CN108148769A - A kind of room temperature preserves culture medium of Arthrinium fungi strain and its preparation method and application - Google Patents
A kind of room temperature preserves culture medium of Arthrinium fungi strain and its preparation method and application Download PDFInfo
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- CN108148769A CN108148769A CN201810225936.2A CN201810225936A CN108148769A CN 108148769 A CN108148769 A CN 108148769A CN 201810225936 A CN201810225936 A CN 201810225936A CN 108148769 A CN108148769 A CN 108148769A
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- arthrinium
- culture medium
- room temperature
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/04—Preserving or maintaining viable microorganisms
Abstract
The invention discloses the culture mediums that a kind of room temperature preserves Arthrinium fungi strain, which is characterized in that is as made from the raw material of following weight percent：Wheat stalk 5 10%, rice straw 5 10%, maize straw 5 10%, organic acid 0.1 1%, bamboo 5 40%, sawdust 5 10%, molasses 1 5%, γ polyglutamic acids 1 5%, konjaku flour 1 5%, glycerine 0.5 2%, adsorbent 1 5%, surplus are taro juice.The invention also discloses the preparation method and application of the culture medium.There is the culture medium of the present invention room temperature to preserve 3 years or more, it is easy to use, energy saving excellent properties, facilitate and preserve Arthrinium fungi strain, picking spore and mycelia is facilitated to be observed or used, culture medium can set different thickness and shape according to specific requirements, can regular replenishment culture medium extended shelf life.
Description
Technical field
The invention belongs to microbial preservation technique fields, and in particular to a kind of room temperature preserves the training of Arthrinium fungi strain
Support base and its preparation method and application.
Background technology
Arthrinium is recorded in the whole world altogether 60 kinds, and the category fungi host range is distributed more widely.Foreign countries are distributed in Great Britain
Zhu Dao, Holland, Germany, Poland, France, Italy, the U.S., Australia, New Zealand, India, Norway, Swaziland etc..In
State is distributed in 13 provinces such as Hebei, Henan, Shandong, Shanxi, Xinjiang, Qinghai, Ningxia, Liaoning.The category fungi can cause mao bamboon
Basal stem rot, Ph. bambusoides droop and Mao bamboo timber go mouldy, and can also cause the skin disease of people, are a kind of fungies to merit attention.More
Importantly, the category fungi can cause sugarcane to go bad nosotoxicosis, from statistics report in 1972, in China's popular harm more than 40
Year, cause great economic loss and casualties invalid.But the category fungi still belongs to beneficial bacterium, can generate a variety of anti-
Raw element and other active materials.Therefore, which has very high research value and value of exploiting and utilizing, and therefore, the category is true
Its germ plasm resource of bacterium becomes research hotspot, studies its new method for preserving and has broad prospects and important realistic meaning.
At present, there are many store method of fungi, mainly including following several：1) inclined-plane low temperature preservation：Strain is inoculated with
It is to be grown to be placed on 4 DEG C of Storage in refrigerator well on suitable slant medium, after carrying out switching culture at regular intervals, then
New inclined-plane is continued into preservation.2) atoleine preserving process：Aseptically, it pours into and has sterilized into the fresh inclined-plane grown
Atoleine, oil reservoir will be higher by inclined-plane upper end 1cm, be allowed to and air exclusion, then vertically be put in preservation in room temperature or refrigerator.
