The cultural method of Nutrition Materials of Spawn For Morchella and preparation method thereof and hickory chick strain
The invention patent claims priority, the patent name of earlier application: Nutrition Materials of Spawn For Morchella and preparation method thereof and
The cultural method of hickory chick strain, number of patent application: 2018110949463, the applying date: 2018-9-19.
Technical field
The invention belongs to hickory chick culture technique fields;More particularly to a kind of strain base that promotion sclerotium of morchella esculenta is formed
Matter;Further relate to the preparation method of above-mentioned strain substrate;Further relate to the hickory chick Spawn incubation method based on above-mentioned strain substrate.
Background technique
Hickory chick is rare Rare edible fungus mushroom, is under the jurisdiction of Eumycota Ascomycetes Pezizale Morchellaceae morchella
(Morchellagenus).Hickory chick delicious flavour, nutritive value are high, rich in essential amino acid, granulose and anti-oxidant
Substance, it is expensive.Acquisition wild toadstool has been unable to satisfy the demand of consumption market.In recent years, hickory chick artificial cultivation skill
Art makes a breakthrough, and realizes mature and stable commercialization field production.
Strain is the first step of mushroom cultivation.The quality of strain quality decides the vigor and physiological status of thallus, thus
Affect the yield of the mushroom (fructification) finally harvested.The current existing result of study about hickory chick cultivation technique and specially
Benefit display promotes it to form sclerotium, peomotes hickory chick fruiting, improves hickory chick in the vegetative growth phase of hickory chick
Yield.For example, being described in the patent " Cultivation of Morchella " (1989) of U.S. Patent number US4866878
Vegetative growth phase forms more sclerotium then more rule of hickory chick fruiting.The patent of U.S. Patent number US6907691B2
" Cultivation of Morchella " (2005) are directly using sclerotium as realizing sheep in strain sowing to cultivation matrix
The factory culture of tripe bacterium discloses sclerotium of morchella esculenta for the importance of formation fructification.China Patent No.
ZL201510532171.3 authorization patent of invention " it is a kind of promote sclerotium of morchella esculenta formed conversion bag and hickory chick cultivation side
Method " provides a kind of conversion bag, and the hickory chick being seeded into agricultural land soil can be promoted to form sclerotium and improve final receive
The yield of the Morchella esculenta (L.) Pers sporophore obtained.But in current China, Australia, New Zealand, Turkey, Romania, France etc.
In the hickory chick crop field agriculture method cultivation mode of state's prevalence, there has been no patents and paper to pass through design and rational hickory chick bacterium culture medium
Matter formula also decomposes without analysis strain substrate formula and hickory chick to promote to form more sclerotium in strain and utilizes nutriment
Enzyme activity between have which kind of connection, have which kind of influence to Sclerotia forming and final fruiting body yield.Publication No.
By to potato-in a kind of application stage patent of invention " efficient method for promoting sclerotium of morchella esculenta to be formed " of CN105779308A
Soil filtrate is added in glucose-agar medium, the Morciiella Esculeuta Mycelia cultivated in laboratory culture dish can be promoted to be formed more
More sclerotium, if but not yet evidence show be added to most widely used bacterium culture medium in current production practices
Whether same facilitation effect is had in matter.Also, the preparation method needs of " soil filtrate " provided in the application for a patent for invention
" soil of high organic content " and " hickory chick stem soil " are used, the easy acquirement of material and supply are limited.Preparation
Step includes " be centrifuged, take supernatant, being filtered with cell filter ", and preparation method is cumbersome.In the application for a patent for invention not
The formation which kind of ingredient in " soil filtrate " promotes sclerotium of morchella esculenta by which kind of effect is analyzed, and various regions soil constituent exists
Difference, the ingredient of " soil filtrate " that causes different regions this method to prepare is there may be difference, the stability of effect and again
Existing property is not easy to guarantee.
Existing hickory chick Spawn incubation material fabrication process and strain production technology patent of invention (contain application at present for other
Inspection phase), such as " screening of strain of steepletop hickory chick and the strain preparation " of publication No. CN1793315A, publication No.
