CN105165389B - A kind of wild ' Ganba ' fungus industrial production method - Google Patents
A kind of wild ' Ganba ' fungus industrial production method Download PDFInfo
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- CN105165389B CN105165389B CN201510418076.0A CN201510418076A CN105165389B CN 105165389 B CN105165389 B CN 105165389B CN 201510418076 A CN201510418076 A CN 201510418076A CN 105165389 B CN105165389 B CN 105165389B
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- 241000233866 Fungi Species 0.000 title claims abstract description 25
- 238000009776 industrial production Methods 0.000 title abstract description 4
- 241000894006 Bacteria Species 0.000 claims abstract description 96
- 239000002689 soil Substances 0.000 claims abstract description 51
- 244000005700 microbiome Species 0.000 claims abstract description 3
- 238000004321 preservation Methods 0.000 claims abstract description 3
- 241000196324 Embryophyta Species 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- 238000003306 harvesting Methods 0.000 claims description 12
- 238000004140 cleaning Methods 0.000 claims description 10
- CSNNHWWHGAXBCP-UHFFFAOYSA-L mgso4 Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 10
- 240000007842 Glycine max Species 0.000 claims description 9
- 235000010469 Glycine max Nutrition 0.000 claims description 9
- 229940068124 Pine tar Drugs 0.000 claims description 9
- 240000008042 Zea mays Species 0.000 claims description 9
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 9
- 238000004458 analytical method Methods 0.000 claims description 9
- 239000011297 pine tar Substances 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 9
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 7
- 229940088594 Vitamin Drugs 0.000 claims description 6
- 235000013343 vitamin Nutrition 0.000 claims description 6
- 239000011782 vitamin Substances 0.000 claims description 6
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 6
- 229930003231 vitamins Natural products 0.000 claims description 6
- 229960004494 Calcium Gluconate Drugs 0.000 claims description 5
- NEEHYRZPVYRGPP-IYEMJOQQSA-L Calcium gluconate Chemical compound [Ca+2].OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O NEEHYRZPVYRGPP-IYEMJOQQSA-L 0.000 claims description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 5
- GNSKLFRGEWLPPA-UHFFFAOYSA-M Monopotassium phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 5
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims description 5
- 239000004227 calcium gluconate Substances 0.000 claims description 5
- 235000013927 calcium gluconate Nutrition 0.000 claims description 5
- 235000013312 flour Nutrition 0.000 claims description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 5
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 5
- 235000009973 maize Nutrition 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 5
- 235000008331 Pinus X rigitaeda Nutrition 0.000 abstract description 51
- 235000011613 Pinus brutia Nutrition 0.000 abstract description 51
- 241000018646 Pinus brutia Species 0.000 abstract description 51
- 235000015097 nutrients Nutrition 0.000 abstract description 31
- 239000001963 growth media Substances 0.000 abstract description 20
- 239000004033 plastic Substances 0.000 abstract description 14
- 238000004519 manufacturing process Methods 0.000 abstract description 11
- 238000001035 drying Methods 0.000 abstract description 6
- 239000003205 fragrance Substances 0.000 abstract description 6
- 230000004083 survival Effects 0.000 abstract description 5
- 235000016709 nutrition Nutrition 0.000 abstract description 2
- 230000035764 nutrition Effects 0.000 abstract description 2
- 239000003415 peat Substances 0.000 description 11
- 230000001954 sterilising Effects 0.000 description 10
- 238000004659 sterilization and disinfection Methods 0.000 description 10
- 238000005286 illumination Methods 0.000 description 8
- 238000009423 ventilation Methods 0.000 description 7
- 241000018652 Pinus yunnanensis Species 0.000 description 6
- 235000011611 Pinus yunnanensis Nutrition 0.000 description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N D-sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 4
