CN108476868A - A kind of cultural method of armillaria mellea accreting with Rhizoma Gastrodiae - Google Patents
A kind of cultural method of armillaria mellea accreting with Rhizoma Gastrodiae Download PDFInfo
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- CN108476868A CN108476868A CN201810563987.6A CN201810563987A CN108476868A CN 108476868 A CN108476868 A CN 108476868A CN 201810563987 A CN201810563987 A CN 201810563987A CN 108476868 A CN108476868 A CN 108476868A
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- rhizoma gastrodiae
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- 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
- A01G18/40—Cultivation of spawn
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- 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
- A01G18/20—Culture media, e.g. compost
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of cultural methods of armillaria mellea accreting with Rhizoma Gastrodiae comprising following steps：1, strain improvement screens halimasch strain female parent, is detached, purification step selects parent species；2, strain expands numerous, parent species are placed in culture dish are cultivated using fluid nutrient medium and be inoculated into the fermentation tank equipped with fluid nutrient medium after seed liquid strain and continue to cultivate into halimasch liquid bacterial strains；3, halimasch liquid bacterial strains solid medium is transplanted to cultivate into cultigen.The present invention be select the growing thickly of artificial culture, the Rhizomorph isolation method that thickness is suitable, growth is strong, top is white out carries out seed selection cultivation, the quality of bacterium source is ensured that from source first, shoestring after seed selection is also needed to by purifying and screening, so it can not only be polluted to avoid remaining mycelia, and expand again after original strain selection and breeding it is numerous made of strain purity be 100%, without any miscellaneous bacteria, ensure that the mycelia health that last cultigen is cultivated is thick and strong in this way, kind is pure.
Description
Technical field
The present invention relates to biotechnology, more particularly to a kind of cultural method of armillaria mellea accreting with Rhizoma Gastrodiae.
Background technology
Armillaria is general medium big in Agaricales, Armillaria, fructification, and there are about more than 40 in the whole world for the category fungi
Kind, also relatively broad in Chinese branch, Central China, southwest and the Northeast are distributed.Halimasch is rare Chinese medicine Rhizoma Gastrodiae
Important fungal component, Rhizoma Gastrodiae can not oneself absorb nutrition, during growth mainly using halimasch with the shape of rhizomorph
Formula provides nutriment.Therefore the quality of strain and the purity etc. of strain all can play prodigious influence to the yield of Rhizoma Gastrodiae, existing
It is usually first to be seeded on solid medium to develop and grow up with halimasch kind that some Rhizoma Gastrodiaes, which commercially produce middle production stage, is waited for
It dresses seed as cultigen and Rhizoma Gastrodiae after development and growth are ripe.The nothing of most attention to training bases and operating process in this process
Bacterium operates, but often has ignored the strain of inoculation itself, and the strain being directly inoculated with is that growth situation differs first, Wu Fabao
The strain that it is high quality is demonstrate,proved, secondly other miscellaneous bacterias may be mixed in it, but is difficult to identify that in the state of strain
Come, unless waiting for that its maturation is grown up can differentiate, is directly inoculated with the clarity for not simply failing to ensure strain, and bacterium using strain
The quality of kind is also irregular, and such breeding strain can not only influence the quality and yield of cultigen at cultigen, but also
Later stage and empty pool rate after Rhizoma Gastrodiae association can be caused high.
Invention content
The technical problem to be solved in the present invention is to provide a kind of cultural methods of armillaria mellea accreting with Rhizoma Gastrodiae, can ensure strain
Purity and quality, the halimasch cultivated is free of any miscellaneous bacteria, and mycelia health is thick and strong, and kind is pure.
The technical scheme is that：
A kind of cultural method of armillaria mellea accreting with Rhizoma Gastrodiae comprising following steps：
(1) strain improvement screens halimasch strain female parent, is detached, purification step selects parent species；
(2) strain expands numerous, and parent species are placed in after culture dish cultivates seed liquid strain using fluid nutrient medium and are inoculated into
Fermentation tank equipped with fluid nutrient medium continues to cultivate into halimasch liquid bacterial strains；
(3) halimasch liquid bacterial strains solid medium is transplanted to cultivate into cultigen.
Further, liquid spawn is transplanted to after solid medium in 22 DEG C -26 DEG C of room temperature, relative humidity 65%-80%
Darkroom cultivate and obtain cultigen in 55-65 days.
Further, fluid nutrient medium is made of the material of following parts by weight：0.1 part of amino acid, 1-3 parts of glucose, phosphorus
0.1-0.2 parts of acid dihydride potassium, 0.1-0.3 parts of trace element, 0.05-0.1 parts of magnesium sulfate, 90-100 parts of water, 1-2 parts of sucrose.
