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Method for disinfecting surface of cinnamomum longepaniculatum and separating and culturing endophytic fungi of cinnamomum longepaniculatum

An endophytic fungi, isolation and culture technology, applied in the direction of fungi, etc.

Inactive Publication Date: 2021-02-12
YIBIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there are no relevant research reports on the surface disinfection of different tissues of camphor and the isolation of endophytic fungi at home and abroad.

Method used

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  • Method for disinfecting surface of cinnamomum longepaniculatum and separating and culturing endophytic fungi of cinnamomum longepaniculatum
  • Method for disinfecting surface of cinnamomum longepaniculatum and separating and culturing endophytic fungi of cinnamomum longepaniculatum
  • Method for disinfecting surface of cinnamomum longepaniculatum and separating and culturing endophytic fungi of cinnamomum longepaniculatum

Examples

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Embodiment 1

[0027] A preferred embodiment of the present invention provides a method for disinfecting the surface of camphor oil and isolating and cultivating endophytic fungi, the specific steps are as follows:

[0028] S1. Select mature healthy camphor, collect the roots, stems, and leaves respectively, and rinse the roots with sterile water for 3 times, soak in 95% ethanol for 1 min, shake and wash with 5 wt% sodium hypochlorite for 5 min, soak in 95% ethanol for 1 min, and wash with sterile water. Rinse 5 times for disinfection, the stems were rinsed 3 times with sterile water, soaked in 95% ethanol for 1 min, oscillatingly washed with 4wt% sodium hypochlorite for 5 min, soaked in 95% ethanol for 1 min and rinsed with sterile water 5 times for disinfection, and the leaves were disinfected in turn. Rinse with sterile water for 3 times, soak in 95% ethanol for 1 min, shake and wash with 4wt% sodium hypochlorite for 4 min, soak in 95% ethanol for 1 min, and rinse with sterile water for 5 ...

Embodiment 2

[0031] A preferred embodiment of the present invention provides a method for disinfecting the surface of camphor oil and isolating and cultivating endophytic fungi, the specific steps are as follows:

[0032] S1. Select mature healthy camphor, collect the roots, stems, and leaves respectively, and rinse the roots with sterile water for 3 times, soak in 95% ethanol for 1 min, shake and wash with 5 wt% sodium hypochlorite for 5 min, soak in 95% ethanol for 1 min, and wash with sterile water. Rinse 5 times for disinfection, the stems were rinsed 3 times with sterile water, soaked in 95% ethanol for 1 min, oscillatingly washed with 4wt% sodium hypochlorite for 5 min, soaked in 95% ethanol for 1 min and rinsed with sterile water 5 times for disinfection, and the leaves were disinfected in turn. Rinse with sterile water for 3 times, soak in 95% ethanol for 1 min, shake and wash with 4wt% sodium hypochlorite for 4 min, soak in 95% ethanol for 1 min, and rinse with sterile water for 5 ...

Embodiment 3

[0035] A preferred embodiment of the present invention provides a method for disinfecting the surface of camphor oil and isolating and cultivating endophytic fungi, the specific steps are as follows:

[0036] S1. Select mature healthy camphor, collect the roots, stems, and leaves respectively, and rinse the roots with sterile water for 3 times, soak in 95% ethanol for 1 min, shake and wash with 5 wt% sodium hypochlorite for 5 min, soak in 95% ethanol for 1 min, and wash with sterile water. Rinse 5 times for disinfection, the stems were rinsed 3 times with sterile water, soaked in 95% ethanol for 1 min, oscillatingly washed with 4wt% sodium hypochlorite for 5 min, soaked in 95% ethanol for 1 min and rinsed with sterile water 5 times for disinfection, and the leaves were disinfected in turn. Rinse with sterile water for 3 times, soak in 95% ethanol for 1 min, shake and wash with 4wt% sodium hypochlorite for 4 min, soak in 95% ethanol for 1 min, and rinse with sterile water for 5 ...

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Abstract

The invention discloses a method for disinfecting the surface of cinnamomum longepaniculatum and separating and culturing endophytic fungi of cinnamomum longepaniculatum, and belongs to the technicalfield of fungus separation and culture. The method comprises the following steps: selecting mature healthy cinnamomum longepaniculatum, respectively collecting roots, stems and leaves, and respectively and sequentially rinsing with water 3-5 times, soaking with 95% ethanol 1-2 minutes, oscillating and washing with 4-5wt% sodium hypochlorite 4-5 minutes, soaking with 95% ethanol for 1-2 minutes andrinsing with water 5-6 times for disinfection treatment; and cutting and discarding surrounding tissues of the disinfected material, and inoculating the rest middle tissues into a PDA culture mediumcontaining chloramphenicol or a PDA culture medium containing chloramphenicol and cinnamomum longepaniculatum leaf leaching liquor or a PDA culture medium containing chloramphenicol and carrot juice or an MEA culture medium containing chloramphenicol for culture. According to the method, the surface of cinnamomum longepaniculatum tissue is disinfected, and endophytic fungi are protected against damage while the disinfection effect is guaranteed.

Description

technical field [0001] The invention belongs to the technical field of fungal isolation and cultivation, and in particular relates to a method for disinfecting the surface of camphor oil and isolating and cultivating endophytic fungi. Background technique [0002] Oil camphor (Cinnamomum longepaniculatum (Gamble) N.Chao) belongs to Lauraceae and Cinnamomum genus. It is a national second-level key protected plant, and it is also a unique economic crop rich in natural aromatic oil in my country. At present, the planting area of ​​Yibin camphor oil exceeds 500,000 mu, accounting for more than 75% of the total resources of oil camphor in the country. The unique geographical features and climate characteristics of this area guarantee and preserve the biological independence of the species. Therefore, there may be potential endophytic fungal species resources completely different from other regions and other species in Yibin camphor. [0003] The isolation of endophytic fungi fro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14
CPCC12N1/14
Inventor 周万海冯瑞章魏琴田忠琼刘雯雯龚云飞杨婷
Owner YIBIN UNIV
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