Preparation and applications of salvia miltiorrhiza endophytic fungi hypha water-soluble extract and fractions of salvia miltiorrhiza endophytic fungi hypha water-soluble extract

A technology for endophytic fungi and water-soluble extracts of Salvia miltiorrhiza, applied in the directions of botanical equipment and methods, chemicals for biological control, applications, etc., can solve the problems of low content of active ingredients, restricting the application of hairy roots, etc.

Inactive Publication Date: 2014-05-21
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the hairy root culture technology of Salvia miltiorrhiza is quite mature, but the low content of active ingredients greatly restricts the application of hairy root

Method used

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  • Preparation and applications of salvia miltiorrhiza endophytic fungi hypha water-soluble extract and fractions of salvia miltiorrhiza endophytic fungi hypha water-soluble extract
  • Preparation and applications of salvia miltiorrhiza endophytic fungi hypha water-soluble extract and fractions of salvia miltiorrhiza endophytic fungi hypha water-soluble extract
  • Preparation and applications of salvia miltiorrhiza endophytic fungi hypha water-soluble extract and fractions of salvia miltiorrhiza endophytic fungi hypha water-soluble extract

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: the fermentation of endogenous Trichoderma dark green D16

[0029] Inoculate endophytic fungus D16 of Salvia miltiorrhiza on 1 / 2B5 solid medium and cultivate at room temperature for 3-5 days (see figure 1 ); then use a hole puncher to inoculate the D16 mycelia on the solid medium into a 250ml Erlenmeyer flask (containing 100ml 1 / 2B5 culture solution), and cultivate at 26°C and 180rpm for 3-5 days to obtain the cultured seeds liquid (see figure 2); Finally, inoculate the culture seed solution into a 5L Erlenmeyer flask (containing 4L 1 / 2B5 culture solution), and cultivate at 26°C and 160rpm for 10 days.

Embodiment 2

[0030] Embodiment 2: Preparation of Salvia miltiorrhiza endophyte mycelia water-soluble extract (WSE)

[0031] The D16 mycelium cultivated for 10 days was collected by suction filtration under reduced pressure, and washed three times with distilled water; then an appropriate amount of distilled water was added to resuspend the D16 mycelium, and treated at 121° C. for 40 minutes under high temperature and high pressure. After natural cooling, the filtrate obtained by suction filtration under reduced pressure is properly concentrated to be endogenous Trichoderma dark green D16 mycelia WSE.

[0032] The sulfuric acid-phenol method was used to determine the concentration of total sugar in the mycelia WSE of endophytic Trichoderma dark greens D16. Draw a standard curve with glucose to obtain a linear regression equation: Y=0.0136X-0.0151, correlation coefficient R 2 =0.9989, the glucose concentration range is 10-50ug / ml.

Embodiment 3

[0033] Example 3: Preparation of the polysaccharide protein fraction (PPF) of the water-soluble extract of the endophytic fungus mycelia of Salvia miltiorrhiza

[0034] Concentrate endogenous Trichoderma dark green D16 mycelia WSE solution (30L) under reduced pressure to an appropriate volume, add 5 times the volume of 95% ethanol and let stand at 4°C for 48 hours, centrifuge at 5000 rpm for 10 minutes to obtain a white precipitate , washed with absolute ethanol for 3 times, the precipitate was collected, and the white powder obtained by vacuum freeze-drying was endophytic Trichoderma dark green D16 mycelia PPF.

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PUM

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Abstract

The invention discloses a salvia miltiorrhiza endophytic fungi hypha water-soluble extract (WSE). The salvia miltiorrhiza endophytic fungi hypha water-soluble extract (WSE) is prepared by following steps: salvia miltiorrhiza endophytic fungi hyphostoma obtained via 9 to 14 days of fermentation is collected; distilled water is added for resuspending; an obtained suspension liquid is sterilized for 30 to 60min at a temperature of 110 to 130 DEG C, and then is subjected to vacuum pumping filtration so as to obtain a filtrate. The invention also discloses polysaccharide protein fractions (PPF) of the salvia miltiorrhiza endophytic fungi hypha water-soluble extract, and polysaccharide fractions (PSF) of the salvia miltiorrhiza endophytic fungi hypha water-soluble extract. The invention further discloses applications of SWE, PPF, and PSF in promoting growth of tissue culture object salvia miltiorrhiza hairy roots of host salvia miltiorrhiza, and increasing tanshinone content of salvia miltiorrhiza hairy roots.

Description

technical field [0001] The present invention relates to a water-soluble extract of endophytic fungus mycelium of Salvia miltiorrhiza and its polysaccharide protein part and polysaccharide part. In addition, the present invention also relates to the water-soluble extract of endophytic fungus mycelia of Danshen and its polysaccharide protein part and polysaccharide part It is used to promote the growth of hairy roots of Salvia miltiorrhiza and increase the content of tanshinone components. Background technique [0002] Salvia miltiorrhiza Bge. is a plant of the genus Salvia in the Labiatae family. Its dry root and rhizome are used as medicine with the same name. It was first recorded in "Shen Nong's Materia Medica" and listed as the top grade. It is a traditional Chinese medicine commonly used in my country. It tastes bitter and is slightly cold in nature. , GUIXIN, Liver Meridian, has blood stasis dispelling and pain relieving, promoting blood circulation to stimulate menstrua...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N63/04A01P21/00A01G7/06
Inventor 明乾良秦路平韩婷张巧艳郑承剑张宏贾敏
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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