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Extraction method and application of A.aequalis endophytic fungi as well as secondary metabolites of A.aequalis endophytic fungi

A technology of secondary metabolites and subspora, applied in the field of microorganisms, can solve problems such as endophytes that have not been reported yet, and no studies have shown endophytes, etc., to improve the problem of early sprouts and reduce the bolting rate. , the effect of increasing the content

Active Publication Date: 2022-07-05
WEST ANHUI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The microbial resources of Peucedanum are extremely rich, and new microorganisms have been isolated from them, but there is no report on the isolation of coumarin-producing endophytes from Peucedanum, and no research has shown that endophytic fungi can inhibit bolting of Peucedanum

Method used

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  • Extraction method and application of A.aequalis endophytic fungi as well as secondary metabolites of A.aequalis endophytic fungi
  • Extraction method and application of A.aequalis endophytic fungi as well as secondary metabolites of A.aequalis endophytic fungi
  • Extraction method and application of A.aequalis endophytic fungi as well as secondary metabolites of A.aequalis endophytic fungi

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Isolation and identification of endophytic fungi of P. chinensis

[0027] 1. Experimental materials

[0028] The root of Baihua Qianhu in Donghekou Qianhu Cultivation Base, Lu'an City, Anhui Province.

[0029] 2. Separation and purification

[0030] Cut the roots of the collected healthy and disease-free Qianhu plants with scissors, rinse with tap water, remove the sediment attached to the surface, and place them in an ultra-clean workbench to dry for surface disinfection: place them in 0.1% mercury chloride for 2-3 minutes, Rinse with sterile water for 4 to 5 times, dry with sterile gauze, soak in 75% ethanol for 2 to 3 minutes, rinse with sterile water for 6 to 7 times, and finally wipe off the water attached to the surface with sterile gauze. Take the sterile water from the last rinse and apply it to the PDA medium to test the disinfection effect on the surface of the sample.

[0031] Tissue sectioning method: Use sterile scissors to cut the surface-ster...

Embodiment 2

[0032] Example 2 Subspora genus endophytic fungi extract secondary strain-producing metabolites

[0033] 1. Preparation of the test product

[0034] The 49 strains of bacteria obtained in Example 1 were inoculated on the PDA medium for activation, and the activated strains were inoculated into the PDB medium, cultured at 28° C., 140 r / min shaker for 5-7 d, to obtain a strain fermentation product, Brucella. The funnel is vacuum-filtered to separate the mycelium and the fermentation broth, rinse the mycelium with distilled water, dry the mycelium at 50±2°C, and grind to obtain the mycelium powder. Precisely add dichloromethane at a weight (g) volume ratio (mL) of 1:20, seal it, ultrasonically treat it for 10 minutes, filter, evaporate to dryness in a 50°C water bath, rinse the residue with methanol, and filter with a 0.22 μm filter membrane to obtain mycelium Extract.

[0035] 2. Chromatographic conditions

[0036] ZORBAX Eclipse Plus C18 (150mm×4.6mm, 5μm) chromatographic co...

Embodiment 3

[0038] Example 3 LC-MS determination of strain secondary metabolites.

[0039] 1. Chromatographic conditions

[0040]C18 chromatographic column: ZORBAX Eclipse plus C18 column (2.1×100 mm, 1.8 μm, Agilent, USA); mobile phase: eluent A is water, and eluent B is methanol. The flow rate was 0.25 ml / min, and the injection volume was 2 μL. Column temperature: 30°C. The gradient program is: 0~3.00min, 30→75%B; 3.00~15.00min, 75%B; 15.00~16.00min, 75→100%B; 16.00~20.00min, 100%B; 20.00~21.00min, 100 →30%; 21.00~22.00min, 30%B.

[0041] 2. Mass spectrometry conditions

[0042] Electrospray ion source (ESI), positive ion scanning; ion source temperature 325°C; capillary voltage 3500V, collision energy 30V, fragmentation voltage 175V, the reference ion of the standard solution is located at m / z [M+Na] + : Amphetamine: 409.1263, Coumarin I: 411.142, Amphetamine: 449.1577 and Amphetamine E: 451.1733.

[0043] 3. The secondary metabolites of the strains of Example 2 were injected int...

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Abstract

The invention belongs to the technical field of microorganisms, and discloses an extraction method and application of A.aequalis endophytic fungi and secondary metabolites of the A.aequalis endophytic fungi. The bacterial strain is named as Didymeria segeticola Q1-5, and the preservation number of the bacterial strain is CCTCC (China Center for Type Culture Collection) NO.M2022032. The invention further discloses a preparation method of the bacterial strain. The Didymellasegetum is an endophytic fungus separated from peucedanum praeruptorum roots, secondary metabolites of the strain contain peucedanum praeruptorum A, peucedanum praeruptorum B and peucedanum praeruptorum E, bolting of the peucedanum praeruptorum can be effectively inhibited, the problem of early bolting of the peucedanum praeruptorum can be solved, and improvement of the quality of the peucedanum praeruptorum medicinal material is facilitated.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to an extraction method and application of an endophyte of the genus Subspora and its secondary metabolites. E element, can inhibit the front bolt bolting. Background technique [0002] Endophytes are a class of microorganisms that can survive in healthy plant tissues at a certain stage without causing obvious pathological changes in host plants, mainly including bacteria and fungi. Endophytes play an indispensable role in the growth and development of plants, and more and more scholars focus on the study of plant-related microorganisms. [0003] Qianhu, the dry root of the Umbelliferae plant Baihua Qianhu, was first recorded in "Famous Doctors", has the effect of antipyretic and phlegm, and is generally used to treat certain respiratory diseases, such as wind-heat cold, cough with excessive phlegm, Asthma etc. Coumarins are the main chemical constituents of genitalia, mainly includ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P17/06C12P17/18A01N63/30A01N43/90A01P21/00C12R1/645
CPCC12N1/14C12P17/06C12P17/181A01N63/30A01N43/90Y02A50/30
Inventor 韩邦兴刘丽马猛华汪学军朱富成王威王芳宋向文王广林陈存武
Owner WEST ANHUI UNIV
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