3) sand tube preservation method：The mould grown on slant medium or actinospore are scraped, is connected in the sandy soil pipe of sterilizing and stirs
It is even, it is placed in and fills drier (such as CaCl2) container in seal, low-temperature preservation.4) lyophil preservation method：At a lower temperature
Make bacterium solution in frozen state, while be evacuated decompression and be allowed to drying.5) nitrogen ultra low temperature freezing process：Culture presevation is in liquid nitrogen freezer
In, temperature can be down to -196 DEG C, this is the new technology of current preservation of bacteria strain.6) silica gel preserving process, etc..More than fungi preservation side
The common ground of method is：Undesirable environment and nutritional condition are created, microorganism is made to be in the state or spore state of suspend mode, makes it
Vital movement is in minimum state, and metabolic activity stops.Such store method require to give barren nutrition, anoxic, drying,
Low temperature, water shortage etc..Countries in the world Culture Collection Center is substantially take more than method remove preservation of bacteria strain, but these are preserved
Method there are it is many shortcomings that：1. some preservation methods expend a large amount of financial resources, material resources and manpower or even can not achieve the big of strain
Batch preserves.For example：Liquid nitrogen storage method.2. some methods are not suitable for the preservation of Arthrinium fungi, for example atoleine preservation
Method, the method anoxic gas；Native pipe preserving process dries water shortage, and strain easily inactivates.In short, the method for preserving of each strain has certainly
The strain or strain state that body is suitble to, that is, each method has the defects of certain property.
Invention content
The present invention provides a kind of room temperature easy to use to preserve Arthrinium (Arthrinium spp.) fungi strain
Culture medium.
The system of the culture medium of Arthrinium (Arthrinium spp.) fungi strain is preserved the present invention also provides the room temperature
Preparation Method.
Answering for the culture medium of Arthrinium (Arthrinium spp.) fungi strain is preserved the present invention also provides the room temperature
With.
The purpose of the present invention is what is be achieved through the following technical solutions：
A kind of room temperature preserves the culture medium of Arthrinium (Arthrinium spp.) fungi strain, is by following weight hundred
Made from the raw material for dividing ratio：Wheat stalk 5-10%, rice straw 5-10%, maize straw 5-10%, organic acid 0.1-1%,
Bamboo 5-40%, sawdust 5-10%, molasses 1-5%, gamma-polyglutamic acid 1-5%, konjaku flour 1-5%, glycerine 0.5-2%, absorption
Agent 1-5%, surplus are taro juice.
Described, organic acid is to shine 3 by citric acid and ammonium lacate：Made from 1 weight ratio.
Described, taro juice is as made from following steps：Fresh taro 200g is weighed, 1cm is cut into peeling3Taro
Block；Taro block is added in the water of 1000ml, turn slow boiling after being heated to boiling, continue to heat 20-30min；Naturally cool to room
Temperature by taro solution filtered through gauze after cooling, removes solids, obtains taro juice.
Described, adsorbent is diatomite.
A kind of room temperature preserves the preparation method of the culture medium of Arthrinium fungi strain, is as made from following steps：
1) wheat stalk, rice straw, maize straw, bamboo and sawdust are crushed, crosses 100-800 mesh sieve, high pressure sterilization
20-45min must expect A；
2) organic acid, molasses, gamma-polyglutamic acid, taro juice, konjaku flour and glycerine with adsorbent are uniformly mixed, gone out
Bacterium 20-30min must expect B；
3) under aseptic technique, material A and material B are uniformly mixed, the pie that thickness is 0.2-0.5cm is pressed into, obtains
Room temperature preserves the culture medium of Arthrinium fungi strain.
A kind of room temperature preserves the application of the culture medium of Arthrinium fungi strain, and the application is the culture medium in Nodule defects
Belong to the application during fungi strain room temperature preserves.
The culture medium that the application preserves Arthrinium fungi strain for room temperature is cut according to the shape of box, after cutting
Culture medium be put into filtering and airing function plexiglass box in, under aseptic technique be inoculated with Arthrinium fungi
Strain, sealing, you can realize that the room temperature of Arthrinium fungi strain preserves.
Arthrinium fungi strain used in the present invention is purchased from China General Microbiological culture presevation administrative center (address：
Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica), deposit number is respectively：CGMCC No
51218th, CGMCC No313755 and CGMCC No38582.
Beneficial effects of the present invention：
1. there is the culture medium that room temperature of the present invention preserves Arthrinium fungi strain room temperature to preserve 3 years or more, easy to use,
Energy saving excellent properties facilitate and preserve Arthrinium fungi strain, picking spore and mycelia is facilitated to be observed or used,
Culture medium can set different thickness and shape according to specific requirements, can regular replenishment culture medium extended shelf life.
