Paris polyphylla var.yunnanensis endogenetic gliomastix for preparing volatile oil and antibiotic activity

A technology of volatile oil and sticky mildew, applied in the field of microbiology

Inactive Publication Date: 2008-09-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, there is no report on the extraction of volati

Method used

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  • Paris polyphylla var.yunnanensis endogenetic gliomastix for preparing volatile oil and antibiotic activity
  • Paris polyphylla var.yunnanensis endogenetic gliomastix for preparing volatile oil and antibiotic activity
  • Paris polyphylla var.yunnanensis endogenetic gliomastix for preparing volatile oil and antibiotic activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Isolation of endophytic fungus Ppf8

[0038]Collect fresh rhizomes of Prunus yunnanensis, first treat the surface with 70% ethanol for 30 seconds, then treat with 0.2% mercuric chloride for 20 minutes to carry out thorough surface sterilization, remove the epidermis under aseptic conditions, and divide the rhizomes into about 0.5 cm×0.5cm×0.5cm in size, placed on PDA medium, 1 piece per plate, cultured at 25°C under light conditions until the 7th to 20th day, pick a small piece from the edge of each colony The hyphae were inoculated on PDA medium for purification, and the purification was performed continuously for 4 to 5 times until the colony morphology was consistent. The purified strains were preserved on the PDA slant, and one of the endophytic fungi was numbered Ppf8.

Embodiment 2

[0039] Embodiment 2: Observation of the morphological characteristics of the endophytic fungus Ppf8 strain

[0040] The endophytic fungus Ppf8 strain was cultured on PDA medium at 25°C. The colony edge was petal-like and gray-green at the initial stage, and the hyphae in the later stage were long and villous, slightly erect, and the middle was raised. The hyphae were colorless to dark, and the conidiophores were long and thin. Septate, branched or unbranched, conidia gather into heads from mucus at the end of the conidiophores, conidia are dark, unicellular, spores are oval, with a diameter of 2.4 μm to 3.3 μm. Morphological characteristics of the endophytic fungus Ppf8 see Figure 1 to Figure 4 . Under culture conditions, no sexual generation of Ppf8 was found.

Embodiment 3

[0041] Example 3: ITS sequence analysis and molecular identification of the endophyte Ppf8 strain

[0042] Firstly, the hyphae of Ppf8 strain activated on the plate were transferred to PDA liquid medium, fresh mycelia were obtained after suspension culture for 5 days, and genomic DNA was extracted by conventional molecular biology methods. Using PCR technology, ITS universal primers ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3') were used for DNA amplification. The primers used for DNA sequencing were ITS1 and ITS4, and the ABI PRISM 3730 sequencer was used for forward (5'→3') and reverse (3'→5') bidirectional sequencing, respectively.

[0043] The reverse sequence was reverse-complemented by DNAMAN software and spliced ​​with the forward sequence to form a 5'→3' complete sequence. The obtained ITS rDNA sequence was submitted to the GenBank database (National Center for Biotechnology Information Website, http: / / www.ncbi.nlm.nih.gov), and the accession ...

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Abstract

The invention relates to a Paris polyphylla var. yunnanensis endogenetic Gliomastix used to produce volatile oil. The endogenetic fungi Ppf8 is acquired from Paris polyphylla var. yunnanensis by the endogenetic fungi isolation technique, and is classified and determined to be Gliomastix. The bacterial strain is preserved in China General Microbiological Culture Collection Center with the registered number of CGMCC No. 2474. Endogenetic Gliomastix used to produce volatile oil is isolated from Paris polyphylla var. yunnanensis for the first time herein, and the chemical compositions and the antibacterial activity of the volatile oil are identified. The invention is to make use of the endogenetic fungi resource of plant to acquire natural active ingredients.

Description

technical field [0001] The invention relates to the technical field of microbes, and relates to the application of an endophytic fungus with antibacterial activity, in particular to a microorganism whose metabolite is volatile oil-endophytic Myxomyces diangiensis. Background technique [0002] Plant endophytic fungi refer to those fungi that live in healthy plant tissues or organs during a certain stage or all stages of their life history, and the host plant does not show external disease symptoms. Plant endophytic fungi are a new microbial resource that can produce metabolites of various structural types such as alkaloids, peptides, steroids, terpenes, phenols, quinones, aliphatics, isocoumarins, etc. , has various biological activities such as antibacterial, antiviral, antitumor, anti-insect, and regulation of plant growth. Because endophytic fungi can produce metabolites with novel structures and unique functions, they become potential resources for new compounds and new...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P7/64A61K36/06A61P31/04A61P31/10C12R1/645
CPCY02A50/30
Inventor 周立刚马占鸿王敬国王明安赵江林黄永富张迎
Owner CHINA AGRI UNIV
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