CN102870598A - Method for screening Pleurotus ostreatus strains capable of efficiently degrading cotton stalk lignin - Google Patents
Method for screening Pleurotus ostreatus strains capable of efficiently degrading cotton stalk lignin Download PDFInfo
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Abstract
The invention discloses a method for screening Pleurotus ostreatus strains capable of efficiently degrading cotton stalk lignin. The preserved Pleurotus ostreatus strains are taken out and activated, and are then inoculated to improved PDA-Bavendamm plate medium and improved PDA-RB plate medium for primary screening the strains high in metachromatism and fast in response. The strains are then inoculated to cotton stalk solid medium for screening again. The Pleurotus ostreatus strains capable of efficiently degrading cotton stalk lignin are obtained by secondary screening. The invention provides an established scientific screening method system. Primary screening is simple. A proper amount of cotton stalk powder is added to common allochroic medium, so that screening is more targeted, and screening accuracy is improved. The cotton stalk solid medium has single carbon source, so that the Pleurotus ostreatus strains capable of efficiently degrading cotton stalk lignin are unique to selection of the carbon source, and accuracy in secondary screening results is guaranteed.
Description
Technical field
The present invention relates to the edible mushroom degraded and utilize stalk resource and fungus growing technique field, be specifically related to the screening technique of the edible mushroom strains of efficient degradation straw lignin.
Background technology
Agricultural crop straw contains a large amount of organic matters, and nitrogen phosphorus potassium and trace element are important living resources.China's year is produced nearly 600,000,000 tons of stalk, but because its utilization that is difficult to be degraded, 1/3 stalk is by open incineration.This is not only the huge waste to resource, has also caused serious environmental pollution and a series of social concern.The main cause of stalk hard degradation is that lignin is combined with the covalent bond form with hemicellulose in the cell wall, and cellulosic molecule is embedded in wherein, and this has formed the natural cover for defense one, has limited the decomposition function of digestive ferment to cell wall and cellular content.The content of the lignin in the cotton stalk has greatly limited the degraded utilization of cotton stalk up to more than 25%, and also to be cotton stalk with other agricultural crop straws compare more is difficult to the main cause utilized.Therefore eliminate the lignin natural cover for defense in the stalk, can promote effective utilization of stalk resource.Wherein screening efficient Degradation by White-rot Fungus lignin is one of significant effort direction.The method of the whiterot fungi bacterial strain of screening efficient degradation straw lignin has multiple, but the shortcomings such as ubiquity complicated operation, poor accuracy, cost costliness should not be used as the screening of bacterial strain on a large scale.Therefore take cotton stalk as composts or fertilisers of cultivating, set up a screening technique system that cover is simple to operate, cost is low, accuracy is good, the oyster cap fungus bacterial strain of the cotton straw lignin of screening efficient degradation, significant to the development and use of stalk resource.Both can be the straw biological degraded effective way was provided, can obtain economic benefit, environmental contamination reduction again simultaneously.
Summary of the invention
The purpose of this invention is to provide a kind of method of screening the oyster cap fungus bacterial strain of the cotton straw lignin of efficient degradation, solve the problem that stalk resource is difficult to develop because of the existence of lignin.
Realize that the goal of the invention technical scheme is as follows:
1, whole screening system is divided into primary dcreening operation and sieved again for two steps, comprises following concrete steps:
(1) the different oyster cap fungus bacterial strains after the activation culture is inoculated into through the PDA-Bavendamm plating medium of improvement with through on the PDA-RB plating medium of improveing, under 25 ℃ ± 1 ℃ condition, cultivate 7d and carry out Preliminary screening, observe the formational situation of variable color circle every day and record developing time and the colour developing loop diameter;
(2) with the inoculation that metachromasia is strong and the reaction time is fast on two kinds of medium that filters out in the step (1) to the cotton stalk solid culture medium, under 25 ℃ ± 1 ℃ condition, cultivate 30d;
(3) matrix after the cultivation that step (2) is obtained adopts Van Soest method to measure Lignin degradation rate and calculate selectivity index, filters out on this basis the oyster cap fungus bacterial strain of the cotton straw lignin of efficient degradation.
Described PDA-Bavendamm plating medium through improvement is: tannic acid to the final concentration that adds separately sterilization in the PDA medium is 0.01g/100ml, is 0.1% to add 60 order sub-sieves, particle diameter less than the cotton stalk dry powder of 0.25mm by W/V again.
