CN101974436A - Lignocellulose degrading bacteria and application thereof - Google Patents
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Abstract
The invention discloses lignocellulose degrading bacteria and application thereof. The lignocellulose degrading bacteria is expanded penicillium W4 with the preservation number of CGMCC (China General Microbiological Culture Collection) No.4077. The stain can generate cellulose and laccase and degrade straw lignocellulose, can be used as a main effective component of microbial fungicides of a mushroom culture medium and has the effects of shortening the biological transformation time of the mushroom culture medium and improving the quality of the mushroom culture medium. Particularly for the biological transformation of a small quantity of packed mushroom culture media of a family workshop, the lignocellulose degrading bacteria has obvious advantage in environment with lower temperature.
Description
Technical field
The present invention relates to the microbiobacterial agent field, specifically, relate to a kind of ligocellulose degradation's microbiobacterial agent of expanding mould that contains.
Background technology
China is a large agricultural country, according to rough Statistics, annual about 800,000,000 tons of the agricultural crop straw that produces, but utilization ratio is very low, both caused the waste of Biological resources, also environment is caused severe contamination, therefore how making full use of these Biological resources has become one of key issue of restriction China agricultural sustainable development.
Twospore Mushroom (Agaricus bisporus) or title white mushroom, its culture material generally are to be formed by agricultural crop straw, feces of livestock and poultry and a small amount of chemical fertilizer mixed preparing.Agricultural crop straw if can be fully used in mushroom produces, will improve the effective rate of utilization of Biological resources greatly, and slow down the pressure of agricultural wastes environment.At present China's bisporous mushroom greatly relies on peasant's individual workship to produce, and often is subjected to therefore that envrionment temperature is low excessively, compost fermentation overlong time, puzzlement that compost quality is low.Shorten the compost fermentation time, improve compost quality and become individual workship's bisporous mushroom production problem demanding prompt solution.
A large amount of studies show that, is used for the culture material of mushroom-cultivating, is (Burton, 1997) that form through the biotransformation that the micropopulation of a complexity is participated in directly.The mushroom mycelium utilization mainly be after the culture material bio-transformation finishes, a kind of dun material at the crop material surface deposition, this mixture derives from the lignin product after the metabolism, wherein contain the fragment that left behind after a large amount of nitrogen and complete microorganism cells composition and part are decomposed, simultaneously, this mixture is except that mushroom mycelium, and other microorganisms or assorted bacterium be difficult utilization (Adams, 2008) nearly all.Finishing of mushroom compost biotransformation is that the acting in conjunction that depends on microflora realizes, the researchist is to appearance, the rule of development of bacterium, actinomycetes, fungi three major types type flora in the stage culture material of banking up, effect characteristics separately and the reciprocity between them, antagonism have all carried out a large amount of research (ginger one-tenth, 2003; Ashraf, 2007).The carbon source that mushroom growth utilized fundamentally is the degraded product from the stalk lignocellulose.Therefore, in the culture material biotransformation, add external source ligocellulose degradation bacterium and have potential quickening culture material fermentation maturity speed, the effect that improves compost quality.
There are a lot of researchists to carry out the research of in agricultural wastes and city compost process, adding external source ligocellulose degradation bacterium at present, though add external source ligocellulose degradation bacterium whether can accelerate compost fermentation become thoroughly decomposed speed, improve on the compost quality and also have dispute, add the research emphasis that the effect of external source ligocellulose degradation bacterium in compost become the researchist.
Because the Mierocrystalline cellulose of crop material and hemicellulose, xylogen form fine and close reticulated structure, and the crystallization of internal height causes it to be difficult to by microorganism as utilization of carbon source, and therefore screening obtains the key that stalk ligocellulose degradation bacterium efficiently becomes this technology.
