Trichoderoma koningii TC-72 and application of trichoderoma koningii TC-72 to biological control of aspergillus flavus
A technology of Trichoderma pseudoconnei, TC-72, applied in microorganism-based methods, chemicals for biological control, applications, etc., can solve chemical resistance and even drug resistance, high cost, easy to pollute the environment And other issues
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Embodiment 1
[0048] Example 1: Solid fermentation culture of Trichoderma TC-72 strain, the following are weight percentages.
[0049] The Trichoderma TC-72 strain involved in the present invention has an accession number of CCTCCNO: M2016015 and a classification named TrichoderomakoningiiTG-72. It is isolated from the peanut root system in Hezhou, Guangxi. It is found through indoor confrontation antagonism experiment, in vivo anti-toxin experiment and field anti-toxin experiment. It is found that the strain has strong antibacterial effect, good antibacterial and detoxification effect and stable, easy to cultivate, no pollution, environment safety.
[0050] The solid fermentation culture and preparation of Trichoderma TC-72 strain are as follows:
[0051] ⑴Slope strain: inoculate the spores or hyphae of Trichoderma TC-72 strain in PDA medium, and cultivate for 5 days at 25-28℃.
[0052] ⑵Eggplant bottle culture: Inoculate the Trichoderma in the test tube slant medium to the PDA medium in the eggp...
Embodiment 2
[0057] Determination of Inhibition Rate of Metabolites of Trichoderma TG-72 Strain
[0058] Preparation of biocontrol bacteria metabolites
[0059] After the biocontrol bacteria TG-72 is activated on the PDA slope, take a loop in the PD culture solution, 100mL / 300mL filling volume, 30℃, 160r / min shaking culture for 72h, prepare the following treatment solution: supernatant: The culture solution was centrifuged at 12000 r / min for 15 minutes, the supernatant was taken, and filtered with a 0.22 μm bacterial filter to obtain a sterile supernatant.
[0060] Bacterial suspension: The culture solution was centrifuged at 12000r / min for 15min, the supernatant was discarded, washed with sterile water for 3 times and then centrifuged, and sterile water was added.
[0061] Crude protein extract: The culture solution was separated at 12000r / min for 15min at 4℃ to discard the precipitate, solid ammonium sulfate was added to the supernatant to 70% saturation, and the solution was allowed to stand ov...
Embodiment 3
[0069] Root colonization experiment of Trichoderma TC-72 strain
[0070] Peanut 8252 is germinated indoors, and germinated seeds with consistent growth are selected for sowing, 10 seeds per pot, and repeated five times. After the two cotyledons of peanut seedlings are flattened, inoculation begins. The concentration of the spore suspension of Trichoderma TC-72 strain is 2×10 6 / mL, the spore suspension was injected into the rhizosphere soil with a syringe, 15mL per hole, and clean water was used as a control. After 4 weeks, the TSM medium was used to determine the Trichoderma TC-72 strain in the rhizosphere soil of the potato plants, which was used as an indicator of the Rhizosphere colonization ability of Trichoderma fungi. The rhizosphere soil of peanut plants not inoculated with Trichoderma was used as a control when counting. The results are shown in Table 6, which shows that whether the strain TG-72 was inoculated alone or mixed with Aspergillus flavus, the survival rate a...
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