CN1952109A - Short term storage method for large-size fungus species - Google Patents

Abstract

The invention disclosed a way of preserving the big fungus strain for a short term. It comprises the following steps: (1) inoculating the strain to the solid medium and cutting the medium into several pieces when the hyphal body nearly covered the whole flat plate; (2) adding sterile distilled water to the container first, then putting the pieces of medium in the container; covering the lid and preserving it at 4 Deg C. The invention can preserve the strain for a short term without variation; besides, it's very simple and economical.

Description

The short term storage method of large-size fungus species

Technical field

The present invention relates to the culture collection process technical field, particularly relate to the short term storage method of large-size fungus species.

Background technology

At present, the short term storage method of large-size fungus species mainly is the inclined-plane low temperature preservation, need to be about to the bacterial classification inoculation of preservation on suitable slant medium, thermophilic is cultivated, when covering with the inclined-plane healthy and strongly, mycelia takes out, be placed on preservation in 3-5 ℃ cryodrying place or 4 ℃ of refrigerators, the refrigerator-freezer, transplanted tube once every 4-6 month.The shortcoming of this store method is (1) consumption consuming time space; (2) the easy dehydration of substratum and make the bacterial classification inactivation; (3) the easy mildew fungi pollution of tampon bacterial classification; (4) switching often bacterial strain easily produce variation.(the U.S. work of Yang Xin, " edible mushrooms research method ", Chinese agriculture press, 1999; Yellow Nian Lai chief editor, " edible fungi of china encyclopaedia ", Chinese agriculture goes out newspaper office, in May, 1993).

The large-size fungus species preservation mechanism of China's national level mainly contains " China Committee for Culture Collection of Microorganisms microbial strains preservation administrative center (CCGMC) ", " 3 of China Committee for Culture Collection of Microorganisms agricultural microbial strains preservation administrative center (ACCC) and " agriculture microbial strains preservation administrative center edible fungus species branch center "; provincial have more than ten; also have some local bacterial classifications to preserve mechanism; it is costly that these centers or mechanism are used for the bacterial classification of preservation macro fungi every year; a kind of large-size fungus species method for preserving of simple economy if can be provided, and that meaning is very great.

Summary of the invention

The objective of the invention is to overcome the deficiency that exists on the short term storage method of existing large-size fungus species, a kind of short term storage method of simple, economic large-size fungus species is provided.

The short term storage method of large-size fungus species of the present invention may further comprise the steps:

(1) will need the bacterial classification of preservation to be inoculated on the solid medium earlier and cultivate, when waiting mycelium to cover with flat board solid medium be cut into some bacterial classification pieces;

(2) in container, add sterile distilled water earlier, put the bacterial classification piece again into, cover tight bottle cap, preserve down in 4 ℃.

The preferred frozen pipe of said vesse.

The short term storage method of large-size fungus species of the present invention can be with culture presevation 1～12 month.

Compared with prior art, the present invention has following beneficial effect: the short term storage method of large-size fungus species of the present invention, easy and economical, bacterial classification is difficult for producing variation.

Description of drawings

Fig. 1 for needle mushroom Fve1 bacterial classification 4 ℃ of mycelial growth situations that are inoculated into after preserving 8 months on the inclined-plane;

Fig. 2 for needle mushroom Fve4 bacterial classification 4 ℃ of mycelial growth situations that are inoculated into after preserving 8 months on the inclined-plane;

Fig. 3 is that needle mushroom Fve1 bacterial classification is inoculated into the fruiting situation in the liquid nutrient medium after 4 ℃ are preserved 8 months;

Fig. 4 is that needle mushroom Fve5 bacterial classification is inoculated into the fruiting situation in the liquid nutrient medium after 4 ℃ are preserved 8 months;

Fig. 5 is that needle mushroom Fve5 bacterial classification is inoculated into the fruiting situation in the solid medium after 4 ℃ are preserved 8 months;

Fig. 6 is that needle mushroom Fve4 bacterial classification is inoculated into the fruiting situation in the solid medium after 4 ℃ are preserved 8 months;

Fig. 7 is that needle mushroom Fve1 bacterial classification is inoculated into the fruiting situation in the solid medium after 4 ℃ are preserved 8 months;

Embodiment

Embodiment 1: the preservation of needle mushroom bacterial classification

1. used substratum is culture medium for golden mushroom commonly used, and their prescription is respectively:

PDSA solid plate substratum: peeling potato 200g, glucose 20g, KH ₂PO ₄3g, MgSO ₄1.5g, vitamins B ₁5-10mg, agar 20g, H ₂O 1L;

2. practise edible mushrooms research centre, wine town for ensaying pin mushroom bacterial strain available from Xishui County, Guizhou city, strain number is Fve1, Fve2, Fve3, Fve4, Fve5.

