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

A kind of Pichia pastoris, volatile oil technology, applied in the field of microorganisms

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 vol...

Method used

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

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Example 1: Isolation of endophytic fungus Ppf9

[0037]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 the number of one endophytic fungus was Ppf9.

Embodiment 2

[0038] Embodiment 2: Observation of the morphological characteristics of the endophytic fungus Ppf9 strain

[0039] The endophytic fungus Ppf9 strain was cultured on PDA medium at 25°C. The colonies were dense, with irregular protrusions, slow growth, wrinkles on the back, single cells, pseudohyphae, oval, oval, colorless, size 2.2 μm~4.8μm×2μm~4.4μm. Morphological characteristics of the endophytic fungus Ppf9 see Figure 1 ~ Figure 3 . Under culture conditions, no sexual generation of Ppf9 was found.

Embodiment 3

[0040] Example 3: ITS sequence analysis and molecular identification of the endophyte Ppf9 strain

[0041] Firstly, the hyphae of Ppf9 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 ABIPRISM 3730 sequencer was used for forward (5'→3') and reverse (3'→5') bidirectional sequencing, respectively.

[0042] 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 n...

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Abstract

The invention relates to a Paris polyphylla var. yunnanensis endogenetic Pichia pastoris used to produce volatile oil. The endogenetic fungi Ppf9 is acquired from Paris polyphylla var. yunnanensis by the endogenetic fungi isolation technique, and is classified and determined to be Pichia. The bacterial strain is preserved in China General Microbiological Culture Collection Center with the registered number of CGMCC No. 2475. 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 microorganisms, and relates to the application of an endophytic fungus with antibacterial activity, in particular to a microorganism whose metabolite is volatile oil-endophytic Pichia diangi. 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, which can produce metabolites with various structural types, and have 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 drugs, and they have important application prospects in the fields of agriculture, medicine, ...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12P7/64A61K36/064A61P31/04A61P31/10C12R1/84
CPCY02A50/30
Inventor 周立刚高希武刘西莉王明安赵江林黄永富蔡晓月
Owner CHINA AGRI UNIV
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