CN114315438A - Walnut green husk biological organic fertilizer and application thereof - Google Patents
Walnut green husk biological organic fertilizer and application thereof Download PDFInfo
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- CN114315438A CN114315438A CN202111474214.9A CN202111474214A CN114315438A CN 114315438 A CN114315438 A CN 114315438A CN 202111474214 A CN202111474214 A CN 202111474214A CN 114315438 A CN114315438 A CN 114315438A
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- walnut green
- organic fertilizer
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- green husk
- lignin
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Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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- Fertilizers (AREA)
Abstract
The invention belongs to the technical field of fertilizer production, and particularly relates to a walnut green husk biological organic fertilizer and application thereof, wherein 4-8 parts by weight of rice bran, 5-10 parts by weight of sawdust, 6-12 parts by weight of fungus sticks, 20-35 parts by weight of walnut green husks, 10-30 parts by weight of oil cakes and 5-10 parts by weight of macadamia nut peels are mixed, microbial suspension liquid with the functions of degrading cellulose and lignin is added into raw materials according to the proportion of suspension liquid (8-10) ml/g of walnut green husks for natural fermentation for 60-65 days, and then the raw materials are turned over once per week in the natural fermentation process to obtain the walnut green husk biological organic fertilizer; according to the invention, the microorganisms with the functions of degrading cellulose and lignin are used for composting, so that the walnut green husk has good degradation activity, the exposure rate of effective components in the walnut green husk can be adjusted, and the effective components can better flow into soil or be absorbed; in addition, the invention greatly improves the composting effect by preferably selecting the microorganisms which are not antagonistic to each other.
Description
Technical Field
The invention belongs to the technical field of fertilizer production, and particularly relates to a walnut green husk biological organic fertilizer and application thereof.
Background
The walnut green husk is also called walnut green dragon skin and is green peel outside the walnut fruit, and after the fruit is harvested, the green peel is often stacked in the field as waste, which not only causes resource waste, but also pollutes the surrounding environment. The variety of chemical components in the pericarpium juglandis is various, and more than 130 chemical components are found, wherein the chemical components such as naphthoquinones, polyphenols, flavonoids, diarylheptanoids, polysaccharides, trace elements and the like are main effective substances of the pericarpium juglandis. In the study on the biological activity of the pericarpium juglandis, the ethanol extract and the ethyl acetate extract of the pericarpium juglandis have obvious inhibition effect on the growth of plant seedlings, and the water extract of the pericarpium juglandis has low promotion and high inhibition effect on the germination rate and the germination potential of wheat and rape; it also shows that: the extract of the pericarpium juglandis has stronger contact activity and antifeedant activity to radish aphids. The ethanol extract of the exocarpium Juglandis Immaturum shows strong poisoning effect on 3 rd larvae of Spodoptera litura, and can prolong the larval stage and pupal stage. The decomposition products of the pericarpium juglandis can effectively enhance the activity of soil enzymes, improve the content of soil nutrients, increase the number of microorganisms and improve the soil fertility. These characteristics provide the possibility and basis for the use of the pericarpium juglandis for fertilizer production.
In addition, most organic matter sources of the existing organic fertilizers are wastes of livestock, poultry excrement and crops in large-scale farms, the excrement is used as a raw material, heavy metals and antibiotics in the fertilizers exceed standards easily, and the wastes are used as the raw material and are insufficient in nutrient content, so that the research and development of the organic fertilizer produced by using the pericarpium juglandis as the raw material are further promoted.
However, the extraction process of the walnut green seedcase is complex, the extraction efficiency is low, the utilization rate is low, and no report on the production of walnut green seedcase organic fertilizer for planting passion fruit is provided.
Disclosure of Invention
The invention provides a walnut green husk biological organic fertilizer and application thereof, aiming at the defects of the prior art.
The method is realized by the following technical scheme:
a walnut green husk biological organic fertilizer is prepared by mixing 4-8 parts by weight of rice bran, 5-10 parts by weight of sawdust, 6-12 parts by weight of fungus sticks, 20-35 parts by weight of walnut green husks, 10-30 parts by weight of oil cake and 5-10 parts by weight of macadamia nut peels, adding a microbial suspension liquid with the functions of degrading cellulose and lignin into raw materials according to the proportion of 8-10 ml of the suspension liquid per gram of the walnut green husks, naturally fermenting for 60-65 days, and turning over once a week in the natural fermentation process.
The microorganism with the function of degrading cellulose and lignin is any one or more of white bacillus Leucobeater sp, Stenotrophomonas rhizophila, Fusarium solani, Penicillium rolfsli and Fungal endophyte sp.
More preferably, the microorganism having the function of degrading cellulose and lignin is any one of the following compositions:
(1) mixing the leucobacter sp and the Fungal endophytic sp in any ratio;
(2) a mixture of Stenotrophomonas rhizophila and Fungal endophyte sp in any ratio;
(3) a mixture of the white bacillus Leucobater sp, Stenotrophomonas rhizophila Stenotrophora rhizophila and Fungal endophyte sp in any ratio.
Preparing the suspension liquid: inoculating microbial strain (3-15ml) with cellulose and lignin degrading function into liquid culture medium, and shake culturing at 30 deg.C and 130rpm for 5 d.
The liquid culture medium is determined according to the condition that microbial strains with the functions of degrading cellulose and lignin are fungi and bacteria, a martin liquid culture medium is adopted when the strains are the fungi, and a beef peptone culture medium is adopted when the strains are the bacteria.
The Martin's medium: k2HPO41.0g，MgSO4·7H20.5g of O, 5g of peptone, 10g of glucose and 1000mL of water, and sterilizing at 121 ℃ for 20 min.
