CN104871829A - Method for culturing Clitocybe maxima from cattail scraps - Google Patents
Method for culturing Clitocybe maxima from cattail scraps Download PDFInfo
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- CN104871829A CN104871829A CN201510323696.6A CN201510323696A CN104871829A CN 104871829 A CN104871829 A CN 104871829A CN 201510323696 A CN201510323696 A CN 201510323696A CN 104871829 A CN104871829 A CN 104871829A
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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
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Abstract
The invention provides a method for culturing Clitocybe maxima from cattail scraps. The method comprises steps as follows: A, preparing and culturing liquid strains; B, preparing and culturing grain strains; C, preparing a culture medium for culture; D, under the sterile condition, inoculating the culture medium prepared in Step C with the grain strains prepared in Step B for culture, fruiting management and harvesting. The method is characterized in that the cattail scraps are contained in the culture medium in Step A and/or Step B and/or Step C. Wetland cattail waste is used, ecological damage to water is reduced, the culture cost is reduced, and the method has significant economic benefit and ecological benefit and has broad application prospect.
Description
Technical field
The invention belongs to the technical field of edible fungus cultivation, be specifically related to a kind of cattail that utilizes and consider the method for cultivating Clitocybe maxima to be worth doing, and the preparation method of special culture material.
Background technology
Clitocybe maxima (Clitocybe maxima) has another name called Clitocybe Maxima, and popular name bamboo shoot mushroom, large funnel mushroom, gain the name because of its fruit body likeness in form funnel or cup.It is the more autochthonal bacterium of wood corruption of a kind of natural distribution.Clitocybe maxima belongs to Basidiomycotina, Hymenomycetes, Agaricales, Tricholomataceae, Clitocybe.This genus has found that in the whole world more than 250 plant, and current China known more than 30 plants.
Clitocybe maxima is the Rare edible fungus of summer high temperature energy in season fruiting, fruit body is nutritious, in Clitocybe maxima cap, the relative total content of essential amino acid is up to 43%, secondly the relative total content of the essential amino acid in mushroom cap is 40.9%, content in Clitocybe maxima stem is slightly 40.3% lower than mushroom cap, and in Clitocybe maxima, the content of essential amino acid is higher than edible mushrooms such as mushroom, mushroom, flat mushrooms.Especially the sulfur amino acid in Clitocybe maxima, the content of methionine are higher, exceed nearly one times than mushroom, straw mushroom, flat mushroom etc.Clitocybe maxima, with its abundant nutrition, tender and crisp quality, pleasant mouthfeel, unique local flavor and excellent outward appearance, is subject to the favor of consumers in general deeply.The three mushroom classes be separated in Clitocybe maxima fruit body or mycelium and polysaccharose substance have develop immunitypty and antitumor action.Therefore, Clitocybe maxima is a kind of food medicine dual-purpose bacterium.
Cattail is the wetland water plants Dominant variety of generally acknowledging in the world, although wait can the consumable portion stem or leaf of cattail in braiding, consumption is limited.Cattail growth and breeding is fast and biomass is large, widely distributed in China, owing to having the advantage such as wide adaptability, strong stress resistance, at present, the amount reproduction of cattail at wetland and then the residual branch the like waste of generation, cause the destruction to water body, and form the endogenous pollution to wetland.
Cellulose is that edible and medicinal fungi grows necessary nutriment, also be the main component of cattail, so utilize cattail to cultivate mushroom, expand the resource evaluation and exploration technology approach of cattail, not only become the new medium of cultivation mushroom, for more opportunity is created in increasing peasant income, and, be conducive to the benign development of Wetland ecological.
