial spectrum, significant control effect, and good antibacterial effect
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Embodiment 1
[0021] Embodiment 1: the isolation and screening of Trichoderma viride ZBS6 bacterial strain
[0022] (1) Separation and Purification of Trichoderma The collected soil sample was dried in the shade and ground, weighed 5 g, and put it into a 150 ml Erlenmeyer flask filled with 45 ml of normal saline. Using the soil dilution separation method, draw 1mL of the diluent and add it to a test tube containing 9mL of water, shake and mix to prepare 10 -2 Soil dilution. Take 0.1ml of the soil dilution and spread it on the PDA medium containing sodium propionate and streptomycin sulfate, and incubate it upside down in a constant temperature incubator at 25°C for 3 days. The isolated Trichoderma colonies were picked, separated and purified from single spores, and then transferred to PDA slant medium. After the mycelia and spores covered the slant, sterilized paraffin oil was added for storage for later use.
[0023] (2) The tested pathogens include Fusarium graminearum, Fusarium oxyspor...
Embodiment 2
[0031] Embodiment 2: Identification of ZBS6 bacterial strain
[0032] (1) Morphological classification and identification Trichoderma was identified by colony morphology observation and microscopic observation, and its species was determined by searching literature.
[0033] (2) Molecular identification Antagonistic Trichoderma genomic DNA was extracted with reference to the DNA extraction method of Wu Fahong and other fungi, and extracted with ddH 2 O dissolved DNA to 500ng / μl. The upstream and downstream primers used to amplify the ITS sequence of Trichoderma were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd., respectively ITS4: 5'-GGAAGTAAAAGTCGTAACAAGG-3' and ITS6: 5'-TCCTCCGCTTATTGATATGC-3'. Using the genomic DNA of the antagonistic Trichoderma as a template, PCR amplification was performed using primers ITS4 and ITS6. The PCR reaction system is 25μl: 10×PCR Buffer 2.5μl, template 1μl, dNTP (10mM each) 0.5μl, primer ITS4 (10mM) 1μl, ITS6 (10...
Embodiment 3
[0045] Embodiment 3: the preparation of a kind of microbial bacterial agent
[0046] (1) Preparation of liquid bacterial agent Transfer the preserved Trichoderma viride strain ZBS6 onto a PDA medium plate, culture in alternating light and dark at 25°C for 4 days, scrape conidia under aseptic conditions, and prepare 1 × 10 with sterile water. 7 The conidia suspension was inoculated according to 1ml spore suspension / 100ml PDB medium, placed at 25°C and shaken at 150rpm for 3 days, stirred with an electric stirrer at 2000rpm for 2min, and mixed with 1.9L phosphate buffer (inner Containing 0.1% auxiliary agent Tween 100 and 0.2% preservative benzoic acid), the liquid biocontrol agent can be obtained.
[0047] (2) Preparation of solid bacterial agent Transfer the preserved Trichoderma viride strain ZBS6 onto a PDA medium plate, culture in alternating light and dark at 25°C for 4 days, scrape conidia under aseptic conditions, and prepare 1×10 with sterile water. 7 The conidia suspe...
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