CN104531542A - Rapid grifola frondosa stock culture germination culture medium and preparation method thereof - Google Patents
Rapid grifola frondosa stock culture germination culture medium and preparation method thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
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- C05G5/00—Fertilisers characterised by their form
- C05G5/40—Fertilisers incorporated into a matrix
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Abstract
The invention provides a rapid grifola frondosa stock culture germination culture medium which is prepared from grifola frondosa entity, water, oak leaves and a coagulator. The invention also provides a preparation method of the medium. The method comprises the following steps: (1) mixing the grifola frondosa entity and the water, regulating the pH value to be 6.0-8.0, filtering after water bath, and collecting filtrate, thereby obtaining a matrix solution; (2) grinding the oak leaves, thereby obtaining auxiliary materials; (3) mixing the auxiliary materials and the matrix solution according to a mass volume ratio of (1 to 20) to (1 to 50), thereby obtaining a mixture; and (4) adding the coagulator, mixing, and regulating the pH value to be 6, thereby obtaining the medium. The invention provides the rapid grifola frondosa stock culture germination culture medium prepared by adopting the preparation method. The culture medium is manufactured by natural raw materials by virtue of proper treatment and is specially used for germination of a grifola frondosa stock culture. Experiments prove that the culture medium contributes to growth of the grifola frondosa stock culture and also contributes to maintaining the activity of grifola frondosa mycelium, and the condition that grifola frondosa grows in a chemical reagent culture medium for a long time so as to cause degeneration is avoided.
Description
Technical field
The present invention relates to field of edible fungus culture, be specifically related to a kind of Grifola frondosa mother plant fast-germination substratum and Grifola frondosa mother plant the making method of fast-germination substratum.
Background technology
Grifola frondosa, also claims dance fine and soft, is a kind of nutritious, with rich flavor and have the edible mushrooms of health value.A polysaccharide fraction " Fraction-D " in Grifola frondosa is proved has the multiple biological activitys such as antitumor, immunity moderation, thus enjoys academia to pay close attention to." Grifola frondosa capsule " (Mai Te disappears) is exactly a kind of is the medicine of main component with " Fraction-D " polysaccharide.Based on the tempting industry development prospect of Grifola frondosa, China manyly From government to private aspect all made corresponding deployment, take the lead and carried out variety certification to excellent Grifola frondosa kind in the such as Agriculture of Zhejiang Province Room in 2013; Within 2014, Fujian Province's Youxi has set up the first industrial cultivation of grifola frondosa base of China.
The making of Grifola frondosa mother culture media is the basic link during Grifola frondosa is produced, and directly affects Grifola frondosa mother and plants growth quality, also have impact on to cascade the economical character such as the output of each stage mycelium in downstream and sporophore, long speed.
In existing cultivation and production technology, the making of Grifola frondosa mother culture media generally adopts comprehensive potato culture or the commercially available conventional nutrients substratum containing the reagent such as peptone, yeast extract.Although these substratum can meet the growth that Grifola frondosa mother plants substantially, Grifola frondosa mother plants that on these substratum, grow long speed comparatively slow, and long-term subculture uses, can make maitake mushroom mycelia get used to being easy to get nutrition environment and produce degeneration gradually.
Summary of the invention
For above deficiency, the invention provides a kind of substratum being conducive to Grifola frondosa mother kind fast-germination.
The present invention is achieved the above object by following scheme:
A kind of Grifola frondosa mother plants fast-germination substratum, is prepared by Grifola Frondosa sporophore, water, oak leaf, peptizer and is obtained.
Preferably, a kind of Grifola frondosa mother plants fast-germination substratum, and described medium pH is 6, comprise matrix solution, auxiliary material and peptizer, described matrix solution is prepared by Grifola Frondosa sporophore and water and is obtained, and described auxiliary material is pulverized by oak leaf and obtains, and peptizer is preferably agar powder.
Preferred further, a kind of Grifola frondosa mother plants fast-germination substratum, described medium pH is 6, comprise matrix solution, auxiliary material and peptizer, described matrix solution is mixed by Grifola Frondosa sporophore and water and filters after water-bath and obtain, described auxiliary material is pulverized by oak leaf and obtains, and add peptizer after described matrix solution mixes with auxiliary material again and prepare substratum, peptizer is preferably agar powder.