The twigs of the chaste tree (Vitex negundo var ineica) are very extensive in the distribution of Hebei hilly region in Taihang Mountains, belong to dominant shrub species seeds.Taihang Mountain, Hebei twigs of the chaste tree aboundresources, widely distributed, many places are in wild state according to investigations, there is no larger utilization; Compile with regard to traditional Land use systems chaste tree, total utilization of the twigs of the chaste tree also not enough stock number 10%, research shows have cellulose, lignin in the twigs of the chaste tree, these edible and medicinal fungis of half fiber grow necessary nutriment, also containing abundant organic matter and the nutritive element required for crop growth.But up to the present, the twigs of the chaste tree are regarded as agriculture and forestry organic waste material always and do not obtain fully and effectively developing, and seriously waste resource.
Summary of the invention
The object of this invention is to provide a kind of cattail that utilizes for the method for major ingredient cultivation Clitocybe maxima, reduce cattail to the destruction of water body.
In order to realize object of the present invention, the invention provides following technical scheme.
This cattail that utilizes considers the method for cultivating Clitocybe maxima to be worth doing, comprises the steps:
A. the preparation of liquid spawn and cultivation;
B. the preparation of grain spawn and cultivation;
C. the preparation of cultivation medium;
D., under aseptic condition, the medium that the grain spawn that prepared by step B access step C prepares, carries out cultivating, management of producing mushroom, to gather;
It is characterized in that: in steps A or/and step B or/and step C medium in containing cattail bits.
The described cattail that utilizes considers the method for cultivating Clitocybe maxima to be worth doing, in steps A Clitocybe maxima liquid spawn culture medium in as substrate for induction thing, add cattail bits in advance; It optimizes liquid spawn culture medium formulation weight percentage: potato 20%, cattail bits 5%, wheat bran 3%, glucose 2%, KH2PO40.3%, MgSO40.15%, VB110mg/L, carboxymethyl cellulose (CMC, lower same) 0.3%, water 1000mL, pH nature.
The described cattail that utilizes considers the method for cultivating Clitocybe maxima to be worth doing, and in steps A, the preparation of liquid spawn is characterised in that:
A. the preparation of level liquid bacterial classification
Under aseptic condition, commercially available mother is planted in access level liquid medium, 26-28 DEG C, 160r/min constant-temperature shaking culture 7d, i.e. obtained level liquid bacterial classification;
Described level liquid bacterium culture medium comprises: by weight percentage, potato 20%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, CMC 0.3%, VB
110mg/L;
B. the preparation of second-class liquid isolate
Under aseptic condition, by level liquid bacterial classification with in the inoculum concentration access secondary liquid medium of 10%, 26-28 DEG C, 160r/min constant-temperature shaking culture 7d, i.e. obtained second-class liquid isolate;
Described second-class liquid isolate culture medium prescription percentage by weight is: potato 20%, cattail 5%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, CMC 0.3%, VB
110mg/L.
The described cattail that utilizes considers the method for cultivating Clitocybe maxima to be worth doing, in step B Clitocybe maxima grain spawn medium in also as substrate for induction thing, add wood chip shape cattail bits in advance; Its grain spawn culture medium prescription optimized: wheat 85%, cattail 7%, twigs of the chaste tree branch bits 7%, quicklime 1%.
The described cattail that utilizes considers the method for cultivating Clitocybe maxima to be worth doing, and in step B, the preparation of grain spawn is characterised in that:
Second-class liquid isolate access grain spawn medium steps A obtained, inoculum concentration is 10%, 26-28 DEG C and is cultured to full, obtains grain spawn.
The described cattail that utilizes considers the method for cultivating Clitocybe maxima to be worth doing, and in step C, cattail bits are as the major ingredient of cultivation Clitocybe maxima, and culture medium for cultivating weight composition comprises: cattail bits 60%, twigs of the chaste tree branch bits 10%, cotton seed hull 10%, wheat bran 19%, quicklime 1%; Cattail, twigs of the chaste tree branch are ground into wood chip shape by preparation method, and be equipped with cotton seed hull, wheat bran and quicklime, material-water ratio is 1: 1.4.