The beef extract peptone culture medium: 3.0g of beef extract, 10g of peptone, 5.0g of NaCl, 15-25g of agar and 1000mL of water, wherein the pH value is 7.4-7.6, the volume is adjusted to 1000mL by using distilled water, and the beef extract is sterilized at 121 ℃ for 20 min.
The application of the walnut green husk bio-organic fertilizer in promotion of radish seed germination comprises the following steps: taking a walnut green husk biological organic fertilizer, adding a functional microbial agent according to the amount of 0.57-1% of the mass of the walnut green husk, uniformly mixing to obtain a combined fertilizer, extracting according to the water-fertilizer ratio of 2:1, filtering, centrifuging the filtrate for 20min, absorbing the supernatant, adding distilled water to prepare a leaching solution with the volume concentration of 50-100%, absorbing 2mL of the leaching solution, placing the leaching solution in a culture dish paved with filter paper, placing 10 radish seeds in a constant-temperature incubator at 25 ℃ and culturing in the dark for 72 h.
The application of the walnut green husk bio-organic fertilizer in passion fruit planting comprises the following steps: taking walnut green husk biological organic fertilizer, adding functional microbial agent according to the amount of 0.57-1% of the mass of the walnut green husk, uniformly mixing, using the obtained product as base fertilizer, applying the base fertilizer into soil, turning over, and standing for 7 days to plant passion fruit seedlings.
The functional microbial agent is any one or more of trichoderma harzianum, bacillus subtilis and bacillus megaterium.
The mixture of the walnut green husk bio-organic fertilizer and the functional microbial agent is used as a base fertilizer for passion fruit planting.
Has the advantages that:
according to the invention, the microorganisms with the functions of degrading cellulose and lignin are used for composting, so that the walnut green husk has good degradation activity, the exposure rate of effective components in the walnut green husk can be adjusted, and the effective components can better flow into soil or be absorbed; in addition, the invention greatly improves the composting effect by preferably selecting the microorganisms which are not antagonistic to each other.
According to the invention, the substance obtained by composting the walnut green husk-containing substance is used as a base fertilizer for passion fruit planting in combination with the functional microbial agent, so that the growth condition and the number of blossoms and fruits of the passion fruit are greatly improved, and the growth promoting effect on the passion fruit is better.
The method realizes large-scale production, and realizes the characteristics of novelty, environmental protection, simplicity, easy implementation, low cost, good fertilizer efficiency and the like.
Screening of microorganism with cellulose and lignin degrading function
1.1 materials and methods
1.1.1 preparation of the culture Medium
CMC culture medium: CMC-Na 10g, K2HPO41g、NH3NO3 1g、CaCl2 0.02g、 MgSO4·7H2O 0.2g、FeCl30.05g of agar, 20g of agar and 1000mL of distilled water, and sterilizing for 20min at 121 ℃;
congo red medium: CMC-Na 5g, K2HPO4 1g、KNO3 3g、KCl 0.5g、 MgSO4·7H20.5g of O, 30.01g of FeCl30, 20g of agar and 1000mL of distilled water, and sterilizing for 20min at 121 ℃;
PCS culture medium: peptone 5g, NaCl 5g, CaCO32g, 1g of yeast powder and 1000mL of distilled water, and sterilizing for 20min at 121 ℃;
azure B medium: 10g of yeast extract, 20g of glucose, 20g of agar and 1000mL of distilled water, preparing 1.0% aniline blue (Azure-B) mother liquor, adding 1.0mL mother liquor into 100mL of a basic culture medium when a plate is prepared, and sterilizing for 20min at 121 ℃;
degrading lignin culture medium: NaNO3 2.5g，K2HPO4 1.0g，MgSO4·7H2O 1.0g，NaCl 1.0g，CaCl20.5g, 0.1g of yeast extract, 3.0g of alkali lignin, 0.1g of aniline blue and 20g of agar, wherein the volume is determined to be 1000mL by using distilled water, and the mixture is sterilized for 20min at 121 ℃;
lignin liquid medium: NaNO3 2.5g，K2HPO4 1.0g，MgSO4·7H2O 1.0g，NaCl 1.0g，CaCl20.5g, 0.1g of yeast extract and 3.0g of alkali lignin, wherein the volume is determined to be 1000mL by distilled water, and the mixture is sterilized for 20min at 121 ℃;
martin's liquid medium: k2HPO4 1.0g，MgSO4·7H20.5g of O, 5g of peptone, 10g of glucose and 1000mL of water, and sterilizing at 121 ℃ for 20 min;
LB culture medium: 10g of tryptone, 5g of yeast extract and 10g of NaCl, adjusting the pH to 7.0 by using 5mol/L NaOH, fixing the volume to 1000mL by using distilled water, and sterilizing for 20min at 121 ℃;
beef extract peptone medium: 3.0g of beef extract, 10g of peptone, 5.0g of NaCl, 15-25g of agar and 1000mL of water, wherein the pH value is 7.4-7.6, the volume is determined to be 1000mL by using distilled water, and the beef extract is sterilized for 20min at 121 ℃;
1.1.2 screening of cellulose-degrading bacteria
Taking out 0-20cm soil from pine forest soil, fir forest soil, walnut forest soil and green husk decomposition product, diluting to 10%-3、10-4、10-5、10-6Coating the suspension on a primary screening CMC culture medium; selecting a single bacterial colony of the obtained bacterial strain, inoculating the single bacterial colony on Congo red culture medium, selecting a bacterial colony which can grow well on the Congo red culture medium and has a transparent ring, transferring the bacterial colony into CMC-Na culture medium, carrying out shake culture at the constant temperature of 28 ℃, and measuring the cellulase activity of each bacterial strain for 3, 5, 7, 9 and 11 days;
