A strain of Trichoderma citrus viride and its use
A technology of Trichoderma citrus viridans and uses, which can be applied to microorganisms, Trichoderma citrus viridans and in the field of plant disease biological control, and can solve the problems of chemical pesticide environmental pollution and the like
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Embodiment 1
[0036] Isolation, purification, identification and preservation of the bacterial strain of embodiment 1
[0037] 1. Single spore isolation and purification of Trichoderma strains
[0038] Drop 75% ethanol and sterile water into the groove of a sterile cuvette, pick a mature subsea and place it in the ethanol in the cuvette for surface disinfection, rinse with sterile water, and rinse with sterile water. Crush the ascus with tweezers, pick the ascus shell containing the ascus into another cuvette, further crush the ascus, stir well, spread the suspension evenly on the water agar medium, and incubate at room temperature. Place them under a dissecting microscope for observation every other day, pick out germinated single spores with a sterile dissecting needle, transfer them to PDA medium containing ampicillin (2mg / L), and place them in dark culture at 25°C until the growth of Trichoderma After the ascospores germinate and colonies are formed, transfer the actively growing part ...
Embodiment 2
[0056] Embodiment 2 confrontation culture method
[0057] Trichoderma aeruginosa P-T 7987 and five target plant pathogens (Botrytis cinerea, Sclerotinia sclerotiorum, Pythium melonum, Phytophthora capsici, Rhizoctonia solani) were activated and cultured on PDA medium, and then samples were taken from the edge of the colony Bacteria blocks with a diameter of 5 mm were inoculated with Trichoderma aurantiaceae P-T 7987 and plant pathogens at symmetrical positions 15 mm from the edge of the dish on both sides on the diameter line. Incubate at a constant temperature of 25°C, and observe the growth of the strains daily. When the petri dish is covered with the control, measure the control growth amount (colony radius) and the treatment growth amount (inhibition growth radius after inoculation of Trichoderma) of the target bacteria, and calculate the bacteriostatic rate.
[0058] Bacterial inhibition rate (%)=(control growth amount-treatment growth amount) / control growth amount×100% ...
Embodiment 3
[0062] Example 3 Heavy Parasitic Observation
[0063] Using the glass slide culture method, use a sterilized scalpel to cut out a 15×10mm PDA film, place it in the center of a sterilized glass slide, and then pick the same amount of hyphae of Trichoderma aurantii P-T 7987 and plant pathogen Inoculate at the midpoint of the two parallel sides of the PDA film, culture at a constant temperature and humidity at 25°C, and observe the interaction between the two bacteria with a microscope every day.
[0064] The reparasitic process of Trichoderma auranthoderma P-T 7987 on plant pathogens was observed by slide culture method combined with optical microscope. The hyphae of Trichoderma auranthoderma P-T 7987 can recognize and tend to grow on five plant pathogens, and form mycelium The adsorption structure of tendrils attaches and gradually wraps around the hyphae of plant pathogens, and then dissolves the host cell wall through the secretion of various enzymes and secondary metabolites...
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