CN103172446B - Fluid nutrient medium for large-scale cultivation of termitomyces albuminosus mycelium and application thereof - Google Patents
Fluid nutrient medium for large-scale cultivation of termitomyces albuminosus mycelium and application thereof Download PDFInfo
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- CN103172446B CN103172446B CN201310098607.3A CN201310098607A CN103172446B CN 103172446 B CN103172446 B CN 103172446B CN 201310098607 A CN201310098607 A CN 201310098607A CN 103172446 B CN103172446 B CN 103172446B
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Abstract
The invention provides fluid nutrient medium for large-scale cultivation of termitomyces albuminosus mycelium and application thereof. The culture medium is prepared from the following materials in parts by weight: 50-70 parts of lignocellulose powder, 30-50 parts of water, 0.0001-0.1 part of monopotassium phosphate, 0.0001-0.1 part of sodium chloride, 0.0001-0.1 part of calcium chloride, 0.0001-0.1 part of cobalt chloride, 0.0001-0.1 part of magnesium sulfate, 0.0001-0.1 part of manganese sulfate, 0.0001-0.1 part of ferrous sulfate, 0.0001-0.1 part of copper sulfate, 0.0001-0.1 part of zinc sulfate, 0.0001-0.1 part of aluminum potassium sulfate, 0.0001-0.1 part of ammonium tartrate, 0.0001-0.1 part of boric acid, 0.0001-0.1 part of citric acid, 0.0001-0.1 part of formic acid, 0.0001-0.1 part of acetic acid, 0.0001-0.1 part of fumaric acid, 0.0001-0.1 part of succinic acid, 0.0001-0.1 part of propylene-1,2,3-tricarboxylic acid, and 0.0001-0.1 part of guaiacol. The termitomyces albuminosus of the fluid nutrient medium disclosed by the invention is customized, and can be used for large-scale indoor cultivation of termitomyces albuminosus mycelium by using lignocelluloses; and the fluid nutrient medium is low in cost and high in yield.
Description
(1) technical field
The present invention relates to a kind of liquid nutrient medium and application thereof of cultivating for the mass-producing of Collybia albuminosa mycelia.
(2) background technology
Collybia albuminosa (Termitomyces albuminosus) is under the jurisdiction of Basidiomycotina Hymenomycetes Agaricales Tricholomataceae, is a kind of famous and precious wild edible fungus.People are to the scientific research of the Collybia albuminosa history of existing more than 200 year, first German natural scientist J.D.K ǒ ning in 1799 is finding on fungus garden to give birth to many fungies that can form jack shape in the termitarium of India eastern region, is namely known as afterwards the fungi of Collybia albuminosa.For termite, the mycelium of Collybia albuminosa is their Major Foods, and they are cultivated or the mycelial object of " cultivation " Collybia albuminosa is also this.The mycelia of Collybia albuminosa extensively distributes in whole ant nest, and in the time that termite goes to search for food, it is replaced by further growth again.Indigested food is piled up the nutrition of growing as Collybia albuminosa on fungus garden by termite, and the fungus garden that termite was infected taking Collybia albuminosa is food.
Research discovery, the fungus garden in termitarium is become to be grouped into other by half digest, wood fibre, soil particle, Collybia albuminosa mycelia, the worker ant secretory product of worker ant.According to analysis, in termite fugus gardens, contain protein 6.44%, fat 0.48%, total reducing sugar 0.24%, robust fibre 13.9%, ash content 26.6%, P
2o
50.28%, Ca1.86%, K
2o1.56%, amino acid/11 8.7%.
Temperature-stable is at 22 ± 2 DEG C in termitarium, and minimum is 15 DEG C, is the highlyest no more than 28 DEG C.Stable temperature is grown and has been created good condition to Collybia albuminosa mycelial growth.Relative air humidity in termitarium is 80%-90%.Fungus garden soil moisture content is 46.75%-58.35%.In addition, the respiratory metabolism activity of termite in nest, has produced the carbonic acid gas of higher concentration, CO
2concentration can reach 3-5%, the CO of high density
2more be conducive to the growth of Collybia albuminosa mycelia, and unfavorable to the growth of its sporophore and other fungi.Meanwhile, a large amount of formic acid of Released by Termites, acetic acid, propylene-1,2,3-tricarboxylic acid, FUMARIC ACID TECH GRADE, succinic acid etc., regulated the pH of fungus garden, makes pH be stabilized in 4-4.5, this has also produced restraining effect to the growth of other fungus and bacteriums in fungus garden, makes Collybia albuminosa mycelia energy Fast Growth.
