CN107177514A - A kind of purity is the preparation method of 100% morchella mother culture - Google Patents
A kind of purity is the preparation method of 100% morchella mother culture Download PDFInfo
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- CN107177514A CN107177514A CN201710548197.6A CN201710548197A CN107177514A CN 107177514 A CN107177514 A CN 107177514A CN 201710548197 A CN201710548197 A CN 201710548197A CN 107177514 A CN107177514 A CN 107177514A
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- culture
- mycelium
- superclean bench
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- culture dish
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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
Abstract
The invention discloses a kind of preparation method of purity for 100% morchella mother culture, it is related to morchella mother culture preparation method field, comprises the following steps：Strain is obtained after the preparation of culture medium, the preparation of Morchella esculenta (L.) Pers sporophore, culture, mycelia observation and mycelium purification, the section that the present invention has carried out mycelia before purification is observed under the microscope, Purification is carried out again after ensureing without other miscellaneous bacteria mycelia and miscellaneous bacteria spore, it is ensured that the purity of hickory chick Spawn incubation 100%.
Description
Technical field
The present invention relates to field of hydrometallurgy, more particularly to a kind of system of purity for 100% morchella mother culture
Make method.
Background technology
Hickory chick is a kind of Rare edible fungus kind, is gained the name because of its cap surface irregularity, shape such as sheep tripe.Hickory chick
(Morchella) it is, also known as morel, a kind of precious edible mushroom and medicinal fungus.Hickory chick was found in 1818.For eating
Pneumatosis is stagnant, abdominal fullness and distention, phlegm stops up the circulation of vital energy in the wrong direction are breathed with cough.Hickory chick is also known as sheep tripe dish, sheep mushroom.Its structure is similar to cup fungi, and top is in pleat
Wrinkle is netted, both as individual honeycomb, also like individual sheep tripe, thus gains the name.Hickory chick yield in 2 to three years after mountain fire is extra-high, because
The picker of this North America can gather hickory chick according to mountain fire.However, after fire is controlled, in same area, it
Growth population can be reduced year in year out.
Bionical domesticating and cultivating is had started at present.Morchella mother culture manufacture be bionical domesticating and cultivating the first step,
It is the key link.The making of morchella mother culture is most important link in whole bionical domesticating and cultivating production process, is also base
Basis in plinth；Due in the manufacturing process of morchella mother culture, not can determine that the mycelia in culture dish is sheep tripe before purification
Bacterium mycelia, can not determine whether that a kind of only material of Morchella esculenta (L.) Pers mycelium is present, institute is in the conventional way it cannot be guaranteed that after purification
Morchella mother culture purity, but the purity of parent species is directly connected to ensuing plantation success.
Therefore a kind of method that degree of making purity is 100% morchella mother culture is searched out particularly important.
The content of the invention
It is an object of the invention to：In order to solve the technical problem that existing morchella mother culture purity is low, the present invention is provided
A kind of purity is the preparation method of 100% morchella mother culture.
The present invention specifically uses following technical scheme to achieve these goals：
A kind of purity is the preparation method of 100% morchella mother culture, it is characterised in that comprised the following steps：
1st, the preparation of culture medium：
A:It is standby superclean bench is put into after blank culture dish autoclaving；
B：Mother culture media autoclaving, superclean bench is put into after bacterium of having gone out, ultraviolet lamp sterilization, 5 minutes is opened
Afterwards, ultraviolet lamp is closed；
C：Superclean bench air supply pattern is opened, quickly the mother of the liquid condition by step B in superclean bench
Culture medium is planted to be dispensed into by each blank culture dish in step A；
2nd, the preparation of Morchella esculenta (L.) Pers sporophore：The new fresh sporophore of ready hickory chick is put into superclean bench, ultraviolet
After sterilization 20~30 minutes, ultraviolet lamp is closed, subsequent step is stand-by；
3rd, cultivate：Open on the inside of the other mode of operations of superclean bench, the hickory chick stem for step 2 of being learnt from else's experience with scalpel
Mung bean grain size fructification block, then bundle solid block is put into the culture medium of the center by the culture dish in step 1
In, mouth is sealed in the edge of culture dish by the lid for then covering culture dish with sealed membrane, is being put into biochemical cultivation case culture 3~4
My god, temperature control system is at 12 DEG C~18 DEG C, and fructification block surrounding grows mycelium；
4th, mycelia is observed：With transfer needle picking, some are put into preprepared cover glass to the mycelium obtained in step 3
On, and add a small amount of sterilized water to drop on mycelium, and with being taken after slide compacting mycelium under microscope, confirmation is hickory chick
Mycelium, whole process will carry out in the case where the other mode of operations of superclean bench are opened；
5th, mycelium is purified：After step 4 observation, without carrying out purification work again after other miscellaneous bacteria mycelia and miscellaneous bacteria spore
Make, the slant tube culture medium that autoclaving is good and solidifies is put into superclean bench, cultured mycelium is put into super
Other inoculating tools, are put into superclean bench by net workbench, are opened after ultraviolet lamp is sterilized 30 minutes and are closed, open ultra-clean work
Make the other mode of operations of platform, the center on the mycelia access solidification Tube propagation base inclined-plane grown in culture dish from fructification surrounding
Behind position, put at 15 DEG C~18 DEG C of people's biochemical cultivation case and cultivate after test tube mouth is obturaged with silica gel plug, invisible spectro mycelia after 7 days
After body is covered with, morchella mother culture is obtained.