- 235000005822 corn Nutrition 0.000 description 4
- 235000005824 corn Nutrition 0.000 description 4
- 238000005520 cutting process Methods 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 239000010440 gypsum Substances 0.000 description 4
- 229910052602 gypsum Inorganic materials 0.000 description 4
- 239000002609 media Substances 0.000 description 4
- 235000015099 wheat brans Nutrition 0.000 description 4
- 230000012010 growth Effects 0.000 description 3
- 239000005416 organic matter Substances 0.000 description 3
- 210000004215 spores Anatomy 0.000 description 3
- OYHQOLUKZRVURQ-IXWMQOLASA-N Linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000003698 anagen phase Effects 0.000 description 2
- 230000001488 breeding Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 229910001710 laterite Inorganic materials 0.000 description 2
- 239000011504 laterite Substances 0.000 description 2
- XJKSTNDFUHDPQJ-UHFFFAOYSA-N 1,4-diphenylbenzene Chemical group C1=CC=CC=C1C1=CC=C(C=2C=CC=CC=2)C=C1 XJKSTNDFUHDPQJ-UHFFFAOYSA-N 0.000 description 1
- 102000020179 B family Human genes 0.000 description 1
- 108091014904 B family Proteins 0.000 description 1
- 241000221198 Basidiomycota Species 0.000 description 1
- 102000011782 Keratins Human genes 0.000 description 1
- 108010076876 Keratins Proteins 0.000 description 1
- 235000005205 Pinus Nutrition 0.000 description 1
- 241000218602 Pinus <genus> Species 0.000 description 1
- 235000008591 Pinus cembroides var edulis Nutrition 0.000 description 1
- 244000003118 Pinus cembroides var. edulis Species 0.000 description 1
- 231100000614 Poison Toxicity 0.000 description 1
- 241000222383 Polyporales Species 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 241000566961 Thelephora Species 0.000 description 1
- 241000101568 Thelephora ganbajun Species 0.000 description 1
- 241000991705 Thelephora vialis Species 0.000 description 1
- 241000566950 Thelephoraceae Species 0.000 description 1
- 240000004942 Trevesia palmata Species 0.000 description 1
- 229940045997 Vitamin A Drugs 0.000 description 1
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 description 1
- 208000010399 Wasting Syndrome Diseases 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003078 antioxidant Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 235000020778 linoleic acid Nutrition 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 229940023569 palmate Drugs 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 229960003471 retinol Drugs 0.000 description 1
- 230000033764 rhythmic process Effects 0.000 description 1
- AHMCFSORHHSTSB-UHFFFAOYSA-N selanylidenecalcium Chemical compound [Se]=[Ca] AHMCFSORHHSTSB-UHFFFAOYSA-N 0.000 description 1
- KEAYESYHFKHZAL-UHFFFAOYSA-N sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
Abstract
The present invention discloses a kind of wild ' Ganba ' fungus industrial production method, and bacterium strain of drying up is in China Committee for Culture Collection of Microorganisms's common micro-organisms center（CGMCC）Preservation, preserving number is：CGMCC NO. 9902.Bulk is longitudinally torn into after wild ' Ganba ' fungus is sterilized, progress inclined-plane culture in culture medium is inoculated in and obtains wizened bacterium strain.The wizened bacterium strain is inoculated into the culture medium of nutrient bag, the pine roots disinfected are inserted in the nutrient bag simultaneously and cultivated, it is separated into being inoculated in respectively after several parts in above-mentioned identical culture medium and cultivates in the same way, then whole plastic bag cultivation kind is in the soil of pine tree root, with certain condition culture until fructification is exposed after soil 20~30 days, wizened bacterium can be plucked；So realize scale, the industrialized production of wild ' Ganba ' fungus.The wizened mushroom entity of gained of the invention ensure that original fragrance and the nutrition of wizened bacterium.The survival rate of the wizened bacterium of present invention production is high, can reach 99%.
Description
Technical field
The invention belongs to wild edible fungus culture technology field, it is related to a kind of wild ' Ganba ' fungus industrial production method.