Due to Collybia albuminosa energy lignocellulose degradation, therefore it has a very important role in bioenergy industry.But although people have done large quantity research to the artificial culture of Collybia albuminosa Termitomyces albuminosus, knowing that Collybia albuminosa is cultivated under artificial condition needs certain CO
2concentration, high humidity and acid condition, but up to now, because the effect in Collybia albuminosa growth and development process is also short in understanding to kind and the concentration, fungus garden symbiotic bacterium etc. of plant composition, species of metal ion and concentration, acid, also there is no a kind of method of lignocellulose as Major Nutrient base large scale culturing Collybia albuminosa mycelia of utilizing.
(3) summary of the invention
In order to realize industrialized application, the invention provides a kind of in indoor liquid nutrient medium and the application thereof that utilizes lignocellulose large scale culturing Collybia albuminosa mycelia.
The technical solution used in the present invention is:
A liquid nutrient medium of cultivating for the mass-producing of Collybia albuminosa mycelia, prepare by following quality proportioning:
Described lignocellulose is 1: 1 mixture of white poplar wood powder and mugwort powder.White poplar Populus tomentosa Carr wood powder obtains from sawmill, and 10 mesh sieves that dried in the sun are for subsequent use; Mugwort powder is the dry powder of pulverizing 10 mesh sieves after the mugwort Artemisia indica Willd plant of field collection is dried under the sun with pulverizer.
In this formula, lignocellulose is grown carbon source and energy matter is provided for mycelial growth, metallic salt is grown essential metal ion is provided for mycelial growth, acid compounds is grown essential sour environment is provided for mycelial growth, methyl catechol provides signal for the Laccase in mycelial growth growth course, and water is grown and essential moisture content is provided and maintains essential humidity for mycelial growth.
Preferably, described substratum is prepared by following quality proportioning:
More preferred, described substratum is prepared by following quality proportioning:
The invention still further relates to the application of described liquid nutrient medium in the mass-producing of Collybia albuminosa mycelia is cultivated.
Concrete, described being applied as: by formulated liquid nutrient medium, add Collybia albuminosa bacterium liquid and fungal component liquid, at 12 DEG C～30 DEG C, cultivate 30 days, obtain Collybia albuminosa fermented liquid, after filtration, filtrate is Collybia albuminosa bacterium liquid to fermented liquid, and this bacterium liquid can be used for the degraded of lignocellulose and the commercialization of Collybia albuminosa is produced.Symbiotic bacterium liquid is from blackwing subterranean fungus garden, to separate, cultivate the symbiotic bacterium obtaining, these bacteriums comprise: 4 kinds of bacterium of Bacillus thuringiensis (Bacillus thuringiensis), bacillus (Bacillus sp.) and clostridium bifermentans (Clostridium bifermentans) etc., and for removing some harmful secondary compound that mycelial growth growth course produces and the growth of growing and promote Collybia albuminosa mycelia that suppresses other fungi.Collybia albuminosa bacterium liquid is the Collybia albuminosa mycelia aqueous solution that utilizes potato dextrose agar to cultivate, and cultivates bacterial classification is provided for mycelia mass-producing.
Beneficial effect of the present invention is mainly reflected in:
(1) directly use the powder mixture of various plants as the carbon source of Collybia albuminosa mycelium culture, cost is economized, and meanwhile, also can promote the utilization of nature and waste resource;
(2) in nutrient solution, add various metals salt compounds simultaneously, for various enzymes in mycelial growth growth course synthetic provides essential metal ion, Collybia albuminosa mycelia can be grown by normal growth on lignocellulose substratum;
(3) in nutrient solution, add multiple acid compounds simultaneously, for the growth of Collybia albuminosa mycelia provides good sour environment, such sour environment has limited the growth of other mushroom on the other hand on the one hand, more on the one hand for the growth of Collybia albuminosa mycelia provides information substance;
(5) in nutrient solution, add methyl catechol, can induce Collybia albuminosa to produce laccase, promote can destroy rapidly the xylogen in plant drymeal mixture as the Collybia albuminosa mycelia of bacterial classification, make Collybia albuminosa mycelia can have that enough nutrition is grown, development and fecundity, thereby realize the quick production of Collybia albuminosa mycelia;
(4) in culturing process, add the symbiotic bacterium liquid being formed by multiple fungus garden symbiotic bacterium, the growth of small insect and the growths of other miscellaneous bacteria such as fruit bat are suppressed on the one hand in culturing process, on the other hand symbiotic bacterium in growth and development process, consumed in Collybia albuminosa mycelial growth growth course, produce to harmful secondary compound that itself grows, make Collybia albuminosa mycelia energy normal growth and growth.