CMCase Activity assay:
drawing a standard curve: drying anhydrous glucose to constant weight at 80 ℃, preparing 1mg/mL standard solution, taking 6 test tubes, respectively adding 0, 0.2, 0.4, 0.6, 0.8 and 1.0mL of the standard solution, adding distilled water to 2mL, adding 1.5mL of DNS reagent, carrying out boiling water bath for 5min, cooling, then setting the volume to 25mL, and measuring the OD value under 540nm of a spectrophotometer;
② preparing a crude enzyme solution: preparing a re-screening culture medium with the volume of 45mL per bottle, inoculating 5mL of seed suspension, placing in a shaking table at 28 ℃ for culturing, respectively taking 1.5mL of the seed suspension in a centrifuge tube for 3, 5, 7, 9 and 11d, and centrifuging at 10000r/min for 10min to obtain a crude enzyme solution;
③ measuring the enzyme activity: 1mL of the crude enzyme solution was centrifuged at 10000rpm for 10 min. Adding 1mL of CMC-Na solution with mass fraction of 1%, hydrolyzing at 45 deg.C for 20min, adding 1.5mL of LDNS, developing, boiling in water bath for 5min, cooling, diluting to 25mL, and measuring OD value (m1) with spectrophotometer at 540 nm. Taking 1mL of supernatant as a control, adding 1mL of distilled water, hydrolyzing at the constant temperature of 45 ℃ for 20min, adding 1.5mL of LDNS for color development, boiling in a water bath for 5min, cooling, metering to 25mL, measuring an OD value (m2) at 540nm of a spectrophotometer, wherein m1-m2 is A, and the enzyme activity is A1000/20 (U/mL);
1.1.3 determination of hemicellulase Activity of Strain
Centrifuging the fermentation liquor of 11d at 8000rpm at 4 deg.C for 15min, and placing the supernatant on ice for testing; sample loading was performed as in table 1 below; mixing the above solutions, developing in boiling water bath for 5min, cooling, sucking 200 μ L into micro cuvette, and measuring absorbance at 540 nm; measure Δ a ═ a measure-a pair, measure Δ a standard ═ a standard-a blank, enzyme activity (U/mL) [ measure Δ a ÷ (measure Δ a ÷ C standard) ] ÷ T ═ 0.13 × Δ a measure ÷ Δ a standard;
TABLE 1 hemicellulase Activity determination sample addition
1.1.4 screening of Lignin-degrading bacteria
Taking out 0-10cm soil from pine forest soil, fir forest soil, walnut forest soil and green husk decomposition product, diluting to 10%-3、10-4、10-5、10-6Coating the suspension on a primary-screening PCS culture medium; selecting a single bacterial colony from the obtained bacterial strains, inoculating the single bacterial colony on an Azure B culture medium, selecting a bacterial colony which can grow better on the Azure B culture medium and can generate a transparent ring, transferring the bacterial colony into a lignin liquid culture medium, carrying out constant-temperature shaking culture at 28 ℃, and measuring the activity of ligninase of each bacterial strain at 11 d;
because laccase is an important member of a lignin degradation enzyme system and is an environment-friendly copper polyphenol oxidase, electrons are usually extracted from lignin to reduce oxygen in the air into water; screening 12 strains with better growth vigor from an Azure B culture medium, inoculating the strains into a Martin liquid culture medium, culturing at the constant temperature of 28 ℃ for 10 days, taking an 11 th sample, measuring the enzyme activity of the sample by using a laccase measurement kit, taking a culture solution of thalli as a measurement sample, and detecting according to the following step 2. After the solution is fully mixed, the solution is quickly placed in a water bath at 45 ℃ for 3min after the absorption value A1 is measured at 420nm, the absorption value A2 is quickly measured at 10s, the absorption value A is measured as A2 and measured as A1, the absorption value A is measured as A2 (blank) -A1 (blank), the absorption value A is measured as delta A and delta A (blank), and the enzyme activity (U/mL) is measured as delta A and epsilon multiplied by d multiplied by V multiplied by 109 and V and T is measured as 61.7 multiplied by delta A;
TABLE 2 laccase Activity determination sample addition
| Reagent | Measuring tube | Blank tube |
| Sample (μ L) | 30 | —— |
| Distilled water (mu L) | —— | 30 |
| Working fluid (mu L) | 170 | 170 |
1.2 results and analysis
1.2.1 determination of cellulase Activity
The results of the glucose standard curve preparation are shown in Table 3, R2When the standard curve is equal to 0.9943, the linearity is good, and the equation of the standard curve of the glucose is obtained, wherein y is equal to 0.521x + 0.0043; a total of 22 strains were screened, and 8 strains capable of degrading cellulose were obtained, among which 4 strains with higher absorbance values (see Table 4). Calculating the enzyme activities of the strains 5X, 7-2Z and 13Z (shown in Table 5), and obtaining that the enzyme activities of 5X and 7-2Z are higher and are respectively 180U/mL and 265U/mL at 11 d; therefore, the cellulase activity of the strains 5X and 7X is relatively high, and particularly after the 7 th day, the cellulase activity of the strains is gradually increased, which shows that the 5X strain and the 7X strain have relatively good cellulose degradation;
TABLE 3 glucose Standard Curve