Further, in the step C in step 1, the thickness for the culture medium being dispensed into culture dish is 1.5mm~4mm.
Further, microscope observes mycelium from 10 times of object lens in step 4.
Beneficial effects of the present invention are as follows：
1st, process of the invention is simple, during traditional hickory chick Spawn incubation, due to not can determine that culture dish before purification
In mycelia be Morciiella Esculeuta Mycelia, can not determine whether that only a kind of material of Morchella esculenta (L.) Pers mycelium is present, institute is in the conventional way
It cannot be guaranteed that the purity of the morchella mother culture after purification, the section that the present invention has carried out mycelia before purification is seen under the microscope
Examine, carry out Purification again after ensureing without other miscellaneous bacteria mycelia and miscellaneous bacteria spore, it is ensured that hickory chick Spawn incubation 100%
Purity.
2nd, increase microscopic section observes this step, is not a sample investigation, we can be in each culture dish
The mycelium for not doing mycelia Purification also is first observed under the microscope, decides whether to also need to again after observation to carry out ensuing
Purification reduces the blindness of work, reduces the workload of technician, reduces what problematic strain came into the market
Possibility.
Embodiment
In order to which those skilled in the art are better understood from the present invention, following examples are further retouched in detail to present invention work
State.
Embodiment 1
The present embodiment is provided
A kind of purity is the preparation method of 100% morchella mother culture, it is characterised in that comprised the following steps：
1st, the preparation of culture medium：
A:It is standby to be put into superclean bench after one group of blank culture dish autoclaving 8；
B：The mother culture media autoclaving of the liquid condition in glass container, superclean bench is put into after bacterium of having gone out,
Ultraviolet lamp sterilization is opened, after 5 minutes, ultraviolet lamp is closed；
C：Superclean bench air supply pattern is opened, quickly the mother of the liquid condition by step B in superclean bench
Culture medium is planted to be dispensed into by each blank culture dish in step A, the thickness for the culture medium being dispensed into culture dish is
4mm；
2nd, the preparation of Morchella esculenta (L.) Pers sporophore：The new fresh sporophore of ready hickory chick is put into superclean bench, ultraviolet
After sterilization 30 minutes, ultraviolet lamp is closed, subsequent step is stand-by；
3rd, cultivate：Open on the inside of the other mode of operations of superclean bench, the hickory chick stem for step 2 of being learnt from else's experience with scalpel
Mung bean grain size fructification block, then bundle solid block is put into the culture medium of the center by the culture dish in step 1
In, mouth is sealed in the edge of culture dish by the lid for then covering culture dish with sealed membrane, is being put into biochemical cultivation case culture 4 days,
Temperature control system is at 18 DEG C, and fructification block surrounding grows mycelium；
4th, mycelia is observed：With transfer needle picking, some are put into preprepared cover glass to the mycelium obtained in step 3
On, and add a small amount of sterilized water to drop on mycelium, and with being taken after slide compacting mycelium under microscope, seen with 10 times of object lens
Examine, confirmation is the mycelium of hickory chick, whole process will be carried out in the case where the other mode of operations of superclean bench are opened；
5th, mycelium is purified：After step 4 observation, without carrying out purification work again after other miscellaneous bacteria mycelia and miscellaneous bacteria spore
Make, the slant tube culture medium that autoclaving is good and solidifies is put into superclean bench, cultured mycelium is put into super
Other inoculating tools, are put into superclean bench by net workbench, are opened after ultraviolet lamp is sterilized 30 minutes and are closed, open ultra-clean work
Make the other mode of operations of platform, the center on the mycelia access solidification Tube propagation base inclined-plane grown in culture dish from fructification surrounding
Behind position, put at 18 DEG C of people's biochemical cultivation case and cultivate after test tube mouth is obturaged with silica gel plug, invisible spectro mycelium is covered with after 7 days
Afterwards, so as to obtain 100% morchella mother culture.