The described cattail that utilizes considers the method for cultivating Clitocybe maxima to be worth doing, and in step C, the preparation of cattail bits medium is characterised in that:
Cattail bits are pulverized in advance as wood chip shape, take each constituent of cattail bits medium, add water after mixing, pile vexed 30min, load in container, sterilizing, for subsequent use after cooling.
The described method utilizing cattail to consider medium cultivation Clitocybe maxima to be worth doing, in step D, the inoculum concentration of grain spawn is 5-10%.
The described method utilizing cattail to consider medium cultivation Clitocybe maxima to be worth doing, in liquid nutrient medium, cattail bits used, pulverize and cross 80 mesh sieves in advance.
The described method utilizing cattail to consider medium cultivation Clitocybe maxima to be worth doing, in step C, the sterilising conditions of cattail bits medium is 100 DEG C of normal-pressure sterilization 36h.
Innovative point of the present invention is that the culture medium for cultivating major ingredient of Clitocybe maxima is cattail bits; Add cattail bits in liquid medium within advance as substrate for induction thing, and have employed the female kind-liquid spawn-grain spawn of Clitocybe maxima PDA-cattail bits fruiting bag technique, the production cycle nearly 1/3rd can be shortened.Technological progress effect is:
1, for edible mushroom, for it provides growth required carbon source;
2, be not yet developed before Typhales, aboundresources, low for Edible Fungi cost, high financial profit;
3, from the angle of ecology, decrease the destruction to water body, for edible fungus culturing provides a kind of cheap culturing raw material, not only avoid the endogenous pollution of wetland, and significantly reduce cultivation cost, solve the urgency of culturing edible fungus resource, there is considerable economic benefit, have a extensive future.
Embodiment
Below in conjunction with specific embodiment, content of the present invention is further described in detail.
Embodiment: utilize cattail to cultivate the method for Clitocybe maxima, Clitocybe maxima bacterial classification is market purchasing.
Steps A. the preparation of liquid spawn
Level liquid Spawn incubation based formulas: potato 20%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, VB
110mg/L, CMC 0.3%, pH nature;
Second-class liquid isolate culture medium prescription: cattail 5%, potato 20%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, VB
110mg/L, CMC 0.3%, pH nature; Wherein cattail end was pulverized 80 mesh sieves and was obtained powdery, and boil 30min altogether with potato, wheat bran, filter and remove residue obtains leaching liquor.
Use 500mL conical flask time prepared by level liquid bacterial classification, liquid amount is 200mL, after 121 DEG C of autoclaving 30min, accesses commercially available PDA slant strains under aseptic condition, and every bottle of inoculum concentration is 4 pieces of 0.5cm × 0.5cm bacterial classification blocks.After inoculation 28 DEG C, under the condition of 160r/min, constant-temperature shaking culture 7d.
Second-class liquid isolate liquid amount specification, sterilising conditions and condition of culture are the same, and inoculum concentration is 10%.
A bacterium velocity contrast of table 1 Clitocybe maxima liquid spawn and PDA bacterial classification
| Bacterial classification | Send out bacterium speed (mm/d) |
| The liquid spawn optimized | 4.18±0.02 |
| Conventional PDA bacterial classification | 2.88±0.01 |
As shown in Table 1, liquid spawn is faster than a bacterium speed of PDA bacterial classification, thus shortens the production cycle.
The preparation of step B. grain spawn
Grain spawn culture medium prescription: wheat 85%, cattail bits 14%, quicklime 1%.
Accurately take each constituent of medium, material-water ratio 1:1.4 carries out spice, and load 17cm × 32cm × 0.05cm polyethylene plastic bag, every packed siccative 300g, prepares 120 bags.The second-class liquid isolate that optimization is obtained and the control group liquid spawn of not being added cattail access grain spawn medium respectively, inoculate 60 bags respectively, measure the long speed of bacterium, and record the purseful time.