The present invention also provides a kind of and makes the method that above-mentioned Grifola frondosa mother plants fast-germination substratum, comprises the following steps:
(1) prepare matrix solution: get Grifola Frondosa sporophore and water, mixing, preferably mixes according to the mass volume ratio of 1:10, regulates pH between 6.0-8.0, is preferably 6.0 or 6.8 or 8.0, filters after water-bath, collects filtrate and obtains matrix solution;
(2) auxiliary material is prepared: get oak leaf and pulverize, as auxiliary material;
(3) mix: auxiliary material and matrix solution are mixed to get mixture according to the ratio of mass volume ratio 1:20-1:50, be preferably 1:50, more preferably 1:20;
(4) add peptizer and regulate pH: adding peptizer, preferred agar powder, and after mixing, regulating pH to 6, obtain substratum.
In a preferred embodiment, above-mentioned water-bath adopts 80 DEG C of water-bath 2h.
Preferably, make in the method for above-mentioned Grifola frondosa mother kind fast-germination substratum a kind of, the adjustment of pH optionally adopts 1N hydrochloric acid soln or 1N sodium hydroxide solution to carry out pH regulator, such as in the preparation matrix solution of step (1), Grifola Frondosa sporophore and water are 6.8 according to the mixed pH value of the mass volume ratio of 1:10,1N hydrochloric acid soln can be adopted to regulate pH to be 6.0,1N sodium hydroxide solution also can be adopted to regulate pH to be 8.0.
In a preferred embodiment, during above-mentioned preparation auxiliary material, get oak leaf and after pulverizing, sieve, such as, crossing 60 mesh sieves, as auxiliary material.
In a preferred embodiment, the amount of above-mentioned interpolation agar powder is added according to the ratio of agar powder and mixture quality volume ratio 1.5:100.
Preferably, above-mentioned Grifola Frondosa sporophore is dry Grifola Frondosa sporophore, and above-mentioned oak leaf is dry Oak Tree fallen leaves.
Preferably, the water of above-mentioned preparation matrix solution is two pure water, and described two pure water are ultrapure water.
After the making method of planting fast-germination substratum by above-mentioned Grifola frondosa mother makes and obtains substratum, will plate is down flat can use after medium sterilization, sterilising conditions optionally adopts 121 DEG C, sterilizing 20min under 0.1MPa normal atmosphere.
In sum, the invention provides a kind of Grifola frondosa mother made as above-mentioned making method and plant fast-germination substratum.This substratum is made through suitably processing with natural matter, be exclusively used in the sprouting that Grifola frondosa mother plants, experiment confirms, this substratum is conducive to the growth that Grifola frondosa mother plants, also be conducive to keeping maitake mushroom mycelia vigor, be unlikely to long term growth and cause degenerating in chemical reagent substratum.This substratum source is natural, and composition is simple, remarkably productive, is worth of widely use.
Accompanying drawing explanation
Fig. 1 is that in each substratum of effect example 1, mycelium sprouts situation.
Fig. 2 is mycelium bacterium colony outward appearance in each substratum of effect example 1.
Fig. 3 is that in each substratum of effect example 2, mycelium sprouts situation.
Fig. 4 is mycelium bacterium colony outward appearance in each substratum of effect example 2.
Embodiment
Below in conjunction with specific embodiment, the present invention is further described.
Embodiment 1
A kind of method making above-mentioned Grifola frondosa mother kind fast-germination substratum:
(1) prepare matrix solution: get Grifola Frondosa sporophore and water, mixing, regulate pH between 6.0-8.0, filter after water-bath, collect filtrate and obtain matrix solution;
(2) auxiliary material is prepared: get oak leaf and pulverize, as auxiliary material;
(3) mix: auxiliary material and matrix solution are mixed to get mixture according to the ratio of mass volume ratio 1:20-1:50;
(4) add peptizer and regulate pH: adding the peptizer that substratum is conventional, and after mixing, regulating pH to 6, obtain substratum.
Embodiment 2:
A kind of method making above-mentioned Grifola frondosa mother kind fast-germination substratum:
(1) prepare matrix solution: get dry Grifola Frondosa sporophore and two pure water, mix according to the mass volume ratio of 1:10, adopt 1N hydrochloric acid soln to regulate pH to be 6.0,80 DEG C of water-bath 2h, filter, collect filtrate and obtain matrix solution;
(2) prepare auxiliary material: get dry oak leaf and pulverize, cross 60 mesh sieves, as auxiliary material;
(3) mix: auxiliary material and matrix solution are mixed to get mixture according to the ratio of mass volume ratio 1:50;
(4) add peptizer agar powder and regulate pH: adding agar powder according to the ratio of agar powder and mixture quality volume ratio 1.5:100, and after mixing, adopting 1N hydrochloric acid soln to regulate pH to 6, obtain substratum.