Background technology
Wizened bacterium（Thelephora ganbajun）Belong to Basidiomycota（Basidiomyeo-ta）Layer bacterium steel
（Hymenomyeetes）Aphyllophorales（Aphyhoporales）Lead fungi section（TheLephoraceae）Lead fungi belongs to
（Thelephora）In species, be commercially referred to as wizened bacterium also includes orange lead fungi（T.aurantiotineta）It is punished for being related to
Lead fungi（T.vialis）Palmate lead fungi（T.palmata）Deng its fructification from the multiple branch of raw coralliform, by the thicker bacterium of base portion
Dry tough after softness is old when meat fiber shape keratin is young, gas fragrance is beautiful to be liked by consumers in general deeply, elder brother widely distributed in Yunnan Province
The area such as bright, Chuxiong, Yuxi, Lijing, the Baoshan, Qujing, Simao is distributed.
Wizened bacterium is mainly grown in 1000~2200m of height above sea level or so pineland, mainly Yunnan pinon pine in coniferous forest,
But also it is distributed in oak-pine mixed forest or pure oakery, it and pine genus plant（Such as pinus yunnanensis and pinus khasys）There is exotrophic mycorrhiza pass
System, laterite soil this growing environment that its unique fragrance is had in Pinus Yunnanensis Forest can just be guaranteed, other pine forests
Or other soil can not then ensure its unique fragrance.Research finds that the wizened bacteria growing phase is longer, is grown to from mushroom flower bud and is adapted to adopt
The fructification plucked needs 20 days or so, and it is aging to grow to fructification, 30~40 days or so is needed, in aerial growth phase, the change of weather
Change to bacterium of drying up grow and pest and disease damage generation important, if the air humidity that do not rain for a long time is less than 70%,
Sporophore growth is bad, easily occurs wasting disease, and one day light rain rain or shine replaces, and keeps air humidity 80% or so, most has
Growth, development beneficial to wizened bacterium, if but it is cloudy and drizzly for days on end for a long time, air humidity is 95% or so, and wizened bacterium easily occurs pink
Single-ended spore end disease, therefore wizened bacterium is a kind of very fragile wild-type strain, the best period of harvesting is after fructification is unearthed
20~30 days, the bacterium master recovery time of drying up was June~September, and the wizened bacterium after October is very rare.
Wizened bacterium contains substantial amounts of nutriment, and the content of wherein protein is that 24.15%, crude fibre is 8.45%, total reducing sugar
Measure as 3.1%, and the vitamin A in bacterium of drying up per 100g is that 16.0mg, B family vitamin are 1.2mg, and wizened bacterium also contains someone
18 kinds of amino acid necessary to body, linoleic acid content highest therein, also containing new p-terphenyl class pigment, with anti-oxidant work
Property, wizened bacterium contains abundant mineral trace element, such as iron, potassium, sodium, phosphorus, calcium selenium.Bacterium of drying up in a word is nutritious, mouth
Feel, all the time all in the situation that supply falls short of demand, wild ' Ganba ' fungus sales volume was reduced year by year in recent years, price is soaring tight
Weight, is up to 1500 yuan or so, it has also become expensive goods per 1000g prices.Factorial praluction is also not carried out now, therefore
Exploitation is a kind of to realize scale, the method for industrialization production wild ' Ganba ' fungus, be to meet the important way needed for people and market
Footpath.
The content of the invention
To realize scale, the industrialization production of wizened bacterium, the present invention provides a kind of wild ' Ganba ' fungus factorial praluction side
Method.
The present invention is realized by following technical proposal：A kind of wizened bacterium strain, the strain is protected in Chinese microorganism strain
Hide administration committee's common micro-organisms center（CGMCC）Preservation, preserving number is：CGMCC NO. 9902, address：Beijing is exposed to the sun
The institute 3 of area North Star West Road 1.