Described PDA-RB plating medium through improvement is: RB light blue to the final concentration that adds separately sterilization in the PDA medium is 625ug/mL, added 60 order sub-sieves, particle diameter less than the cotton stalk dry powder of 0.25mm, the addition of cotton stalk dry powder is that every 100mlPDA medium adds 0.1g again;
Described cotton stalk solid culture medium is: 5g crosses 60 order sub-sieves, particle diameter less than the cotton stalk dry powder of 0.25mm, and 20mL synthesizes culture fluid, and synthetic culture fluid changes the low high mineral salt culture fluid of nitrogen sugar-free into, fill a prescription following (numerical value is volume ratio):
Ammonium tartrate liquid: macroelement liquid: liquid microelement: VB1 liquid: water=15:15:3:16.Wherein, ammonium tartrate (22.0g/L) is nitrogenous source, and every liter of macroelement liquid contains 20gKH
2PO
4, 13.8gMgSO
47H
2O, 1.0gCaCl
2And 0.6gNaCl, every liter of liquid microelement contains 0.35gMnSO
4H
2O, 60mgFeSO
47H2O, 110mgCoCl
26H
2O, 60mgZnSO
47H
2O, 95mgCuSO
45H
2O, 6mgAlK (SO
4)
212H
2O, 6mgH
3BO
3And 6mgNa
2MoO
42H
2O, VB
1Mass fraction be 100mg/L.
The potato extract prepares as follows: potato scrapes off tertia, removes eye, accurately takes by weighing 200g, is cut into fragment and puts in the pot, add water 1000ml heating and boil, during constantly stir; Boil 20～25min again after water boils, filter with double gauze, gained filtrate is the potato extract.
The present invention compared with the prior art, its beneficial effect is embodied in:
(1) the present invention has set up a whole set of screening system, is divided into primary dcreening operation and sieves again two steps.Preliminary screening is simple to operate, fast and easy and with low cost, when the oyster cap fungus bacterial strain of the cotton straw lignin of Large-scale Screening efficient degradation, can save time, save cost and reduce quite a few workload: multiple sieve is quantitatively accurately, accuracy is good, the present invention makes up two kinds of screening technique science, and is both simple to operate, saves cost, again accurately and reliably, set up one and overlapped the more screening system of science.
(2) in the primary dcreening operation process of the present invention, by in common variable color medium, adding an amount of cotton stalk powder, make screening have more specific aim, guaranteed the accuracy of the selection result.
(3) again in the sieve process, need in the cotton stalk solid culture medium to add in addition the low high mineral nutrition liquid of nitrogen sugar-free, thereby carbon source can only be the single degraded from lignocellulose in the cotton stalk, guarantee multiple sieve result's accuracy.
Description of drawings
Fig. 1 is that embodiment sieves each bacterial strain Lignin degradation rate comparison in the experiment again.
Fig. 2 is that each bacterial strain selectivity index compares in the multiple sieve experiment.
Below by embodiment technical solution of the present invention is described further.
Embodiment
Embodiment:
1.1 experiment material
1.1.1 experimental strain
From Eight Strains of Pleurotus Ostreatus totally 12 strains that experiment is adopted, wherein flat No. 1 of Anhui, P928, P3 are provided by Academy of Agri-Science and Technology Anhui Province gardening; Flat No. 1 of farming, P17, sky reach 300 to be provided for Agricultural University Of Anhui's forestry and gardens institute; No. 1, Su Ping, No. 3, Su Ping, newly flat 400 be that academy of agricultural sciences, Jiangsu Province Vegetable Research Institute provides; North-1, early, black flat A provides by using fungus research institute of Hua Zhong Agriculture University.
1.1.2 medium
PDA synthetic medium: potato extract 1000ml, glucose 20g, KH
2PO
43.0g, MgSO
4H
2O1.5g, vitamin B1 trace, agar 15.0g.
The PDA-Bavendamm medium: tannic acid to the final concentration that adds separately sterilization in the PDA medium is 0.01g/100ml, added 60 order sub-sieves, particle diameter less than the cotton stalk dry powder of 0.25mm, the addition of cotton stalk dry powder is that every 100mlPDA medium adds 0.1g again.
PDA-RB light blue medium: (be thunder agate azoles light blue, English by name: Remazol Brilliant Blue R, chemical formula is C to add the RB light blue of sterilizing separately in the PDA medium
22H
16N
2O
11S
3) to final concentration be 625ug/mL, added again 60 order sub-sieves, particle diameter less than the cotton stalk dry powder of 0.25mm, the addition of cotton stalk dry powder is that every 100mlPDA medium adds 0.1g.
