CN107129938A - It is a kind of to improve the method that morchella mother culture manufactures success rate - Google Patents
It is a kind of to improve the method that morchella mother culture manufactures success rate Download PDFInfo
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- CN107129938A CN107129938A CN201710547037.XA CN201710547037A CN107129938A CN 107129938 A CN107129938 A CN 107129938A CN 201710547037 A CN201710547037 A CN 201710547037A CN 107129938 A CN107129938 A CN 107129938A
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- 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
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Abstract
The method that morchella mother culture manufactures success rate is improved the invention discloses a kind of, is related to morchella mother culture preparation method field, comprises the following steps：The preparation of culture medium, the preparation of inoculating tool, the preparation of fructification, culture and mycelia purification, the present invention turns down the cultivation temperature of Morchella esculenta (L.) Pers sporophore tissue block in culture dish, effectively inhibit the growth of miscellaneous bacteria mycelia and miscellaneous bacteria spore, effectively raise the success rate of parent species making, the method of invention to turn out the probability increase by 80% that the mycelia come is entirely Morciiella Esculeuta Mycelia, the success rate of morchella mother culture making is so also just improved, morchella mother culture power can be fabricated to and bring up to 60% or so.
Description
Technical field
The present invention relates to biological study field, success rate is manufactured more particularly to a kind of raising morchella mother culture
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.
In recent years, as hickory chick industry is by the understanding and concern of increasing people, also with planting technology increasingly
It is perfect, therefore there is many enterprises or self-employed worker to be added to hickory chick growing industry, in order to which the plantation for reducing oneself is put into
This, many enterprises or self-employed worker think oneself production hickory chick strain, but but often in this ring of morchella mother culture production
Just it have failed in section, they are during parent species are made, and the Morchella esculenta (L.) Pers sporophore fritter that (1) can have access test tube is not sprouted
Situation, it is existing make be during morchella mother culture：Whole Morchella esculenta (L.) Pers sporophore is cut with scalpel in an aseptic environment,
Stem part is left and taken, stem is indulged face and scratched with scalpel, stem inner hollow, the stem inner side and outer side after scratching all is used
75% alcohol swab wipes the Morchella esculenta (L.) Pers sporophore for carrying out cutting 0.2 centimetre square after sterilization processing, alcohol wipe with scalpel
Tissue block, which is put into after culture dish center, sealing, is put into the degree of biochemical cultivation case 23-25 culture 3 days, in such making parent species
Under mode of operation, the tissue block of access can be caused not sprout, because the process of alcohol wipe sterilization is killing the same of other miscellaneous bacterias
When can also kill the tissue block of hickory chick, the hickory chick tissue block of access culture dish is lost activity, directly result in making failure；
(2) it there is also and be linked into the Morchella esculenta (L.) Pers sporophore tissue block of test tube and sprout and have issued mycelium, but be not singly
Morciiella Esculeuta Mycelia also has other miscellaneous bacteria mycelia, after Morciiella Esculeuta Mycelia is together with miscellaneous bacteria mycelial growth, be difficult to they are independent
Separate, just have failed in the production first step, their parent species are fabricated to power generally only 1% or lower.
The content of the invention
It is an object of the invention to：In order to solve the pure low success rate of technical problem of existing morchella mother culture system, the present invention
A kind of method that raising morchella mother culture manufactures success rate is provided.
The present invention specifically uses following technical scheme to achieve these goals：
It is a kind of to improve the method that morchella mother culture manufactures success rate, it is characterised in that to comprise the following steps：
1st, the preparation of culture medium：Mother culture media is made, mother culture media is put into after autoclaving, cooling and solidification
In culture dish, then that culture dish is put into superclean bench is standby；
2nd, the preparation of inoculating tool：It is standby that the various inoculating tools such as bag scalpel are put into superclean bench；
3rd, the preparation of fructification：The new fresh sporophore of ready hickory chick is put into superclean bench, ultraviolet sterilization 20
It is standby after~30 minutes；
4th, cultivate：In an aseptic environment, the whole hickory chick in step 3 is cut with the scalpel in step 2, leaves and takes stem
Part, scratches stem with scalpel, and the tissue block for taking stem inside length to be 0.2 centimetre is put into the culture dish in step 1
Heart position, is put at 12 DEG C~18 DEG C of biochemical cultivation case after culture dish sealing and cultivates 3~4 days, tissue block surrounding grows mycelia, when
During the long 0.8cm~1cm of mycelia in culture dish around hickory chick tissue block, next step purification is carried out；
5th, mycelia purifies：Culture dish Jing Guo step 4 is put into superclean bench, other inoculating tools are put into ultra-clean work
Make platform, open after ultraviolet lamp is sterilized 20~30 minutes and close, the other mode of operations of superclean bench are opened, sheep in culture dish
Behind the center on the mycelia length access solidification Tube propagation base inclined-plane around tripe hyphostroma block, test tube mouthful is obturaged with silica gel plug
After put and cultivated at 12 DEG C~18 DEG C of people's biochemical cultivation case, observe whether the mycelium that covers with is the mycelia of hickory chick, so as to obtain
Morchella mother culture.