Another object of the present invention is to provide a kind of preparation method of wizened bacterium strain, by following each step：
It is that the cleaning of 75% alcohol is dry with volumetric concentration after A, the fresh sturdy wild ' Ganba ' fungus of harvesting, cleaning weeds and soil
Bar bacterium surface, aseptically longitudinal direction is torn into bulk afterwards；
B, block wizened bacterium obtained by step A is inoculated in following media：800~1200 parts of maize flour, analysis for soybean powder
800~1200 parts, 300~600 parts of pine tar, 0.2~0.8 part of vitamin B1,0.5~1.5 part of magnesium sulfate, mass concentration be 50%
450~650 parts of glucose solution, mass concentration for 10% 180~300 parts of calcium gluconate solution, potassium dihydrogen phosphate 0.2~
0.8 part, 40~60 parts of yeast, in carrying out inclined-plane culture at 22~25 DEG C 20~25 days, obtain wizened bacterium strain, its preserving number
For：CGMCC NO. 9902。
Block wizened bacterium length is 3~5cm, 1~2cm of thickness in the step A.
The inclined-plane culture of the step B is completed in test tube.
A further object of the present invention is to provide a kind of with the wizened bacterium strain industrialized production wild ' Ganba ' fungus
Method, comprises the following steps：
（1）It is by preserving number：CGMCC NO. 9902 wizened bacterium strain, is inoculated into the culture medium of nutrient bag, simultaneously
In the culture medium that the pine roots disinfected are inserted to the nutrient bag, in being cultivated 25~30 days at 22~25 DEG C, cultivation obtains the
Two generations wizened bacterium strain；The culture medium of the nutrient bag is made up of the component of following masses part：80~120 parts of pine, pine needle 20
~40 parts, 20~40 parts of corn, 20~40 parts of analysis for soybean powder, 5~15 parts of wheat bran, 0.01~0.05 part of gypsum, 5~15 parts of white sugar,
5~15 parts of pine tar, 0.05~0.15 part of yeast；
（2）Take out step（1）The wizened bacterium strain of the gained second generation is divided into after several parts, and the culture medium of nutrient bag is inoculated in respectively
In, and insert the pine roots disinfected in every bag of nutrient bag, in being cultivated 25~30 days at 22~25 DEG C, obtain expanding it is numerous after
The wizened bacterium strain of the third generation；The same step of culture medium of the nutrient bag（1）；
（3）By step（2）Expand the wizened bacterium strain of numerous gained third generation, whole plastic bag cultivation kind is in the soil of pine tree root, plantation
Depth be below 10cm, after plantation with sterilization after peat cover, and control temperature for 17~18 DEG C, humidity 70~80%, wind
1~2 grade of power, intensity of illumination are 400~600lux, until fructification is exposed after soil 20~30 days, can pluck wizened bacterium；
（4）With the wizened bacterium of ground under the concordant ground horizontal cutting of knife during harvesting, the wizened bacterium in underground continues to leave, and controls temperature
Degree is 17~18 DEG C, humidity 70~80%, 1~2 grade of wind-force, intensity of illumination are 400~600lux, until fructification exposes soil
20~30 days afterwards, and wizened bacterium can be plucked；So realize scale, the industrialized production of wild ' Ganba ' fungus.
The step（1）、（2）In the pine roots disinfected refer to 25~35cm survive, long, 0.5~1cm of diameter
Pine tree tree root be cut into inclined-plane, bifurcated is removed, then cleared up and sterilized for 75% alcohol with volumetric concentration, up to without soil
Attachment, then inserts 10cm in nutrient bag by tree root, and seals, and keeps dry behaviour's ventilation.
The step（2）Second generation bacterium strain of drying up be divided into several parts and refer to：Second will covered with each bag of nutrient bag
In generation, wizened bacterium strain was divided into 15~25 bags.
The step（3）Peat after middle sterilization is to be planted with mass concentration for the corruption after 3~5 ‰ acetic acid solution sprinkling
Soil.