Embodiment 3
A kind of method making above-mentioned Grifola frondosa mother kind fast-germination substratum:
(1) prepare matrix solution: get dry Grifola Frondosa sporophore and two pure water, mix according to the mass volume ratio of 1:10, measuring pH is 6.8, does not need to regulate, 80 DEG C of water-bath 2h, filters, and collects filtrate and obtains matrix solution;
(2) prepare auxiliary material: get dry oak leaf and pulverize, cross 60 mesh sieves, as auxiliary material;
(3) mix: auxiliary material and matrix solution are mixed to get mixture according to the ratio of mass volume ratio 1:50;
(4) add peptizer agar powder and regulate pH: adding agar powder according to the ratio of agar powder and mixture quality volume ratio 1.5:100, and after mixing, adopting 1N hydrochloric acid soln to regulate pH to 6, obtain substratum.
Embodiment 4
A kind of method making above-mentioned Grifola frondosa mother kind fast-germination substratum:
(1) prepare matrix solution: get dry Grifola Frondosa sporophore and two pure water, mix according to the mass volume ratio of 1:10, adopt 1N sodium hydroxide solution to regulate pH to be 8.0,80 DEG C of water-bath 2h, filter, collect filtrate and obtain matrix solution;
(2) prepare auxiliary material: get dry oak leaf and pulverize, cross 60 mesh sieves, as auxiliary material;
(3) mix: auxiliary material and matrix solution are mixed to get mixture according to the ratio of mass volume ratio 1:50;
(4) add peptizer agar powder and regulate pH: adding agar powder according to the ratio of agar powder and mixture quality volume ratio 1.5:100, and after mixing, adopting 1N hydrochloric acid soln to regulate pH to 6, obtain substratum.
Embodiment 5
A kind of method making above-mentioned Grifola frondosa mother kind fast-germination substratum:
(1) prepare matrix solution: get dry Grifola Frondosa sporophore and two pure water, mix according to the mass volume ratio of 1:10, adopt 1N hydrochloric acid soln to regulate pH to be 6.0,80 DEG C of water-bath 2h, filter, collect filtrate and obtain matrix solution;
(2) prepare auxiliary material: get dry oak leaf and pulverize, cross 60 mesh sieves, as auxiliary material;
(3) mix: auxiliary material and matrix solution are mixed to get mixture according to the ratio of mass volume ratio 1:20;
(4) add peptizer agar powder and regulate pH: adding agar powder according to the ratio of agar powder and mixture quality volume ratio 1.5:100, and after mixing, adopting 1N hydrochloric acid soln to regulate pH to 6, obtain substratum.
Embodiment 6
A kind of method making above-mentioned Grifola frondosa mother kind fast-germination substratum:
(1) prepare matrix solution: get dry Grifola Frondosa sporophore and two pure water, mix according to the mass volume ratio of 1:10, measuring pH is 6.8, does not need to regulate, 80 DEG C of water-bath 2h, filters, and collects filtrate and obtains matrix solution;
(2) prepare auxiliary material: get dry oak leaf and pulverize, cross 60 mesh sieves, as auxiliary material;
(3) mix: auxiliary material and matrix solution are mixed to get mixture according to the ratio of mass volume ratio 1:20;
(4) add peptizer agar powder and regulate pH: adding agar powder according to the ratio of agar powder and mixture quality volume ratio 1.5:100, and after mixing, adopting 1N hydrochloric acid soln to regulate pH to 6, obtain substratum.
Embodiment 7
A kind of method making above-mentioned Grifola frondosa mother kind fast-germination substratum:
(1) prepare matrix solution: get dry Grifola Frondosa sporophore and two pure water, mix according to the mass volume ratio of 1:10, adopt 1N sodium hydroxide solution to regulate pH to be 8.0,80 DEG C of water-bath 2h, filter, collect filtrate and obtain matrix solution;
(2) prepare auxiliary material: get dry oak leaf and pulverize, cross 60 mesh sieves, as auxiliary material;
(3) mix: auxiliary material and matrix solution are mixed to get mixture according to the ratio of mass volume ratio 1:20;
(4) add peptizer agar powder and regulate pH: adding agar powder according to the ratio of agar powder and mixture quality volume ratio 1.5:100, and after mixing, adopting 1N hydrochloric acid soln to regulate pH to 6, obtain substratum.
