CN103947454A - Artificial culturing method of lepista sordida mycelium and culturing medium thereof - Google Patents

Abstract

The invention discloses an artificial culturing method of a lepista sordida mycelium and a culturing medium thereof, wherein the culturing medium comprise the following components according to parts by weight: 30-50 parts of pine needle, 5-12 parts of peptone, 6-12 parts of yeast extract powder, 2.5-10 parts of glucose, and 15-20 parts of agar; the artificial culturing method comprises cleaning and boiling the pine needle in water for 20-30 minutes, filtering to extract the juice, metering 1000ml of pine needle juice, then sequentially adding the peptone, the yeast extract powder, the glucose and the agar, boiling and mixing uniformly, subpackaging the prepared culturing medium into a test tube, covering a rubber plug, sterilizing under the temperature of 120-130 DEG C for 20-30 minutes, then taking out the culturing medium, naturally cooling to 20-30 DEG C, making an inclined plane culturing medium, inoculating a purified lepista sordida strain under the aseptic condition, putting the inoculated culturing medium in a constant temperature incubator, culturing for 5-10 days at the temperature of 20-30 DEG C until the mycelium grows fully in the test tube, and then refrigerating the mycelium. According to the artificial culturing method and the culturing medium, the operation is simple, the culturing period is shortened, no pollutants or wastes are produced, and the yield of the mycelium is high.

Description

The mycelial artificial culture method of a kind of Lepista sordida and medium thereof

 

Technical field:

The present invention relates to the mycelial artificial culture method of a kind of Lepista sordida and medium thereof, belong to pharmacognosy technical field.

Background technology:

Lepista sordida belongs to mycota Fungi, Basidiomycota Basidiomycota, agaric guiding principle Agaricomycetes, Agaricales Agaricales, Tricholomataceae Tricholomataceae, Lepista lentinus lepistamacro fungi.It is fresh and tender sweet, aromatic flavour, and mouthfeel is fabulous, have the good reputation of " a Shiqi taste is heard its perfume (or spice) for ten ".In Europe, the pouplarity of Lepista sordida can be equally celebrated for their achievements with matsutake and bolete, in France, is regarded as first-class food materials, has very high edibility.

In addition, Lepista sordida is delicious flavour not only, and the vitamin that contains many needed by human and amino acid and rich in protein, for example threonine, alanine, glutamine, serine, glutamic acid, valine, leucine, asparagine acid, asparatate and glycine ^[8].In addition the fruit body of Lepista sordida also contains mannitol, arabite, glucose, glycerine, fructose and trehalose, and wherein the content of trehalose is the highest.

Aspect medical value, < < day according to the Yuan Dynasty is recorded through > > by the < < book on Chinese herbal medicine art that draft > > and the yellow official of the Qing Dynasty embroider, long-term edible Lepista sordida can prevent vascular sclerosis effectively, can treat cirrhosis, diabetes, kidney trouble, gall stone etc.Up to the present, the research of the external medical value about Lepista sordida mainly still concentrates on its antioxidation activity and antibacterial activity two aspects.The fruit body of Lepista sordida also contains abundant vitamin B1, can regulate the normal glycometabolism of organism, promotes nerve conduction, and prevention beriberi is had to good effect.The hot water extract of Lepista sordida fruit body has significant active anticancer, and its pharmacological testing proof Lepista sordida fruiting body extract is respectively 100% and 90% to the inhibiting rate of small white mouse Emhorn knurl and sarcoma S180.And the juice that fruit body decocts can help to treat abscess, sand holes and wound.