The step（3）Pine tree refer to transplant pine tree in greenhouse, and soil comes from pine forests soil, concrete operations
It is：Soil movement from pine forests is digged into greenhouse and tiling forms plastic shed soil, between root device will be controlled routinely
It is placed in every being vertically placed on plastic shed soil, then by pine tree in control root device, the soil from pine forests is inserted and covered in control root device
Lid and fixed pine tree root, and routinely managing and protecting makes pine tree grow more than 1 year.
The pine tree is pinus yunnanensis, and the age of tree is 5~25 years.
The soil is height above sea level in the pine forests that 1500~2200m, pH value are that more than 4.5, organic matter is more than 1.5
Soil.
The height of the control root device is 0.8~1m, a diameter of 0.8~1.2m.The third generation dry up bacterium strain whole plastic bag cultivation kind when
Bag number determined depending on the root size of pine tree.
Advantage and effect that the present invention possesses：The present invention can be buried under pine tree root by strain, long-term to preserve old
Strain produces spore, utilizes the natural growth rhythm for manually building the Nature；Or wizened bacterium is built into by transplanting breeding
The environment that can be grown naturally, adjusts temperature, humidity, luminosity, demeanour all by Automated condtrol, with 5~25 years using warmhouse booth
Pinus yunnanensis and Yunnan laterite soil are condition, build the same sandstone soil of mountain forest, then train with breeding strain spore or inclined-plane
The mode educated largely obtains wizened bacterium.It will not be made a variation using wizened mushroom entity obtained by the method for the present invention, mouthfeel fragrance and open country
Raw wizened bacterium is identical, it is ensured that original fragrance of wizened bacterium and nutrition.The survival rate of the wizened bacterium of present invention production is high,
99% can be reached, and without using agriculture chemical, picking time is short, can pluck once within every 20~30 days, pass through manual control environment
Condition, can be achieved scale, the wizened bacterium of industrialization production, and can harvest throughout the year.
Embodiment
Below by embodiment, the present invention will be further described.
Embodiment 1
It is that the cleaning of 75% alcohol is dry with volumetric concentration after A, the fresh sturdy wild ' Ganba ' fungus of harvesting, cleaning weeds and soil
Bar bacterium surface, aseptically longitudinal direction is torn into length for 3~5cm, 1~2cm of thickness bulk afterwards；
B, block wizened bacterium obtained by step A is inoculated in following media：1000 parts of maize flour, analysis for soybean powder 1000
Part, 550 parts of the glucose solution that 500 parts of pine tar, 0.5 part of vitamin B1,1 part of magnesium sulfate, mass concentration are 50%, mass concentration
For 10% 200 parts of calcium gluconate solution, 0.5 part of potassium dihydrogen phosphate, 50 parts of yeast, in inclined-plane training is carried out at 23 DEG C in test tube
Support 22 days, obtain wizened bacterium strain, its preserving number is：CGMCC NO. 9902；
C, by preserving number it is：CGMCC NO. 9902 wizened bacterium strain, is inoculated into the culture medium of nutrient bag, will survive
, long 30cm, diameter 1cm pine tree tree root be cut into inclined-plane, remove bifurcated, then cleared up for 75% alcohol with volumetric concentration
And sterilization, until without soil attachment, tree root then is inserted into 10cm in nutrient bag, and seal, dry behaviour's ventilation is kept, at 25 DEG C
Culture 28 days, cultivation obtains the wizened bacterium strain of the second generation；The culture medium of the nutrient bag is made up of the component of following masses part：Pine
100 parts of wood, 30 parts of pine needle, 30 parts of corn, 30 parts of analysis for soybean powder, 10 parts of wheat bran, 0.03 part of gypsum, 10 parts of white sugar, 10 parts of pine tar, ferment
It is female 0.1 part；
