CN105779310A - Morchella strain isolation method - Google Patents
Morchella strain isolation method Download PDFInfo
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- CN105779310A CN105779310A CN201610324807.XA CN201610324807A CN105779310A CN 105779310 A CN105779310 A CN 105779310A CN 201610324807 A CN201610324807 A CN 201610324807A CN 105779310 A CN105779310 A CN 105779310A
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- morchella
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- morchella esculenta
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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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Abstract
The invention discloses a morchella strain isolation method, belonging to the technical field of strain cultivation. The method comprises the following steps: preparing a disposable cup and a toothpick and performing disinfection and sterilization treatment; preparing a potato medium, antibiotic medium or water agar medium and morchella in a mature stage; cutting the morchella root, flushing the surface with sterile water and sucking moisture on the surface; transversely piercing the morchella stem with the toothpick; hanging the morchella upside down in the disposable cup; reversing the disposable up on the prepared medium; changing the medium every 6-8h until no spore is sprayed; and culturing the changed medium at 15-25 DEG C to isolate the morchella strain. The morchella strain isolation method is implemented by use of the toothpick and disposable cup, and the morchella strain can be isolated through tool disinfection, morchella cleaning, spore collection and spore culture; and the method has the advantages of simple tools, high operability, high germination rate and low cost and provides a new scheme for artificial cultivation of morchella.
Description
Technical field
The present invention relates to breeding strain technical field, be specially a kind of Morchella esculenta (L.) Pers strain separating method.
Background technology
Morchella esculenta (L.) Pers is a kind of Rare edible fungus kind, gains the name because of its cap surface irregularity, shape such as Gaster caprae seu Ovis.Morchella esculenta (L.) Pers
Being foremost delicious food bacterium in ascomycetes, its cap part contains isoleucine, leucine, lysine, methionine, phenylpropyl alcohol ammonia
Acid, threonine and the aminoacid of 7 kinds of needed by human of valine, sweet cold nontoxic, useful the intestines and stomach, phlegm-eliminiating and qi-regulating drug effect.Morchella esculenta (L.) Pers
Nutrition is the abundantest, and according to surveying and determination, Morchella esculenta (L.) Pers contains crude protein 20%, crude fat 26%, carbohydrate 38.1%, possibly together with multiple ammonia
Base acid, particularly content of glutamic acid is up to 1.76%.Therefore, Morchella esculenta (L.) Pers is the best food protein source, and has " in element
Meat or fish " laudatory title.Protein in human body is made up of the collocation of 20 kinds of aminoacid, and Morchella esculenta (L.) Pers just contains 18 kinds, and wherein 8
Planting aminoacid is that human body can not manufacture.
Owing to Morchella esculenta (L.) Pers exists technical difficulty in strain separating and purge process, duplicating property of artificial culture is poor, yield
Performance low, inferior, is difficult to produce promote at present.
Summary of the invention
It is an object of the invention to provide a kind of simple, easily operation, Morchella esculenta (L.) Pers strain separating method that efficiency is high.
For reaching above-mentioned purpose, the technical scheme of employing is: a kind of Morchella esculenta (L.) Pers strain separating method, first prepares disposable
Water tumbler and toothpick, and carry out disinfection, sterilization processing；Make potato culture, Antibiotic medium or water agar；With
And the Morchella esculenta (L.) Pers in period of maturation；
Morchella esculenta (L.) Pers is removed root, uses aseptic water washing surface, then blot surface moisture；Morchella esculenta (L.) Pers stem is crossed, Gaster caprae seu Ovis with toothpick
Bacterium hangs upside down in disposable water cup, and disposable water cup is placed upside down in ready-made culture medium, changes a subculture every 6-8h, until
Not fill-before-fire spore, is placed in 15-25 DEG C of cultivation by the culture medium changed, can isolated Morchella esculenta (L.) Pers strain.
Described disposable water cup and toothpick use 75% alcohol disinfecting, then superclean bench medium ultraviolet sterilization 15-25min.
The manufacture method of described potato culture is: Rhizoma Solani tuber osi 200g, and 20-25min is boiled in peeling, 8 layers of filtered through gauze
Take filtrate, be subsequently adding 20g glucose or sucrose, 20g agar, be settled to 1L, high temperature 121 DEG C, high pressure 0. 1MPa sterilizing
25min, treats that culture medium is cooled to 50-60 DEG C, pours sterilizing flat board or disposable sterilized flat board into.
The manufacture method of described Antibiotic medium is: Rhizoma Solani tuber osi 200g, and 20-25min is boiled in peeling, 8 layers of filtered through gauze
Take filtrate, be subsequently adding 20g glucose or sucrose, 20g agar, be settled to 1L, high temperature 121 DEG C, high pressure 0. 1MPa sterilizing
25min, treats that culture medium is cooled to 50 DEG C and adds streptomycin 30U/mL and penicillin 30U/mL, is subsequently poured into sterilizing flat board or once
The aseptic flat board of property.