The stalk solid culture medium: take by weighing 5g cross 60 order sub-sieves, particle diameter less than the cotton stalk dry powder of 0.25mm in the 250mL conical flask, add the synthetic culture fluid of 22mL, fill in 8 layers of gauze, other ties up with two layers of newspapers covering and with fine rule.121-125 ℃ of high-temp steam sterilizing 30min.Synthetic culture fluid changes the low high mineral salt culture fluid of nitrogen sugar-free into, fill a prescription following (numerical value is volume ratio):
Synthetic culture fluid: ammonium tartrate liquid: macroelement liquid: liquid microelement: VB1 liquid: water=1:15:15:3:16.Wherein, ammonium tartrate (22.0g/L) is nitrogenous source, every liter of macroelement liquid contains 20gKH2PO4,13.8gMgSO47H2O, 1.0gCaCl2 and 0.6gNaCl, every liter of liquid microelement contains 0.35gMnSO4H2O, 60mgFeSO47H2O, 110mgCoCl26H2O, 60mgZnSO47H2O, 95mgCuSO45H2O, 6mgAlK (SO4) 212H2O, 6mgH3BO3 and 6mgNa2MoO42H2O, and the mass fraction of VB1 is 100mg/L.
The potato extract prepares as follows: potato scrapes off tertia, removes eye, accurately takes by weighing 200g, is cut into fragment and puts in the pot, add water 1000ml heating and boil, during constantly stir; Boil 20～25min again after water boils, filter with double gauze, gained filtrate is the potato extract.
Culture medium for cultivating: experimental group adopts cotton stalk 70%, wheat bran 26%, lime 3%, sucrose 1%; Control group is cotton seed hulls 70%, wheat bran 26%, lime 3%, sucrose 1%.
1.2 experimental technique
1.2.1 the activation culture of bacterial classification
The female kind of bacterial strain to be tried taken out under preservation state, respectively inclined plane inoculating activation culture on the PDA medium.The bacterial classification inoculation that then will activate on the PDA culture medium flat plate, 25 ℃ ± 0.5 ℃ lower cultivate behind the 7d for subsequent use.
1.2.2 the Preliminary screening of efficient degradation lignin bacterial strain
It is that the bacterium piece of 3.5mm * 3.5mm is inoculated in respectively on PDA-Bavendamm culture medium flat plate and the PDA-RB culture medium flat plate that the bacterial strain flat board of common PDA being cultivated seven ages in days is beaten the diameter of learning from else's experience with card punch, cultivate 7d under 25 ℃ ± 0.5 ℃ condition, each bacterial strain is done 3 repetitions.Observe the formational situation of variable color circle every day and record the variable color time, draw two orthogonal lines centered by bacterium colony, the mean value that increases along the colony diameter of two lines is got in the measurement of variable color loop diameter, and mycelial density adopts intuitively relatively to be judged and record.Select the bacterial classification that variable color is fast, the variable color circle is large to treat the experiment of lower step.
1.2.3 lignin degradation experiment (multiple sieve)
The From Eight Strains of Pleurotus Ostreatus seven age in days PDA plating mediums that Preliminary screening is gone out are got 5.0mm * 5.0mm bacterium piece with card punch and are inoculated on the attached body medium of stalk, cultivate 30d under 25 ℃ ± 0.5 ℃ condition.Each bacterial classification is established three repetitions.
1.2.4 the calculating of the mensuration of degradation rate and selectivity index (SF) (adopting the FIWE6 fiber type analyzer of Italian VELP company)
Adopt Van Soest method to survey lignin, cellulose and hemicellulose level.
Lignin in degradation rate=lignin (or cellulose, hemicellulose) degraded total amount/raw sample (or cellulose, hemicellulose) total amount * 100%
Selectivity index (SF)=Lignin degradation rate/holocellulose (cellulose+hemicellulose) degradation rate * 100%.
1.2.5 experiment in cultivation
Adopt the bottle hydroponics, each bacterial strain is by prescription, control group and experimental group respectively fill 10 bottles, after the sealing, in 121 ℃ of high pressure moist heat sterilization 120min, access each bacterial strain cultivated species in 5% inoculum concentration (with weight in wet base) after the cooling, place 25 ℃ of constant incubators to be cultured to the mycelia purseful after, move in the mushroom room and cultivate according to a conventional method, pluck to cultivate after the 2 damp fruit bodys and finish.