Embodiment 2
A kind of purity is the preparation method of 100% morchella mother culture, it is characterised in that comprised the following steps：
1st, the preparation of culture medium：
A:It is standby to be put into superclean bench after one group of blank culture dish autoclaving 8；
B：The mother culture media autoclaving of the liquid condition in glass container, superclean bench is put into after bacterium of having gone out,
Ultraviolet lamp sterilization is opened, after 4 minutes, ultraviolet lamp is closed；
C：Superclean bench air supply pattern is opened, quickly the mother of the liquid condition by step B in superclean bench
Culture medium is planted to be dispensed into by each blank culture dish in step A, the thickness for the culture medium being dispensed into culture dish is
1.5mm；
2nd, the preparation of Morchella esculenta (L.) Pers sporophore：The new fresh sporophore of ready hickory chick is put into superclean bench, ultraviolet
After sterilization 20 minutes, ultraviolet lamp is closed, subsequent step is stand-by；
3rd, cultivate：Open on the inside of the other mode of operations of superclean bench, the hickory chick stem for step 2 of being learnt from else's experience with scalpel
Mung bean grain size fructification block, then bundle solid block is put into the culture medium of the center by the culture dish in step 1
In, mouth is sealed in the edge of culture dish by the lid for then covering culture dish with sealed membrane, is being put into biochemical cultivation case culture 3 days,
Temperature control system is at 12 DEG C, and fructification block surrounding grows mycelium；
4th, mycelia is observed：With transfer needle picking, some are put into preprepared cover glass to the mycelium obtained in step 3
On, and add a small amount of sterilized water to drop on mycelium, and with being taken after slide compacting mycelium under microscope, seen with 10 times of object lens
Examine, confirmation is the mycelium of hickory chick, whole process will be carried out in the case where the other mode of operations of superclean bench are opened；
5th, mycelium is purified：After step 4 observation, without carrying out purification work again after other miscellaneous bacteria mycelia and miscellaneous bacteria spore
Make, the slant tube culture medium that autoclaving is good and solidifies is put into superclean bench, cultured mycelium is put into super
Other inoculating tools, are put into superclean bench by net workbench, are opened after ultraviolet lamp is sterilized 20 minutes and are closed, open ultra-clean work
Make the other mode of operations of platform, the center on the mycelia access solidification Tube propagation base inclined-plane grown in culture dish from fructification surrounding
Behind position, put at 15 DEG C of people's biochemical cultivation case and cultivate after test tube mouth is obturaged with silica gel plug, invisible spectro mycelium is covered with after 7 days
Afterwards, so as to obtain 100% morchella mother culture.
Embodiment 3
A kind of purity is the preparation method of 100% morchella mother culture, it is characterised in that comprised the following steps：
1st, the preparation of culture medium：
A:It is standby to be put into superclean bench after one group of blank culture dish autoclaving 8；
B：The mother culture media autoclaving of the liquid condition in glass container, superclean bench is put into after bacterium of having gone out,
Ultraviolet lamp sterilization is opened, after 4.5 minutes, ultraviolet lamp is closed；
C：Superclean bench air supply pattern is opened, quickly the mother of the liquid condition by step B in superclean bench
Culture medium is planted to be dispensed into by each blank culture dish in step A, the thickness for the culture medium being dispensed into culture dish is
3.2mm；
2nd, the preparation of Morchella esculenta (L.) Pers sporophore：The new fresh sporophore of ready hickory chick is put into superclean bench, ultraviolet
After sterilization 25 minutes, ultraviolet lamp is closed, subsequent step is stand-by；
3rd, cultivate：Open on the inside of the other mode of operations of superclean bench, the hickory chick stem for step 2 of being learnt from else's experience with scalpel
Mung bean grain size fructification block, then bundle solid block is put into the culture medium of the center by the culture dish in step 1
In, mouth is sealed in the edge of culture dish by the lid for then covering culture dish with sealed membrane, is being put into biochemical cultivation case culture 3.5
My god, temperature control system is at 15 DEG C, and fructification block surrounding grows mycelium；
4th, mycelia is observed：With transfer needle picking, some are put into preprepared cover glass to the mycelium obtained in step 3
On, and add a small amount of sterilized water to drop on mycelium, and with being taken after slide compacting mycelium under microscope, seen with 10 times of object lens
Examine, confirmation is the mycelium of hickory chick, whole process will be carried out in the case where the other mode of operations of superclean bench are opened；
5th, mycelium is purified：After step 4 observation, without carrying out purification work again after other miscellaneous bacteria mycelia and miscellaneous bacteria spore
Make, the slant tube culture medium that autoclaving is good and solidifies is put into superclean bench, cultured mycelium is put into super
Other inoculating tools, are put into superclean bench by net workbench, are opened after ultraviolet lamp is sterilized 25 minutes and are closed, open ultra-clean work
Make the other mode of operations of platform, the center on the mycelia access solidification Tube propagation base inclined-plane grown in culture dish from fructification surrounding
Behind position, put at 16 DEG C of people's biochemical cultivation case and cultivate after test tube mouth is obturaged with silica gel plug, invisible spectro mycelium is covered with after 7 days
Afterwards, so as to obtain 100% morchella mother culture.
