CN104381009A - Artificial bag cultivation process for Phellinus igniarius - Google Patents
Artificial bag cultivation process for Phellinus igniarius Download PDFInfo
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Abstract
The invention relates to a cultivation method for fungi, especially to an artificial bag cultivation process for Phellinus igniarius. The process comprises the following steps: (1) purifying a strain isolated from wild Phellinus igniarius sporocarp, then inoculating the purified strain to a mother culture medium for cultivation so as to obtain a mother Phellinus igniarius strain, inoculating the mother Phellinus igniarius strain to a stock culture medium for cultivation so as to obtain a Phellinus igniarius stock and inoculating the Phellinus igniarius stock to a cultivar culture medium for cultivation so as to obtain a Phellinus igniarius cultivar; (2) inoculating the Phellinus igniarius cultivar to a mushroom-stick and carrying out cultivation for Ganoderma germination; and (3) managing germinated Ganoderma. According to results of considerable cultivation practice, it is proved that the artificial bag cultivation process for Phellinus igniarius in the invention can realize artificial cultivation of real Phellinus igniarius; moreover, the size of individual sporocarp can be artificially controlled by controlling the size of an opening. Through artificial bag cultivation of Phellinus igniarius, artificial short-term regeneration of wild rare resources and resources needing a long regeneration period is realized.
Description
Technical field
The present invention relates to a kind of breeding method of mushroom, particularly technique planted by the artificial bag of a kind of Phellinus.
Background technology
Phellinus, Chinese medicine name, is shown in " Chinese medicine voluminous dictionary ".Classification belongs to mycota (Fungi), Basidiomycota (Basigiomycota), thorn lead fungi order (Hymenochaxtales) sting lead fungi order (Hymenochaxtaceae), being the bracket fungus of hard, is the medicinal fungi that the ground such as China, Japan and Korea are famous.Existing " Phellinus " cultivation method fruiting is more difficult, and artificial culture growing state is bad, and productive rate is low.The artificial cultivation method of a kind of Phellinus (Phellinus baumiiPil á t) of application number 200910077398.8, applying date 2009-02-19, comprising: (1) is inoculated in mother culture media to cultivate and obtains Phellinus mother kind after the bacterial strain purifying that Phellinus fruit body is separated; (2) Phellinus mother plants to be inoculated on pedigree seed culture medium and cultivates acquisition Phellinus original seed; (3) Phellinus original seed is inoculated in the Cultivar culture medium in cultivation bag and cultivates; (4) cultivation bag covering with mycelia is transferred to mushroom producing room and carries out mushroom producing culture; Wherein, described mushroom producing culture is 85 ~ 90% at relative air humidity, and temperature is carry out mushroom producing culture under the environmental condition of 22 ~ 25 DEG C.This method provide a kind of new Phellinus cultivation method, but due to its process comparatively complicated, not easily promote the use of in peasant household, due to current to Phellinus large area produce in the urgent need to, need to improve existing cultivation method.
Summary of the invention
The invention provides one cultivation relatively simple, management is relatively easy, goes out sesame relative to high yield, the Phellinus artificial bag cultivation technique can carrying out large area greenhouse cultivation.
The technical solution adopted for the present invention to solve the technical problems is:
Technique planted by the artificial bag of a kind of Phellinus, and this technique comprises the steps:
(1) be inoculated into after the bacterial strain be separated from wild Phellinus fruit body is purified in mother culture media to cultivate and obtain Phellinus mother and plant, Phellinus mother plants to be inoculated into pedigree seed culture medium cultivates and obtains Phellinus original seed; Phellinus original seed is inoculated in Cultivar culture medium and cultivates acquisition Phellinus cultivated species;
Pedigree seed culture medium and Cultivar culture medium are mainly mixed by ramulus mori wood chip, wheat skin, corn flour and land plaster; Further preferably, the percentage by weight of each raw material components of pedigree seed culture medium and Cultivar culture medium is: wheat skin 10-12%, corn flour 10-12%, land plaster 1%, and surplus is ramulus mori wood chip.Wheat skin also claims wheat bran.