The bacterial species of lignocellulose degradation is a lot, actinomycetes are the stronger class filamentous bacteriums of degradation capability of generally acknowledging, comprise streptomycete (Streptomyces), Arthrobacter (Arthrobacter), small single-cell bacteria (Micromonospora), Nocardia bacteria (Li, 1997 such as (Nocardia); Liu Dongbo, 1990).Actinomycetes mainly are to increase the water-soluble of it to the Degradation of xylogen.Because actinomycetes can penetrate insoluble matrix such as lignocellulose, therefore in neutrality, slight alkalinity soil or compost, actinomycetes participate in organic initial degraded and humify.Crawford people such as (1983) studies the streptomycete in the actinomycetes, finds that this bacterium mycelia branch is many and output is big, and the bacterium colony quality is tight on Gauss's substratum, combines with substratum firmly to be difficult for provoking.The elementary metabolism stage this bacterium the lignin degrading ability is arranged, mainly be by the fracture of demethylation, aromatic ring and three kinds of approach lignin degradings of oxide side chain.Suppress other microbial growth though can produce microbiotic in the streptomycete process of growth, be test strain preferably, and this causes it to be difficult to and other wooden rotten microorganism Synergistic degradation xylogen, and it is very limited on industrial application.
The non-filamentous bacterium of lignin degrading also can cause the degraded of xylogen to a certain extent.For example: the anaerobism clostridium (Clostridium) in the bacterium, pseudomonas (Pseudomonas), acinetobacter calcoaceticus (Acinetobacter), genus bacillus (Bacillus), Flavobacterium (Flavobacterium) and Xanthomonas campestris (Xanthomonas).The thermotolerance of the rotten bacterium of wood is higher than wood-rotting fungi, and the foreign scholar separates under 65～82 ℃ of conditions from envrionment temperature and obtains the rotten bacterium of multiple wood (Beffa, 1996).Because wooden rotten bacterium thermotolerance is strong, can adapt to the high temperature in the compost internal medium, has begun to utilize the domestic refuse Degradation and Transformation that these bacteriums will contain xylogen to be fertilizer (Tuomela, 2000 abroad; Yang Xiaohuan, 2007).At present isolating bacterium with cellulose degradation ability is more, wherein belongs to Gram-positive Pseudomonas (G
+) Cytophaga (Cytophaga) arranged, hot rod Pseudomonas (Caldibacillus), bacillus (Bacillus) gives birth to spore and has a liking for fiber bacterium (Sporacytophga) etc.; Belong to Gram-negative Pseudomonas (G
-) Pseudomonas (Pseudomonas), erwinia (Erwinia), fiber monospore Pseudomonas (Cellulomonas), Cellfalcicula (Schwarz, 2001 such as (Cellfacicula) are arranged; Lynd, 2002; Kenyon, 2005).But because the secreted lignocellulose degrading enzyme of these prokaryotic organism all is an intracellular enzyme, this has just determined it to be in an accessory relatively status in the research of lignin-degrading bacteria.
The plain enzyme of born of the same parents' outer fiber of fungi is lived stronger, especially with Penicillium (Penicillium), Trichoderma (Trichderma), Chaetomium (Chaetomium) Aspergillus (Aspergillus) and Rhizopus (Rhizopus), more with the research of Trichoderma (Trichderma), aspergillus niger (Aspergillus niger) wherein, they often are used in the production of industrial enzyme, to improve resource conversion rate (Wood, 1994 of cellulose substances;
1999; Lynd, 2002).According to the type of foxy, the fungi of lignin degrading can be divided into brown rot fungus, whiterot fungi and soft-rot bacterium three classes, and the above two belong to basidiomycetes, and soft-rot bacterium belongs to imperfect fungi.Brown rot fungus is more more responsive than xylogen to polysaccharide (as Mierocrystalline cellulose).It at first the filemot pigment of degraded cellulose justacrine make timber be brown, part lignin degrading slowly just then; Soft-rot bacterium makes decay of wood under the condition of humidity, main degraded cellulose more slowly and not thorough, is gained the name because of making the wood surface deliquescing to the degraded of xylogen; Whiterot fungi is a kind of filamentous fungus, and the ability of lignin degrading is better than the ability of degraded cellulose, this class bacterium at first make the xylogen in the timber take place degraded and not chromogenesis, make the wooden white (Wu Kun, 2000) that still keeps.