2. the culture medium raw material that room temperature of the present invention preserves Arthrinium fungi strain derives from a wealth of sources, cheap and easy to get, at low cost,
Holding time is long, and stability is good, while has certain ornamental value, significantly improves the survival rate of Arthrinium fungi strain and numerous
Grow rate.
3. the preparation method of the present invention is simple, easy to operate, large area culture is not take up, is used manpower and material resources sparingly.
Specific embodiment
With reference to specific embodiment, the present invention will be further described.
Embodiment 1
A kind of room temperature preserves the culture medium of Arthrinium fungi strain, is as made from the raw material of following weight percent：
Wheat stalk 10%, rice straw 10%, maize straw 10%, organic acid 1%, bamboo 5%, sawdust 10%, molasses 5%, γ-
Polyglutamic acid 5%, konjaku flour 5%, glycerine 0.5%, adsorbent 5%, surplus are taro juice.
Described, organic acid is to shine 3 by citric acid and ammonium lacate：Made from 1 weight ratio.
Described, taro juice is as made from following steps：Fresh taro 200g is weighed, 1cm is cut into peeling3Taro
Block；Taro block is added in the water of 1000ml, turn slow boiling after being heated to boiling, continue to heat 30min；Cooled to room temperature,
By taro solution filtered through gauze after cooling, solids is removed, obtains taro juice.
Described, adsorbent is diatomite.
A kind of room temperature preserves the preparation method of the culture medium of Arthrinium fungi strain, is as made from following steps：
1) wheat stalk, rice straw, maize straw, bamboo and sawdust are crushed, crosses 100-800 mesh sieve, high pressure sterilization
20min must expect A；
2) organic acid, molasses, gamma-polyglutamic acid, taro juice, konjaku flour and glycerine with adsorbent are uniformly mixed, gone out
Bacterium 30min must expect B；
3) under aseptic technique, material A and material B are uniformly mixed, the pie that thickness is 0.2-0.5cm is pressed into, obtains
Room temperature preserves the culture medium of Arthrinium fungi strain.
A kind of room temperature preserves the application of the culture medium of Arthrinium fungi strain, and the application is the culture medium in Nodule defects
Belong to the application during fungi strain room temperature preserves.
The culture medium that the application preserves Arthrinium fungi strain for room temperature is cut according to the shape of box, after cutting
Culture medium be put into filtering and airing function plexiglass box in, under aseptic technique be inoculated with Arthrinium fungi
Strain, sealing, you can realize that the room temperature of Arthrinium fungi strain preserves.
Embodiment 2
A kind of room temperature preserves the culture medium of Arthrinium fungi strain, is as made from the raw material of following weight percent：
Wheat stalk 8%, rice straw 7%, maize straw 8%, organic acid 0.5%, bamboo 20%, sawdust 8%, molasses 3%, γ-
Polyglutamic acid 3%, konjaku flour 3%, glycerine 1%, adsorbent 3%, surplus are taro juice.
Described, organic acid is to shine 3 by citric acid and ammonium lacate：Made from 1 weight ratio.
Described, taro juice is as made from following steps：Fresh taro 200g is weighed, 1cm is cut into peeling3Taro
Block；Taro block is added in the water of 1000ml, turn slow boiling after being heated to boiling, continue to heat 25min；Cooled to room temperature,
By taro solution filtered through gauze after cooling, solids is removed, obtains taro juice.
Described, adsorbent is diatomite.
A kind of room temperature preserves the preparation method of the culture medium of Arthrinium fungi strain, is as made from following steps：
1) wheat stalk, rice straw, maize straw, bamboo and sawdust are crushed, crosses 100-800 mesh sieve, high pressure sterilization
30min must expect A；
2) organic acid, molasses, gamma-polyglutamic acid, taro juice, konjaku flour and glycerine with adsorbent are uniformly mixed, gone out
Bacterium 25min must expect B；
3) under aseptic technique, material A and material B are uniformly mixed, the pie that thickness is 0.2-0.5cm is pressed into, obtains
Room temperature preserves the culture medium of Arthrinium fungi strain.