| Glucose content (mg) | 0 | 0.2 | 0.4 | 0.6 | 0.8 | 1 |
| Absorbance (A) | 0.014 | 0.092 | 0.211 | 0.317 | 0.444 | 0.511 |
TABLE 4 determination of the absorbance values of cellulase Activity of the strains
Table 5 cellulase Activity of strains
1.2.2 measurement of hemicellulase Activity
The absorbance values and enzyme activities of the hemicellulose of the strain are shown in table 6; as can be seen from Table 6, the hemicellulase activities of strains 13Z, 10Z, 7X and 4Z were high, and were 0.036, 0.034, 0.021 and 0.017U/mL, respectively;
TABLE 6 determination of hemicellulase activity of the strains
1.2.3 measurement of Lignin Activity
TABLE 7 Strain laccase assay
| Sample(s) | 10s (absorbance) | 3min10s (absorbance) | Enzyme Activity (U/mL) |
| CK | 0.033 | 0.036 | —— |
| 10Z | 0.086 | 0.085 | —— |
| 5X | 0.221 | 0.243 | 1.172 |
| 13Z | 0.036 | 0.041 | 0.123 |
| 9X | 0.194 | 0.186 | —— |
| 1Z | 0.181 | 0.192 | 0.494 |
| 7X | 0.305 | 0.323 | 0.926 |
| 4Z | 0.171 | 0.171 | —— |
| 17 | 0.119 | 0.192 | 4.319 |
| 12 | 0.135 | 0.126 | —— |
| 7-1 | 0.816 | 0.821 | 0.123 |
| 7-2 | 0.810 | 0.828 | 0.925 |
| 11 | 0.22 | 0.23 | 0.432 |
2 identification of the Strain
Extracting total DNA of pathogenic bacteria by using a fungus and bacterium DNA extraction kit (Shanghai life Co., Ltd.), and extracting the total DNA by using fungus universal primers ITS1: TCCGTAGGTGAACCTTGCG; ITS4: TCCTCCGCTTATTGATATGC and bacterial universal primer 27F: AGAGTTTGATCCTGGCTCAG; 1492R GGTTACCTTGTTACGACTT, 25 μ L reaction system: 10 XPCR buffer (containing Mg)2+) mu.L of each 2.5. mu.L, 10mol/L dNTP 0.5. mu.L, 10. mu. mol/L primer 0.5. mu.L, template DNA 1. mu. L, Taq enzyme 0.3. mu.L, plus ddH2O to the total volume of 25 mu L; reaction conditions are as follows: pre-denaturation at 94 ℃ for 2min, denaturation at 94 ℃ for 30s, annealing at 56 ℃ for 30s, extension at 72 ℃ for 1min, and 35 cycles; the obtained PCR product is subjected to purification, gel cutting and recovery, and then is sent to a company Limited in bioengineering (Shanghai) for sequencing, the measured sequence is subjected to homology comparison on NCBI, and the comparison result shows that 5X is Leuconobater sp; 7X is Stenotrophomonas rhizophila rhizotropis rhizozophila; 7-2Z and 13Z are both Fusarium solani Fusarium solani.e.; 4Z is Penicillium rolfsli; 17Z is Fungal endophytic sp;
antagonistic test of 3 strains
Inoculating bacteria and fungi to an agar culture medium by adopting a scribing method and a fungus cake inoculation method for culture, observing the antagonistic condition between strains every two, and confirming that no antagonistic exists when the strains are close to each other in growth vigor, wherein the antagonistic action is shown when the growth vigor is gradually far away; results of antagonism are shown in table 8; as can be seen from table 8: 5X does not antagonize other 3 bacteria, and 7X, 17Z and 4Z antagonize the bacteria, so 5 combinations are designed for the comprehensive degradation efficiency of the strains on cellulose and lignin: 5X +7X, 5X +17Z, 5X +4Z, 7X +17Z and 5X +7X +17Z are compounded; transferring the strains into an enrichment medium according to the equal volume, culturing for 7 days, and inoculating into a prepared compost raw material for fermentation;
TABLE 8 Strain mutual antagonism
| Strain numbering | 13Z | 17Z | 4Z | 7-2Z | 5X | 7X |
| 13Z | 0 | - | - | - | + | + |
| 17Z | - | 0 | - | - | + | + |
| 4Z | - | - | 0 | - | + | + |
| 7-2Z | - | - | - | 0 | + | + |
| 5X | + | + | + | + | 0 | + |
| 7X | - | + | - | + | + | 0 |
Note: "+" indicates no antagonism; "-" indicates antagonism;
4 establishment of efficient degradation model of strain
4.1 materials and methods
Green tangerine orange peel culture medium: peptone 2g, yeast extract 0.5g, K2HPO4 1g、MgSO4·7H2O 0.5g、NaCl 0.5g、CaCl20.2g, PH 7.0, 1000mL of distilled water; adding 100mL of culture medium into each triangular flask, weighing 0.5g of walnut green husk powder into each triangular flask respectively, and sterilizing at 121 deg.C for 20 min;
inoculating strains into the culture medium, culturing at 28 ℃ for 72h, and measuring the weight loss rate of the green tangerine peel; the method comprises the following steps: drying the filter paper at 105 ℃ to constant weight, and recording the weight of the filter paper; centrifuging the bacterial liquid at 5000r/min, pouring out supernatant, washing with mixed solution of acetic acid and nitric acid to eliminate thallus (prepared by mixing acetic acid and nitric acid solution with 150mL of 80% acetic acid and 15mL of concentrated nitric acid), centrifuging, washing with clear water until the pH is neutral, filtering with filter paper with known weight, drying at 105 ℃, weighing, and calculating weight loss and weight loss rate;
4.2 results and analysis
4.2.1 determination of Green Tangerine Peel degradation Effect of Individual Strain
The degradation effect of the single strain on green tangerine peels is shown in table 9; as can be seen from Table 9, the weight loss ratios of the green seedcase of 4Z, 17Z, 5X and 7X are higher, namely 18.34%, 15.28%, 14.40% and 14.36%, so that the bacterial strains have good degradation effects on the green seedcase of the walnut;
TABLE 9 weight loss ratio of walnut green husk of single strain
| Strain numbering | Green husk weight loss (%) |
| 4Z | 18.34 |
| 5X | 14.40 |
| 7X | 14.36 |
| 17Z | 15.28 |
5.2.2 establishment of efficient degradation model of Strain