Because most ligocellulose degradation fungi can secrete lignin-degrading enzymes, cellulose degrading enzyme and hemicellulose degrading enzyme simultaneously outside born of the same parents in vegetative activity, existing studies show that is applied to fungi in the compost, lignocellulose in the degrading straw effectively, lignocellulose is converted into soil ulmin (Wang Lian Zhen, 2003).(2005) such as yellow lead lotuses add white-rot fungi in the agricultural crop straw compost after, the degradation rate of xylogen reaches 43.86%, degradation rate far above xylogen in the compost of not inoculating microbial inoculum, show inoculation white-rot fungi microbial inoculum in compost, can quicken composting process, make the more thorough of lignin degradation in the compost.Along with the further investigation of scientific research personnel to mushroom production matrix compost, find in compost, to add thermophilic fungus with the plain degradation enzyme system of eccrine fiber, can improve compost quality, and mushroom yield (Ceng Wei, 1996; Sabatini, 2006).
In sum, the ligocellulose degradation bacterium is an effective way that improves mushroom compost bio-transformation speed and Mushroom planting substrate quality.Existing at present about utilizing the ligocellulose degradation bacterium to promote the research of agricultural wastes and city compost, but the ligocellulose degradation bacterium is added in the mushroom compost, and the Study on Quality that is used to improve the Mushroom planting substrate of the bio-transformation speed of culture material and acquisition thereof is not appeared in the newspapers; Have the existing report of screening of bacterial strain of the Penicillium of ligocellulose degradation's ability, but the research with expansion mould bacteria agent of efficient ligocellulose degradation effect does not appear in the newspapers.
Summary of the invention
The purpose of this invention is to provide a kind of ligocellulose degradation bacterium (expansion mould W4) and the application in the microbiobacterial agent of preparation mushroom compost biological treatment thereof.
In order to realize the object of the invention, expansion mould W4 of the present invention, its classification called after Penicillium expansum, now be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, BeiChen West Road, Chaoyang District, BeiJing City, address institute of microbiology of the Chinese Academy of Sciences, deposit number CGMCC NO.4077, preservation date on August 9th, 2010.
Expansion mould W4 strains separation of the present invention is from the septic stalk of Beijing suburb.
The morphological feature of expansion mould W4 of the present invention is: bacterial strain W4 30 ℃ of constant temperature culture 2d on the PDA substratum can see white colony clearly.Bacterium colony is carried out, and is velvet-like, and the top layer gives birth to one deck sap green conidial powder, and the strain growth later stage back side is observed and is brown.Conidiophore is smooth, and the asymmetric 3-6 of branch, to arrange closely, its top gives birth to 5-8 short and small phialide, 8.7-10.6 * 3.0 μ m, conidium is many, ovalize, it is inferior spherical, smooth that part becomes, major diameter 3.1-3.5 μ m.
The cultural method of expansion mould W4 bacterial strain of the present invention is: bacterial strain joined in the PDA liquid nutrient medium, and 30 ℃, 170rmin
-1Shaking culture 3d.
PDA liquid culture based component and consumption (g/L): potato 200 (peeling), glucose 20, prepare with water pH7.2～7.4.
The present invention also provides expansion mould W4 application in lignocellulose degradation.
The microbiobacterial agent of mushroom compost of the present invention comprises the component of following weight part: expansion mould W4 1-2 part; Rice chaff and/or wheat bran 10-15 part.Effective constituent is mainly expansion mould W4.
The application of the microbiobacterial agent of mushroom compost of the present invention in the mushroom compost biotransformation, main by the agricultural crop straw in the degraded culture material, described agricultural crop straw comprises wheat straw and/or straw etc.
The present invention isolates the expansion mould W4 with the effect of efficient ligocellulose degradation first, and deposit number is CGMCC NO.4077.This bacterial strain can produce cellulase and laccase, can the degrading straw lignocellulose, it as the main effective constituent in the microbiobacterial agent of mushroom compost, is had the effect of shortening the mushroom compost bio-transformation time, improving the Mushroom planting substrate quality.Especially the bio-transformation of a small amount of mushroom compost of banking up at the family workshop under the lower environment of temperature, has remarkable advantages.
Description of drawings
Fig. 1 is the influence that incubation time is lived to the plain enzyme of bacterial strain W4 born of the same parents outer fiber.
