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The endophytic fungus strain hubu0122 of Dirt grass and its application

A HUBU0122, a technology of endophytic fungi, which is applied in the fields of application, fungi, biocides, etc., and can solve the problems of undisclosed endogenous microorganisms of Dieuthenia chinensis

Active Publication Date: 2022-03-29
HUBEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the prior art does not disclose that the related endophytic microorganisms

Method used

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  • The endophytic fungus strain hubu0122 of Dirt grass and its application
  • The endophytic fungus strain hubu0122 of Dirt grass and its application
  • The endophytic fungus strain hubu0122 of Dirt grass and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1. Isolation and purification of endophytic fungus HUBU0122

[0022] 1. Plant sample collection and pretreatment

[0023] The fresh and healthy whole plant of C. chinensis was collected from Qichun County, Huanggang City, Hubei Province in September 2019. It grows well and has no obvious diseases and insect pests. Rinse the healthy plant samples of C. chinensis with pure water or tap water to remove possible impurities on the surface of the sample tissue. After rinsing, cut off the poor quality tissue of the sample plant in time, cut into small pieces of tissue with an incision of about 0.5×1 cm and put them Blot dry on clean filter paper. After the surface liquid is blotted, carry out surface disinfection. Under the aseptic environment in the ultra-clean workbench, immerse and disinfect with 75% ultra-clean filtered ethanol for 3 minutes. Be careful not to directly contact the tissue incision with ethanol, and then use sterile ultra-pure The tissue sections w...

Embodiment 2

[0030] Example 2, identification of the endophytic fungus HUBU0122 of Dirtwort

[0031] The isolated and purified endophytic fungus HUBU0122 was inoculated in PDA medium, and cultured upside down in a constant temperature fungal incubator at 28°C for 7 days. After the fungal DNA was extracted, the extracted genomic DNA was subjected to PCR amplification. Then measure its ITS sequence, and compare the measured sequence with the known strain genome DNA sequence in the database to obtain the determined strain species.

[0032] 1. Extraction of endophytic fungus HUBU0122 DNA

[0033] Fungal genomic DNA was extracted using a fungal genomic DNA extraction kit (Cat. No. D2300, Solarbio). The whole extraction process was carried out at room temperature, specifically:

[0034] ① Take 50-100 mg of mycelia, add 200 μL of solution A, grind and disperse the mycelium with a glass grinder, add 20 μL of RNase A, and then add 100 mg of glass beads, shake on a high-speed oscillator for about 3...

Embodiment 3

[0056] Example 3, Antibacterial Activity Screening of Methanol Extracts from Endophytic Fungus HUBU0122 Metabolites

[0057] Resuscitation and activation of strains: inoculate the fungi stored in the refrigerator at 4°C into fresh PDA solid medium containing penicillin and streptomycin, place them in a 28°C fungal constant temperature incubator and cultivate them for 7 days to activate, and activate them twice to restore the fungi to normal growth.

[0058] Preparation of methanol extracts of strain metabolites: the strains were inoculated on fresh PDA plates containing penicillin and streptomycin, and cultured in a 28°C fungal incubator for 7 days. The cultured strains and their metabolites in the PDA medium were soaked in 200 mL of methanol for 6-8 hours, extracted three times, and ultrasonicated at 40 °C for 2-3 hours. The crude methanol extracts of each strain were filtered with a suction filter bottle, and the filtered filtrate was dried under reduced pressure using a ro...

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Abstract

The present invention relates to the endophytic fungus strain HUBU0122 of Dirtwort and its application. The endophytic fungus named Alternaria ( Alternaria sp. ) HUBU0122, whose strain preservation unit is the China Type Culture Collection Center, the preservation address is No. 299 Bayi Road, Wuhan City, Hubei Province, the preservation date is April 19, 2021, and the preservation number is CCTCC No.M2021414. The bacterium has antibacterial effect on Salmonella and Xanthomonas oryzae, and can be used as a utilization resource of plant endophytic fungi, laying the foundation for obtaining new agricultural antibiotic natural ingredients that resist plant pathogenic fungal diseases.

Description

technical field [0001] The invention relates to the technical field of endophytic fungus of Diomydia chinensis, in particular to endophytic fungus strain HUBU0122 of Diomynia chinensis and application thereof. Background technique [0002] Ground Ear Grass ( Hypericum japonicum Thunb. ex Murray), also known as vanilla and Tianjihuang, is one of the commonly used medicinal plants. Dirtgrass is widely distributed. In China, it is mainly distributed in Liaoning, Shandong and the provinces south of the Yangtze River. It grows on relatively humid land below 2,800 meters above sea level, including fields, ditches, grasslands, and wastelands. All its tissues and organs can be made into medicines, which can clear away heat and stop bleeding, detoxify and reduce swelling, and have significant effects on the treatment of chronic hepatitis, bruises and infectious sores. [0003] According to the latest research results of modern pharmacology, Dirtwort mainly contains a variety of fla...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14A01N63/30A01P1/00C12R1/645
CPCC12N1/14A01N63/30
Inventor 胡琳珍江雯胡萍童周胡天卉
Owner HUBEI UNIV
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