The manufacture method of described water agar is: 1L water adds 20g agar, and high temperature 121 DEG C, high pressure 0. 1MPa goes out
Bacterium 25min, treats that culture medium is cooled to 50-60 DEG C, pours sterilizing flat board or disposable sterilized flat board into.High temperature 121 DEG C, high pressure
0. 1MPa sterilizing 25min, treats that culture medium is cooled to 50-60 DEG C, pours sterilizing flat board or disposable sterilized flat board into.
Use such scheme has the beneficial effect that this Morchella esculenta (L.) Pers strain separating method uses toothpick and disposable water cup to enter
OK, just recruitment can be made with isolated Morchella esculenta (L.) Pers strain through instrument sterilization, Morchella esculenta (L.) Pers cleaning, collection spore and cultivation spore
Tool is simple, workable, and germination rate is high, and with low cost, the artificial growth for Morchella esculenta (L.) Pers provides new departure.
Accompanying drawing explanation
Fig. 1 is the structural representation of Morchella esculenta (L.) Pers strain separating device of the present invention.
Fig. 2 is the Morchella esculenta (L.) Pers. Mycelium of isolated of the present invention.
Fig. 3 is the microstructure of Morchella esculenta (L.) Pers ascospore 400 times.
In figure, 1-disposable water cup, 2-toothpick, 3-thalline, 4-culture plate.
Detailed description of the invention
It is further described the present invention below in conjunction with embodiment and accompanying drawing, but the present invention is not limited only to following embodiment, permissible
Those skilled in the art are in the case of combining prior art in prediction, and performance may produce many variations.
Such as Fig. 1, for the structural representation of Morchella esculenta (L.) Pers strain separating device of the present invention.
Morchella esculenta (L.) Pers strain separating method, first prepares disposable water cup 1 and toothpick 2, and carry out disinfection, sterilization processing；System
Make potato culture, Antibiotic medium or water agar；And the Gaster caprae seu Ovis thalline thalline 3 in period of maturation.
Morchella esculenta (L.) Pers thalline 3 is removed root, uses aseptic water washing surface, then blot surface moisture；Thalline 3 is crossed with toothpick 2
Stem, hangs upside down thalline 3 in disposable water cup 1, and disposable water cup 1 is placed upside down on the culture plate 4 installing culture medium, every 6-
8h changes a subculture, continues about 3 days, until not fill-before-fire spore, the culture medium changed is placed in 15-25 DEG C of cultivation,
Can isolated Morchella esculenta (L.) Pers strain.
Described disposable water cup and toothpick use 75% alcohol disinfecting, then superclean bench medium ultraviolet sterilization 15-25min.
The manufacture method of described potato culture is: Rhizoma Solani tuber osi 200g, and 20-25min is boiled in peeling, 8 layers of filtered through gauze
Take filtrate, be subsequently adding 20g glucose or sucrose, 20g agar, be settled to 1L, high temperature 121 DEG C, high pressure 0. 1MPa sterilizing
25min, treats that culture medium is cooled to 50-60 DEG C, pours sterilizing flat board or disposable sterilized flat board into.
The manufacture method of described Antibiotic medium is: Rhizoma Solani tuber osi 200g, and 20-25min is boiled in peeling, 8 layers of filtered through gauze
Take filtrate, be subsequently adding 20g glucose or sucrose, 20g agar, be settled to 1L, high temperature 121 DEG C, high pressure 0. 1MPa sterilizing
25min, treats that culture medium is cooled to 50 DEG C and adds streptomycin 30U/mL and penicillin 30U/mL, is subsequently poured into sterilizing flat board or once
The aseptic flat board of property.
The manufacture method of described water agar is: 1L water adds 20g agar, and high temperature 121 DEG C, high pressure 0. 1MPa goes out
Bacterium 25min, treats that culture medium is cooled to 50-60 DEG C, pours sterilizing flat board or disposable sterilized flat board into.High temperature 121 DEG C, high pressure
0. 1MPa sterilizing 25min, treats that culture medium is cooled to 50-60 DEG C, pours sterilizing flat board or disposable sterilized flat board into.
Such as the Morchella esculenta (L.) Pers. Mycelium that Fig. 2 is isolated of the present invention.
Such as the microstructure that Fig. 3 is Morchella esculenta (L.) Pers ascospore 400 times.
The present invention does not carries out alcohol disinfecting to Morchella esculenta (L.) Pers sporophore, can prevent ethanol from immersing thalline tissue, with traditional clock
Cover method separates and compares little, the strong operability that takes up space, and can high-volume separate simultaneously, and efficiency is obviously improved.Because Morchella esculenta (L.) Pers is raw
Long rapid, the mycelia of isolated needs timely picking to be purified.