(2) Phellinus cultivated species is inoculated in bacterium rod carry out sesame cultivate;
The preparation of bacterium rod: ramulus mori wood chip is carried out water spray comprehensively and prewet, humid control, at 60%-70%, adds wheat skin, corn flour and land plaster and carries out spice after fully prewetting, mix rear dress plastic sack, bag diameter is 15-17cm, length≤35cm, obtains the bacterium rod for cultivating Phellinus; Bacterium rod adopts Phellinus cultivated species to inoculate after sterilizing; Postvaccinal bacterium rod is put into culturing room's lucifuge and is cultivated, and temperature controls at 20-30 DEG C, and humid control, at 40-50%, cultivates 60-90 days;
(3) sesame management is gone out:
A, when Phellinus bacterium rod start annesl yellowing or brown color time, move into greenhouse cultivation, carry out scattered light cultivation, loosen the soil to placing bacterium rod place in booth before immigration, more than pine soil depth 10cm, carries out disinfection with limewash after loosening the soil, then bacterium rod is uprightly put, spacing is at 10cm × 10cm ~ 15cm × 15cm, and greenhouse temperature controls at 15-32 DEG C, and humid control is at 80-90%;
B, cultivation more than 10 days, after treating the complete annesl yellowly of bacterium rod or brown color, carry out sealing excision work (being breathed freely in bacterium rod top) one by one, then uprightly put back to same as before in soil by bacterium rod;
C, continuation cultivation, after 4 days, are treated that the mycelia of bacterium rod sealing incision covers with bacterium rod sack again, are carried out bacterium rod open-work; First, the sealing of bacterium rod placed down, each bacterium rod outside wall surface opening 3-4, is then buried in the earth, length of embedment 3-4 centimetre; Booth ventilates 2-3 time every day, each 30-40 minute; Cultivate and obtain Phellinus fruit body after 60-90 days.
As preferably, bacterium rod carries out high pressure or normal-pressure sterilization, and during autoclaving, pressure is 3-5 atmospheric pressure, temperature 120 DEG C, and sterilization time is 6-8 hour; During normal-pressure sterilization, pressure is 1 atmospheric pressure, temperature 100 DEG C, and sterilization time is 24-36 hour; More than 18 hours are cooled to room temperature after sterilizing.
As preferably, in step (2), the seeded process of bacterium rod is as follows:
A, before inoculation starts, inoculating hood to be cleaned, then aerial fog disinfectant is carried out lighting sterilization according to 8 grams every cubic metre;
B, cooled for sterilizing bacterium rod and Phellinus cultivated species to be placed in the inoculating hood of having sterilized, carries out disinfection with bacterium aerial fog disinfectant 8g in inoculating hood, aerosol is inoculated after disappearing;
C, open cultivated species and bacterium rod sack, get cultivated species diameter 3-4cm, thickness 0.5-1cm puts into bacterium rod sack, clutches bacterium rod sack with hand, by cultivated species in bacterium rod bag by solid, then put the collar and collar shroud sealing.
As preferably, in the c of step (3), the cylinder of bacterium rod is divided into four limits, respectively opens 1 mouth on relative both sides, opening is apart from 6 centimetres, ground, and both sides are apart from ground 12 centimeters opening in addition, make 4 interleaved openings.
As preferably, the curved mouth of described opening, the two ends of arc port upward.Fruit body is made to grow up to postforming attractive in appearance.
As preferably, the percentage by weight of each raw material components of pedigree seed culture medium and Cultivar culture medium is: wheat skin 10%, corn flour 10%, land plaster 1%, and surplus is ramulus mori wood chip.Further, the granularity of described corn flour is 0.1-0.5mm, and the granularity of ramulus mori wood chip is 0.5-1.0cm.