(4) embodiment
Below in conjunction with specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in this:
Embodiment 1:
Bacterial classification:
Take blackwing subterranean nest from field, fungus garden is taken back indoor.It is upper that moved into slant medium (potato dextrose agar) by jack bacterium (vegetative phase of Collybia albuminosa) same day, and good inclined-plane moves pipe purifying to select growing way, and the bacterial classification after purifying gives over to female kind;
The preparation of potato dextrose agar: peeling potato 200g, glucose 20g, agar 15～20g, distilled water 1000ml.Peeling potato is cut into small pieces, and water boil 30min, by 8 layers of filtered through gauze, adds glucose and agar in filtrate, and water is settled to 1000ml, pours in Glass tubing for spawn culture and purifying after autoclaving.
The acquisition of Collybia albuminosa bacterium liquid: by the Collybia albuminosa silk of potato dextrose agar and upper cultivation thereof, according to the solid-liquid mass ratio of 1:1, join in sterilized water, fully stir, leave standstill after 1 hour, suspension is poured out, this suspension is Collybia albuminosa bacterium liquid.
The acquisition of fungal component liquid: pulverize dig the blackwing subterranean fungus garden instrument that nest obtains from field, then according to fungus garden: the mass ratio of sterilized water=1:9 mixes, stir, leave standstill 2 hours, get supernatant liquor and be fungal component liquid.
Culture medium prescription is (mass parts):
Production stage is:
(1) accurately take various salts (potassium primary phosphate, sodium-chlor, calcium chloride, cobalt chloride, magnesium sulfate, manganous sulfate, ferrous sulfate, copper sulfate, zinc sulfate, potassium aluminium sulfate, ammonium tartrate) with scale, then water is dissolved, be made into the metallic salt aqueous solution, for subsequent use.
(2) accurately take various acid (boric acid, citric acid, formic acid, acetic acid, FUMARIC ACID TECH GRADE, succinic acid, propylene-1,2,3-tricarboxylic acid) with scale, be then dissolved in water, be made into acid solution, for subsequent use.
(3) accurately take symbiotic bacterium liquid with scale, be put in bucket, for subsequent use.
(4) accurately take Termitomyces albuminosus solution and methyl catechol with scale, place it in bucket and stir, for subsequent use.
(5) accurately take lignocellulose with scale, place it in container, then add the metallic salt aqueous solution, stir; Add again acid solution, stir; Add again symbiotic bacterium liquid, stir; The mixed solution that adds again Collybia albuminosa and methyl catechol, stirs; Finally, will fill the container cover upper cover of said mixture, be placed in the environment of 12 DEG C～30 DEG C.
After (6) 30 days, remove the lid on container, the fermented product in container is filtered, the base-material of filter residue for further cultivating Collybia albuminosa mycelia or cultivating as earthworm, cockroach, filtrate is Collybia albuminosa bacterium liquid, and this bacterium liquid can be used for the degraded of lignocellulose and the commercialization of Collybia albuminosa is produced.
Cultivate according to above method, not only can obtain than potato dextrose agar and cultivate the low more than 20% a large amount of mycelia stostes of cost, and can directly be used as through the wood powder of mycelia processing the raw material that earthworm, cockroach are cultivated, thereby provide the one new way of getting rich for hill farmer.Meanwhile, the wood powder repeatedly infecting through Collybia albuminosa, also can be used as the feed of ox, sheep, for ox, sheep cultivation.In addition, utilize this formula to cultivate, the mycelia amount of acquisition increases by 4500 times of left and right.
Embodiment 2:
Substratum quality formula is as follows, and other are with embodiment 1:
Utilize this formula to cultivate, the mycelia amount of acquisition increases by 3800 times of left and right.
Embodiment 3:
Substratum quality formula is as follows, and other are with embodiment 1:
Utilize this formula to cultivate, the mycelia amount of acquisition increases by 3000 times of left and right.
Embodiment 4:
Substratum quality formula is as follows, and other are with embodiment 1:
Utilize this formula to cultivate, the mycelia amount of acquisition increases by 3500 times of left and right.
Embodiment 5:
Substratum quality formula is as follows, and other are with embodiment 1:
Utilize this formula to cultivate, the mycelia amount of acquisition increases by 4000 times of left and right.
Claims (4)
1. a liquid nutrient medium of cultivating for the mass-producing of Collybia albuminosa mycelia, prepare by following quality proportioning:
Described lignocellulose is 1: 1 mixture of white poplar wood powder and mugwort powder.
2. liquid nutrient medium as claimed in claim 1, is characterized in that described substratum prepares by following quality proportioning:
3. liquid nutrient medium as claimed in claim 1, is characterized in that described substratum prepares by following quality proportioning:
4. the application of the liquid nutrient medium as described in one of claim 1～3 in the mass-producing of Collybia albuminosa mycelia is cultivated.
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