Effect example 1
The Grifola frondosa mother that the present embodiment adopts plants as Grifola frondosa kind Gf-3 (purchased from Fujian Province); Adopt the substratum of embodiment 2-7 as test group, in Fig. 1-Fig. 2 and table 1, respectively be 1-6, adopt commercial fungal substratum as a control group (CK), be CK in Fig. 1-Fig. 2 and table 1; The formula of commercial fungal substratum is: peptone 5g/L, yeast extract powder 2g/L, glucose 20g/L, dipotassium hydrogen phosphate 1g/L, magnesium sulfate 0.5g/L.
1, get commercial fungal substratum and illustratively prepare, regulating pH to be 6.0, and sterilizing, this substratum is (CK) in contrast.
2, aseptically, get the substratum that Grifola frondosa kind Gf-3 bacterial classification block is inoculated in control medium and embodiment 2-7 respectively, inoculation piece size is consistent.
3, in 25 ± 1 DEG C of incubators, lucifuge is cultivated.
4, observe inoculation ferfas filament and sprout situation and mycelial growth situation etc.; Wherein, respectively as depicted in figs. 1 and 2, Mycelium growth rate is in table 1 for mycelium sprouting situation and mycelium bacterium colony outward appearance.
5, result and analysis:
(1) mycelium germination situation
In 25 ± 1 DEG C of incubators, lucifuge is cultivated after 48h, each formula inoculation of medium Grifola frondosa Gf-3 mycelial sprouting situation as shown in Figure 1, each media samples two repetition.
As can be seen from Figure 1, compared with commercial fungal substratum (CK), the substratum of embodiment 2-7 (1-6 in Fig. 1) all makes inoculation block accelerate Germination Strain filament.
(2) Mycelium growth rate
In culturing process, by every day, line record is carried out to Grifola frondosa Gf-3 mycelium front end, measure the per day growth distance of mycelia, be the speed of growth of mycelium in each formula substratum, the speed of growth the results are shown in Table 1, each media samples three repetition.
As seen from Table 1, in the substratum of embodiment 2-4 (1-3, the i.e. substratum of the formula of three pH value matrix solutions of 5% auxiliary material amount interpolation), mycelium on average long speed is greater than commercial fungal culture medium C K, and significant difference (LSD, α=0.05); And in the substratum (4-6, the i.e. substratum of the formula of three pH value matrix solutions of 2% auxiliary material amount interpolation) of embodiment 5-7, mycelium average growth rate and commercial fungal culture medium C K no significant difference.
Mycelium growth rate result in each substratum of table 1
(3) mycelium bacterium colony outward appearance
In 25 ± 1 DEG C of incubators, lucifuge is cultivated after 10d, the mycelium bacterium colony outward appearance of each substratum as shown in Figure 2, each media samples two repetition.
As can be seen from Fig. 2, compared with commercial fungal substratum, Gf-3 colonial morphology in six kinds of substratum is undistorted.
Effect example 2
The Grifola frondosa mother that the present embodiment adopts plants as Grifola frondosa kind CZ (purchased from Zhejiang Province); Adopt the substratum of embodiment 2-7 as test group, in Fig. 3-Fig. 4 and table 2, respectively be 1-6, adopt commercial fungal substratum as a control group (CK), be CK in Fig. 3-Fig. 4 and table 2; The formula of commercial fungal substratum is: peptone 5g/L, yeast extract powder 2g/L, glucose 20g/L, dipotassium hydrogen phosphate 1g/L, magnesium sulfate 0.5g/L.
1, get commercial fungal substratum and illustratively prepare, regulating pH to be 6.0, and sterilizing, this substratum is (CK) in contrast.
2, aseptically, get the substratum that Grifola frondosa kind CZ bacterial classification block is inoculated in control medium and embodiment 2-7 respectively, inoculation piece size is consistent.
3, in 25 ± 1 DEG C of incubators, lucifuge is cultivated.
4, observe inoculation ferfas filament and sprout situation and mycelial growth situation etc.; Wherein, respectively as shown in Figure 3 and Figure 4, Mycelium growth rate is in table 2 for mycelium sprouting situation and mycelium bacterium colony outward appearance.
5, result and analysis:
(1) mycelium germination situation
In 25 ± 1 DEG C of incubators, lucifuge is cultivated after 48h, each formula inoculation of medium Grifola frondosa CZ mycelial sprouting situation as shown in Figure 3, each media samples two repetition.
As can be seen from Figure 3, compared with commercial fungal substratum (CK), the substratum of embodiment 2-7 (1-6 in Fig. 3) all makes inoculation block accelerate Germination Strain filament.