The invention has the beneficial effects as follows: artificial bag of the present invention is planted technique and be facts have proved through a large amount of cultivations; the artificial cultivation of real Phellinus can be realized; and single fruit body size can according to openings of sizes manual control; the artificial bag of Phellinus is planted; wild scarce resource, regenerate very long resource, artificial short period regeneration.And artificial bag young plant entity component content is suitable with the yellow component content of Wild Mulberry.Current cultivation scale is about 35 mu, 250,000 bags, grows up to that Phellinus becomes fan-shaped, the shape of a hoof, and sporiferous layer (the fruit body outside of belly) has cavernous structure, surface in golden yellow or brown color, becomes yellowish-brown time old.Yellow, the brown or vernicose wooden look of bacterial context, darker in cap near surface color.Sporiferous layer be glossiness wooden look to dark-brown, be often divided into two-layer or multilayer.
Accompanying drawing explanation
Fig. 1 is the glucose standard curve that polyoses content is analyzed;
Fig. 2 is the content of the polysaccharide of 12 kinds of separate sources Phellinus;
Fig. 3 is the amino acid canonical plotting that water extraction is analyzed;
Fig. 4 is the amino acid content figure of various sources Phellinus;
Fig. 5 is the schematic diagram of bacterium of the present invention rod processing procedure.
Embodiment
Below by specific embodiment, technical scheme of the present invention is described in further detail.Should be appreciated that enforcement of the present invention is not limited to the following examples, any pro forma accommodation make the present invention and/or change all will fall into scope.
In the present invention, if not refer in particular to, all parts, percentage are unit of weight, and all equipment and raw material etc. all can be buied from market or the industry is conventional.
The present invention's Phellinus bacterial classification (Inonotus Sanghuang) used, from China Committee for Culture Collection of Microorganisms's common micro-organisms center, is numbered CGMCC NO:8854.
Embodiment 1:
The making of a, bacterial classification:
(1), Phellinus bacterial classification can be divided into production bacterial classification and preservation bacterial classification by application target; Female kind (test tube kind, one-level kind can be claimed), original seed (secondary kind) and cultivated species (three grades of kinds) 3 ranks can be divided into by Reproduction program.
(2), female kind refers to by spore separation, organizes separation and the phellinus liteus pure culture obtained through purifying, cultivation.Container is test tube, therefore is also called test tube kind or one-level kind.It had both been suitable for transplanting with test tube slant, and expanding propagation is again upper for production, is suitable for again purebred preservation.
Phellinus bacterium Inonotus sanghuang Sheng H.Wu provided by the invention; T.Hatt. & Y.C.Dai is from Chunan, Zhejiang Province Thousand Islands "Hur" mulberry edible mushroom Specialty Co-operative Organizations, is to obtain through tissue separation, rejuvenation of going down to posterity in the wild Phellinus fruit body of parasitism from the mulberry tree of Thousand-Island Lake.This Pseudomonas Basidiomycota (Basidiomycota), thorn lead fungi order (Hymenochaetales), Hymenochaetales (Hymenochaetaceae).Its biological property is: phellinus liteus growth optimum temperature is 28 DEG C, and optimum pH is 6.5, and female kind growth optimum medium is PDA medium, and mycelia the most suitable growth Bag Material medium is mulberry tree sawdust medium.Be inoculated in mulberry tree sawdust medium with this thalline and cultivate resulting bottle entity morphology for cap is fan-shaped, shell-like, the shape of a hoof, upper surface is without wart like structure and crackle, and cap is in golden yellow or brown color.Other physiological and biochemical property is: moisture must not cross 10.0%; Total ash must not cross 6%; Water-soluble extractives content must not be less than 15%; Polysaccharide is with DEXTROSE ANHYDROUS (C
6h
12o
6) meter, must not 2.60% be less than.This bacterial strain in the preservation of China General Microbiological culture presevation administrative center, is numbered CGMCC No.8854 on March 07th, 2014.Address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica's postcode 100101.The biomaterial that preservation of the present invention provides is mulberry all No. 1, and Classification And Nomenclature is: Phellinus Inonotus sanghuang.