Fig. 2 is the influence of incubation time to bacterial strain W4 laccase activity.
Fig. 3 is the pester variation characteristic of microflora in the straw culture material of the present invention, and ◇ does not add microbial inoculum, and ■ adds microbial inoculum.
Fig. 4 is the pester variation characteristic of microflora in the wheat straw culture material of the present invention, and ◇ does not add microbial inoculum, and ■ adds microbial inoculum.
Embodiment
Following examples are used to illustrate the present invention, but are not used for limiting the scope of the invention.
The preparation of embodiment mushroom compost microbiobacterial agent
The fermentor cultivation liquid formula is (weight percent): KH
2PO
40.1%, NaCl 0.01%, MgSO
47H
2O 0.03%, NaNO
30.25%, FeCl
30.001%, CaCl
20.01%, straw powder 0.5%, pH is 7.2～7.4, prepares with water.
The W4 inoculation on the PDA substratum, is cultivated 72h for 28-30 ℃, insert the 500mL triangular flask then, at 30 ℃, static cultivation 48h.Be linked in the 15L seeding tank by 1% inoculum size then, pH7.2, under the air flow 0.5vvm, cultivate 48h after, be encased in the fermentor tank of 100L by 5% inoculum size again, pH7.2 under the air flow 0.6vvm, cultivates 5d.
After fermentation was finished, the weight ratio according to 2: 10～15 joined bacterial strain W4 in the rice chaff or wheat bran of the bacterium of having gone out, and aseptic pack is sealed, and normal temperature is preserved.
Experimental example 1 bacterial strain W4 produces the enzyme activity experiment
Liquid fermentation medium (weight percent): KH
2PO
41.0g, NaCl 0.1g, MgSO
47H
2O 0.3g, NaNO
32.5g, FeCl
30.01g, CaCl
20.1g straw powder 20g, pH are 7.2～7.4, prepare with water.
The preparation of Mierocrystalline cellulose crude enzyme liquid: the bacterial classification spore that will cultivate on PDA is transferred in the physiological saline, with the blood counting chamber counting, by 1% (10
8Individual spore amount) inoculum size is produced in the enzyme substratum to 300ml, 28 ℃, the 170rmp liquid fermentation and culture, the cellulosic enzyme activity of every 24h sampling and measuring, cultured liquid fermenting is produced the enzyme substratum use earlier two-layer filtered through gauze, filtrate is again in 4 ℃, and the centrifugal 10min of 5000rmp gets the crude enzyme liquid that supernatant liquor is preparation.
The mensuration that holoenzyme is lived: get the enzyme liquid 0.5ml after the suitable dilution, join in the tool plug scale test tube that contains no starch Xinhua's filter paper that 50mg handled and 1.5ml 0.05M pH5.0 citrate buffer solution, behind the insulation 1h, press the DNS method immediately and measure reducing sugar content in 50 ℃ of water-baths.The mensuration that excision enzyme is lived: will not have starch Xinhua filter paper and change absorbent cotton into, the mensuration that other is lived with holoenzyme.The mensuration of restriction endonuclease vigor: get the enzyme liquid 0.5ml after the suitable dilution, join in the tool plug scale test tube that contains 1.5ml 1%CMC citrate buffer solution, behind insulation 30 min, press DNS method mensuration reducing sugar content immediately in 50 ℃ of water-baths.The mensuration of beta-glucoside enzyme activity: change the 1%CMC citrate buffer solution into 1% saligenin acetate buffer solution, the mensuration that other is lived with restriction endonuclease.
Above enzyme activity unit (U/ml) is defined as: every ml enzyme liquid makes degradation of substrates generate the required enzyme amount of 1 μ mol glucose under above-mentioned reaction conditions in 1min.The conversion formula of enzyme activity is as follows:
Wherein, 5.56 is the μ mol number of 1mg glucose.
The plain enzyme of born of the same parents' outer fiber of bacterial strain W4 is lived, and comprises filter paper enzyme activity, exoglucanase is lived, endoglucanase is lived and beta-glucosidase is lived measurement result as shown in Figure 1.