Concrete steps are as follows:

(1) prepares the solid plate substratum according to a conventional method, in Bechtop, inoculate 0.5cm respectively ²5 needle mushroom inclined-plane mothers of size plant in dull and stereotyped middle, and inoculation is placed under 21 ℃ of temperature cultivates;

When (2) one weeks, the back mycelia was just covered with culture dish, utilize scalper to cut apart flat board gently in Bechtop, making becomes 0.5cm ²Square bacterial classification piece; Or the hollow stainless steel column that utilizes design voluntarily is cut into the disk bacterial classification piece of diameter 0.5cm.

(1) gets the aseptic frozen pipe of 5 1.8mL, in Bechtop, add the sterile distilled water of 0.5-1.3mL, put the 0.5cm of Fve1, Fve2, Fve3, Fve4 and the Fve5 of 3-8 piece respectively into ²Square bacterial classification piece or diameter be 0.5cm disk bacterial classification piece, cover tight bottle cap, write label, in 4 ℃ of refrigerators, preserve.Each bacterial classification is preserved 12 pipes.

Embodiment 2: needle mushroom is preserved the detection of bacterial classification

1. the dull and stereotyped mycelia experiment in inclined-plane:

(1) prepares the PDSA substratum routinely, be made into flat board and inclined-plane respectively.

(2) from 4 ℃ of refrigerators, take out the needle mushroom bacterial classification that embodiment 1 is preserved every 1 month, be inoculated in respectively on the inclined-plane, place 21 ℃ constant incubator to cultivate afterwards.

Normal and the growth conditions good (seeing Fig. 1, Fig. 2) of mycelia form of the bacterial classification in 1～October is preserved in the back observation of (3) one weeks, discovery, descends to some extent in the bacterial classification vitality of preservation after 11 months.

(4) in Bechtop, receiving flat board from the bacterial classification of 4 ℃ of preservations, the mycelia of bacterium colony does not produce the antagonism line when contacting, and not variation of bacterial classification is described.

2. liquid nutrient medium fruiting experiment:

(1) prepares the PDSB liquid nutrient medium routinely, be sub-packed in the triangular flask of 250mL, the bottled 50mL of each triangle.

(2) in Bechtop, get the needle mushroom bacterial classification that embodiment 1 preserved and be inoculated in the triangular flask, place 21 ℃ constant incubator to cultivate.

After (3) 3 weeks, bacterial classification all grows normal sporophore and (sees Fig. 3, Fig. 4).

3. solid medium fruiting experiment:

Solid culture based formulas: cotton seed hulls 85%, wheat bran 10%, sugar 1%, Semen Maydis powder 3%, lime carbonate 1%.

(1) accurately take by weighing each composition by the solid culture based formulas, with 1: 1.5 material-water ratio, the mixing substratum.

(2) get the polypropylene plastics pocket of 17 * 30cm, carefully put into substratum to 1/2 bag, the substratum degree of tightness is suitable, puts the collar, has filled in tampon.

(3) culture medium bag that installs is placed 121 ℃, 1.5kg/cm ²Down sterilization 1.5 hours of pressure.

(4) treat substratum cooling after, in Bechtop, get the needle mushroom that embodiment 1 preserved and be inoculated in the culture bag, place 21 ℃ constant incubator to cultivate afterwards.

After (5) 3 weeks, the mycelia of bacterial classification is covered with the bacterium bag, enters the management of producing mushroom stage, and the management of producing mushroom of needle mushroom is undertaken by the method and the step of " Chinese needle mushroom production " (Guo Meiying chief editor, Chinese agriculture press, 2000).The fruiting situation of bacterial classification (Fig. 5, Fig. 6 and Fig. 7).

Embodiment 3

Method for preserving of the present invention and existing slant preservation method are compared, specifically see Table 1.

Table 1 the inventive method and inclined-plane low temperature preservation are relatively

Method	The preservation container	Cost of container (unit /)	100cm 3The bacterial classification quantity (individual) of preserving	Substratum	Bacterial classification variation situation	The bacterial classification pollution condition	Shelf time (moon)
								Inclined-plane low temperature preservation (existing method)	35mL (1.5 * 15.0cm) glass test tube	1.0	?3	Need	Easily produce variation	Easily pollute	1-8
Bacterium piece low temperature preservation (the inventive method)	1.8mL the frozen pipe of plastics	0.35	?55	Sterilized water	Be difficult for producing variation	Be difficult for polluting	1-12

Claims (3)

1, the short term storage method of large-size fungus species is characterized in that comprising the steps:

(1) will need the bacterial classification of preservation to be inoculated on the solid medium earlier and cultivate, when waiting mycelium to cover with flat board solid medium be cut into some bacterial classification pieces;

(2) in container, add sterile distilled water earlier, put the bacterial classification piece again into, cover tight bottle cap, preserve down in 4 ℃.

2, method for preserving as claimed in claim 1 is characterized in that described container is frozen pipe.

3, method for preserving as claimed in claim 1 is characterized in that described bacterial classification is needle mushroom, mushroom, Grifola frondosa, glossy ganoderma or Hericium erinaceus (Bull. Ex Fr.) Pers..

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