Mother culture based formulas (PDA medium): potato 200 grams, sucrose 20 grams, 20 grams, agar, 1000 milliliters, water.
(3), original seed be by mother plant expand cultivate bacterial classification, Phellinus original seed of the present invention is solid original seed.Container mostly is 750 milliliters of colourless small-bore seed bottle or polyethylene plastic bag.This bacterial classification must close inspection, and the pure culture without living contaminants can be for the production of.
The percentage by weight of each raw material components of bacterium culture medium is: wheat skin 10%, corn flour 10% (granularity 0.1-0.5mm), land plaster 1%, and surplus is ramulus mori wood chip (granularity 0.5-1.0cm).
(4), cultivated species is by the bacterial classification of original seed expanding propagation.Medium is identical with original seed with container.
The preparation of b, bacterium rod:
The same pedigree seed culture medium of composition of bacterium rod, ramulus mori wood chip is carried out water spray comprehensively and prewet, humid control is at 60%-70%; Add wheat skin 10% next day, corn flour 10% (granularity 0.1-0.5mm), land plaster 1%, carries out spice.Medium mixes rear dress Polythene Bag (bag diameter is 15-17cm, length≤35cm), obtains the bacterium rod for cultivating Phellinus.
C, the sterilizing of bacterium rod:
Bacterium rod carries out high pressure or normal-pressure sterilization, and during autoclaving, pressure is 3-5 atmospheric pressure, temperature 120 DEG C, and sterilization time is 6-8 hour; During normal-pressure sterilization, pressure is 1 atmospheric pressure, temperature 100 DEG C, and sterilization time is 24-36 hour; More than 18 hours are cooled to room temperature after sterilizing.
D, inoculation:
(1), before inoculation starts, to clean inoculating hood, then aerial fog disinfectant according to 8 grams every cubic metre light sterilization.
(2), by cooled for sterilizing bacterium rod, Phellinus cultivated species be placed in the inoculating hood of having sterilized, carries out disinfection with bacterium aerial fog disinfectant 8g in inoculating hood, about half an hour, aerosol is inoculated after disappearing.
(3), before inoculation personnel prepare inoculation, the water of two manual cleaning is cleaned, and stretches in inoculating hood, cleans both hands, then medical stainless steel tweezers are carried out alcohol disinfecting sterilizing with the alcohol swab of 70%-75%.After open cultivated species, bacterium rod sack, get cultivated species diameter 3-4cm with tweezers, thickness 0.5-1cm puts into bacterium rod sack, clutches bacterium rod sack with hand, by cultivated species in bacterium rod bag by solid, then put the collar and collar shroud sealing, inoculate one by one.
(4), postvaccinal bacterium rod puts into culturing room lucifuge and cultivates, and temperature controls at 20-30 DEG C, and humid control, at 40-50%, cultivates 60-90 days.
E, go out sesame management:
(1), when the bacterium rod being vaccinated with Phellinus starts annesl yellowing or brown color, move into greenhouse cultivation, carry out scattered light cultivation.Loosen the soil to placing bacterium rod place in booth before immigration, more than pine soil depth 10cm, carries out disinfection with limewash after loosening the soil.When putting, bacterium rod uprightly should be put, the sealing of bacterium rod is placed upward, and spacing is at 10cm × 10cm.Greenhouse temperature controls at 15-32 DEG C, and humid control is at 80-90%.
(2), cultivate about 10 days, after treating the complete annesl yellowly of bacterium rod or brown color, bacterium rod is carried out sealing excision work one by one, and then former state is uprightly put back on soil.Namely excise the position of sealing upward after excision sealing, see Fig. 5.