1.2.5.1 fruiting body yield is measured
Fruiting body yield adopts single bottle of output to calculate, i.e. single bottle entity fresh weight of 10 repetitions of weighing successively, record and calculating mean value;
1.2.5.2 ligocellulose degradation leads mensuration in the cultivation matrix
Assay method is with the mensuration of 1.2.4 degradation rate
2 results and analysis
2.1 the Preliminary screening of efficient degradation lignin bacterial strain
The dull and stereotyped colour developing of each bacterial strain PDA-Bavendamm of table 1 result
Annotate :-expression chromogenic reaction is negative; ± expression chromogenic reaction just begins; ﹢, ﹢ ﹢, ﹢ ﹢ ﹢ represent that chromogenic reaction strengthens gradually; ﹢ ﹢ ﹢ ﹢ represents that chromogenic reaction is very thorough; Lowercase alphabet differential different significantly (P＜0.05) in the table, capitalization represents difference extremely significantly (P＜0.01).
The dull and stereotyped decolouring of each bacterial strain RB-light blue of table 2 result
Annotate :-expression decoloring reaction is negative; ± expression decoloring reaction just begins; ﹢, ﹢ ﹢, ﹢ ﹢ ﹢ represent that decoloring reaction strengthens gradually; ﹢ ﹢ ﹢ ﹢ represents that decoloring reaction is very thorough; Lowercase alphabet differential different significantly (P＜0.05) in the table, capitalization represents difference extremely significantly (P＜0.01).
Many studies show that, the edible mushrooms such as flat mushroom mainly are three kinds of extracellular enzyme classes that rely on its mycelium secretion, i.e. peroxidase, manganese peroxidase and laccase to the degraded utilization of lignin in the stalk.The principle of the dull and stereotyped colour developing of Bavendamm is exactly to utilize the secreted laccase of flat mushroom can make the phenolic compound polymerization, forms the sepia oxidized zone in periphery of bacterial colonies, presents positive reaction.Peroxidase can make dyestuff Remazol Blue(RB) the light blue decolouring, in the PDA medium that adds RB, the bacterial strain that produces peroxidase can form the decolouring torus in periphery of bacterial colonies.These two kinds of metachromatic powers and required time all depend on genetic background and the condition of culture of bacterial classification.And flat mushroom secreted these three kinds to wooden have degradation enzyme all belong to induced enzyme, therefore, in the situation that there is a certain amount of cotton stalk lignin inducing action in the medium, we can be according to the diameter of variable color circle, the depth degree of variable color and the height that generation time is judged the strain enzyme-producing performance, and then judge bacterial strain to the degradation capability of cotton stalk lignin, filter out the higher bacterial strain of degraded cotton stalk lignin ability.
Adopt the SPSS statistical software to carry out data statistic analysis.Experimental data represented with mean ± standard deviation between each was processed, and patted average everywhere and adopted the Tukey method in the variance analysis to carry out significance analysis.The dull and stereotyped colour developing of PDA-Bavendamm experimental result is as shown in table 1.In the experiment, flat No. 1 of Anhui, early, No. 1, Su Ping, No. 3 these four strains of Su Ping just begin to present obvious sepia at postvaccinal first day.All the other most of bacterial strains also begin colour developing at postvaccinal second day.North-1 chromogenic reaction begins the latest, and the 3rd talent has obvious variable color circle to form.Relatively cultivate the variable color loop diameter after seven days, we find except No. 1, Su Ping, north-1 and P928 significant difference, and all the other bacterial strains colour developing diameters do not have obvious difference in statistics.No. 1 variable color loop diameter of Su Ping is significantly greater than other bacterial strains, and color developing effect is very dark simultaneously, it is described under the inducing of cotton stalk lignin, and Laccase is very capable.North-1 and P928 colour developing loop diameter are minimum, and aspect color developing effect, its color developing effect also is the most shallow, illustrates that its secretion laccase ability is more weak simultaneously.
As known from Table 2, the difference of each bacterial strain is comparatively obvious among the dull and stereotyped decolouring of the PDA-RB light blue result.Can find out by relatively bleaching time and decolouring circle size, No. 1, bacterial strain Su Ping, Su Ping No. 3, days reach 300, the bleaching time of Anhui flat No. 1 and P17 begins early, decolouring thoroughly, the decolouring loop diameter is large and obviously, secretion peroxidase ability is stronger; Secondly be morning, north-1, P928, newly put down 400; Farming is put down No. 1 and the P3 bleaching time begins the latest, and the decolouring loop diameter is minimum, and the ability of secretion peroxidase is the most weak.