(2) Mycelium growth rate
In culturing process, carry out line record by every day to Grifola frondosa CZ mycelium front end, measure the per day growth distance of mycelia, be the speed of growth of mycelium in each formula substratum, the speed of growth the results are shown in Table 2, each media samples three repetition.
As seen from Table 2, in the substratum of embodiment 2-4 (1-3, the i.e. substratum of the formula of three pH value matrix solutions of 5% auxiliary material amount interpolation), mycelium on average long speed is greater than commercial fungal culture medium C K, and significant difference (LSD, α=0.05); And in the substratum of embodiment 5-7 (4-6, the i.e. substratum of the formula of three pH value matrix solutions of 2% auxiliary material amount interpolation), mycelium average growth rate and commercial fungal culture medium C K no significant difference.
Mycelium growth rate result in each substratum of table 2
| Numbering | 1 | 2 | 3 | CK | 4 | 5 | 6 |
| Long speed | 3.6±0.3 | 3.8±0.2 | 3.8±0.2 | 3.3±0.2 | 4.3±0.2* | 4.5±0.3* | 4.0±0.2* |
| (mm/d) |
(3) mycelium bacterium colony outward appearance
After in 25 ± 1 DEG C of incubators, lucifuge cultivates 10d, the mycelium bacterium colony outward appearance of each substratum as shown in Figure 4.Each media samples two repetition.
As can be seen from Fig. 4, compared with commercial fungal substratum, CZ colonial morphology in six kinds of substratum is undistorted.
From above result, the substratum of above embodiment is applicable to the growth of maitake mushroom mycelia all very much, the inoculation block can accelerating Grifola frondosa bacterial classification sprouts mycelia, special when the addition of auxiliary material is 5% (ratio of the mass volume ratio 1:20 of auxiliary material and matrix solution), be particularly conducive to the mycelial growth rate accelerating Grifola frondosa bacterial classification.
The above; be only preferably specific embodiment of the present invention; but protection scope of the present invention is not limited thereto; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses; be equal to according to technical scheme of the present invention and design thereof and replace or change, all should be encompassed in protection scope of the present invention.
Claims (10)
1. Grifola frondosa mother plants a fast-germination substratum, it is characterized in that, comprises Grifola Frondosa sporophore, water, oak leaf, peptizer preparation and obtaining.
2. a kind of Grifola frondosa mother as claimed in claim 1 plants fast-germination substratum, it is characterized in that, described medium pH is 6, comprises matrix solution, auxiliary material and peptizer, described matrix solution is prepared by Grifola Frondosa sporophore and water and is obtained, and described auxiliary material is pulverized by oak leaf and obtains.
3. the Grifola frondosa mother as described in claim 1-2 plants a making method for fast-germination substratum, it is characterized in that, comprises the following steps:
(1) prepare matrix solution: get Grifola Frondosa sporophore and water, mixing, regulate pH between 6.0-8.0, filter after water-bath, collect filtrate and obtain matrix solution;
(2) auxiliary material is prepared: get oak leaf and pulverize, as auxiliary material;
(3) mix: auxiliary material and matrix solution are mixed to get mixture according to the ratio of mass volume ratio 1:20-1:50;
(4) add peptizer and regulate pH: add peptizer and after mixing, regulate pH to 6, obtain substratum.
4. a kind of Grifola frondosa mother as claimed in claim 3 plants the making method of fast-germination substratum, and it is characterized in that, the pH regulator of described step 1 is 6.0, or 6.8, or 8.0.
5. a kind of Grifola frondosa mother as claimed in claim 3 plants the making method of fast-germination substratum, and it is characterized in that, described Grifola Frondosa sporophore and water mix according to the mass volume ratio of 1:10.
6. a kind of Grifola frondosa mother as claimed in claim 3 plants the making method of fast-germination substratum, and it is characterized in that, described water-bath is 80 DEG C of water-bath 2h.
7. a kind of Grifola frondosa mother as claimed in claim 3 plants the making method of fast-germination substratum, and it is characterized in that, described adjustment pH adopts 1N hydrochloric acid soln or 1N sodium hydroxide solution to carry out pH regulator.
8. a kind of Grifola frondosa mother as claimed in claim 3 plants the making method of fast-germination substratum, and it is characterized in that, described peptizer is agar powder.
9. a kind of Grifola frondosa mother as claimed in claim 8 plants the making method of fast-germination substratum, and it is characterized in that, the blending ratio of described agar powder and mixture is mass volume ratio 1.5:100.
10. the Grifola frondosa mother of the making method making as described in claim arbitrary in claim 3-9 plants fast-germination substratum.
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