(3), continue cultivation after 4 days, treat that the mycelia of bacterium rod incision covers with bacterium rod sack again, carry out bacterium rod open-work.First, by bacterium rod otch down, each bacterium rod opening 4, the cylinder of bacterium rod is divided into four limits, respectively opens 1 mouth on relative both sides, opening, apart from 6 centimetres, ground, is shown in Fig. 5.Both sides, apart from ground 12 centimeters opening, make 4 openings be Chinese character pin-shaped staggered relatively in addition.Opening requirement: upwards arc port is opened to bacterium rod outer plastic bag, makes fruit body grow up to postforming attractive in appearance.Then, by upright for bacterium rod, bacterium rod incision is buried in the earth, length of embedment 3-4 centimetre.
(4), booth must ventilate 2-3 time every day, each 30-40 minute.Cultivate and obtain Phellinus fruit body in more than 60 days.
Phellinus cultivation mouth place is appeared at, the curved mouth in cross section when receiving, beautiful appearance according to the Phellinus fruit body that the inventive method obtains.The weight of single fruit body is larger.Long 6-12 centimetre, thick 3-6 centimetre, single fruit body weighs 15 grams-50 grams.
The cultivation result of Phellinus fruit body: fresh goods fruit-body color is golden yellow, dry product fruit-body color brown color, fruit body stockless, and shape is fan-shaped, a little in the shape of a hoof, every bag can be produced Phellinus fruit body dry product 15-30g.
When gathering, get 3 fruit bodys at random and measure:
Fruit body fresh goods is long is 12cm, wide 6cm, and thick 3cm, weight is 30g.
Fruit body fresh goods is long is 9cm, wide 5cm, and thick 2.6cm, weight is 24g.
Fruit body fresh goods is long is 6cm, wide 3.5cm, and thick 2cm, weight is 15g.
Embodiment 2:
Concrete grammar is with embodiment 2, and difference is, the percentage by weight of the pedigree seed culture medium of employing and each raw material components of Cultivar culture medium is: wheat skin 12%, corn flour 12%, land plaster 1%, and surplus is ramulus mori wood chip.
Artificial bag of the present invention is planted technique and be facts have proved through a large amount of cultivations; can realize the artificial cultivation of real Phellinus, and single fruit body size can according to openings of sizes manual control, the artificial bag of Phellinus is planted; wild scarce resource, regenerate very long resource, artificial short period regeneration.And artificial bag young plant entity component content is suitable with the yellow component content of Wild Mulberry.Current cultivation scale is about 35 mu, 250,000 bags, grows up to that Phellinus becomes fan-shaped, the shape of a hoof, and sporiferous layer (the fruit body outside of belly) has cavernous structure, surface in golden yellow or brown color, becomes yellowish-brown time old.Yellow, the brown or vernicose wooden look of bacterial context, darker in cap near surface color.Sporiferous layer be glossiness wooden look to dark-brown, be often divided into two-layer or multilayer.
The Phellinus analysis of effective component of separate sources
What below experiment adopted is that the present invention cultivates the Phellinus fruit body (referred to as mulberry No. 1) obtained; the Phellinus of other kinds is all purchased from market; wherein, Jinzhai County's Phellinus purchased from Jinzhai County Shao Xin edible and medical fungi research institute of Anhui Province, mulberry all No. 2-4 gather from Thousand-Island Lake mulberry tree for the present inventor.
One, water analysis
The test sample that sample pretreatment measures, 80 mesh sieves are crossed in general first fragmentation.
Aquametry gets test sample 2 ~ 5g, and be laid in and be dried in the flat measuring cup of constant weight, thickness is no more than 5mm; Loose test sample is no more than 10mm; Accurately weighed, open bottle cap 100 ~ 105 DEG C of dryings 5 hours, bottle cap is built, in dislocation drier, cool 30 minutes, accurately weighed, then said temperature drying 1 hour, cooling, weighed, to double difference of weighing is no more than 5mg.According to the weight of less loss, calculate water content (%) in test sample.Result is as shown in table 1.
