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Gentiana manshurica kitag endophytic fungi and application thereof

A technology of endophytic fungi and gentian, applied in the fields of application, fungi, biocides, etc., can solve the problems of no biological control methods, unclear disease occurrence rules, affecting the quality and safety of medicinal materials, etc.

Active Publication Date: 2013-09-11
都晓伟
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, chemical control methods are mainly used in the cultivation and production of gentian, but the pesticide residues affect the quality and safety of medicinal materials
Since the relevant research on gentian diseases is not yet perfect, important issues such as the law of occurrence of the diseases are still unclear, and there is no practical and feasible biological control method

Method used

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  • Gentiana manshurica kitag endophytic fungi and application thereof
  • Gentiana manshurica kitag endophytic fungi and application thereof
  • Gentiana manshurica kitag endophytic fungi and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Isolation, purification and preservation of endophytic fungus LD421

[0023] Sample: Roots of artificially planted gentian, collected from the medicine garden of Heilongjiang University of Traditional Chinese Medicine, the original plant was identified as Gentiana manshurica Kitag. by Professor Du Xiaowei of Heilongjiang University of Traditional Chinese Medicine.

[0024] 1. Isolation of Endophytic Fungi

[0025] 1.1 Surface disinfection of plant samples

[0026] Rinse the collected samples under running water to remove the dirt and dust on the surface, and then put them into the sterile operating table. After the surface of the sample is naturally dried, cut the sample into small pieces about 5 cm long, and disinfect the surface according to the following procedures:

[0027] Rinse with 75% ethanol for 1 min → rinse with sterile water for 2 min → soak in 3.33% sodium hypochlorite solution for 3-5 min (or sterilize with 0.1% mercury solution for 1-3 min) → ...

Embodiment 2

[0047] The identification of embodiment 2 gentian endophytes LD421

[0048] 1. Morphological identification of strain LD421

[0049] Mainly based on the characteristics of the isolated endophytic fungal colonies, observe the fungal spores and sporulation structure, and carry out the classic classification and identification of the fungi.

[0050] 1.1 Colony morphology observation

[0051] The activated strain was inoculated on a PDA plate, cultured at a constant temperature of 28°C, the growth rate of the strain was recorded, and the characteristics of the colony were observed.

[0052] 1.2 Morphological observation of fungi

[0053] Use the pick-up observation method: add a drop of lactic acid-Medan stain on the slide, use an inoculation needle to pick the fungal hyphae with spores from the plate where the fungus grows, put it into the drop on the slide, and Scatter them with a needle tip, cover with a cover glass to obtain slide specimens, and observe under a microscope. ...

experiment example 2

[0124] Experimental Example 2 Study on Biological Control of Fermentation Liquid of Bacterial Strain LD421

[0125] 1. Experimental method

[0126] 1.1 Preparation of strain LD421 fermentation broth

[0127] Activate the preserved strains for culture, and when the colonies grow to 2 / 3 of the plate, cut out the fungus cakes and put them in PD liquid medium, culture them on a shaker at 28°C and 120r / min for 4-8 days. After the fermentation product is filtered through gauze, it is sterilized and refrigerated for later use. It is taken out and placed at room temperature for 2 hours before spraying.

[0128] 1.2 Treatment of seeds

[0129] About 15 days before sowing, take the seeds, prepare a 150ppm solution with 0.15g of gibberellin mixed with 1L of water, soak the seeds for 24 hours, remove them, rinse them with water for several times, then soak them in 75% ethanol for 30 seconds, take them out and rinse them with water for 4 ~5 times.

[0130] 1.3 Germination of seeds

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Abstract

The invention relates to an endophytic fungi (Metarrhizium) LD421 which is separated from gentiana manshurica kitag. The microbial preservation number of the endophytic fungi preserved on August 17th, 2012 is CGMCC NO.6454; the preservation address is No.3 institute 1, Beichen west road, Chaoyang district, Beijing, institute of microbiology, Chinese academy of sciences; the preservation unit is the China general microbiological culture collection center. The endophytic fungi can be used for treating gentiana manshurica kitag leaf blight, and the bacteria inhibitive rate of the endophytic fungi to gentiana manshurica kitag leaf blight pathogenic bacteria is increased along with the increasing of the liquid bacteria.

Description

technical field [0001] The present invention relates to plant endophytic fungi, in particular to endophytic fungi derived from gentian plants. The present invention also relates to the application of the endophytic fungi in the prevention and treatment of gentian leaf blight, which belongs to the biological control of plant endophytic fungi and their metabolites field of application. Background technique [0002] Gentian is the dried root of Gentiana scabra Bge., Gentiana manshurica Kitag., Gentiana triflora pall. or Gentiana rigescens Franch. And rhizomes, which are traditional Chinese medicines commonly used in my country. In terms of medicinal materials, the first three are commonly called "Guan Gentian", and the latter are commonly called "Dian Gentian". Gentiana is bitter and cold in nature, returns to the liver and gallbladder meridians, has the effect of clearing away heat and dampness, and purging liver and gallbladder fire. Inflammation, cystitis, urethritis, damp...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01N63/04A01P3/00C12R1/645
Inventor 都晓伟周云婷尚晓琳刘多
Owner 都晓伟
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