Described above, only presently preferred embodiments of the present invention is not intended to limit the invention, patent protection model of the invention
Enclose and be defined by claims, the equivalent structure that the content of every specification with the present invention is made changes, and all should similarly wrap
Containing within the scope of the present invention.
Claims (3)
1. a kind of purity is the preparation method of 100% morchella mother culture, it is characterised in that comprised the following steps：
(1) preparation of culture medium：
A:It is standby superclean bench is put into after blank culture dish autoclaving；
B：Superclean bench is put into after mother culture media autoclaving, bacterium of having gone out, ultraviolet lamp sterilization, 4 minutes~5 minutes is opened
Afterwards, ultraviolet lamp is closed；
C：Superclean bench air supply pattern is opened, quickly the mother culture media for passing through step B is dispensed into superclean bench
By in the blank culture dish in step A；
(2) preparation of Morchella esculenta (L.) Pers sporophore：The new fresh sporophore of ready hickory chick is put into superclean bench, ultraviolet sterilization
After 20 minutes~30 minutes, ultraviolet lamp is closed, subsequent step is stand-by；
(3) cultivate：Open on the inside of the other mode of operations of superclean bench, the hickory chick stem for step 2 of being learnt from else's experience with scalpel
The fructification block of mung bean grain size, then bundle solid block are put into the culture medium by the center of the culture dish in step 1,
Mouth is sealed in the edge of culture dish by the lid for then covering culture dish with sealed membrane, is being put into biochemical cultivation case culture 3~4 days,
Temperature control system is at 12 DEG C~18 DEG C, and fructification block surrounding grows mycelium；
(4) mycelia is observed：With transfer needle picking, some are put on preprepared cover glass the mycelium obtained in step 3,
And add a small amount of sterilized water to drop on mycelium, and with being taken after slide compacting mycelium under microscope, confirmation is hickory chick
Mycelium, whole process will be carried out in the case where the other mode of operations of superclean bench are opened；
(5) mycelium is purified：After step 4 observation, without carrying out Purification again after other miscellaneous bacteria mycelia and miscellaneous bacteria spore,
The slant tube culture medium that autoclaving is good and solidifies is put into superclean bench, cultured mycelium is put into ultra-clean work
Make platform, other inoculating tools are put into superclean bench, open after ultraviolet lamp is sterilized 20 minutes~30 minutes and close, open super
The other mode of operations of net workbench, the mycelia access solidification Tube propagation base inclined-plane grown in culture dish from fructification surrounding
Behind center, put at 15 DEG C~18 DEG C of people's biochemical cultivation case and cultivate after test tube mouth is obturaged with silica gel plug, it is invisible spectro after 7 days
After mycelium is covered with, morchella mother culture is obtained.
2. purity according to claim 1 is the preparation method of 100% morchella mother culture, it is characterised in that in step 1
Step C in, the thickness of the culture medium being dispensed into culture dish is 1.5mm~4mm.
3. purity according to claim 1 is the preparation method of 100% morchella mother culture, it is characterised in that in step 4
Microscope is from 10 times of object lens observation mycelium.
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| CN201710548197.6A CN107177514A (en) | 2017-07-06 | 2017-07-06 | A kind of purity is the preparation method of 100% morchella mother culture |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107810786A (en) * | 2017-10-20 | 2018-03-20 | 四川三点水生物科技有限公司 | A kind of disposable method of purification of simple hickory chick strain |
| CN108384711A (en) * | 2018-05-29 | 2018-08-10 | 四川省农业科学院植物保护研究所 | A kind of efficient hickory chick strain separating device and method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4594809A (en) * | 1985-04-29 | 1986-06-17 | Neogen Corporation | Cultivation of morchella |
| CN102242183A (en) * | 2011-06-16 | 2011-11-16 | 河南科技大学 | Morel strain identifying method |
-
2017
- 2017-07-06 CN CN201710548197.6A patent/CN107177514A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4594809A (en) * | 1985-04-29 | 1986-06-17 | Neogen Corporation | Cultivation of morchella |
| CN102242183A (en) * | 2011-06-16 | 2011-11-16 | 河南科技大学 | Morel strain identifying method |
Non-Patent Citations (1)
| Title |
|---|
| 王永斌: "组织分离法制作羊肚菌母种的关键环节", 《北方园艺》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107810786A (en) * | 2017-10-20 | 2018-03-20 | 四川三点水生物科技有限公司 | A kind of disposable method of purification of simple hickory chick strain |
| CN108384711A (en) * | 2018-05-29 | 2018-08-10 | 四川省农业科学院植物保护研究所 | A kind of efficient hickory chick strain separating device and method |
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