Further, amino acid is glutamic acid, glycine or alanine.
Further, trace element is that calcium monohydrogen phosphate, potassium sulfate, potassium nitrate, copper oxide and calcium nitrate press equivalent weight part
It mixes.
Further, solid medium is made of the material of following parts by weight：Sawdust 80-85 parts of Qinggang, 10 parts of gypsum, fine jade
15-20 parts of 2-4 parts of fat, rice bran or wheat bran, 5-8 parts of corn flour, 0.2-0.4 parts of magnesium sulfate.
The invention has the beneficial effects that：
The strain that the present invention is cultivated by seed selection can ensure the quality of strain and excellent compared with directly using strain inoculation
It is benign, it is the growing thickly of artificial culture, the Rhizomorph isolation method that thickness is suitable, growth is strong, top is white out carries out seed selection cultivation,
The quality of bacterium source is ensured that from source first, so the cultigen upgrowth situation cultivated is more good, Hyphal form ratio
More healthy and stronger, empty pool rate is reduced to by 20%-15% 10% hereinafter, and by that can ensure strain after purification after association Rhizoma Gastrodiae
Shoestring is inoculated on the test tube slants PDA culture medium, in constant temperature 25 by clarity by the shoestring after seed selection by sterile working regulation
DEG C, it is cultivated under the conditions of relative humidity 80% 10 days, the tender shoestring of children that then selection is grown into culture medium is purified, screening birth
The highly consistent shoestring of long form so can not only pollute, but also original strain selection and breeding as strain to avoid remaining mycelia
Expand again afterwards it is numerous made of strain purity be 100%, be free of any miscellaneous bacteria, ensure that the bacterium that last cultigen is cultivated in this way
Silk health is thick and strong, and kind is pure.
Specific implementation mode
The specific implementation mode of the present invention is described further below.It should be noted that for these implementations
The explanation of mode is used to help understand the present invention, but does not constitute limitation of the invention.In addition, invention described below
Involved technical characteristic can be combined with each other as long as they do not conflict with each other in each embodiment.
Embodiment 1：
(1) Qinggang branch of strain improvement, the halimasch for selecting artificial culture and a diameter of 40-60mm are planted after being stirred
In the soil for entering hayashishita, covers broad-leaved leaf and sandy soil are cultivated into bacterium bavin, take out bacterium bavin after 14-20 months, taken on bacterium bavin
Lower halimasch is maternal as halimasch strain, filter out grow thickly, the shoestring that thickness is suitable, growth is strong, top is white, by these bacterium
Rope is separated several sections and is placed in culture dish, and shoestring is then inoculated into the test tube slants PDA culture medium by sterile working regulation
On, it is cultivated under the conditions of 25 DEG C of constant temperature, relative humidity 60% 6 days, then selection grows into the tender shoestring of children in culture medium, this step
Suddenly miscellaneous bacteria can be picked out, strain is purified, and filters out the highly consistent shoestring of growthform as parent species.
(2) strain expands numerous, and parent species are placed in after culture dish cultivates seed liquid strain using fluid nutrient medium and are inoculated into
Fermentation tank equipped with fluid nutrient medium continues to cultivate into halimasch liquid bacterial strains, and fluid nutrient medium is 0.1 part of amino acid, glucose
1 part, 0.1 part of potassium dihydrogen phosphate, trace element 0.1 part, 0.05 part of magnesium sulfate, 90 parts of water, 1 part of sucrose, in 25 DEG C of constant temperature, relatively
It is cultivated 25 days under the conditions of humidity 60%.
(3) halimasch liquid bacterial strains solid medium is transplanted to cultivate into cultigen, it is green by the material of following parts by weight
15 parts of 80 parts of ridge sawdust, 10 parts of gypsum, 2-4 parts of agar, rice bran or wheat bran, 5 parts of corn flour, 0.2 part of magnesium sulfate plus water configuration are solid
Halimasch liquid bacterial strains, are then inoculated on solid medium under sterile working, in room temperature by the laggard luggage bottle of body culture medium
22 DEG C, the darkroom of relative humidity 65%, which is cultivated 55 days, obtains cultigen.
Embodiment 2：
(1) strain improvement is removed when being planted from Rhizoma Gastrodiae on bacterium bavin ripe or on the solid medium of cultigen
Halimasch is maternal as halimasch strain, filter out grow thickly, the shoestring that thickness is suitable, growth is strong, top is white, by these shoestrings
It separates several sections to be placed in culture dish, then shoestring is inoculated on the test tube slants PDA culture medium by sterile working regulation,
It is cultivated under the conditions of 25 DEG C of constant temperature, relative humidity 80% 10 days, then selection grows into the tender shoestring of children in culture medium, the step
Miscellaneous bacteria can be picked out, strain is purified, and filters out the highly consistent shoestring of growthform as parent species.