The wizened bacterium strain of the second generation obtained by D, taking-up step C, by the wizened bacterium bacterium of the second generation covered with each bag of nutrient bag
Plant and be divided into after 20 bags, be inoculated in respectively in the culture medium of nutrient bag, 30cm survive, long, diameter 1cm pine tree tree root are cut into
Inclined-plane, removes bifurcated, then is cleared up and sterilized for 75% alcohol with volumetric concentration, until without soil attachment, then by tree root
Insert 10cm in nutrient bag, and seal, keep dry behaviour's ventilation, in being cultivated 28 days at 25 DEG C, obtain expanding it is numerous after the third generation dry up
Bacterium strain；The culture medium of the nutrient bag is with step C；
E, the soil movement from pine forests is digged into greenhouse and tiling forms plastic shed soil, be by height
1m, a diameter of 1m control root device are routinely spaced and are vertically placed on plastic shed soil, then the pine tree that the age of tree is 8 years is placed in into control root
In device, the soil from pine forests is inserted into covering and fixed pine tree root in control root device, and routinely managing and protecting grows pine tree
More than 1 year；Step D is expanded into the numerous gained third generation to dry up bacterium strain, whole plastic bag cultivation kind in control root device in the soil of pine tree root,
Planting depth is 15cm, plant 2 bags, after plantation with sterilization after peat cover, and control temperature for 18 DEG C, humidity 80%, wind
1~2 grade of power, intensity of illumination are 500lux, until fructification is exposed after soil 20~30 days, can pluck wizened bacterium；It is described to disappear
Peat after poison is the peat after being sprayed with mass concentration for 4 ‰ acetic acid solution；
With the wizened bacterium of ground under the concordant ground horizontal cutting of knife when F, harvesting, the wizened bacterium in underground continues to leave, and controls temperature
Degree is 18 DEG C, humidity 80%, 1~2 grade of wind-force, intensity of illumination are 500lux, until fructification exposes after soil 20~30 days, again
Can the wizened bacterium of harvesting；So realize scale, the industrialized production of wild ' Ganba ' fungus.Survival rate 99%.
Embodiment 2
It is that the cleaning of 75% alcohol is dry with volumetric concentration after A, the fresh sturdy wild ' Ganba ' fungus of harvesting, cleaning weeds and soil
Bar bacterium surface, aseptically longitudinal direction is torn into length for 3cm, thickness 1cm bulk afterwards；
B, block wizened bacterium obtained by step A is inoculated in following media：800 parts of maize flour, 800 parts of analysis for soybean powder,
450 parts of glucose solution, mass concentration that 600 parts of pine tar, 0.2 part of vitamin B1,1.5 parts of magnesium sulfate, mass concentration are 50%
For 10% 300 parts of calcium gluconate solution, 0.2 part of potassium dihydrogen phosphate, 40 parts of yeast, in inclined-plane training is carried out at 22 DEG C in test tube
Support 25 days, obtain wizened bacterium strain, its preserving number is：CGMCC NO. 9902；
C, by preserving number it is：CGMCC NO. 9902 wizened bacterium strain, is inoculated into the culture medium of nutrient bag, will survive
, long 25cm, diameter 0.8cm pine tree tree root be cut into inclined-plane, remove bifurcated, then carried out clearly with the alcohol that volumetric concentration is 75%
Reason and sterilization, until without soil attachment, tree root then is inserted into 10cm in nutrient bag, and are sealed, and keep doing behaviour's ventilation, in 23 DEG C
Lower culture 30 days, cultivation obtains the wizened bacterium strain of the second generation；The culture medium of the nutrient bag is made up of the component of following masses part：
80 parts of pine, 20 parts of pine needle, 40 parts of corn, 20 parts of analysis for soybean powder, 5 parts of wheat bran, 0.05 part of gypsum, 5 parts of white sugar, 15 parts of pine tar, ferment
It is female 0.05 part；
The wizened bacterium strain of the second generation obtained by D, taking-up step C, by the wizened bacterium bacterium of the second generation covered with each bag of nutrient bag