In order to investigate the advantage of grain spawn medium of the present invention, invention has been comparative trial, operating the same, control group medium is wheat medium: wheat 99%, quicklime 1%;
The advantage comparative test result of grain spawn medium is as shown in table 2:
The purseful time of table 2 Clitocybe maxima grain spawn
As shown in Table 2, mycelia is faster than the long speed of pure wheat medium in " wheat+cattail bits " grain culture medium, this is because wheat gap, along with the interpolation of cattail bits, more be conducive to mycelial growth, and in liquid medium within, add cattail bits, enable the faster Adaptable growth substrate of mycelia, thus accelerate to send out bacterium speed, shorten the production cycle and enhance productivity.
The preparation of step C. cattail composts or fertilisers of cultivating
In mass ratio, take cattail bits 60%, twigs of the chaste tree branch bits 10%, cotton seed hull 10%, wheat bran 19%, quicklime 1%, material-water ratio is that 1 ︰ 1.4 carries out spice, after mixing, loads in container and carry out sterilizing and inoculation work after piling vexed 30min.
After cattail is pulverized, powder is thinner, as mushroom cultivation matrix, affect respiration, if admixed as auxiliary material by twigs of the chaste tree bits, not only can supplementary carbon source, because of its physical form, the gas permeability of cattail major ingredient can also be improved, be used for cultivating Clitocybe maxima, can reduce costs, increase economic efficiency, can recycling economy be realized again.
Loaded in Polythene Bag, Polypropylene Bag, chest or vial by above-mentioned composts or fertilisers of cultivating, bore is not limit, conventional tying, and normal-pressure sterilization, generally with 100 DEG C of sterilizings, keeps 36-48h; Inoculation after cooling.
The consumption of cattail has carried out analysis of experiments, specific as follows:
Accurately take each constituent by shown in table 3, mixed, add water, material-water ratio is 1 ︰ 1.4, and the composts or fertilisers of cultivating configured is loaded test tube, and each formula repeats 5 times, 100 DEG C of normal-pressure sterilization 36-48h.Grain spawn described in step B is accessed respectively in the test tube containing composts or fertilisers of cultivating, carry out test tube and to nourish and grow test.
Table 3 culture medium prescription (siccative, %)
| Formula | Cattail is considered to be worth doing | Cotton seed hull | The twigs of the chaste tree are considered to be worth doing | Wheat bran | Quicklime |
| P0 | 0 | 80 | 0 | 19 | 1 |
| P1 | 30 | 40 | 10 | 19 | 1 |
| P2 | 40 | 30 | 10 | 19 | 1 |
| P3 | 50 | 20 | 10 | 19 | 1 |
| P4 | 60 | 10 | 10 | 19 | 1 |
| P5 | 70 | 0 | 10 | 19 | 1 |
Observe long speed and the growing way situation of mycelia in a test tube every 2 days, measure the long speed of mycelia with slide measure (accuracy 0.02mm).Growth rata and potential of hypha result is as shown in table 4.
The mycelial growth situation of table 4 Clitocybe maxima
| Formula | Mycelium length (mm/d) | Mycelium growth vigor |
| P0 | 4.25±0.01 | +++ |
| P4 | 4.19±0.02 | +++ |
| P5 | 4.10±0.01 | +++ |
| P3 | 4.07±0.03 | ++ |
| P2 | 3.91±0.04 | ++ |
| P1 | 3.83±0.01 | ++ |
(note: list long speed and be mean value ± standard error; +++ represent that growth is fine and close, ++ represent that growth is finer and close ,+represent that growth is general.) as can be seen from Table 4, during increase along with cattail bits addition, its growing way is better, and mycelia is pure white, finer and close, has comparativity with pure cotton seed hull medium, and mycelia grows the fastest on formula P0, and P4 group is faster than pure cattail bits group leader P5 speed.
Take each composition by the formula in table 3, mix, material-water ratio 1 ︰ 1.4 carries out spice, and load 17cm × 32cm × 0.05cm polyethylene plastic bag, every packed siccative is about 300g, observes and records the growing state of each formula and calculate biological transformation ratio.Result of the test is as shown in table 5.
