Trichoderma longibrachiatum with broad-spectrum antibacterial performance and application thereof
A technology of Trichoderma longibranches and strains, applied in the direction of application, fungicides, fungi, etc., can solve problems such as fertility decline, imbalance of soil nutrient ratio, environmental pollution, etc., and achieve increased germination length, significant salt tolerance, and significant control effect of effect
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Embodiment 1
[0018] Embodiment 1, the isolation and identification of Trichoderma longibrachiae strain MF-3
[0019] 1. Strain isolation and purification
[0020] The medium formula (w / v) used in the experiment of the present invention:
[0021] PDA medium: 20% potato, 2% glucose, 2% agar, natural pH.
[0022] PDB medium: potato 20%, glucose 2%, pH natural.
[0023] (1) Strain isolation
[0024] Soil samples were collected from the rhizosphere soil of non-infested plants in different farming areas. Using the 10-fold gradient dilution method, 10g of soil was weighed, mixed thoroughly and ground, then poured into a triangular flask filled with 90ml of sterile water, and fully shaken to mix. homogeneously, take the supernatant and dilute to a concentration of 10 -3 、10 -4 , 10 -5 、10 -6 soil suspension. The fungi were isolated using the dilution coating plate method, and 200 μL of the dilution was drawn with a pipette gun and coated on the Bengal Red solid medium plate. After culturi...
Embodiment 2
[0035] Embodiment 2, Trichoderma longibrachiae MF-3 growth-promoting ability assay
[0036] Choose plump cucumber seeds of the same size and soak them in 2% (m / V) sodium hypochlorite solution for 15 minutes, disinfect them, then clean them with deionized water, and carry out the seed dressing treatment of fermented liquid. Cucumber seeds were placed in the fermentation broth, and the control was treated with an equal volume of sterile fermentation broth, 28°C, 180r / min, shaking culture for 4 hours, then the seeds were taken out and placed in a 9cm petri dish. Put 2 layers of filter paper in the petri dish, soak it with sterile water, and finally cover the seeds with a layer of filter paper. Each plate contained 20 grains, each treatment was repeated 3 times, and cultured in the dark at 28°C for 72 hours, during which periodical supplementary sterile water was used to maintain moisture. The germination rate and germination length of the seeds were determined. The germination ...
Embodiment 3
[0039] Embodiment 3, the mensuration of Trichoderma longibrachiae MF-3 salt tolerance
[0040] Add different amounts of NaCl to 100mL of PDB liquid medium to configure medium with different salt concentrations (0%, 1%, 5% NaCl), and use a sterilized hole puncher on a plate covered with fungal hyphae Drill a hole, then pick up the agar piece with tweezers and put it into NaCl medium, culture with shaking at 28°C and 180r / min for 72h, and measure the OD600 value of the bacterial solution.
[0041] The results show that if image 3 As shown, as the NaCl concentration increased, the OD600 value of the strain did not decrease significantly, indicating that the strain had a certain salt-tolerant ability, and could resist Rhizoctonia solani, Fusarium oxysporum and Fusarium graminearum in salinized soil. It has great application potential in the control of broad-spectrum pathogenic fungi.
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