" the terraced rib hickory chick of " screening of balck vein hickory chick bacterial strain and strain preparation " of CN1793316A, publication No. CN103141302A
The production method of cultivar ", publication No. CN104686196A " a kind of fructification of drying in the shade that passes through saves and separation hickory chick strain
Method ", " a kind of manufacturing method for morchella mother culture " of publication No. CN101946634A, publication No. CN105154342A " one
" a kind of hickory chick Cultivar culture medium and its system of the cultural method of kind of hickory chick liquid strain ", publication No. CN102757291A
Preparation Method ", " a kind of using dendrobium fiber as the method for preparing morchella mycelium of matrix " of publication No. CN101743854A etc.
The strain substrate formula and preparation method of offer are not directed to be formed with sclerotium of morchella esculenta no facilitation.
Summary of the invention
The present invention provides the cultural methods of a kind of Nutrition Materials of Spawn For Morchella and preparation method thereof and hickory chick strain;The base
Matter can activate the activity of the laccase of hickory chick, manganese peroxidase and lignin peroxidase, make Nutrition Materials of Spawn For Morchella
In Morciiella Esculeuta Mycelia growth it is more vigorous, form more sclerotium, improve the fruiting body yield finally harvested.
The technical scheme is that Nutrition Materials of Spawn For Morchella, is made of rape stalk fragment and wheat berry, by mass parts
Number, rape stalk fragment are 60-75 parts, and wheat berry is 25-40 parts.
Wherein the length of rape stalk fragment is 0.5-5cm, width 1-4mm.
It is of the invention another solution is that the preparation method of above-mentioned Nutrition Materials of Spawn For Morchella, step S1, by mass fraction
60-75 parts of rape stalk fragment are taken, 25-40 parts of wheat berry, rape stalk fragment is impregnated with water, is 1%- with mass concentration
Wheat berry is impregnated with by 1.5% limewash;Step S2 mixes the rape stalk fragment and wheat berry that are impregnated with obtained in step 1
Conjunction is uniformly fitted into strain bag, and the 2-2.5h that sterilizes at 121 DEG C, obtains strain substrate.
Wherein the length of rape stalk fragment is 0.5-5cm, width 1-4mm.
Wherein rape stalk fragment is impregnated into 3-6h in step S1 in water, wheat berry is impregnated into 8-14h in limewash.
It is of the invention another solution is that the cultural method of hickory chick strain, forms in above-mentioned promotion sclerotium of morchella esculenta
Strain substrate the obtained strain substrate of preparation method on be inoculated with morchella mother culture or original seed, obtain hickory chick original seed accordingly
Or cultivar.
Compared with prior art, simple easy the beneficial effects of the present invention are: primary raw material is rape stalk in the matrix
, cost is relatively low, and can reduce rape producing region crop straw burning, promote agricultural wastes resource recycling, has good
Ecological benefits.Laccase, manganese peroxidase and the wood of the hickory chick thallus grown in the matrix energy General Promotion strain substrate
The activity of lignin peroxidase.Without using soil, so that strain substrate production method is simple, raw material easily obtains, mitigate strain
The weight of matrix reduces transportation cost.The Morciiella Esculeuta Mycelia in strain substrate can be made to grow vigorous, the more sclerotium of formation, to most
The Morchella esculenta (L.) Pers sporophore yield harvested eventually has promotion effect.
Specific embodiment
Technical solution of the present invention is further illustrated below with reference to principle and specific embodiment.
In technical solution of the present invention research process, by be grown in a series of crop material materials (including wheat,
Rice, corn, rape, soybean, the stalk of cotton and husk, wheat bran, cotton seed hulls) on hickory chick thallus growing way carry out
Observation, and the activity of its cellulase, hemicellulase, laccase, manganese peroxidase, lignin peroxidase is carried out
Measurement.It was found that the activity height and the growing way quality of Morciiella Esculeuta Mycelia of laccase, manganese peroxidase, lignin peroxidase have
Biggish positive correlation.The wherein stalk of Morciiella Esculeuta Mycelia growing way performance preferably rape, wheat, rice, corn, but arrive
Business strain substrate formula popular in the market is not significantly better than until this step yet.
Carry out simultaneously research and analyse it has also been found that, the amylolytic enzyme of hickory chick has 2 kinds: alpha-amylase and gamma amylase.