Biological transformation ratio (%)=(fruit body fresh weight/composts or fertilisers of cultivating dry weight) × 100%
Above-mentioned Clitocybe maxima biological transformation ratio result of the test is as shown in table 5.
The biological transformation ratio of table 5 Clitocybe maxima
| Medium is numbered | Average yield (g) | Average organism efficiency (%) |
| P0 | 226.05 | 75.35 |
| P4 | 223.35 | 74.45 |
| P5 | 220.11 | 73.37 |
| P3 | 219.33 | 72.11 |
| P2 | 217.44 | 71.48 |
| P1 | 216.48 | 70.16 |
As can be seen from Table 5, when with cattail bits for primary carbon source reaches 60%, the biological transformation ratio of Clitocybe maxima reaches 74.45% respectively, far away higher than the biological transformation ratio 73.37% that pure cattail is considered to be worth doing, although do not reach the biological transformation ratio 75.35% of the pure cotton seed hull of planting material major ingredient, there is comparativity.In sum, cattail bits can become the major ingredient in Clitocybe maxima plantation, and cattail is considered this ecological pollution thing to be worth doing and be have also been obtained recycling, significant to improvement of the ecological environment.
D. inoculate, cultivate, gather
By cooled for sterilizing fruiting bag, according to the inoculum concentration access step B gained grain spawn of 5-10% under sterile working.
Mycelia walked full cultivation bag after 10 days, and just can open a bag fruiting when temperature is stabilized in more than 20 DEG C.Remove the collar one by one, separate opened mouth, at cultivation charge level earthing.Thickness of earth covering is 3-4cm, can select and burn soil, native, the vegetable garden soil in field is cover soil material, and grogs diameter is shine to turning white under first earthing should being placed in the sun before 1.5-2.0cm. uses, and then regulates grogs humidity with adding water.Do not fall apart with pinch flat of grogs for degree.Bacterium bag top after earthing is down rolled over, makes the unearthed face 2-3cm of sack edge height, and the bacterium bag handled well is vertically arranged on indoor fruiting bedstead equably.
After earthing, note keeping earthing humidity, and many pass door and windows or many epiphragmas, stimulate former base to break up early.After general earthing, the former base of 7-10d can expose native face.After former base is unearthed, place relative air humidity should be noted to control at 80-95%. relative air humidity too low, easily chaps in former base top, causes cap to break up; Add forced ventilation simultaneously, keep place air fresh, and notice that there is certain scattered light in place.Whole fruiting stage place temperature should control between 23-32 DEG C.Injection flow rate is specifically grasped according to the humidity of mushroom size, earthing and climatic condition, controls ventilation flexibly according to mushroom bulk-growth different phase; Stem is unearthed, cap is formed, grows stronger ventilation amount successively of each stage, and mushroom room relative air humidity keeps about 90%.Gather in time when mushroom body maturation, every damp mushroom should fill earthing after having adopted in time, carries out lower damp management of producing mushroom after cutting off the water bacteria 3-5 days.
Fruit body reaches eight or nine points of maturations, in funnel-form, edge is involute, should gather in time when spore not yet launches.As long as mushroom body is cut with the mushroom handle place of scissors in the cleaning of native face when gathering.Can adopt when gathering stay greatly little, but note do not injure the immature fruit body of periphery.Often adopt next fruit body, the mushroom handle remained in soil should be pinched by timely hand, rotate gently and pull out, avoid mushroom handle in soil, rot to cause damage by disease and insect and occur.
In sum, cattail of the present invention can be used for cultivating Clitocybe maxima, therefore, using cattail as new raw material, for industrialization development, has important innovative significance.
Claims (10)
1. utilize cattail to consider a method of cultivating Clitocybe maxima to be worth doing, comprise the steps:
A. the preparation of liquid spawn and cultivation;
B. the preparation of grain spawn and cultivation;
C. the preparation of cultivation medium;
D., under aseptic condition, the medium that the grain spawn that prepared by step B access step C prepares, carries out cultivating, management of producing mushroom, to gather;
It is characterized in that: in steps A or/and step B or/and step C medium in containing cattail bits.