The result shows, bacterial strain W4 has the plain enzyme activity of stronger born of the same parents' outer fiber, the enzyme that the plain enzyme work of its born of the same parents' outer fiber continued at 5-8 days to keep higher is lived, enzyme activity reached the highest in the 6th day, exoglucanase 49.75U/ml alive, beta-glucosidase 33.29U/ml alive, endoglucanase 14.25U/ml alive and filter paper enzyme activity 21.27U/ml.
The mensuration of laccase activity: total reaction volume 3ml contains 5mM ABTS 100 μ l, and 50mM sodium tartrate damping fluid (pH3.0) 2.5ml adds enzyme liquid 400 μ l initial actions, and 420nm measures the variation of light absorption value down.Per minute causes that 1 absorbancy increases required enzyme amount and is defined as 1 unit of activity, U/L in every liter of reaction solution.
Lignoenzyme mainly comprises: laccase (Lac), lignin peroxidase (LiP) and manganese peroxidase (MnP).Wherein bacterial strain W4 has stronger laccase activity, and the enzymic activity of LiP and MnP is lower.The laccase activity that Fig. 2 demonstrates bacterial strain W4 is situation over time, and the result shows that bacterial strain W4 reaches the highest at the 8th day laccase activity in the liquid medium within, is 780U/L.
Experimental example 2 bacterial strain W4 are to the degradation experiment of stalk
Stalk fermentation substratum: in 1000ml water, add stalk 20g, and add nutritive salt KH
2PO
41.0g, NaCl 0.1g, MgSO
47H
2O 0.3g, NaNO
32.5g, FeCl
30.01g, CaCl
20.1g regulating pH is 7.2～7.4, with water preparation, 120 ℃ of autoclavings, 20min.
The mensuration of stalk weightlessness: inoculating strain W4 prepares bacterium liquid in liquid seed culture medium, the bacterium liquid that inoculation 1ml prepares is in the stalk fermentation substratum, carrying out isothermal vibration cultivates, after cultivating 10d, use the filter paper filtering fermented liquid, 80 ℃ of oven dry of residue are weighed, calculate the wheat straw rate of weight loss with the loss of weight method.
The mensuration of stalk rate of decomposition: the method for (1987) such as employing Wang Yuwans is carried out the mensuration of Mierocrystalline cellulose, hemicellulose, content of lignin.Cellulosic rate of decomposition is calculated according to following formula:
(control sample content of cellulose * example weight-residual body content of cellulose * residual body weight)/(control sample content of cellulose * example weight) * 100%
The method of calculation of the rate of decomposition of hemicellulose and xylogen are carried out according to cellulosic method of calculation.
The weight of stalk and each components contents are as shown in table 1.
Table 1 bacterial strain W4 degraded each components contents of stalk after 10 days
The result shows that behind the cultivation 5d, the stalk lignocellulose of bacterial strain W4 decomposes to some extent, and the stalk rate of weight loss is 25.6%.After cultivating 10d, the stalk rate of weight loss of bacterial strain W4 has had significant increase, reaches 56.3%.The preceding 5d of straw degradative, the content of hemicellulose obviously reduces in the lignocellulose, and half-and-half cellulosic relative degradation rate is 62.68%, and in the later stage, cellulosic relative content reduces rapidly, its degradation rate reaches 59.06%.Generally speaking, bacterial strain W4 degraded mean rate to the stalk lignocellulose in 10 days is 56.3mg/ days.