Flat mushroom is not to rely on a kind of enzyme effect to the degraded of lignin, but the coefficient result of several enzyme mainly works in coordination with to finish by these 3 kinds of enzymes of lignin peroxidase, manganese peroxidase and laccase mutually.Therefore comprehensive two kinds of plate screening methods, we have selected bacterial strains of the efficient degradation cotton lignin that goes out as Preliminary screening of very strong 8 From Eight Strains of Pleurotus Ostreatus of laccase and peroxidase secretion capacity, put in the multiple sieve experiment of next round.
As can be seen from Figure 1, the Lignin degradation rate of No. 1 bacterial strain of Su Ping is the highest in the multiple sieve experiment, and in the cultivation of 30d, its Lignin degradation rate has reached 14.82%; Next is flat No. 1 and black flat A of Anhui, and Lignin degradation rate is respectively 13.79% and 13.65%; Minimum is that early Lignin degradation rate is 10.81%
We can find out by Fig. 2, and the bacterial strain that selectivity index is the highest is No. 1, black flat A and Su Ping, secondly be that put down No. 1 in No. 3, Su Ping and Anhui, and the selectivity index of all the other four bacterial strains is all less than 1.Studies show that, what the height of selectivity index reflected is whiterot fungi Selective lignin-degradation and brown cellulosic ability, and the From Eight Strains of Pleurotus Ostreatus of high-efficiency lignin degrading should be to utilize under the brown cellulosic prerequisite reducing as far as possible degraded, and the preferential degraded of doing large limit utilizes straw lignin.In sum, in the experiment of multiple sieve, we take each bacterial strain to the Lignin degradation rate of cotton stalk as Main Basis, selectivity is referred to element as important references, filtered out No. 1, the From Eight Strains of Pleurotus Ostreatus-Su Ping of a high-efficiency degradation cotton stalk lignin.
The 8 strain oyster cap fungus that just filter out are carried out experiment in cultivation, and relatively the degradation rate of their lignin in multiple sieve and cultivation just changes, and experimental result data adopts the mean value of three repeated experiments to represent, standard deviation is all less than 1.The result shows, No. 1, the oyster cap fungus bacterial strain-Su Ping of the cotton straw lignin of the efficient degradation that the present invention filters out, Lignin degradation rate and selectivity index degradation rate still are the highest in experiment in cultivation, and each oyster cap fungus bacterial strain of filtering out of the present invention just to change at multiple sieve and Lignin degradation rate in the experiment in cultivation be consistent (table 3).Illustrate that the oyster cap fungus bacterial strain that the present invention is used for screening the cotton straw lignin of efficient degradation is feasible.
Lignin degradation rate relatively in the multiple sieve of table 3 and the cultivation
Simultaneously in the correlation test of the Lignin degradation rate measured of multiple sieve and cotton stalk cultivated fruitbody output (table 4), the correlation coefficient r of sample
2=0.787, illustrate that the oyster cap fungus bacterial strain that utilizes decomposition rate of lignin to screen the cotton stalk of efficient degradation is feasible.
Each bacterial strain fruiting body yield in the cultivation of table 4 cotton stalk
If the Lignin degradation rate that multiple sieve is measured is X, the output of oyster cap fungus on cotton stalk is Y, calculates the one-variable linear regression equation:
Y=3.494X-2.489
The correlation coefficient r of sample
2=0.787, r=0.887
By cultivation experiments, we will sieve again with cultivation experiments in the bacterial strain Lignin degradation rate that respectively employs measured compare, find that both just change is consistent.The Lignin degradation rate of measuring in the screening experiment has reflected that accurately each bacterial strain is to the lignin degradation situation of cotton stalk matrix in the actual cultivation process.The correlation test that sieves again simultaneously between Lignin degradation rate and the fruiting body yield also shows cotton straw lignin degradation rate and fruiting body yield significant positive correlation.
Therefore, our this method of overlapping screening efficient degradation cotton stalk lignin From Eight Strains of Pleurotus Ostreatus of foundation is that science is accurate and feasible.