Performing antagonism test to obtain strains without antagonism, and combining the strains to establish 5 combinations; inoculating the screened strain without antagonism into a liquid culture medium, carrying out shake culture on fungi by using a marting liquid culture medium and bacteria by using a beef extract peptone culture medium at 30 ℃ and 130rpm for 5 d; accurately weighing 2g of walnut green husks dried at 105 ℃ for 2h (numbering each bottle, recording the weighed amount m, and accurately weighing 4 positions after counting a decimal point), adding the green husks into triangular bottles, adding 20mL of bacterial suspension into each triangular bottle (10 mL of each strain if the number is 2), culturing at the constant temperature of 28 ℃, preparing tin pans for weighing (m1) after 20d, preparing filter paper for weighing (m2), pouring the culture in the triangular bottles into the tin pans weighed at first, repeatedly washing the rest part in the bottles by using distilled water, filtering by using the filter paper with weighed weight, putting the filter paper and filter residues into the tin pans, drying at 105 ℃ to constant weight (generally drying for more than 3 h), then weighing the filter paper and the filter residues in the pans (m3), and calculating the weight loss rate; control was 20mL of sterile water, 3 replicates per treatment setup; comparing the degradation effect of a plurality of groups of compound strains on the walnut green husk through the weight loss rate;
TABLE 10 combination pattern of strains and weight loss rate of walnut green husk
| Combined strain | Weight loss w (%) |
| 5X+7X | 19.72 |
| 5X+17Z | 26.27 |
| 5X+4Z | 18.48 |
| 7X+17Z | 23.31 |
| 5X+7X+17Z | 21.49 |
| Sterility control | 13.76 |
As can be seen from Table 10, the combination of 5X and 17Z showed the best degradation of green tangerine peel with a degradation rate of 26.27%, the combination of 7X +17Z and the combination of 5X +7X +17Z with a degradation rate of 23.31% and 21.49, and therefore, these three combinations were selected as green tangerine peel fermentors for the compost fermentation test.
5 walnut Green husk compost test
5.1 materials and methods
Walnut green husk powder, vermiculite, peat soil, ferrous sulfate and urea.
And (3) determination of humic acid content: the content of humic acid in the organic fertilizer is measured by adopting the standard of DB 21/T1322-2004 determination of humic acid content in the organic fertilizer;
because the walnut green seedcase contains a large amount of phenols and flavonoids and has certain bactericidal activity, a composting test without adding bacteria to the walnut green seedcase is carried out in advance, and the fermentation condition of the walnut green case under the condition of adding no degradation bacteria is observed; mixing the materials according to the formula in Table 11Mixing the skin with various materials, and placing into a container with a diameter of 1m3Fermenting in a foam box for a small test, adjusting the water content to 60-70%, adjusting the C: N to 25:1, turning the pile once every week, and observing and recording the change conditions of temperature, PH, humic acid, microorganisms and the like;
TABLE 11 fermentation formula of walnut green husk
| Recipe number | The contents of the formula |
| Formulation I | Walnut green husk: peat soil: vermiculite: ferrous sulfate: urea 25: 5: 2.5: 1: 1 |
| Formulation II | Walnut green husk: peat soil is 1: 1 |
| Formulation III | Walnut green husk: peat soil: urea 50: 2.5: 1 |
| Formulation IV | Walnut green husk: vermiculite: urea 30: 1: 1 |
5.2 results and analysis
As can be seen from fig. 1, the first formula (walnut green husk: peat soil: vermiculite: ferrous sulfate: urea: 25: 5: 2.5: 1: 1) has a good temperature rise effect, the temperature can reach more than 60 ℃ on the 5 th day of fermentation, and the second formula (walnut green husk: vermiculite: urea: 30: 1: 1) has a similar change trend to the first formula, but the fermentation temperature is not higher than the first formula; as can be seen from figure 2, the humic acid change in the formula I is firstly reduced and then increased, and finally reaches 8.929 percent, which meets the index that the content of humic acid in the organic fertilizer is more than 8 percent.
Drawings
FIG. 1: the fermentation temperature of the walnut green seedcase is changed in the walnut green case composting test;
FIG. 2: in the walnut green husk compost test, the content of humic acid in the formula I is changed;
FIG. 3: the temperature change condition of the walnut green husk biological organic fertilizer in the embodiment 1-3 in the natural fermentation process;
FIG. 4: the walnut green husk bio-organic fertilizer of the embodiment 1-3 contains tax rate change situation in the natural fermentation process;
FIG. 5: the change condition of the acid-base degree of the walnut green husk biological organic fertilizer in the natural fermentation process of the embodiment 1-3;
FIG. 6: the EC value of the walnut green husk biological organic fertilizer in the examples 1-3 changes in the natural fermentation process.
Detailed Description
The following is a detailed description of the embodiments of the present invention, but the present invention is not limited to these embodiments, and any modifications or substitutions in the basic spirit of the embodiments are included in the scope of the present invention as claimed in the claims.