Further, in step 4, as the long 1cm of the mycelia in culture dish around hickory chick tissue block, next step is carried out
Purification.
Further, the formula of mother culture media is that the formula of mother culture media in step 1 is in step 1：Potato 200
Gram, 20 grams of glucose, 20 grams of agar, 5 grams of peptone, 3 grams of potassium dihydrogen phosphate, 1.5 grams of magnesium sulfate, 1000 milliliters of water.
The compound method of morchella mother culture culture medium：200g potatos are weighed, peeling chopping is cleaned, the 1000ml that adds water boils
Half an hour, filtered through gauze rejoins potato filtrate in pot, moisturizing to 1000ml, continues to add after ebuillition of heated
20g agar, continue slow fire boil to agar be completely dissolved after again moisturizing to 1000 milliliters, agar dissolving after add glucose
20 grams, 3 grams of potassium dihydrogen phosphate, 1.5 grams of magnesium sulfate, 5 grams of peptone.Test tube is dispensed, per test tube about 5-10ml, depending on test tube size
It is fixed, the test tube of silica gel plug that is stoppered dispensed is put into portable pressure cooker in sterilizing, sterilization process first first emptying pressure cooker
Interior cold air, rises to whne temperature and starts timing when 122-124 spends, and sterilizing terminates after 30 minutes, test tube pendulum inclined-plane is taken out, after cooling
Storage is standby.
Beneficial effects of the present invention are as follows：
1st, the present invention according to hickory chick is a kind of mushroom of suitable Low- temperature culture, and the suitable culture season in its outdoor crop field is
The period in autumn to spring, thus Morciiella Esculeuta Mycelia can be grown in a low temperature of certain it is fine, and miscellaneous bacteria mycelia is in height
The speed of growth is accelerated under warm condition of culture, and growth is slowed down under cryogenic, or is stopped growing, using such principle, adjusts
The cultivation temperature of Morchella esculenta (L.) Pers sporophore tissue block in low culture dish, effectively inhibits the growth of miscellaneous bacteria mycelia and miscellaneous bacteria spore,
Effectively raise the success rate of parent species making, by the method for the present invention to turn out come mycelia be entirely hickory chick bacterium
The probability increase by 80% of silk, so also just improves the success rate of morchella mother culture making, morchella mother culture can be successful
Rate brings up to 60% or so.
2nd, as the long 1cm of the mycelia in culture dish around hickory chick tissue block, next step purification is carried out, is that one kind more has choosing
Select, morchella mother culture can be made to be fabricated to power and bring up to 61.5% or so.
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 provides a kind of method that raising morchella mother culture manufactures success rate, comprises the following steps：
1st, the preparation of culture medium：Mother culture media is made, mother culture media is put into after autoclaving, cooling and solidification
In culture dish, then that culture dish is put into superclean bench is standby；
2nd, the preparation of inoculating tool：It is standby that the various inoculating tools such as bag scalpel are put into superclean bench；
3rd, the preparation of fructification：The new fresh sporophore of ready hickory chick is put into superclean bench, ultraviolet sterilization 20
It is standby after minute；
4th, cultivate：In an aseptic environment, the whole hickory chick in step 3 is cut with the scalpel in step 2, leaves and takes stem
Part, scratches stem with scalpel, and the tissue block for taking stem inside length to be 0.2 centimetre is put into the culture dish in step 1
Heart position, is put at 12 DEG C of biochemical cultivation case after culture dish sealing and cultivates 3 days, tissue block surrounding grows mycelia, when in culture dish
During the long 0.8cm of mycelia around hickory chick tissue block, next step purification is carried out；
5th, mycelia purifies：Culture dish Jing Guo step 4 is put into superclean bench, other inoculating tools are put into ultra-clean work
Make platform, open after ultraviolet lamp is sterilized 20 minutes and close, the other mode of operations of superclean bench are opened, hickory chick in culture dish
Behind the center on the mycelia length access solidification Tube propagation base inclined-plane around tissue block, put after test tube mouth is obturaged with silica gel plug
Cultivated at 12 DEG C of people's biochemical cultivation case, observe whether the mycelium covered with is the mycelia of hickory chick, so as to obtain morchella mother culture.