In recent years, Lepista sordida is gradually by understanding that common people are familiar with, the market demand to it constantly increases, thereby cause wild resource to reduce year by year, be difficult to meet again the demand in market, Lepista sordida is carried out to artificial domesticating cultivation imperative, in Lepista sordida Mycelium culture process, growth rate is slow, mycelia is sparse, unhealthy and strong, is a technical barrier of restriction Lepista sordida artificial domesticating cultivation.At present, the mycelial artificial cultivation of fragrant mushroom class mostly adopts potato dextrose agar (PDA); Potato synthetic medium (CPDA); Wheat bran glucose agar medium (wheat bran 50-70 g, glucose 20 g, agar 18-20 g); Corn flour synthetic medium (corn flour 20-30 g, glucose 20 g, potassium dihydrogen phosphate 2 g, magnesium sulfate 0. 5 g, agar 18-20 g); Compost leaching solution medium [compost (doing) 100 g, glucose 20 g, agar 18-20 g] etc., these medium are for Lepista sordida cultural hypha process, and growth rate is slow, and it is more, sparse that mycelia end produces branch mycelia.

Summary of the invention:

The object of the invention is to overcome the deficiency of above-mentioned prior art and provide a kind of simple to operate, shortening cultivation cycle short, pollution-free without discarded object, the mycelial artificial culture method of Lepista sordida and medium thereof that mycelium production is high.

Object of the present invention can reach by following measure: a kind of for cultivating the mycelial medium of Lepista sordida, it is characterized in that its recipe ingredient is by weight: pine needle 30-50 part, peptone 5-12 part, yeast soak powder 6-12 part, glucose 2.5-10 part, agar: 15-20 part.

In order further to realize object of the present invention, its recipe ingredient is by weight: 36 parts, pine needle, 8 parts of peptones, yeast soak 6 parts, powder, 4 parts of glucose, agar: 18 parts.

The mycelial artificial culture method of Lepista sordida, is characterized in that concrete technology step is:

(1) pine needle pre-treatment:

The pine needle of collecting is cleaned, put into 1000ml water, boil 20-30 minute, cross leaching juice;

(2) medium preparation:

Loose juice is settled to 1000ml, then adds successively peptone, yeast to soak powder, glucose, agar, boil and mix, then carry out packing;

(3) medium packing, sterilizing:

The medium preparing is sub-packed in test tube, covers plug, sterilizing 20-30 minute under 120-130 ℃ of condition;

(4) cooling, inoculation:

Gone out after bacterium and taken out medium, naturally cooled to 20 ℃-30 ℃, bevel medium, under aseptic condition, accesses purified Lepista sordida bacterial classification;

(5) cultivate:

By connecting the medium of planting, be placed in constant incubator, cultivate 5-10 days for 20-30 ℃, until mycelia is covered with test tube;

(6) culture presevation: cultured bacterial classification is put in the refrigerator of 4 ℃ and carries out preservation.

The present invention can produce following good effect compared with the prior art: Lepista sordida is a kind of rare, high-quality wild-type strain, and under wild state, quantity is few, yield poorly but its color and luster grace, smell giving off a strong fragrance, taste is fresh and tender, nutritive value and medical value are high, and expensive.At present, be applicable to the research of Lepista sordida bacterial classification excellent culture medium very few, the raw material that medium of the present invention is used is pollution-free, can not produce any rubbish, simple and practical, shorten cultivation cycle, generally shorten about 4 days, and long dense, sturdy compared with other medium mycelium, be lavender, vitality is vigorous, can impel Lepista sordida mycelium Fast Growth, and the development that realizes large-scale industrial production and medicinal career for Lepista sordida all has very important significance.

Embodiment:

Below the specific embodiment of the present invention is elaborated:

Embodiment 1:

(1) pine needle pre-treatment:

The 30g pine needle of collecting is cleaned, put into 1000ml water, boil 20-30 minute, cross leaching juice;

(2) medium preparation:

Loose juice is settled to 1000ml, then adds successively 5g peptone, 6g yeast to soak powder, 2.5g glucose, 15g agar, boil and mix, then carry out packing;

(3) medium packing, sterilizing:

The medium preparing is sub-packed in test tube, covers plug, sterilizing 20-30 minute under 120-130 ℃ of condition;

(4) cooling, inoculation:

Gone out after bacterium and taken out medium, naturally cooled to 20 ℃-30 ℃, bevel medium, under aseptic condition, accesses purified Lepista sordida bacterial classification;

(5) cultivate:

By connecting the medium of planting, be placed in constant incubator, cultivate 5-10 days for 20-30 ℃, until mycelia is covered with test tube;

(6) culture presevation: cultured bacterial classification is put in the refrigerator of 4 ℃ and carries out preservation.

