CN1844350A

CN1844350A - Pleurotus ostreatus KZH-1 and its preparing process

Info

Publication number: CN1844350A
Application number: CN 200610010845
Authority: CN
Inventors: 张陶; 桂明英; 弓力伟; 罗孝坤
Original assignee: Kunming Edible Fungus Institute Of China Federation Of Supply And Marketing Cooperatives
Current assignee: Kunming Edible Fungus Institute Of China Federation Of Supply And Marketing Cooperatives
Priority date: 2006-04-25
Filing date: 2006-04-25
Publication date: 2006-10-11
Anticipated expiration: 2026-04-25
Also published as: CN100462431C

Abstract

The invention relates to a process for preparing Pleurotus osteatus KZH-1 preserved in China General Microbiological Culture Collection Center (CGMCC) with a docket number of CGMCC NO.1600. The solid bacterial prepared from the bacterial strain can be used for the cultivation of pleurotu ostreatus.

Description

Flat mushroom bacterial strain KZH-1 and preparation method

Technical field

The invention belongs to microbial technology field, be specifically related to flat mushroom bacterial strain KZH-1 and preparation method.

Background technology

Flat mushroom Pleurotus ostreatus is the edible fungus variety of present cultivation amount maximum, existing flat mushroom kind is along with the prolongation of cultivation and duration of service, individual cell age is increasing, the metabolism function lowers gradually, lose anti-adversity ability, gradually lose original good character, the commodity performance reduces, so that lose its use value.Bacterial classification heredity changes; Bacterial classification is parasitic by virus; Culture condition is not suitable for, and can not satisfy its life requirement, makes it lose capacity of self-regulation, and losing factor such as normal physiological function is to cause the reason of spawn degeneration, is the gradual change result of quantitative change to qualitative change, also is a kind of morbid state and old and feeble general performance.At present mainly cultivate new bacterial strain and former bacterial strain proterties is carried out rejuvenation, guarantee the good characteristic of bacterial classification by methods such as natural seed selection, cross-breeding, selection by mutation, bacterial classification detoxifications.The nature seed selection does not need special equipment, but will carry out a large amount of simultaneous tests by collecting the bacterial strain of growing under a large amount of different conditions, and the time is long, and workload is big; Cross-breeding, selection by mutation, bacterial classification detoxification then need specific installation to finish, and complex process is subjected to multiple factor affecting, unstable result.

Summary of the invention:

The present invention has overcome the deficiency of aforesaid method, utilizes wild flat mushroom resource, by sporophore separation, purifying, fruiting experiment screening, cultivate that growth is strong, adaptability strong, the good strain excellent of commodity performance tool.

The fungi that the present invention adopts is flat mushroom Pleurotus ostreatus KZH-1; Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center; Address: China. Beijing dashes closes the village; Preservation date: on 01 25th, 2006; The numbering CGMCC NO.1600 that preservation is registered on the books.

This mushroom carpophore is medium big, and bacteria cover diameter 3-14 centimetre, chocolate, flat semisphere, open and flat back is fan-shaped, and is recessed near shank; Bacterial context white is thick slightly; Lamella white, close slightly; Stem adnation, weak point, interior real, white, long 1-3 centimetre, thick 0.5-1.5 centimetre; Spore print white, spore are closely cylindrical, and is smooth colourless.22-26 ℃ of mycelium culture temperature, 5-28 ℃ of fruiting temperature.This bacterial strain adaptability is strong, high yield and high quality, and stable performance is fit to multiple culture material cultivation.

Technical scheme of the present invention:

1. gather wild mushroom carpophore;

2. separate tissue;

3. purifying bacterial strain and identification of strains;

4. fruiting experiment (biological characteristics relatively reach the production traits relatively);

5. determine strain excellent;

6. bacterial classification production and bacterial classification are used;

Through repeated screening, obtain the flat mushroom bacterial strain KZH-1 of output height, strong stress resistance.

Fungi Pleurotus ostreatusKZH-1 cultural method of the present invention:

Strain separating purifying substratum: (PDA substratum)

Potato 200 grams, glucose 20 grams, agar 20 grams are converted 1000 milliliters in water.

Flat mushroom strains separation and authentication method:

1, the mushroom carpophore tissue block is inoculated on the above-mentioned test tube nutrient agar inclined-plane, cultivated 5-7 days down in 25 ℃, to test tube observed and recorded every day of separation and Culture, the test tube that rejecting has in time been polluted, pick out bacterial strain pollution-free, that grow fine, obtain flat mushroom and separate the test tube kind.

2, will separate test tube kind inoculated by hypha block and to the dull and stereotyped culture dish of above-mentioned nutrient agar, carry out the bacterial classification purifying, get most advanced and sophisticated inoculated by hypha block after 2 days on medium slant, cultivate 7 days down, obtain flat mushroom purifying test tube kind in 25 ℃.

3, with flat mushroom purifying test tube kind, make the secondary kind, produce cultivar, carry out fruiting experiment, obtain sporophore, determine that separating the pure strain that obtains is flat mushroom strain.

The preparation method of flat mushroom strain of the present invention is divided into: two kinds of solid spawn and liquid spawns.

The solid spawn preparation method:

The solid culture based formulas is: 1. wheat or buckwheat grain 83%, weed tree sawdust 15%, terra alba or lime carbonate 2%.

2. weed tree sawdust 78%, wheat bran 20%, sucrose 1%, gypsum 1%, water content 60%, PH nature.

3. cotton seed hulls 35%, wood chip 45%, wheat bran 18%, sucrose 1%, gypsum 1%, water content 60%, PH nature.

1, the mycelium with flat mushroom is inoculated on the test tube nutrient agar inclined-plane, and culture medium prescription is the PDA substratum, cultivates 7-8 days down in 25 ℃, obtains test tube strains.