Morchella mother culture can be fabricated to power and bring up to 62% by the present embodiment.
Embodiment 2
The present embodiment provides a kind of method that raising morchella mother culture manufactures success rate, comprises the following steps：
1st, the preparation of culture medium：Mother culture media is made, mother culture media is put into after autoclaving, cooling and solidification
In culture dish, then that culture dish is put into superclean bench is standby；
2nd, the preparation of inoculating tool：It is standby that the various inoculating tools such as bag scalpel are put into superclean bench；
3rd, the preparation of fructification：The new fresh sporophore of ready hickory chick is put into superclean bench, ultraviolet sterilization 30
It is standby after minute；
4th, cultivate：In an aseptic environment, the whole hickory chick in step 3 is cut with the scalpel in step 2, leaves and takes stem
Part, scratches stem with scalpel, and the tissue block for taking stem inside length to be 0.2 centimetre is put into the culture dish in step 1
Heart position, is put at 18 DEG C of biochemical cultivation case after culture dish sealing and cultivates 4 days, tissue block surrounding grows mycelia, when in culture dish
During the long 1cm of mycelia around hickory chick tissue block, next step purification is carried out；
5th, mycelia purifies：Culture dish Jing Guo step 4 is put into superclean bench, other inoculating tools are put into ultra-clean work
Make platform, open after ultraviolet lamp is sterilized 30 minutes and close, the other mode of operations of superclean bench are opened, hickory chick in culture dish
Behind the center on the mycelia length access solidification Tube propagation base inclined-plane around tissue block, put after test tube mouth is obturaged with silica gel plug
Cultivated at 18 DEG C of people's biochemical cultivation case, observe whether the mycelium covered with is the mycelia of hickory chick, so as to obtain morchella mother culture.
Morchella mother culture can be fabricated to power and bring up to 61.3% by the present embodiment.
Implement 3
The present embodiment provides a kind of method that raising morchella mother culture manufactures success rate, comprises the following steps：
1st, the preparation of culture medium：Mother culture media is made, mother culture media is put into after autoclaving, cooling and solidification
In culture dish, then that culture dish is put into superclean bench is standby；
2nd, the preparation of inoculating tool：It is standby that the various inoculating tools such as bag scalpel are put into superclean bench；
3rd, the preparation of fructification：The new fresh sporophore of ready hickory chick is put into superclean bench, ultraviolet sterilization 25
It is standby after minute；
4th, cultivate：In an aseptic environment, the whole hickory chick in step 3 is cut with the scalpel in step 2, leaves and takes stem
Part, scratches stem with scalpel, and the tissue block for taking stem inside length to be 0.2 centimetre is put into the culture dish in step 1
Heart position, is put at 16 DEG C of biochemical cultivation case after culture dish sealing and cultivates 3.5 days, tissue block surrounding grows mycelia, works as culture dish
In mycelia around hickory chick tissue block long 0.9cm when, carry out next step purification；
5th, mycelia purifies：Culture dish Jing Guo step 4 is put into superclean bench, other inoculating tools are put into ultra-clean work
Make platform, open after ultraviolet lamp is sterilized 25 minutes and close, the other mode of operations of superclean bench are opened, hickory chick in culture dish
Behind the center on the mycelia length access solidification Tube propagation base inclined-plane around tissue block, put after test tube mouth is obturaged with silica gel plug
Cultivated at 16 DEG C of people's biochemical cultivation case, observe whether the mycelium covered with is the mycelia of hickory chick, so as to obtain morchella mother culture.
Embodiment 4
The formula of mother culture media is：200 grams of potato, 20 grams of glucose, 20 grams of agar, 5 grams of peptone, biphosphate
3 grams of potassium, 1.5 grams of magnesium sulfate, 1000 milliliters of water.