Claims (5)
1. a Morchella esculenta (L.) Pers strain separating method, is characterized in that:
Prepare disposable water cup and toothpick, and carry out disinfection, sterilization processing；Make potato culture, Antibiotic medium or
Water agar；And the Morchella esculenta (L.) Pers in period of maturation；
Morchella esculenta (L.) Pers is removed root, uses aseptic water washing surface, then blot surface moisture；Morchella esculenta (L.) Pers stem is crossed, Gaster caprae seu Ovis with toothpick
Bacterium hangs upside down in disposable water cup, and disposable water cup is placed upside down in ready-made culture medium, changes a subculture every 6-8h, until
Not fill-before-fire spore, is placed in 15-25 DEG C of cultivation by the culture medium changed, can isolated Morchella esculenta (L.) Pers strain.
Morchella esculenta (L.) Pers strain separating method the most according to claim 1, is characterized in that: described disposable water cup and toothpick use
75% alcohol disinfecting, then superclean bench medium ultraviolet sterilization 15-25min.
Morchella esculenta (L.) Pers strain separating method the most according to claim 1, is characterized in that: the making side of described potato culture
Method is: Rhizoma Solani tuber osi 200g, and 20-25min is boiled in peeling, and 8 layers of filtered through gauze take filtrate, are subsequently adding 20g glucose or sucrose,
20g agar, is settled to 1L, high temperature 121 DEG C, high pressure 0. 1MPa sterilizing 25min, treats that culture medium is cooled to 50-60 DEG C, pour into and go out
Bacterium flat board or disposable sterilized flat board.
Morchella esculenta (L.) Pers strain separating method the most according to claim 1, is characterized in that: the making side of described Antibiotic medium
Method is: Rhizoma Solani tuber osi 200g, and 20-25min is boiled in peeling, and 8 layers of filtered through gauze take filtrate, are subsequently adding 20g glucose or sucrose,
20g agar, is settled to 1L, high temperature 121 DEG C, high pressure 0. 1MPa sterilizing 25min, treats that culture medium is cooled to 50 DEG C and adds streptomycin
30U/mL and penicillin 30U/mL, is subsequently poured into sterilizing flat board or disposable sterilized flat board.
Morchella esculenta (L.) Pers strain separating method the most according to claim 1, is characterized in that: the making side of described water agar
Method is: 1L water adds 20g agar, high temperature 121 DEG C, high pressure 0. 1MPa sterilizing 25min, treats that culture medium is cooled to 50-60 DEG C, falls
Enter sterilizing flat board or disposable sterilized flat board；High temperature 121 DEG C, high pressure 0. 1MPa sterilizing 25min, treats that culture medium is cooled to
50-60 DEG C, pour sterilizing flat board or disposable sterilized flat board into.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610324807.XA CN105779310A (en) | 2016-05-17 | 2016-05-17 | Morchella strain isolation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610324807.XA CN105779310A (en) | 2016-05-17 | 2016-05-17 | Morchella strain isolation method |
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| CN105779310A true CN105779310A (en) | 2016-07-20 |
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| CN201610324807.XA Pending CN105779310A (en) | 2016-05-17 | 2016-05-17 | Morchella strain isolation method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109122049A (en) * | 2018-08-03 | 2019-01-04 | 内蒙古农业大学 | A kind of more spore separators of Cordyceps militaris and method |
| CN111448939A (en) * | 2020-05-12 | 2020-07-28 | 毕节市农业科学研究所 | Wild morchella planting method |
| CN112449988A (en) * | 2020-10-20 | 2021-03-09 | 安徽乐永园农业科技有限公司 | Breeding and transferring method suitable for morchella |
| CN112655468A (en) * | 2020-10-20 | 2021-04-16 | 安徽乐永园农业科技有限公司 | Process suitable for fungus picking spores to prevent mixed bacteria |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104928195A (en) * | 2015-07-10 | 2015-09-23 | 青川县智农农业开发有限公司 | Toadstool strain isolation method |
-
2016
- 2016-05-17 CN CN201610324807.XA patent/CN105779310A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104928195A (en) * | 2015-07-10 | 2015-09-23 | 青川县智农农业开发有限公司 | Toadstool strain isolation method |
Non-Patent Citations (3)
| Title |
|---|
| 刘胜责等: "一种简易的抱子采集方法", 《食用菌》 * |
| 汤光华等: "食用菌菌种的抱子分离法", 《植物杂志》 * |
| 麻兵继: "《药食兼用真菌重要次生代谢产物及其生物活性》", 30 November 2011 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109122049A (en) * | 2018-08-03 | 2019-01-04 | 内蒙古农业大学 | A kind of more spore separators of Cordyceps militaris and method |
| CN111448939A (en) * | 2020-05-12 | 2020-07-28 | 毕节市农业科学研究所 | Wild morchella planting method |
| CN112449988A (en) * | 2020-10-20 | 2021-03-09 | 安徽乐永园农业科技有限公司 | Breeding and transferring method suitable for morchella |
| CN112655468A (en) * | 2020-10-20 | 2021-04-16 | 安徽乐永园农业科技有限公司 | Process suitable for fungus picking spores to prevent mixed bacteria |
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Application publication date: 20160720 |