Alpha-amylase can decompose starch and generate maltose, and maltose is further decomposed into alpha-D-glucose by alpha-glucosidase.γ-shallow lake
Powder enzymically decomposed starch directly generates β-D-Glucose, needs not move through maltose intermediate product.β-D-Glucose is easy by glucose
Oxydasis generates gluconic acid and hydrogen peroxide, and alpha-D-glucose is not easy by glucose oxidase.Hickory chick
Gamma amylase activity is much higher than alpha-amylase, and alpha-glucosidase activity is weaker, cause amylolytic most of product with
β-D-Glucose form exists, and is easy to form hydrogen peroxide by glucose oxidase.Therefore, it is suitably added in formula
Starch materials can increase the supply of glucose and hydrogen peroxide in katabolism approach.
Hydrogen peroxide be laccase, manganese peroxidase, lignin peroxidase catalysis react necessary to electronics by
Body.The increase of hydrogen peroxide facilitates the oxidation of laccase, manganese peroxidase, lignin peroxidase to lignin in stalk
It decomposes, but lignin decomposes there are three types of the small molecule products generated: vanillic aldehyde, syringaldehyde, parahydroxyben-zaldehyde.Different stalks
In lignin degradation products in three kinds of monomers portfolio ratio it is different, may be to cellulase, hemicellulase, laccase, manganese
There is feedback inhibition (downward) or feedback to activate (up-regulation) for one or more of peroxidase, lignin peroxidase
Effect.
By further experiment, wheat berry, mass ratio model is added in discovery in wheat, rice, corn, rape stalk
It encloses for stalk: wheat berry=(60~75): (25~40);Then it is inoculated with hickory chick.Made with corresponding pure stalk inoculation hickory chick
For comparison.The result shows that: wheat berry, which is added, can make to be adjusted to 2.2 times, lignin peroxidase in the laccase activity of wheat stalk
6.5 times are adjusted in enzymatic activity, but manganese peroxidase enzymatic activity is declined slightly;Make to be adjusted in the laccase activity of rice straw 6.8 times,
Manganese peroxidase enzymatic activity is raised to 2.2 times, but lignin peroxidase activity drops to 50%;Make the manganese mistake of corn stover
It is adjusted to 6 times on peroxidase activity, is adjusted to 4 times on lignin peroxidase activity, but laccase activity drops to 47%;Only
Three kinds of enzymatic activitys of rape stalk all raise, and 4.8 times are wherein adjusted in laccase activity, manganese peroxidase enzymatic activity raises to 6
Again, 3.3 times are adjusted on lignin peroxidase activity.Meanwhile be added the rape stalk of wheat berry than be added wheat berry its
The speed of growth of Morciiella Esculeuta Mycelia is faster on his three kinds of stalks, mycelia is more healthy and stronger, sclerotium is more.
Therefore, rape stalk and wheat berry are according to (60~75): the mass ratio of (25~40) collocation, can make laccase,
The activity of key enzyme needed for these three lignin degradations of manganese peroxidase, lignin peroxidase is comprehensively activated
(up-regulation) promotes the decomposition of lignin, is conducive to the growth of mycelia and the formation of sclerotium.And the collocation of other stalks and wheat berry
Can only activate wherein 2 kinds of enzymes but inhibit remaining a kind of enzyme, effect is not so good as the combination of rape stalk and wheat berry.
The present invention provides a kind of strain substrates that promotion sclerotium of morchella esculenta is formed, and the strain substrate is by rape stalk and small
Wheat composition, by mass percentage, wherein rape stalk fragment is 60-75%, wheat berry 25-40%, and two kinds of substances
The sum of mass percent is 100%, and rape stalk fragment and wheat berry are drying regime;Specific rape stalk fragment
Length is 0.5-5cm, width 1-4mm.
The present invention also provides the preparation methods of above-mentioned Nutrition Materials of Spawn For Morchella comprising following embodiment:
Embodiment 1
The wheat berry of 60% rape stalk fragment and 40% is taken by mass percentage, and wherein the length of rape stalk fragment is
0.5-1.5cm, width 1-2mm, be soaked in water rape stalk fragment 3h, and rape stalk fragment is impregnated with, mass concentration is used
Wheat berry is impregnated into 8h for 1% limewash, wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into polypropylene strain bag after uniformly, sterilize 2h at 121 DEG C, obtains strain substrate.
It is inoculated with morchella mother culture on the strain substrate, obtains hickory chick original seed.