Claims (2)
1. a method of screening the rough skin side bacterial strain of the cotton straw lignin of efficient degradation is characterized in that, specifically comprises the steps:
(1) with the female respectively inclined plane inoculating activation culture on the PDA medium of planting of bacterial strain to be screened, the bacterial classification inoculation that then will activate on the PDA culture medium flat plate, 25 ℃ ± 0.5 ℃ lower cultivate behind the 7d for subsequent use; Described PDA medium is pressed following component proportion: potato extract 1000ml, glucose 20g, KH
2PO
43.0g, MgSO
4H
2O1.5g, Cobastab
10.1g, agar 15.0g;
It is that the bacterium piece of 3.5mm * 3.5mm is inoculated in respectively on PDA-Bavendamm culture medium flat plate and the PDA-RB culture medium flat plate that the bacterial strain flat board of seven ages in days of (2) step 1) being cultivated is beaten the diameter of learning from else's experience with card punch, cultivate 7d under 25 ℃ ± 0.5 ℃ condition, observe the formational situation of variable color circle every day and record the variable color time and the variable color loop diameter;
Described PDA-Bavendamm medium is that tannic acid to the final concentration that adds separately sterilization in the PDA medium of step 1) is 0.01g/100ml, added 60 order sub-sieves, particle diameter less than the cotton stalk dry powder of 0.25mm, the addition of described cotton stalk dry powder is that every 100mlPDA medium adds 0.1g again;
Described PDA-RB medium is that RB light blue to the final concentration that adds separately sterilization in the PDA medium of step 1) is 625ug/mL, added 60 order sub-sieves, particle diameter less than the cotton stalk dry powder of 0.25mm, the addition of described cotton stalk dry powder is that every 100mlPDA medium adds 0.1g again;
(3) with filter out in the step (2) in metachromasia is strong and the reaction time is fast on two kinds of medium inoculation to cotton stalk solid culture medium, under 25 ℃ ± 1 ℃ condition, cultivate 30d;
Described cotton stalk solid culture medium is crossed 60 order sub-sieves, particle diameter less than the cotton stalk dry powder of 0.25mm by following component proportion: 5g, 20mL synthesizes culture fluid, and by volume its prescription of described synthetic culture fluid is: ammonium tartrate liquid: macroelement liquid: liquid microelement: VB1 liquid: water=15:15:3:16; Wherein, the ammonium tartrate solution concentration is 22.0g/L, and every liter of macroelement liquid contains 20gKH
2PO
4, 13.8gMgSO
47H
2O, 1.0gCaCl
2And 0.6gNaCl, every liter of liquid microelement contains 0.35gMnSO
4H
2O, 60mgFeSO
47H2O, 110mgCoCl
26H
2O, 60mgZnSO
47H
2O, 95mgCuSO
45H
2O, 6mgAlK (SO
4)
212H
2O, 6mgH
3BO
3And 6mgNa
2MoO
42H
2O, VB
1Mass fraction be 100mg/L;
(4) matrix after the cultivation that step (3) is obtained adopts Van Soest method to measure Lignin degradation rate and calculate selectivity index, filters out on this basis the oyster cap fungus bacterial strain of the cotton straw lignin of efficient degradation.
2. a kind of method of screening the rough skin side bacterial strain of the cotton straw lignin of efficient degradation according to claim 1, it is characterized in that, described potato extract prepares as follows: potato scrapes off tertia, remove eye, accurately take by weighing 200g, be cut into fragment and put in the pot, add water 1000ml heating and boil, during constantly stir; Boil 20～25min again after water boils, filter with double gauze, gained filtrate is the potato extract.
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| CN103583230A (en) * | 2013-10-17 | 2014-02-19 | 固镇县玉鹏蔬菜开发有限责任公司 | Cotton straw based bag cultivation method for oyster mushroom |
| CN107177637A (en) * | 2017-06-15 | 2017-09-19 | 沈阳农业大学 | The methods and applications pre-processed using microorganism to maize straw hydrolysis and saccharification |
| CN107815459A (en) * | 2017-12-06 | 2018-03-20 | 安徽农业大学 | A kind of oyster cap fungus manganese peroxidase enzyme gene and its application |
| CN108419607A (en) * | 2018-03-07 | 2018-08-21 | 安徽农业大学 | A kind of process parameter optimizing method of oyster cap fungus degradation cotton stalk |
| CN113416655A (en) * | 2021-07-21 | 2021-09-21 | 嘉兴学院 | Efficient fast-rotting straw strain, screening process and application thereof |
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