Experimental example 3 bacterial strain W4 are to the promotion experiment 1 of stalk culture material compost
Outside temperature improves the influence of mushroom compost biotransformation to microbial inoculum:
Get the raw materials ready: press 100m
2Cultivated area is calculated: wheat straw/straw 2000kg, do cow dung 1500kg, corn cob 50kg, urea 5kg, ammonium sulfate 10kg, lime carbonate 40kg, calcium superphosphate 10kg, terra alba 10kg.Build heap: carry out mid-April, piles preceding 2 days to wheat straw, straw, and cow dung is prewetted, and humidity is: hold cow dung, webs have 4～5 to drip; When wheat straw/straw being twisted, have under the water droplet with hand.Wide 2.0m is piled in the stockpile north-south, long 2.0m, high 1.8m, totally 7 layers.The 1st layer of elder generation spreads the thick wheat straw/straw of 30cm on the ground, and piss is plain then, and regulating C/N is 33, sprinkle 150kg cow dung again, the 2nd～7 layer of wheat straw/straw thickness is 20cm, and the cow dung amount is every layer of 100kg for 2～6 layers, residue is all covered at the 7th layer, and every layer of consumption of urea is with the 1st layer.The turning time is the 7th, 14,19,24,28,31,35 day.In the fermentation reactor system process, regularly the heap body being sprayed water remains between 60%～70% the water content of heap body.Regularly detect the pH value of heap body, be adjusted in about pH7.5 with liming.The difference that test is handled is: the processing of adding microbial inoculum is to add microbial inoculum with the ratio of 10g/kg culture material.Whole culture material bio-transformation test period is 35d.Table 2 is the physico-chemical property of compost initial-stage culture material.
The time of banking up of culture material is the last ten-days period in mid-April to May, and outside air temperature is about 15-28 ℃.Culture material 2 begin the time of banking up be mid or late July to mid or late August, outside air temperature is about 25-35 ℃.
The culture material character that table 2 test is adopted
Mushroom compost is finished the apparent characteristic that bio-transformation can be used as Mushroom planting substrate: heap temperature is near external environment; Material stack volume obviously dwindles, about 60% when only building heap; Culture material is brown, the adularescent bacterial plaque; Straw/wheat straw original shape still exists, and sponginess draws promptly disconnectedly with have gentle hands, but be not broken mashed; Expect moisturely about 65%, when firmly holding, water is arranged between webs but do not drip.Simultaneously culture material does not have peculiar smell such as ammonia, smelly, acid, slightly cookie flavor or mouldy slightly flavor.
The result is as shown in table 3, and this result shows: 1. add the bio-transformation speed that microbial inoculum can obviously shorten culture material, especially under low temperature environment, can with bio-transformation time of mushroom straw culture material by 34 days, shorten to 29 days; 2. add microbial inoculum and can increase the humus content that the mushroom compost bio-transformation generates, illustrate that the quality of Mushroom planting substrate increases after the interpolation microbial inoculum; 3. compared to straw, wheat straw is as the main ingredient of mushroom compost, and not only the transformation time of culture material is longer, and the quality of cultivation matrix also is lower than straw, illustrates that straw is more suitable for as mushroom compost; 4. outside temperature has bigger influence to the biotransformation of culture material, culture material transformation efficiency and changing effect all are lower than higher external temperature environment under the low temperature environment, but by adding the defective that microbial inoculum can remedy low temperature environment to a certain extent and caused.
Table 3 ambient temperature and microbial inoculum are to the influence of culture material bio-transformation time and effect
Experimental example 4 bacterial strain W4 are to the promotion experiment 2 of stalk culture material compost
The variation of microflora in the culture material biotransformation:
Mushroom compost bank up and biotransformation as described in the experimental example 3, Fig. 3 and Figure 4 shows that mushroom compost carries out bacterium in the biotransformation, fungi and actinomycetic number change feature under low temperature environment.
The result shows, the number change trend of bacterium and fungi is consistent in straw culture material and the wheat straw culture material, bacterium has the back downward trend that raises earlier at the culture material bio-transformation initial stage, maintain on the relative constant amount after then being increased to certain quantity, and after the interpolation microbial inoculum, bacteria content increases to some extent in the culture material biotransformation, but when finishing near bio-transformation, its quantity is consistent with the effect of the culture material that does not add microbial inoculum; The quantity of fungi presents tangible downtrending in the culture material biotransformation, maintain 10 substantially latter stage to transforming
3About; After adding microbial inoculum, fungi quantity the starting stage with respect to the culture material showed increased of not adding microbial inoculum, but its variation tendency is consistent with the number change trend of fungi in the culture material that does not add microbial inoculum, and its quantity also maintains 10 when the culture material bio-transformation is finished
3About.Actinomycetic variation tendency is different in straw culture material and the wheat straw culture material, mainly be in the wheat straw culture material actinomycetic quantity the bio-transformation initial stage many a trend of rising, but no matter be wheat straw culture material or straw culture material, actinomycetic quantity all descends to some extent after the interpolation microbial inoculum.