1. Determination of physical and chemical indexes of soil and walnut green husk
1.1 materials and methods
Materials: taking a soil sample of a Lurong county Xiuyihu village passion fruit planting base in Zhefeng county, walnut green skin, macadamia nut skin, fungus stick, rice bran, oil cake and wood dust for detection;
the method comprises the following steps: adopting a 5-point Z-shaped sampling method to sample soil in a passion fruit planting base of Lucapacity countryside Xiu village in Zhengfeng county, uniformly mixing the soil sample, and determining the conditions of nitrogen, phosphorus, potassium, organic matters, PH, water content and the like in the soil sample according to a forestry industry standard LY/T1270-: LY/T1228-2015 forest soil nitrogen determination; LY/T1232-2015 determination of forest soil phosphorus; LY/T1234-2015 determination of forest soil potassium; determining the PH value of the forest soil by LY/T1239-1999; LY/T1237-1999 forest soil organic matter determination and carbon-nitrogen ratio calculation; LY/T1215-1999 determination of forest soil moisture-physical properties;
the nutrient content of the walnut green seedcase and other raw materials is detected by adopting the following method: the total nitrogen is measured according to a Kjeldahl method for measuring crude protein in GB/T6432-2018 feed and a method for measuring protein in GB-5009.5-2016 food safety national standard food; measuring total phosphorus by spectrophotometry according to soil agro-chemical analysis method, and measuring total potassium by atomic absorption method according to soil agro-chemical analysis method; the lignin, cellulose and hemicellulose are measured according to the measurement method of acidic washed lignin (ADL) in GBT 20805-;
1.2 results and analysis
1.2.1 soil test results of passion fruit planting base
Determining the nutrient condition of the passion fruit planting soil, wherein the content of the nutrient is shown in table 12; according to the judgment of the national soil nutrient content grading standard table, the content of organic matters in the passion fruit soil is moderate, the content of total nitrogen and alkaline hydrolysis nitrogen is low, the content of total phosphorus and available phosphorus is extremely low, and the content of total potassium and quick-acting potassium is high.
TABLE 12 Passion fruit soil nutrient content
1.2.2 measurement results of walnut Green husk nutrient
The nutrient conditions of the walnut green seedcase and other raw materials are determined, and the content of the nutrients is shown in table 13; as can be seen from Table 13, the content of potassium in the green husk of walnut is the highest, reaching 49.11g/kg, the content of nitrogen is moderate, and the content of phosphorus is low; the oil cake has high nitrogen content and phosphorus content, and can complement with pericarpium Citri Reticulatae viride to balance nitrogen, phosphorus, and potassium components required by plants. In the three-element determination, the highest lignin content of the walnut green husk reaches 30.12%, which is similar to the literature report, while the cellulose content is 24.60%, which is in a medium level, and the hemicellulose content is lower, which is only 4.36%, therefore, in order to rapidly degrade the walnut green husk, a strain capable of degrading lignin and cellulose needs to be screened.
TABLE 13 determination of nutrient content of raw materials
Example 1
A walnut green husk biological organic fertilizer is prepared by mixing 4kg of rice bran, 5kg of sawdust, 6kg of fungus sticks, 20kg of walnut green husks, 10kg of oil cake and 5kg of macadamia nut peels, adding a microbial suspension liquid with the functions of degrading cellulose and lignin into raw materials according to the proportion of 10ml of suspension liquid per gram of walnut green husks, naturally fermenting for 62 days, and turning over once a week in the natural fermentation process;
the microorganism with the function of degrading cellulose and lignin is a mixed bacterium of a leucobacter Leucobarater sp and a Fungal endophytic sp in equal mass ratio;
preparing the suspension liquid: inoculating 10ml of microbial strains with the functions of degrading cellulose and lignin into a liquid culture medium, and carrying out shake culture at 30 ℃ and 130rpm for 5 d; the liquid culture medium is determined according to that microbial strains with functions of degrading cellulose and lignin are fungi and bacteria, a martin liquid culture medium is adopted when the strains are the fungi, and a beef peptone culture medium is adopted when the strains are the bacteria; the Martin's medium: k2HPO4 1.0g，MgSO4·7H20.5g of O, 5g of peptone, 10g of glucose and 1000mL of water, and sterilizing at 121 ℃ for 20 min; the beef extract peptone culture medium: 3.0g of beef extract, 10g of peptone, 5.0g of NaCl, 15-25g of agar and 1000mL of water, wherein the pH value is 7.4-7.6, the volume is adjusted to 1000mL by using distilled water, and the beef extract is sterilized at 121 ℃ for 20 min.
Example 2
A walnut green husk biological organic fertilizer is prepared by mixing 8kg of rice bran, 10kg of sawdust, 12kg of fungus sticks, 35kg of walnut green husks, 25kg of oil cake and 10kg of macadamia nut peels, adding a microbial suspension liquid with the functions of degrading cellulose and lignin into raw materials according to the proportion of 10ml of suspension liquid per gram of walnut green husks, naturally fermenting for 62 days, and turning over once a week in the natural fermentation process;
the microorganism with the function of degrading cellulose and lignin is a mixed bacterium of Stenotrophomonas rhizophila and Fungal endophyte Fungal endopyte sp in equal mass ratio;
preparing the suspension liquid: inoculating 10ml of microbial strains with the functions of degrading cellulose and lignin into a liquid culture medium, and carrying out shake culture at 30 ℃ and 130rpm for 5 d; the liquid culture medium is determined according to that microbial strains with functions of degrading cellulose and lignin are fungi and bacteria, a martin liquid culture medium is adopted when the strains are the fungi, and a beef peptone culture medium is adopted when the strains are the bacteria; the Martin's medium: k2HPO4 1.0g，MgSO4·7H20.5g of O, 5g of peptone, 10g of glucose and 1000mL of water, and sterilizing at 121 ℃ for 20 min; the beef extract peptone culture medium: 3.0g of beef extract, 10g of peptone, 5.0g of NaCl, 15-25g of agar and 1000mL of water, wherein the pH value is 7.4-7.6, the volume is adjusted to 1000mL by using distilled water, and the beef extract is sterilized at 121 ℃ for 20 min.