(2) strain expands numerous, and parent species are placed in after culture dish cultivates seed liquid strain using fluid nutrient medium and are inoculated into
Fermentation tank equipped with fluid nutrient medium continues to cultivate into halimasch liquid bacterial strains, fluid nutrient medium be amino acid 0.1 divide, glucose
3 parts, 0.2 part of potassium dihydrogen phosphate, trace element 0.3 part, 0.1 part of magnesium sulfate, 100 parts of water, 2 parts of sucrose, in 25 DEG C of constant temperature, relatively
It is cultivated 25 days under the conditions of humidity 80%.
(3) halimasch liquid bacterial strains solid medium is transplanted to cultivate into cultigen, it is green by the material of following parts by weight
20 parts of 85 parts of ridge sawdust, 10 parts of gypsum, 4 parts of agar, rice bran or wheat bran, 8 parts of corn flour, 0.4 part of magnesium sulfate plus water configure solid
Halimasch liquid bacterial strains, are then inoculated on solid medium under sterile working, in room temperature 26 by the laggard luggage bottle of culture medium
DEG C, the darkroom of relative humidity 80%, which is cultivated 65 days, obtains cultigen.
Amino acid can select glutamic acid, glycine or alanine in above-described embodiment.
Trace element can select calcium monohydrogen phosphate, potassium sulfate, potassium nitrate, copper oxide and calcium nitrate to be mixed by equivalent weight part
It forms, naturally it is also possible to select other feasible formulas, enough trace elements are provided i.e. for the development of strain as long as can guarantee
It can.
The strain cultivated by seed selection is compared with the quality and Optimality that can directly ensure strain using strain inoculation,
Be select the growing thickly of artificial culture, the Rhizomorph isolation method that thickness is suitable, growth is strong, top is white out carries out seed selection cultivation, first
The quality of bacterium source is first ensured that from source, so the cultigen upgrowth situation cultivated is more good, Hyphal form ratio is more
Stalwartness, empty pool rate is reduced to by 20%-15% 10% hereinafter, and by that can ensure the pure of strain after purification after association Rhizoma Gastrodiae
Shoestring is inoculated on the test tube slants PDA culture medium by cleanliness by the shoestring after seed selection by sterile working regulation, in 25 DEG C of constant temperature,
It is cultivated under the conditions of relative humidity 80% 10 days, then selection grows into the tender shoestring of children in culture medium, and the step can be picked out
Miscellaneous bacteria purifies strain, and filters out the highly consistent shoestring of growthform as strain, so not only can be to avoid
Remaining mycelia pollute, and expand again after original strain selection and breeding it is numerous made of strain purity be 100%, be free of any miscellaneous bacteria, this
Sample ensures that the mycelia health that last cultigen is cultivated is thick and strong, and kind is pure.And it traditional direct is connect using strain
Kind does not simply fail to ensure the clarity of strain, and the quality of strain is also irregular, and such breeding strain is at cultigen
The quality and yield of cultigen can be not only influenced, but also later stage and empty pool rate after Rhizoma Gastrodiae association can be caused high.
Embodiments of the present invention are explained in detail above, but the present invention is not limited to described embodiments.It is right
For those skilled in the art, in the case where not departing from the principle of the invention and spirit, these embodiments are carried out more
Kind change, modification, replacement and modification, still fall in protection scope of the present invention.
Claims (6)
1. a kind of cultural method of armillaria mellea accreting with Rhizoma Gastrodiae, it is characterised in that include the following steps：
(1) strain improvement screens halimasch strain female parent, is detached, purification step selects parent species；
(2) strain expands numerous, parent species are placed in culture dish cultivate to be inoculated into after seed liquid strain using fluid nutrient medium and be equipped with
The fermentation tank of fluid nutrient medium continues to cultivate into halimasch liquid bacterial strains；
(3) halimasch liquid bacterial strains solid medium is transplanted to cultivate into cultigen.
2. the cultural method of armillaria mellea accreting with Rhizoma Gastrodiae according to claim 1, it is characterised in that：The liquid spawn is transplanted to solid
In 22 DEG C -26 DEG C of room temperature after body culture medium, the darkroom of relative humidity 65%-80%, which is cultivated 55-65 days, obtains cultigen.
3. the cultural method of armillaria mellea accreting with Rhizoma Gastrodiae according to claim 1, which is characterized in that the fluid nutrient medium is by following
The material of parts by weight forms：0.1 part of amino acid, 1-3 parts of glucose, 0.1-0.2 parts of potassium dihydrogen phosphate, trace element 0.1-0.3
Part, 0.05-0.1 parts of magnesium sulfate, 90-100 parts of water, 1-2 parts of sucrose.