Moisture table in table 1 five kinds of separate sources Phellinus
| Phellinus is originated | Mulberry all No. 1 | Thousand-Island Lake is wild | Yunnan Wild | Thousand-Island Lake willow | Jinzhai County |
| Moisture (%) | 8.24 | 10.75 | 10.79 | 11.14 | 8.27 |
As shown in Table 1, mulberry all the moisture of No. 1 all lower than other bacterial classifications, be easier to store.
Two, ash analysis
The test sample that sample pretreatment measures, 80 mesh sieves are crossed in general first fragmentation.
The test sample that total ash determination method measures must be pulverized, enablely pass through No. two sieves, after mixing, get test sample 2 ~ 3g (as must acid-insoluble ash be measured, desirable test sample 3 ~ 5g), put in the crucible of ignition to constant weight, weighed weight (accurately to 0.01g), slowly red-hot, note avoiding burning, to when carbonizing completely, raised temperature to 500 ~ 600 DEG C gradually, make complete ashing and to constant weight.According to residue weight, calculate the content (%) of total ash in test sample.
As test sample not easily ashing, crucible can be let cool, heating water or 10% iron nitrate solution 2ml, make residue moistening, then put evaporate to dryness in water-bath, and residue is blazing according to front method, to the complete ashing of crucible contents.
Content of ashes table in table 2 five kinds of separate sources Phellinus
| Phellinus is originated | Mulberry all No. 1 | Thousand-Island Lake is wild | Yunnan Wild | Thousand-Island Lake willow | Jinzhai County |
| Content of ashes (%) | 4.95 | 3.73 | 4.86 | 1.95 | 2.87 |
The mulberry all content of ashes of No. 1 is relatively high, this illustrate mulberry all in No. 1 Phellinus inorganic salts ingredients content higher, more eutrophy.
Three, water extraction analysis
The test sample that sample pretreatment measures, 80 mesh sieves are crossed in general first fragmentation.
Hot dipping measures water lixivium method and gets test sample about 2 ~ 4g, accurately weighed, puts in the conical flask of 100 ~ 250ml, and precision adds water 50 ~ 100ml, close plug, weighed weight, left standstill after 1 hour, connect reflux condensing tube, be heated to boiling, and keep micro-and boil 1 hour.After letting cool, take off conical flask, close plug, more weighed weight, the weight of less loss is supplied with water, shake up, filter with dry filter, precision measures filtrate 25ml, put and to be dried in the evaporating dish of constant weight, in water-bath after evaporate to dryness, in 105 DEG C of dryings 3 hours, put in drier and cool 30 minutes, the close weighed weight of rapid nationality.Unless otherwise specified, the content (%) of water-soluble extractives in test sample is calculated with dry product.Result is as shown in table 3.
In table 3 five kinds of separate sources Phellinus, water extraction is containing scale
The mulberry all water extraction content of No. 1, higher than other bacterial classifications nearly 190%, illustrates that when carrying out traditional Chinese medicine preparation (boiling) as Chinese herbal medicine, active principle is more easily separated, and is more conducive to people and utilizes.
Four, polyoses content analysis
The test sample that sample pretreatment measures, 80 mesh sieves are crossed in general first fragmentation.
The preparation of reference substance solution gets glucose control product in right amount, accurately weighed, adds water and makes the solution of every 1ml containing 0.1mg, to obtain final product.
The preparation standard curve of calibration curve: draw Standard glucose solution 0,0.05,0.1,0.15,0.2,0.25,0.3,0.35,0.4mL is in test tube and add water to 0.5mL, add 5% phenol solution 0.5mL again, shake up, add rapidly the 2.5mL concentrated sulfuric acid again, mix latter 50 degrees Celsius and place 30min, measure light absorption value in 490nm place.Take light absorption value as ordinate, concentration of glucose is abscissa drawing standard curve.