Experimental example 5 bacterial strain W4 are to the promotion experiment 3 of stalk culture material compost
The variation of stalk lignocellulose in the culture material biotransformation:
Mushroom compost bank up and biotransformation as described in the experimental example 3, table 4 carries out lignocellulose substances content in the Mushroom planting substrate that bio-transformation generates for mushroom compost under low temperature environment.
The content of lignocellulose material in table 4 Mushroom planting substrate
The result shows, no matter mushroom compost is that straw or its wood fibre cellulose content of wheat straw have had bigger variation through bio-transformation, especially after adding microbial inoculum, the content of Mierocrystalline cellulose and hemicellulose significantly reduces in the stalk, and the content of xylogen increases relatively to some extent.
Experimental example 7 different mushroom composts are to the influence experiment of mushroom yield
After the windrow that ferments carried out secondary " sweating ", in thermostatic chamber, experimentize whole sub-district area 6m
2Windrow is completed by 20cm is thick, inoculation, and inoculum size is 3 bottles/m
3, every bottle is 250ml wheat kind, and layer is broadcast, and disposable earthing, thickness of earth covering are 2cm.Pester and do not gather when not parachute-opening and mycoderm draw back, two tides of gathering.
Bisporous mushroom growth and rate ratio are between table 5 culture material
M: by the Mushroom planting substrate of straw/wheat straw (+microbial inoculum) culture material preparation.A, b, c, d represent the significant difference of same column number certificate.
Adopt SPSS software, the LSD method is analyzed, P＜0.5.
Table 5 has provided bisporous mushroom growth and the yield result by the Mushroom planting substrate acquisition of different culture material preparations, though the result shows that the Mushroom planting substrate that adds microbial inoculum is little to the weight in average influence of single mushroom, the weight in average of single mushroom just slightly increases, but can obviously improve the average speed of growth of mycelia, significantly improve the output of bisporous mushroom; External environment is not remarkable to the quality influence of the Mushroom planting substrate of final acquisition; The cultivation matrix that the straw culture material obtains will be far superior to wheat straw, so straw more is applicable to the preparation Mushroom planting substrate.
Though above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (9)
1. expand mould W4 (Penicillium expansum) bacterial strain, deposit number is CGMCC NO.4077.
2. the cultural method of the described bacterial strain of claim 1, it is that bacterial strain is joined in the PDA liquid nutrient medium, 30 ℃, 170rmin
-1Shaking culture 3d;
Wherein, the PDA liquid nutrient medium is: peeling potato 200g/L, glucose 20g/L, pH7.2～7.4.
3. the application of the described bacterial strain of claim 1 in lignocellulose degradation.
4. the application of the described bacterial strain of claim 1 in the mushroom compost biotransformation.
5. the application of the described bacterial strain of claim 1 in the microbiobacterial agent of preparation mushroom compost.
6. application according to claim 5 is characterized in that, the microbiobacterial agent of described mushroom compost comprises the component of following weight part: expansion mould W4 1-2 part; Rice chaff and/or wheat bran 10-15 part.
7. according to each described application of claim 4-6, it is characterized in that described mushroom compost is mainly agricultural crop straw.
8. application as claimed in claim 7 is characterized in that, described agricultural crop straw is wheat straw and/or straw.
9. the microbiobacterial agent of a mushroom compost, it comprises the component of following weight part: expansion mould W4 1-2 part; Rice chaff and/or wheat bran 10-15 part.
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| CN102391948A (en) * | 2011-09-02 | 2012-03-28 | 单茂莹 | Straw fermentation complex microbial inoculant and application thereof |
| CN102533563A (en) * | 2011-11-08 | 2012-07-04 | 中国科学院微生物研究所 | Celluase producing bacterium and application thereof |
| CN103667075A (en) * | 2013-04-17 | 2014-03-26 | 山东省食品发酵工业研究设计院 | Penicillium expansum strain for degrading liquor distillers' grain cellulose |
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| KR20170012253A (en) * | 2014-05-27 | 2017-02-02 | 노보자임스 에이/에스 | Methods for mushroom cultivation |
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