4. the cultural method of armillaria mellea accreting with Rhizoma Gastrodiae according to claim 3, it is characterised in that：The amino acid be glutamic acid,
Glycine or alanine.
5. the cultural method of armillaria mellea accreting with Rhizoma Gastrodiae according to claim 3, it is characterised in that：The trace element is phosphoric acid hydrogen
Calcium, potassium sulfate, potassium nitrate, copper oxide and calcium nitrate are mixed by equivalent weight part.
6. according to the cultural method of claim 1-5 any one of them armillaria mellea accreting with Rhizoma Gastrodiae, which is characterized in that the solid culture
Base is made of the material of following parts by weight：Sawdust 80-85 parts of Qinggang, 10 parts of gypsum, 2-4 parts of agar, rice bran or wheat bran 15-20
Part, 5-8 parts of corn flour, 0.2-0.4 parts of magnesium sulfate.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109220559A (en) * | 2018-09-30 | 2019-01-18 | 上海市农业科学院 | A kind of production method and growth condition of Stropharia rugoso-annulata reduction strain |
| CN111019835A (en) * | 2019-11-01 | 2020-04-17 | 汉中市汉台区汉麓生物科技有限公司 | Method for separating Armillaria mellea from ambary endophytic fungi |
| CN111109009A (en) * | 2019-12-11 | 2020-05-08 | 西南林业大学 | Armillaria mellea SWFU-09 and application thereof |
| CN113475316A (en) * | 2021-06-04 | 2021-10-08 | 李俊 | Preparation method of stable-yield high-yield liquid armillaria mellea strain |
| CN114287286A (en) * | 2022-02-22 | 2022-04-08 | 康县宁康生物科技有限公司 | Cultivation method of gastrodia elata armillaria mellea |
| CN115250826A (en) * | 2022-07-29 | 2022-11-01 | 陕西理工大学 | Production method of selenium-rich gastrodia elata |
| CN115336503A (en) * | 2022-08-30 | 2022-11-15 | 西南林业大学 | Preparation method of substitute bacterial material for gastrodia elata cultivation |
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| CN106010975A (en) * | 2016-05-23 | 2016-10-12 | 湖北森源生态科技股份有限公司 | Method for producing armillaria strains |
| CN106416744A (en) * | 2016-03-18 | 2017-02-22 | 中国科学院昆明植物研究所 | Armillaria cepistipes Velen. strain and method for preparing same |
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| CN103798055A (en) * | 2013-05-26 | 2014-05-21 | 程立君 | Method for producing rhizoma gastrodiae companion armillaria mellea through liquid submerged fermentation |
| CN103798056A (en) * | 2013-06-09 | 2014-05-21 | 程立君 | Production method of gastrodia elata seed germinating strains |
| CN106416744A (en) * | 2016-03-18 | 2017-02-22 | 中国科学院昆明植物研究所 | Armillaria cepistipes Velen. strain and method for preparing same |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109220559A (en) * | 2018-09-30 | 2019-01-18 | 上海市农业科学院 | A kind of production method and growth condition of Stropharia rugoso-annulata reduction strain |
| CN111019835A (en) * | 2019-11-01 | 2020-04-17 | 汉中市汉台区汉麓生物科技有限公司 | Method for separating Armillaria mellea from ambary endophytic fungi |
| CN111109009A (en) * | 2019-12-11 | 2020-05-08 | 西南林业大学 | Armillaria mellea SWFU-09 and application thereof |
| CN113475316A (en) * | 2021-06-04 | 2021-10-08 | 李俊 | Preparation method of stable-yield high-yield liquid armillaria mellea strain |
| CN114287286A (en) * | 2022-02-22 | 2022-04-08 | 康县宁康生物科技有限公司 | Cultivation method of gastrodia elata armillaria mellea |
| CN115250826A (en) * | 2022-07-29 | 2022-11-01 | 陕西理工大学 | Production method of selenium-rich gastrodia elata |
| CN115250826B (en) * | 2022-07-29 | 2024-04-09 | 陕西理工大学 | Production method of selenium-enriched gastrodia elata |
| CN115336503A (en) * | 2022-08-30 | 2022-11-15 | 西南林业大学 | Preparation method of substitute bacterial material for gastrodia elata cultivation |
| CN115336503B (en) * | 2022-08-30 | 2024-01-16 | 西南林业大学 | Preparation method of substitute bacteria for gastrodia elata cultivation |
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Application publication date: 20180904 |