The preparation of need testing solution is got this product powder and is about 2g, and accurately weighed, add water 30ml, and 90 degrees Celsius of ultrasonic assistant extract 4 hours, 5000rpm 15min, gets supernatant by calibration curve preparation method and get final product.As said method obtains calibration curve, as shown in Figure 1.As shown in Figure 1, gained linear relation is good.As calibration curve, contained by obtaining in 12 samples, polyoses content as shown in Figure 2.Wherein: F represents: Jinzhai County Phellinus J represents: mulberry is No. 1 K representative all: mulberry is No. 3 L representatives all: mulberry is No. 2 M representatives all: mulberry is No. 4 N representatives all: birch Phellinus O represents: the wild Phellinus P in Thousand-Island Lake represents: willow Phellinus Q represents: Yunnan mulberry tree Phellinus R represents: Yulin willow Phellinus S represents: smelly pine tree Phellinus T represents: Syringa amurensis tree Phellinus.
The data of Fig. 2 illustrate, the mulberry all polyoses content of No. 1 is higher, exceeds other bacterial classifications more than about 1 times, with the obvious advantage.
Five, free aminoacid content measures
Extract various sources Phellinus Free Amino Acids, accurately draw test solution 1mL, inject the volumetric flask of 25mL, add 0.5mL pH8.0 phosphate buffer and 0.5mL2% ninhydrin solution, in boiling water bath, heat 15min.Add water after cooling and be settled to 25mL.To place after 10min with 5mm cuvette at 570nm place, do reference with blank reagent solution, mensuration absorbance (A).
The making of amino acid calibration curve
Draw 0.0 respectively, 1.0,1.5,2.0,2.5,3.0mL amino acid working solution is in one group of 25mL volumetric flask, respectively add water 4mL, pH8.0 phosphate buffer 0.5mL and 2% ninhydrin solution 0.5mL, 15min is heated in boiling water bath, add water after cooling and be settled to 25mL, measure absorbance (A570).By the absorbance that records and corresponding aminoglutaric acid concentration drawing standard curve.
Result calculates
Phellinus Determination of Free Amino Acids is with dry mass fraction representation, and calculating formula is as follows
Free amino acid total amount (glutamic acid meter) (%)=[(C/1000) * (L1/L2)] * 100/M0*m
L1---test solution total amount, mL;
L2---mensuration test solution amount, mL;
M0---sample size, g;
C---according to the milligram number of the glutamic acid that the absorbance measured checks in from calibration curve;
M---sample dry matter content.
According to this method Preparation of amino acid calibration curve, result as shown in Figure 3.And then carry out amino acid whose mensuration in the Phellinus of various sources, result is as shown in Figure 4.Bionical Phellinus in figure and mulberry of the present invention all No. 1, all the amino acid content of No. 1 is comparatively outstanding can to find out mulberry, is about 5 times of other bacterial classifications.
The above example is only one of the present invention preferably scheme, but technical characteristic of the present invention is not limited thereto, and any those skilled in the art is in the field of the invention, and the change done or modification are all encompassed among the scope of the claims of the present invention.