Example 3
A walnut green husk biological organic fertilizer is prepared by mixing 8kg of rice bran, 10kg of sawdust, 12kg of fungus sticks, 30kg of walnut green husks, 30kg of oil cakes and 10kg of macadamia nut peels, adding a microbial suspension liquid with the functions of degrading cellulose and lignin into raw materials according to the proportion of 10ml of suspension liquid per gram of walnut green husks, naturally fermenting for 62 days, and turning over once a week in the natural fermentation process;
the microorganism with the function of degrading cellulose and lignin is a mixed bacterium of white bacillus Leucobarater sp, Stenotrophomonas rhizophila Stenotrophorus rhizophila and Fungal endophytic sp in equal mass ratio;
preparing the suspension liquid: inoculating 10ml of microbial strains with the functions of degrading cellulose and lignin into a liquid culture medium, and carrying out shake culture at 30 ℃ and 130rpm for 5 d; the liquid culture medium rootAccording to the method, a microorganism strain with the functions of degrading cellulose and lignin is fungus and bacteria, a Martin liquid culture medium is adopted when the strain is fungus, and a beef peptone culture medium is adopted when the strain is bacteria; the Martin's medium: k2HPO4 1.0g，MgSO4·7H20.5g of O, 5g of peptone, 10g of glucose and 1000mL of water, and sterilizing at 121 ℃ for 20 min; the beef extract peptone culture medium: 3.0g of beef extract, 10g of peptone, 5.0g of NaCl, 15-25g of agar and 1000mL of water, wherein the pH value is 7.4-7.6, the volume is adjusted to 1000mL by using distilled water, and the beef extract is sterilized at 121 ℃ for 20 min.
In the examples 1-3, the temperature, humidity, pH and moisture content change conditions of the walnut green husk biological organic fertilizer (sequentially marked as No. 1, No. 2 and No. 3) are shown in the figures 3-6; as can be seen from FIG. 3, the temperature change trends of the compost No. 1, No. 2 and No. 3 are similar, the temperature reaches more than 50 ℃ from 31d, and the temperature above 50 ℃ can be maintained for 14d, so that the fermentation is relatively thorough and the organic fertilizer maturity standard is reached; as can be seen from FIG. 4, the moisture content of the composts No. 1 and No. 3 has the same trend, and the change is smaller firstly, while the moisture content of the compost No. 2 is reduced firstly and then increased, and the change trend of the compost No. 3 is consistent finally, which is probably caused by the activity of microorganisms in the composts; as can be seen from FIG. 5, the pH values of the composts No. 1 and No. 2 change to decrease first and then increase, the composts No. 3 increase first and then decrease, the change trends are consistent, and finally the pH values of the 3 composts are all between 7 and 8, which meet the pH value correlation regulation in the biofertilizer standard; as can be seen from FIG. 6, the compost Nos. 1, 2 and 3 showed consistent tendency of early change, and slightly different late change, but the final EC value was between 5 and 7 ms/mL.
Example 4
A walnut green husk biological organic fertilizer is prepared by mixing 6kg of rice bran, 7kg of sawdust, 9kg of fungus sticks, 25kg of walnut green husks, 20kg of oil cake and 8kg of macadamia nut peels, adding a microbial suspension liquid with the functions of degrading cellulose and lignin into raw materials according to the proportion of 10ml of suspension liquid per gram of walnut green husks, naturally fermenting for 60-65 days, and turning over once a week in the natural fermentation process;
the microorganism with the function of degrading cellulose and lignin is a mixed bacterium of a leucobacter Leucobarater sp and a Fungal endophytic sp in equal mass ratio;
preparing the suspension liquid: inoculating 3ml of microbial strains with the functions of degrading cellulose and lignin into a liquid culture medium, and carrying out shake culture at 30 ℃ and 130rpm for 5 d; the liquid culture medium is determined according to that microbial strains with functions of degrading cellulose and lignin are fungi and bacteria, a martin liquid culture medium is adopted when the strains are the fungi, and a beef peptone culture medium is adopted when the strains are the bacteria; the Martin's medium: k2HPO4 1.0g，MgSO4·7H20.5g of O, 5g of peptone, 10g of glucose and 1000mL of water, and sterilizing at 121 ℃ for 20 min; the beef extract peptone culture medium: 3.0g of beef extract, 10g of peptone, 5.0g of NaCl, 15-25g of agar and 1000mL of water, wherein the pH value is 7.4-7.6, the volume is adjusted to 1000mL by using distilled water, and the beef extract is sterilized at 121 ℃ for 20 min.