Embodiment 2
The wheat berry of 75% rape stalk fragment and 25% is taken by mass percentage, and wherein the length of rape stalk fragment is
0.5-1.5cm, width 1-2mm, be soaked in water rape stalk fragment 4h, and rape stalk fragment is impregnated with, mass concentration is used
Wheat berry is impregnated into 9h for 1.1% limewash, wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into plastics seed bottle after closing uniformly, is sterilized at 121 DEG C 2.2h using high-pressure steam sterilizing pan, obtain strain substrate.
It is inoculated with morchella mother culture on the strain substrate, obtains hickory chick original seed.
Embodiment 3
The wheat berry of 70% rape stalk fragment and 30% is taken by mass percentage, and wherein the length of rape stalk fragment is
0.1-1.5cm, width 2-3mm, be soaked in water rape stalk fragment 6h, and rape stalk fragment is impregnated with, mass concentration is used
Wheat berry is impregnated into 12h for 1.5% limewash, wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that will be impregnated with
It is fitted into glass seed bottle after mixing, sterilize 2.5h at 121 DEG C, obtains strain substrate.
It is inoculated with hickory chick original seed on the strain substrate, obtains hickory chick cultivar.
Embodiment 4
The wheat berry of 65% rape stalk fragment and 35% is taken by mass percentage, and wherein the length of rape stalk fragment is
4-5cm, width 2-3mm, be soaked in water rape stalk fragment 5h, and rape stalk fragment is impregnated with, is using mass concentration
Wheat berry is impregnated 11h by 1.4% limewash, and wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into glass seed bottle after closing uniformly, sterilize 2.4h at 121 DEG C, obtains strain substrate.
It is inoculated with hickory chick original seed on the strain substrate, obtains hickory chick cultivar.
Embodiment 5
The wheat berry of 69% rape stalk fragment and 31% is taken by mass percentage, and wherein the length of rape stalk fragment is
2-3cm, width 2.5-3.5mm, be soaked in water rape stalk fragment 4.5h, and rape stalk fragment is impregnated with, dense using quality
Wheat berry is impregnated 10h for 1.2% limewash by degree, and wheat berry is impregnated with;Then the rape stalk fragment and wheat that will be impregnated with
Grain is fitted into after mixing in glass seed bottle, and sterilize 2.2h at 121 DEG C, obtains strain substrate.
It is inoculated with hickory chick original seed on the strain substrate, obtains hickory chick cultivar.
Embodiment 6
The wheat berry of 66% rape stalk fragment and 34% is taken by mass percentage, and wherein the length of rape stalk fragment is
1-3cm, width 1-3mm, be soaked in water rape stalk fragment 3.5h, and rape stalk fragment is impregnated with, is using mass concentration
Wheat berry is impregnated 14h by 1% limewash, and wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into polypropylene strain bag after uniformly, sterilize 2h at 121 DEG C, obtains strain substrate.
It is inoculated with hickory chick original seed on the strain substrate, obtains hickory chick cultivar.
Embodiment 7
The wheat berry of 63% rape stalk fragment and 37% is taken by mass percentage, and wherein the length of rape stalk fragment is
1-2cm, width 2-3mm, be soaked in water rape stalk fragment 4h, and rape stalk fragment is impregnated with, is using mass concentration
Wheat berry is impregnated 8.5h by 1.5% limewash, and wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into plastics seed bottle after closing uniformly, sterilize 2.5h at 121 DEG C, obtains strain substrate.
It is inoculated with morchella mother culture on the strain substrate, obtains hickory chick original seed.
Embodiment 8
The wheat berry of 73% rape stalk fragment and 27% is taken by weight percent, and wherein the length of rape stalk fragment is
1-2cm, width 2-3mm, be soaked in water rape stalk fragment 5.5h, and rape stalk fragment is impregnated with, is using mass concentration
Wheat berry is impregnated 12h by 1.5% limewash, and wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into plastics seed bottle after closing uniformly, sterilize 2.3h at 121 DEG C, obtains strain substrate.
It is inoculated with morchella mother culture on the strain substrate, obtains hickory chick original seed.