Claims (8)
1. a technique planted by the artificial bag of Phellinus, it is characterized in that this technique comprises the steps:
(1) be inoculated into after the bacterial strain be separated from wild Phellinus fruit body is purified in mother culture media to cultivate and obtain Phellinus mother and plant, Phellinus mother plants to be inoculated into pedigree seed culture medium cultivates and obtains Phellinus original seed; Phellinus original seed is inoculated in Cultivar culture medium and cultivates acquisition Phellinus cultivated species; Pedigree seed culture medium and Cultivar culture medium are mainly mixed by ramulus mori wood chip, wheat skin, corn flour and land plaster;
(2) Phellinus cultivated species is inoculated in bacterium rod carry out sesame cultivate;
The preparation of bacterium rod: the same pedigree seed culture medium of composition of bacterium rod, ramulus mori wood chip is carried out water spray comprehensively to prewet, humid control is at 60%-70%, add wheat skin, corn flour and land plaster after fully prewetting and carry out spice, mix rear dress plastic sack, bag diameter is 15-17cm, length≤35cm, obtains the bacterium rod for cultivating Phellinus; Bacterium rod adopts Phellinus cultivated species to inoculate after sterilizing; Postvaccinal bacterium rod is put into culturing room's lucifuge and is cultivated, and temperature controls at 20-30 DEG C, and humid control, at 40-50%, cultivates 60-90 days;
(3) sesame management is gone out:
A, when Phellinus bacterium rod start annesl yellowing or brown color time, move into greenhouse cultivation, carry out scattered light cultivation, loosen the soil to placing bacterium rod place in booth before immigration, more than pine soil depth 10cm, carries out disinfection with limewash after loosening the soil, then bacterium rod is uprightly put, spacing is at 10cm × 10cm ~ 15cm × 15cm, and greenhouse temperature controls at 15-32 DEG C, and humid control is at 80-90%;
B, cultivation more than 10 days, after treating the complete annesl yellowly of bacterium rod or brown color, carry out sealing excision work one by one by bacterium rod, then uprightly put back in soil same as before;
C, continuation cultivation, after 4 days, are treated that the mycelia of bacterium rod sealing incision covers with bacterium rod sack again, are carried out bacterium rod open-work; First, the sealing of bacterium rod placed down, each bacterium rod outside wall surface opening 3-4, is then buried in the earth, length of embedment 3-4 centimetre; Booth ventilates 2-3 time every day, each 30-40 minute; Cultivate and obtain Phellinus fruit body after 60-90 days.
2. technique planted by the artificial bag of Phellinus according to claim 1, and it is characterized in that: bacterium rod carries out high pressure or normal-pressure sterilization, during autoclaving, pressure is 3-5 atmospheric pressure, temperature 120 DEG C, and sterilization time is 6-8 hour; During normal-pressure sterilization, pressure is 1 atmospheric pressure, temperature 100 DEG C, and sterilization time is 24-36 hour; More than 18 hours are cooled to room temperature after sterilizing.
3. technique planted by the artificial bag of Phellinus according to claim 1 and 2, it is characterized in that: in step (2), the seeded process of bacterium rod is as follows:
A, before inoculation starts, inoculating hood to be cleaned, then aerial fog disinfectant is carried out lighting sterilization according to 8 grams every cubic metre;
B, cooled for sterilizing bacterium rod and Phellinus cultivated species to be placed in the inoculating hood of having sterilized, carries out disinfection with bacterium aerial fog disinfectant 8g in inoculating hood, aerosol is inoculated after disappearing;
C, open cultivated species and bacterium rod sack, get cultivated species diameter 3-4cm, thickness 0.5-1cm puts into bacterium rod sack, clutches bacterium rod sack with hand, by cultivated species in bacterium rod bag by solid, then put the collar and collar shroud sealing.
4. technique planted by the artificial bag of Phellinus according to claim 1 and 2, it is characterized in that: in the c of step (3), the cylinder of bacterium rod is divided into four limits; 1 mouth is respectively opened on relative both sides; opening is apart from 6 centimetres, ground, and both sides are apart from ground 12 centimeters opening in addition, make 4 interleaved openings.
5. technique planted by the artificial bag of Phellinus according to claim 4, it is characterized in that: the curved mouth of described opening, the two ends of arc port upward.
6. technique planted by the artificial bag of Phellinus according to claim 1 and 2, and it is characterized in that: the percentage by weight of each raw material components of pedigree seed culture medium and Cultivar culture medium is: wheat skin 10-12%, corn flour 10-12%, land plaster 1%, surplus is ramulus mori wood chip.
7. technique planted by the artificial bag of Phellinus according to claim 6, and it is characterized in that: the percentage by weight of each raw material components of pedigree seed culture medium and Cultivar culture medium is: wheat skin 10%, corn flour 10%, land plaster 1%, surplus is ramulus mori wood chip.
8. technique planted by the artificial bag of Phellinus according to claim 6, and it is characterized in that: the granularity of described corn flour is 0.1-0.5mm, the granularity of ramulus mori wood chip is 0.5-1.0 cm.
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