A kind of room temperature preserves the application of the culture medium of Arthrinium fungi strain, and the application is the culture medium in Nodule defects
Belong to the application during fungi strain room temperature preserves.
The culture medium that the application preserves Arthrinium fungi strain for room temperature is cut according to the shape of box, after cutting
Culture medium be put into filtering and airing function plexiglass box in, under aseptic technique be inoculated with Arthrinium fungi
Strain, sealing, you can realize that the room temperature of Arthrinium fungi strain preserves.
Embodiment 3
A kind of room temperature preserves the culture medium of Arthrinium fungi strain, is as made from the raw material of following weight percent：
Wheat stalk 5%, rice straw 5%, maize straw 5%, organic acid 0.1%, bamboo 40%, sawdust 5%, molasses 1%, γ-
Polyglutamic acid 1%, konjaku flour 1%, glycerine 2%, adsorbent 1%, surplus are taro juice.
Described, organic acid is to shine 3 by citric acid and ammonium lacate：Made from 1 weight ratio.
Described, taro juice is as made from following steps：Fresh taro 200g is weighed, 1cm is cut into peeling3Taro
Block；Taro block is added in the water of 1000ml, turn slow boiling after being heated to boiling, continue to heat 20min；Cooled to room temperature,
By taro solution filtered through gauze after cooling, solids is removed, obtains taro juice.
Described, adsorbent is diatomite.
A kind of room temperature preserves the preparation method of the culture medium of Arthrinium fungi strain, is as made from following steps：
1) wheat stalk, rice straw, maize straw, bamboo and sawdust are crushed, crosses 100-800 mesh sieve, high pressure sterilization
45min must expect A；
2) organic acid, molasses, gamma-polyglutamic acid, taro juice, konjaku flour and glycerine with adsorbent are uniformly mixed, gone out
Bacterium 20min must expect B；
3) under aseptic technique, material A and material B are uniformly mixed, the pie that thickness is 0.2-0.5cm is pressed into, obtains
Room temperature preserves the culture medium of Arthrinium fungi strain.
A kind of room temperature preserves the application of the culture medium of Arthrinium fungi strain, and the application is the culture medium in Nodule defects
Belong to the application during fungi strain room temperature preserves.
The culture medium that the application preserves Arthrinium fungi strain for room temperature is cut according to the shape of box, after cutting
Culture medium be put into filtering and airing function plexiglass box in, under aseptic technique be inoculated with Arthrinium fungi
Strain, sealing, you can realize that the room temperature of Arthrinium fungi strain preserves.
Survival rate is tested
By Arthrinium fungi used in the present invention：CGMCC No 51218, CGMCC No313755 and CGMCC No38582
Survival rate experiment is carried out, respectively progress culture medium preservation of the present invention, inclined-plane Cord blood, Liquid nitrogen, freeze-drying
It preserves, glycerine preserves and glycerol liquor preserves.In each store method, each bacterial strain sets 4 test groups, each test group setting
3 repetitions.It is observed continuously 3 years, counts the switching survival rate of each store method, the results are shown in Table 1.
The switching survival rate result of the test of table 1 (100%)
As can be seen from Table 1,3 bacterial strains switching survival rate that culture medium of the present invention preserves is superior to other methods.It follows that
The culture medium of the present invention can be used for room temperature to preserve Arthrinium fungi strain.
Pollution condition is tested
By Arthrinium fungi used in the present invention：CGMCC No 51218, CGMCC No313755 and CGMCC No38582
Pollution condition experiment is carried out, progress culture medium preservation of the present invention, inclined-plane Cord blood, Liquid nitrogen, freezing are dry respectively
Dry preservation, glycerine preserve and glycerol liquor preserves.In each store method, each bacterial strain sets 4 test groups, and each test group is set
Put 3 repetitions.It is observed continuously 3 years, counts the switching pollution rate of each store method, the results are shown in Table 2.