Embodiment 2:

(1) pine needle pre-treatment:

The 36g pine needle of collecting is cleaned, put into 1000ml water, boil 20-30 minute, cross leaching juice;

(2) medium preparation:

Loose juice is settled to 1000ml, then adds successively 8g peptone, 6g yeast to soak powder, 4g glucose, 18g agar, boil and mix, then carry out packing;

(3) medium packing, sterilizing:

The medium preparing is sub-packed in test tube, covers plug, sterilizing 20-30 minute under 120-130 ℃ of condition;

(4) cooling, inoculation:

Gone out after bacterium and taken out medium, naturally cooled to 20 ℃-30 ℃, bevel medium, under aseptic condition, accesses purified Lepista sordida bacterial classification;

(5) cultivate:

By connecting the medium of planting, be placed in constant incubator, cultivate 5-10 days for 20-30 ℃, until mycelia is covered with test tube;

(6) culture presevation: cultured bacterial classification is put in the refrigerator of 4 ℃ and carries out preservation.

Embodiment 3:

(1) pine needle pre-treatment:

The 50g pine needle of collecting is cleaned, put into 1000ml water, boil 20-30 minute, cross leaching juice;

(2) medium preparation:

Loose juice is settled to 1000ml, then adds successively 12g peptone, 12g yeast to soak powder, 10g glucose, 20g agar, boil and mix, then carry out packing;

(3) medium packing, sterilizing:

The medium preparing is sub-packed in test tube, covers plug, sterilizing 20-30 minute under 120-130 ℃ of condition;

(4) cooling, inoculation:

Gone out after bacterium and taken out medium, naturally cooled to 20 ℃-30 ℃, bevel medium, under aseptic condition, accesses purified Lepista sordida bacterial classification;

(5) cultivate:

By connecting the medium of planting, be placed in constant incubator, cultivate 5-10 days for 20-30 ℃, until mycelia is covered with test tube;

(6) culture presevation: cultured bacterial classification is put in the refrigerator of 4 ℃ and carries out preservation.

Above embodiment is described the preferred embodiment of the present invention; not scope of the present invention is limited; design under the prerequisite of spirit not departing from the present invention; various distortion and improvement that the common engineers and technicians in this area make technical scheme of the present invention, all should fall in the definite protection domain of claims of the present invention.

Claims (3)

1. for cultivating the mycelial medium of Lepista sordida, it is characterized in that its recipe ingredient is by weight: pine needle 30-50 part, peptone 5-12 part, yeast soak powder 6-12 part, glucose 2.5-10 part, agar: 15-20 part.

2. according to claim 1 a kind of for cultivating the mycelial medium of Lepista sordida, it is characterized in that its recipe ingredient is by weight: 36 parts, pine needle, 8 parts of peptones, yeast soak 6 parts, powder, 4 parts of glucose, agar: 18 parts.

3. the mycelial artificial culture method of Lepista sordida, is characterized in that concrete technology step is:

(1) pine needle pre-treatment:

The pine needle of collecting is cleaned, put into 1000ml water, boil 20-30 minute, cross leaching juice;

(2) medium preparation:

Loose juice is settled to 1000ml, then adds successively peptone, yeast to soak powder, glucose, agar, boil and mix, then carry out packing;

(3) medium packing, sterilizing:

The medium preparing is sub-packed in test tube, covers plug, sterilizing 20-30 minute under 120-130 ℃ of condition;

(4) cooling, inoculation:

Gone out after bacterium and taken out medium, naturally cooled to 20 ℃-30 ℃, bevel medium, under aseptic condition, accesses purified Lepista sordida bacterial classification;

(5) cultivate:

By connecting the medium of planting, be placed in constant incubator, cultivate 5-10 days for 20-30 ℃, until mycelia is covered with test tube;

(6) culture presevation: cultured bacterial classification is put in the refrigerator of 4 ℃ and carries out preservation.