2, adopt aforementioned solid culture based formulas.After culture material mixed, after adding water and mixing evenly, in the jar of the 750ml that packs into, compress and seal the back,, insert test tube strains, cultivated 25-30 days in 25 ℃, cover with up to mycelia in the sterilisable chamber cooling 121 ℃ of sterilizations 120 minutes.

The liquid spawn preparation method:

The liquid culture based formulas is: potato 20%, glucose 2%, peptone 0.2%, potassium primary phosphate 0.05%, sal epsom 0.05%, remainder is a water, the PH nature.

2, nutrient solution is packed into (every bottled 100ml) in the 500ml triangular flask, culture medium prescription is the aforementioned liquids culture medium prescription, seals with tampon and kraft paper, and sterilization is 30 minutes under 0.15 MPa pressure, takes out, and is chilled to inoculation in 30 o'clock.

3, the test tube kind is inoculated in 500ml triangular flask (the every bottled 100ml) liquid nutrient medium, cultivates in 25 ℃ of following shaking tables, rotating speed is 150r/min, incubation time 5-7 days.

4, will shake bottle bacterial classification inoculation in the fermentor tank of the automatic fermentative production line of 25L, air flow is 1: 0.8v/ (vmin); Mixing speed is 5% for the 150r/min inoculum size; Tank pressure: 0.04Mpa; PH is 6.0; Temperature is 25 ℃; Aerated culture 72 hours obtains the flat mushroom liquid spawn.

Embodiment:

Embodiment one:

1, wild mushroom carpophore tissue block is inoculated on the above-mentioned strain separating purifying medium slant, cultivated 7 days down in 25 ℃, to test tube observed and recorded every day of separation and Culture, the test tube that rejecting has in time been polluted, pick out bacterial strain pollution-free, that grow fine, obtain flat mushroom and separate the test tube kind.

4, the mycelium with Pleurotus ostreatusKZH-1 is inoculated on the test tube nutrient agar inclined-plane, and culture medium prescription is the PDA substratum, cultivates 7 days down, obtains the test tube kind for 25 ℃.

5, adopt solid culture based formulas wheat or foster grain 83%, weed tree sawdust 15%, terra alba or lime carbonate 2%.Wheat or buckwheat grain clear water wash clean, soaked about 15 hours earlier, leach with 2% liming; boiled 15 minutes, and poured out drainage, add weed tree sawdust; terra alba or lime carbonate are mixed thoroughly, and 750 milliliters the special bacteria bottle of packing into seals; sterilized 120 minutes down at 121 ℃; in the sterilisable chamber cooling, insert test tube strains, cultivated 30 days in 25 ℃; cover with up to mycelia, obtain bacterial classification.

Embodiment two:

5, adopt aforementioned solid culture based formulas weed tree sawdust 78%, wheat bran 20%, sucrose 1%, gypsum 1%, water content 60%, PH nature.Culture material is mixed, after adding water and mixing evenly, in the jar of the 750ml that packs into, compress and seal the back,, insert test tube strains, cultivated 30 days in 25 ℃, cover with, obtain the flat mushroom solid spawn up to mycelia in the sterilisable chamber cooling 121 ℃ of sterilizations 120 minutes.

Embodiment three:

5, adopt aforementioned solid culture based formulas cotton seed hulls 35%, wood chip 45%, wheat bran 18%, sucrose 1%, gypsum 1%, water content 60%, PH nature.Culture material is mixed, after adding water and mixing evenly, in the jar of the 750ml that packs into, compress and seal the back,, insert test tube strains, cultivated 30 days in 25 ℃, cover with, obtain the flat mushroom solid spawn up to mycelia in the sterilisable chamber cooling 121 ℃ of sterilizations 120 minutes.

Embodiment four:

4, the mycelium with Pleurotus.ostreatusKZH-1 is inoculated on the test tube nutrient agar inclined-plane, and culture medium prescription is the PDA substratum, cultivates 7 days down, obtains the test tube kind for 25 ℃.

5, the test tube kind is inoculated in the 500ml triangular flask in (every bottled 100ml) liquid nutrient medium, culture medium prescription is: potato 20%, glucose 2%, peptone 0.2%, potassium primary phosphate 0.05%, sal epsom 0.05%, remainder is a water, the PH nature.Cultivate in 25 ℃ of following shaking tables, rotating speed is 150rpm, and incubation time 7 days obtains the level liquid bacterial classification.

In the fermentor tank of the automatic fermentative production line of 25L, air flow is 1 with the level liquid bacterial classification inoculation: 0.8v/ (vmin); Mixing speed is 150r/min; Inoculum size is 5%; Tank pressure: 0.04Mpa; PH is 6.0; Temperature is 25 ℃; Aerated culture 72 hours obtains the flat mushroom liquid spawn.

Claims

1, a kind of flat mushroom bacterial strain KZH-1 and preparation method is characterized in that described bacterial strain is (Pleurotus ostreatus) KZH-1, and the preserving number of this bacterial strain is CGMCC NO.1600.

2, flat mushroom bacterial strain KZH-1 according to claim 1 and preparation method, described bacterial strain obtains through conventional solid culture, it is characterized in that described solid culture based formulas and liquid culture based formulas are as follows:

The solid culture based formulas is: (1) wheat or buckwheat grain 83%, weed tree sawdust 15%, terra alba or lime carbonate 2%;

Or (2) weed tree sawdust 78%, wheat bran 20%, sucrose 1%, gypsum 1%, water content 60%, PH nature;

Or (3) cotton seed hulls 35%, wood chip 45%, wheat bran 18%, sucrose 1%, gypsum 1%, water content 60%, PH nature;