The switching pollution rate result of the test of table 2
As can be seen from Table 2,3 bacterial strains switching pollution rate that culture medium of the present invention preserves be up to 3.45% and liquid nitrogen it is ultralow
Temperature preservation is identical, less than other store methods.It follows that the culture medium of the present invention can be used for room temperature to preserve Arthrinium fungi
Strain, preservation effect is suitable with Liquid nitrogen, but the present invention culture medium can realize room temperature preserve Arthrinium it is true
Bacterium strain convenient for storage, does not consume the energy, switching survival rate is high, and switching pollution rate is low, and picking spore and mycelia is facilitated to be seen
It examines or uses.
Claims (7)
1. a kind of room temperature preserves the culture medium of Arthrinium fungi strain, which is characterized in that is by the original of following weight percent
Made from material：Wheat stalk 5-10%, rice straw 5-10%, maize straw 5-10%, organic acid 0.1-1%, bamboo 5-
40%, sawdust 5-10%, molasses 1-5%, gamma-polyglutamic acid 1-5%, konjaku flour 1-5%, glycerine 0.5-2%, adsorbent 1-
5%, surplus is taro juice.
2. room temperature preserves the culture medium of Arthrinium fungi strain according to claim 1, which is characterized in that the organic acid
It is to shine 3 by citric acid and ammonium lacate：Made from 1 weight ratio.
3. room temperature preserves the culture medium of Arthrinium fungi strain according to claim 1, which is characterized in that the taro liquid
Juice is as made from following steps：Fresh taro 200g is weighed, 1cm is cut into peeling3Taro block；Taro block is added into 1000ml
Water in, be heated to boiling after turn slow boiling, continue heat 20-30min；Cooled to room temperature, by taro solution after cooling
With filtered through gauze, solids is removed, obtains taro juice.
4. room temperature preserves the culture medium of Arthrinium fungi strain according to claim 1, which is characterized in that the adsorbent
For diatomite.
5. room temperature described in a kind of claim 1-4 preserves the preparation method of the culture medium of Arthrinium fungi strain, feature exists
In being as made from following steps：
1) wheat stalk, rice straw, maize straw, bamboo and sawdust are crushed, crosses 100-800 mesh sieve, high pressure sterilization 20-
45min must expect A；
2) organic acid, molasses, gamma-polyglutamic acid, taro juice, konjaku flour and glycerine are uniformly mixed with adsorbent, sterilize 20-
30min must expect B；
3) under aseptic technique, material A and material B are uniformly mixed, the pie that thickness is 0.2-0.5cm is pressed into, obtains room temperature
Preserve the culture medium of Arthrinium fungi strain.
6. a kind of any one of claim 1-5 room temperature preserves the application of the culture medium of Arthrinium fungi strain, feature exists
In application of the application for the culture medium in the preservation of Arthrinium fungi strain room temperature.
7. room temperature preserves the application of the culture medium of Arthrinium fungi strain according to claim 6, which is characterized in that described
It is cut using the culture medium that Arthrinium fungi strain is preserved for room temperature according to the shape of box, the culture medium after cutting is put into
In plexiglass box with filtering and airing function, Arthrinium fungi strain is inoculated under aseptic technique, is sealed, i.e.,
The room temperature that Arthrinium fungi strain can be achieved preserves.
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| CN201810225936.2A CN108148769B (en) | 2018-03-19 | 2018-03-19 | Culture medium for normal-temperature preservation of arthrobotrys fungus strains and preparation method and application thereof |
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| CN110106091A (en) * | 2019-03-13 | 2019-08-09 | 广西大学 | A method of timber is dyed using Li Baojia shell section SHL-1 fungi |
| CN112239729A (en) * | 2020-10-30 | 2021-01-19 | 广西壮族自治区农业科学院 | Culture medium and method for promoting efficient spore production of Arctostaphylos |
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| CN110106091B (en) * | 2019-03-13 | 2021-05-18 | 广西大学 | Method for dyeing wood by using Pseudocercospora pyricularis SHL-1 fungus |
| CN112239729A (en) * | 2020-10-30 | 2021-01-19 | 广西壮族自治区农业科学院 | Culture medium and method for promoting efficient spore production of Arctostaphylos |
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| CN108148769B (en) | 2020-11-27 |
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