2. according to claim 1ly utilize cattail to consider to be worth doing to cultivate the method for Clitocybe maxima, its feature comprises: in steps A Clitocybe maxima liquid spawn culture medium in as substrate for induction thing, add cattail bits in advance; It optimizes liquid spawn culture medium formulation weight percentage: potato 20%, cattail bits 5%, wheat bran 3%, glucose 2%, KH2PO4 0.3%, MgSO4 0.15%, VB1 10mg/L, carboxymethyl cellulose (CMC, lower same) 0.3%, water 1000mL, pH nature.
3. utilize cattail to consider the method for cultivating Clitocybe maxima to be worth doing according to claim 2, in described steps A, the preparation of liquid spawn is characterised in that:
A. the preparation of level liquid bacterial classification
Under aseptic condition, commercially available mother is planted in access level liquid medium, 26-28 DEG C, 160r/min constant-temperature shaking culture 7d, i.e. obtained level liquid bacterial classification;
Described level liquid bacterium culture medium comprises: by weight percentage, potato 20%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, CMC 0.3%, VB
110mg/L;
B. the preparation of second-class liquid isolate
Under aseptic condition, by level liquid bacterial classification with in the inoculum concentration access secondary liquid medium of 10%, 26-28 DEG C, 160r/min constant-temperature shaking culture 7d, i.e. obtained second-class liquid isolate;
Described second-class liquid isolate culture medium prescription percentage by weight is: potato 20%, cattail 5%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, CMC 0.3%, VB
110mg/L.
4. according to claim 1ly utilize cattail to consider to be worth doing to cultivate the method for Clitocybe maxima, its feature comprises: in step B Clitocybe maxima grain spawn medium in also as substrate for induction thing, add wood chip shape cattail bits in advance; Its grain spawn culture medium prescription optimized: wheat 85%, cattail 7%, twigs of the chaste tree branch bits 7%, quicklime 1%.
5. utilize cattail to consider the method for cultivating Clitocybe maxima to be worth doing according to claim 4, in described step B, the preparation of grain spawn is characterised in that:
Second-class liquid isolate access grain spawn medium steps A obtained, inoculum concentration is 10%, 26-28 DEG C and is cultured to full, obtains grain spawn.
6. the cattail that utilizes according to claim 1 considers the method for cultivating Clitocybe maxima to be worth doing, its feature comprises: in step C, cattail bits are as the major ingredient of cultivation Clitocybe maxima, and culture medium for cultivating weight composition comprises: cattail bits 60%, twigs of the chaste tree branch bits 10%, cotton seed hull 10%, wheat bran 19%, quicklime 1%; Cattail, twigs of the chaste tree branch are ground into wood chip shape by preparation method, and be equipped with cotton seed hull, wheat bran and quicklime, material-water ratio is 1: 1.4.
7. utilize cattail to consider the method for cultivating Clitocybe maxima to be worth doing according to claim 6, in described step C, the preparation of cattail bits medium is characterised in that:
Cattail bits are pulverized in advance as wood chip shape, take each constituent of cattail bits medium, add water after mixing, pile vexed 30min, load in container, sterilizing, for subsequent use after cooling.
8. utilize cattail to consider the method for medium cultivation Clitocybe maxima to be worth doing according to claim 1, it is characterized in that, in step D, the inoculum concentration of grain spawn is 5-10%.
9. utilize cattail to consider the method for medium cultivation Clitocybe maxima to be worth doing according to claim 2, it is characterized in that, in liquid nutrient medium, cattail bits used, pulverize and cross 80 mesh sieves in advance.
10. utilize cattail to consider the method for medium cultivation Clitocybe maxima to be worth doing according to claim 1, it is characterized in that, in step C, the sterilising conditions of cattail bits medium is 100 DEG C of normal-pressure sterilization 36h.
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Application publication date: 20150902 |