The compound method of morchella mother culture culture medium：200g potatos are weighed, peeling chopping is cleaned, the 1000ml that adds water boils
Half an hour, filtered through gauze rejoins potato filtrate in pot, moisturizing to 1000ml, continues to add after ebuillition of heated
20g agar, continue slow fire boil to agar be completely dissolved after again moisturizing to 1000 milliliters, agar dissolving after add glucose
20 grams, 3 grams of potassium dihydrogen phosphate, 1.5 grams of magnesium sulfate, 5 grams of peptone.Test tube is dispensed, per test tube about 5-10ml, depending on test tube size
It is fixed, the test tube of silica gel plug that is stoppered dispensed is put into portable pressure cooker in sterilizing, sterilization process first first emptying pressure cooker
Interior cold air, rises to whne temperature and starts timing when 122-124 spends, and sterilizing terminates after 30 minutes, test tube pendulum inclined-plane is taken out, after cooling
Storage is standby.
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 improve the method that morchella mother culture manufactures success rate, it is characterised in that comprises the following steps：
(1) preparation of culture medium：Mother culture media is made, mother culture media is put into training after autoclaving, cooling and solidification
Support in ware, then that culture dish is put into superclean bench is standby；
(2) preparation of inoculating tool：It is standby that the various inoculating tools such as bag scalpel are put into superclean bench；
(3) preparation of fructification：The new fresh sporophore of ready hickory chick is put into superclean bench, ultraviolet sterilization 20~30
It is standby after minute；
(4) cultivate：In an aseptic environment, the whole hickory chick in step 3 is cut with the scalpel in step 2, leaves and takes stem portion
Point, stem is scratched with scalpel, the tissue block for taking stem inside length to be 0.2 centimetre is put into the culture dish center in step 1
Position, is put at 12 DEG C~18 DEG C of biochemical cultivation case after culture dish sealing and cultivates 3~4 days, tissue block surrounding grows mycelia, works as training
When supporting the long 0.8cm~1cm of mycelia in ware around hickory chick tissue block, next step purification is carried out；
(5) mycelia purifies：Culture dish Jing Guo step 4 is put into superclean bench, other inoculating tools are put into ultra-clean work
Platform, opens after ultraviolet lamp is sterilized 20~30 minutes and closes, the other mode of operations of superclean bench are opened, sheep tripe in culture dish
Behind the center on the mycelia length access solidification Tube propagation base inclined-plane around hyphostroma block, after test tube mouth is obturaged with silica gel plug
Put and cultivated at 12 DEG C~18 DEG C of people's biochemical cultivation case, observe whether the mycelium covered with is the mycelia of hickory chick, so as to obtain sheep
Tripe starter kind.
2. the method that morchella mother culture manufactures success rate is improved, it is characterised in that in step 4, when hickory chick is organized in culture dish
During the long 1cm of mycelia around block, next step purification is carried out.
3. improve the method that morchella mother culture manufactures success rate, it is characterised in that the formula of mother culture media is in step 1：Horse
200 grams of bell potato, 20 grams of glucose, 20 grams of agar, 5 grams of peptone, 3 grams of potassium dihydrogen phosphate, 1.5 grams of magnesium sulfate, 1000 milliliters of water.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710547037.XA CN107129938A (en) | 2017-07-06 | 2017-07-06 | It is a kind of to improve the method that morchella mother culture manufactures success rate |
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Cited By (3)
| 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 |
| CN108812079A (en) * | 2018-05-30 | 2018-11-16 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of hickory chick strain separating method |
| CN111937676A (en) * | 2020-08-11 | 2020-11-17 | 南京康之春生物科技有限公司 | Morchella mycelium culture medium and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4594809A (en) * | 1985-04-29 | 1986-06-17 | Neogen Corporation | Cultivation of morchella |
| CN106544279A (en) * | 2016-11-25 | 2017-03-29 | 秦小波 | A kind of Morchella esculenta (L.) Perss cultivation strain and its induction and cultural method |
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2017
- 2017-07-06 CN CN201710547037.XA patent/CN107129938A/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 |
| CN106544279A (en) * | 2016-11-25 | 2017-03-29 | 秦小波 | A kind of Morchella esculenta (L.) Perss cultivation strain and its induction and cultural method |
Non-Patent Citations (2)
| Title |
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| 侯军: "高羊肚菌MH1菌株母种培养基的正交优化", 《食用菌》 * |
| 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 (3)
| 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 |
| CN108812079A (en) * | 2018-05-30 | 2018-11-16 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of hickory chick strain separating method |
| CN111937676A (en) * | 2020-08-11 | 2020-11-17 | 南京康之春生物科技有限公司 | Morchella mycelium culture medium and preparation method thereof |
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