Embodiment 9
The wheat berry of 72% rape stalk fragment and 28% is taken by weight percent, and wherein the length of rape stalk fragment is
3.5-5cm, width 3-4mm, be soaked in water rape stalk fragment 4h, and rape stalk fragment is impregnated with, is using mass concentration
Wheat berry is impregnated 8h by 1.2% limewash, and wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into plastics seed bottle after uniformly, sterilize 2h at 121 DEG C, obtains strain substrate.
It is inoculated with morchella mother culture on the strain substrate, obtains hickory chick original seed.
Embodiment 10
The wheat berry of 73% rape stalk fragment and 27% is taken by weight percent, and wherein the length of rape stalk fragment is
3-4.5cm, width 3-4mm, be soaked in water rape stalk fragment 3h, and rape stalk fragment is impregnated with, is using mass concentration
Wheat berry is impregnated 8h by 1.2% limewash, and wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into plastics seed bottle after uniformly, sterilize 2h at 121 DEG C, obtains strain substrate.
It is inoculated with hickory chick original seed on the strain substrate, obtains hickory chick cultivar.
Embodiment 11
The wheat berry of 69% rape stalk fragment and 31% is taken by weight percent, and wherein the length of rape stalk fragment is
3-4.5cm, width 1-2mm, be soaked in water rape stalk fragment 5h, and rape stalk fragment is impregnated with, is using mass concentration
Wheat berry is impregnated 9h by 1.3% limewash, and wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into plastics seed bottle after uniformly, sterilize 2h at 121 DEG C, obtains strain substrate.
It is inoculated with hickory chick original seed on the strain substrate, obtains hickory chick cultivar.
Embodiment 12
The wheat berry of 72% rape stalk fragment and 28% is taken by weight percent, and wherein the length of rape stalk fragment is
2-3cm, width 1-2mm, be soaked in water rape stalk fragment 4h, and rape stalk fragment is impregnated with, is using mass concentration
Wheat berry is impregnated 10h by 1.3% limewash, and wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into glass seed bottle after closing uniformly, sterilize 2.5h at 121 DEG C, obtains strain substrate.
It is inoculated with morchella mother culture on the strain substrate, obtains hickory chick original seed.
Embodiment 13
The wheat berry of 68% rape stalk fragment and 32% is taken by weight percent, and wherein the length of rape stalk fragment is
2-3cm, width 1-2mm, be soaked in water rape stalk fragment 4h, and rape stalk fragment is impregnated with, is using mass concentration
Wheat berry is impregnated 14h by 1.3% limewash, and wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into glass seed bottle after closing uniformly, sterilize 2.5h at 121 DEG C, obtains strain substrate.
It is inoculated with morchella mother culture on the strain substrate, obtains hickory chick original seed.
Embodiment 14
The wheat berry of 67% rape stalk fragment and 33% is taken by weight percent, and wherein the length of rape stalk fragment is
3-4.5cm, width 3-4mm, be soaked in water rape stalk fragment 3h, and rape stalk fragment is impregnated with, is using mass concentration
Wheat berry is impregnated 8h by 1.2% limewash, and wheat berry is impregnated with;Then the rape stalk fragment and wheat berry that are impregnated with are mixed
It is fitted into plastics seed bottle after uniformly, sterilize 2h at 121 DEG C, obtains strain substrate.
It is inoculated with hickory chick original seed on the strain substrate, obtains hickory chick cultivar.
Embodiment 15
The wheat berry of 68% rape stalk fragment and 32% is taken by weight percent, and wherein the length of rape stalk fragment is
2-3cm, width 2.5-3.5mm, be soaked in water rape stalk fragment 4.5h, and rape stalk fragment is impregnated with, dense using quality
Wheat berry is impregnated 10h for 1.2% limewash by degree, and wheat berry is impregnated with;Then the rape stalk fragment and wheat that will be impregnated with
Grain is fitted into after mixing in glass seed bottle, and sterilize 2.2h at 121 DEG C, obtains strain substrate.
It is inoculated with hickory chick original seed on the strain substrate, obtains hickory chick cultivar.
The invention also discloses the bacterium that the preparation method for the strain substrate for using above-mentioned promotion sclerotium of morchella esculenta to be formed obtains
Kind matrix carries out inoculation morchella mother culture or hickory chick original seed, wherein hickory chick original seed is obtained when being inoculated with morchella mother culture, when
Hickory chick cultivar is obtained when being inoculated with hickory chick original seed.