Plant and be divided into after 15 bags, be inoculated in respectively in the culture medium of nutrient bag, 25cm survive, long, diameter 0.8cm pine tree tree root are cut
Into inclined-plane, bifurcated is removed, then is cleared up and is sterilized for 75% alcohol with volumetric concentration, until without soil attachment, then will tree
10cm in root insertion nutrient bag, and is sealed, and keeps dry behaviour's ventilation, in being cultivated 30 days at 22 DEG C, obtain expanding it is numerous after the third generation do
Bar bacterium strain；The culture medium of the nutrient bag is with step C；
E, by height above sea level in 1500~2200m, pH value it is the soil from pine forests that more than 4.5, organic matter is more than 1.5
Movement is digged into greenhouse and tiling forms plastic shed soil, will height for 0.8m, a diameter of 1.2m control root device routinely
Interval is vertically placed on plastic shed soil, then the age of tree was placed in control root device for the pinus yunnanensis of 5 years, by the soil from pine forests
Covering and fixed pine tree root in control root device are inserted, and routinely managing and protecting makes pine tree grow more than 1 year；Step D is expanded into numerous gained
The wizened bacterium strain of the third generation, whole plastic bag cultivation kind is in control root device in the soil of pine tree root, and planting depth is 12cm, plants 1 bag, plants
Covered after kind with the peat after sterilization, and to control temperature be 17 DEG C, humidity 70%, 1 grade of wind-force, intensity of illumination are 400lux, directly
Expose to fructification after soil 20 days, wizened bacterium can be plucked；Peat after the sterilization is with the second that mass concentration is 3 ‰
Peat after acid solution sprinkling；
With the wizened bacterium of ground under the concordant ground horizontal cutting of knife when F, harvesting, the wizened bacterium in underground continues to leave, and controls temperature
Degree is 17 DEG C, humidity 70%, 1 grade of wind-force, intensity of illumination are 400lux, until fructification is exposed after soil 20 days, and can pluck and do
Bar bacterium；So realize scale, the industrialized production of wild ' Ganba ' fungus.Survival rate 98.9%.
Embodiment 3
It is that the cleaning of 75% alcohol is dry with volumetric concentration after A, the fresh sturdy wild ' Ganba ' fungus of harvesting, cleaning weeds and soil
Bar bacterium surface, aseptically longitudinal direction is torn into length for 5cm, thickness 2cm bulk afterwards；
B, block wizened bacterium obtained by step A is inoculated in following media：1200 parts of maize flour, analysis for soybean powder 1200
Part, 650 parts of the glucose solution that 300 parts of pine tar, 0.8 part of vitamin B1,0.5 part of magnesium sulfate, mass concentration are 50%, quality are dense
180 parts of the calcium gluconate solution, 0.8 part of potassium dihydrogen phosphate, 60 parts of yeast for 10% are spent, in carrying out inclined-plane at 25 DEG C in test tube
Culture 20 days, obtains wizened bacterium strain, and its preserving number is：CGMCC NO. 9902；
C, by preserving number it is：CGMCC NO. 9902 wizened bacterium strain, is inoculated into the culture medium of nutrient bag, will survive
, long 35cm, diameter 0.5cm pine tree tree root be cut into inclined-plane, remove bifurcated, then carried out clearly with the alcohol that volumetric concentration is 75%
Reason and sterilization, until without soil attachment, tree root then is inserted into 10cm in nutrient bag, and are sealed, and keep doing behaviour's ventilation, in 22 DEG C
Lower culture 25 days, cultivation obtains the wizened bacterium strain of the second generation；The culture medium of the nutrient bag is made up of the component of following masses part：
120 parts of pine, 40 parts of pine needle, 20 parts of corn, 40 parts of analysis for soybean powder, 15 parts of wheat bran, 0.01 part of gypsum, 15 parts of white sugar, 5 parts of pine tar,
0.15 part of yeast；
The wizened bacterium strain of the second generation obtained by D, taking-up step C, by the wizened bacterium bacterium of the second generation covered with each bag of nutrient bag
Plant and be divided into after 25 bags, be inoculated in respectively in the culture medium of nutrient bag, 35cm survive, long, diameter 0.5cm pine tree tree root are cut
Into inclined-plane, bifurcated is removed, then is cleared up and is sterilized for 75% alcohol with volumetric concentration, until without soil attachment, then will tree
10cm in root insertion nutrient bag, and is sealed, and keeps dry behaviour's ventilation, in being cultivated 25 days at 23 DEG C, obtain expanding it is numerous after the third generation do
Bar bacterium strain；The culture medium of the nutrient bag is with step C；
E, by height above sea level 1500~2200m, pH value be 6, organic matter be 2 the soil movement from pine forests into greenhouse
Digged and tiling form plastic shed soil, will height be 1m, a diameter of 0.8m control root device is routinely spaced and is vertically placed on
On plastic shed soil, then the age of tree is placed in control root device for the pinus yunnanensis of 15~25 years, the soil from pine forests is inserted into control root
Covered in device and fixed pine tree root, and routinely managing and protecting makes pine tree grow more than 1 year；Step D is expanded into numerous gained third generation to do
Bar bacterium strain, whole plastic bag cultivation kind is in control root device in the soil of pine tree root, and planting depth is below 10cm, plants 3 bags, after plantation
Covered with the peat after sterilization, and to control temperature be 17~18 DEG C, humidity 75%, 2 grades of wind-force, intensity of illumination are 600lux, directly
Expose to fructification after soil 30 days, wizened bacterium can be plucked；Peat after the sterilization is with the second that mass concentration is 5 ‰
Peat after acid solution sprinkling；
With the wizened bacterium of ground under the concordant ground horizontal cutting of knife when F, harvesting, the wizened bacterium in underground continues to leave, and controls temperature
Degree is 17~18 DEG C, humidity 75%, 2 grades of wind-force, intensity of illumination are 600lux, until fructification is exposed after soil 30 days, can be adopted again
Pluck wizened bacterium；So realize scale, the industrialized production of wild ' Ganba ' fungus.Survival rate 99.2%.
Claims (3)
1. a kind of wizened bacterium strain, it is characterised in that：The strain is commonly micro- in China Committee for Culture Collection of Microorganisms
Bio-Centers（CGMCC）Preservation, preserving number is：CGMCC NO. 9902；
The wizened bacterium strain is prepared through the following steps：
It is the wizened bacterium of 75% alcohol cleaning with volumetric concentration after A, the fresh sturdy wild ' Ganba ' fungus of harvesting, cleaning weeds and soil
Surface, aseptically longitudinal direction is torn into bulk afterwards；
B, block wizened bacterium obtained by step A is inoculated in following media：800~1200 parts of maize flour, analysis for soybean powder 800~
1200 parts, 300~600 parts of pine tar, 0.2~0.8 part of vitamin B1,0.5~1.5 part of magnesium sulfate, mass concentration be 50% Portugal
180~300 parts of calcium gluconate solution, the potassium dihydrogen phosphate 0.2~0.8 of 450~650 parts of grape sugar juice, mass concentration for 10%
Part, 40~60 parts of yeast, in carrying out inclined-plane culture at 22~25 DEG C 20~25 days, obtain wizened bacterium strain, its preserving number is：
CGMCC NO. 9902。
2. wizened bacterium strain according to claim 1, it is characterised in that：Block wizened bacterium length is 3 in the step A
~5cm, 1~2cm of thickness.
3. wizened bacterium strain according to claim 1, it is characterised in that：The inclined-plane culture of the step B is in test tube
Complete.
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Effective date of registration: 20180717 Address after: 650307 new spring village, Anning City, Kunming, Yunnan Patentee after: Kunming golden Moore edible fungus planting Co., Ltd. Address before: 650307 Anning hot spring new house village in Kunming, Yunnan Patentee before: Tang Yuanzhi |