Example 5
A walnut green husk biological organic fertilizer is prepared by mixing 5kg of rice bran, 8kg of sawdust, 10 parts by weight of fungus sticks, 25kg of walnut green husks, 15kg of oil cake and 7kg of macadamia nut peels, adding a microbial suspension liquid with the functions of degrading cellulose and lignin into the raw materials according to the proportion of 8ml of suspension liquid per gram of walnut green husks, naturally fermenting for 62 days, and turning over once a week in the natural fermentation process;
the microorganism with the function of degrading cellulose and lignin is a mixed bacterium of a leucobacter Leucobarater sp and a Fungal endophytic sp in equal mass ratio;
preparing the suspension liquid: inoculating 15ml of microbial strains with the functions of degrading cellulose and lignin into a liquid culture medium, and carrying out shake culture at 30 ℃ and 130rpm for 5 d; the liquid culture medium is determined according to that microbial strains with functions of degrading cellulose and lignin are fungi and bacteria, a martin liquid culture medium is adopted when the strains are the fungi, and a beef peptone culture medium is adopted when the strains are the bacteria; the Martin's medium: k2HPO4 1.0g，MgSO4·7H20.5g of O, 5g of peptone, 10g of glucose and 1000mL of water, and sterilizing at 121 ℃ for 20 min; the beef extract eggWhite peptone medium: 3.0g of beef extract, 10g of peptone, 5.0g of NaCl, 15-25g of agar and 1000mL of water, wherein the pH value is 7.4-7.6, the volume is adjusted to 1000mL by using distilled water, and the beef extract is sterilized at 121 ℃ for 20 min.
Example 6
The produced walnut green husk biological organic fertilizer is tested and demonstrated in Luo-Li-Xiu-village Luo family Baixiang orchard (N:25.393 degrees, E:105.788 degrees and elevation 475.5m) in Zhengfeng county in 4 months in 2021, the farmyard manure and the walnut green husk biological organic fertilizer are used as a reference, the walnut green husk biological organic fertilizer in the example 1-3 is taken, 200g of functional microbial agent (shown in Table 14) is added and uniformly mixed, the obtained substance is used as base fertilizer and is applied to soil, and after ploughing, the mixture is placed for 7 days, and then the passion fruit seedlings can be planted;
surface 14 functional microorganism and walnut green husk biological organic fertilizer combination
| Trichoderma harzianum (a) | Bacillus subtilis (b) | Bacillus megaterium (c) | |
| Example 1(A) | Aa | Ab | Ac |
| Example 2(B) | Ba | Bb | Bc |
| Example 3(C) | Ca | Cb | Cc |
Setting 4 passion fruit seedlings in each treatment group for 3 times of repetition, performing non-fertilization treatment as CK comparison, and applying the same tending measures in the later period; the growth conditions (ground diameter, seedling height, leaf number, flowering number and fruiting number) of the passion fruit seedlings are measured once a month, and the fruiting conditions are measured at 9 months in 2021 as follows:
TABLE 15 average ground diameter (mm) growth of Passion fruit treated by each group
| | Day | 0 | 45 days | 80 days | 120 days | 150 days |
| Aa | 3.25 | 4.89 | 12.09 | 16.48 | 19.19 | |
| Ba | 3.42 | 5.04 | 11.30 | 17.12 | 18.60 | |
| Ca | 3.81 | 5.11 | 11.84 | 16.36 | 16.65 | |
| Ab | 3.19 | 5.24 | 12.98 | 18.74 | 18.83 | |
| Bb | 3.45 | 4.94 | 12.08 | 15.39 | 16.41 | |
| Cb | 3.42 | 4.89 | 12.66 | 19.00 | 20.02 | |
| Ac | 3.36 | 4.94 | 11.92 | 18.35 | 20.59 | |
| Bc | 3.58 | 5.19 | 11.87 | 17.16 | 18.11 | |
| Cc | 3.39 | 4.70 | 10.55 | 14.81 | 16.53 | |
| Farm manure | 3.11 | 5.19 | 13.48 | 18.73 | 19.45 | |
| Organic fertilizer | 3.46 | 5.05 | 10.82 | 14.75 | 15.97 | |
| Blank control | 3.77 | 4.73 | 8.73 | 15.02 | 18.66 |
TABLE 16 average plant height (cm) growth of Passion fruit treated by each group
TABLE 17 average number of leaves of passion fruit treated in each group
| | Day | 0 | 45 days | 80 days | 120 |
| Aa | |||||
| 6 | 18 | 77 | 221 | ||
| |
6 | 18 | 60 | 171 | |
| |
7 | 21 | 69 | 260 | |
| |
6 | 23 | 92 | 254 | |
| |
8 | 22 | 91 | 283 | |
| |
5 | 19 | 69 | 255 | |
| |
7 | 18 | 69 | 214 | |
| |
7 | 18 | 57 | 192 | |
| |
6 | 18 | 56 | 169 | |
| |
7 | 19 | 83 | 225 | |
| |
9 | 19 | 58 | 201 | |
| |
9 | 12 | 12 | 167 |
TABLE 18 average flowering number of Passion fruit treated by each group
| Numbering | 45 days | 80 days | 120 days |
| Aa | 3.56 | 3.33 | 12.55 |
| Ba | 4.50 | 5.33 | 6.67 |
| Ca | 5.70 | 4.64 | 6.92 |
| Ab | 3.25 | 3.50 | 14.00 |
| Bb | 3.88 | 4.50 | 9.27 |
| Cb | 4.00 | 4.00 | 5.67 |
| Ac | 4.30 | 3.50 | 4.88 |
| Bc | 5.29 | 2.38 | 5.22 |
| Cc | 4.33 | 3.80 | 3.63 |
| Farm manure | 3.43 | 3.60 | 10.33 |
| Organic fertilizer | 5.67 | 2.25 | 7.56 |
| Blank control | 5.33 | 6.78 | 8.57 |
TABLE 19 average number of passion fruit treatments for each group
| Numbering | 80 days | 120 days | 150 days |
| Aa | 1.67 | 12.55 | 82.00 |
| Ba | 1.50 | 6.67 | 97.00 |
| Ca | 2.50 | 6.92 | 66.00 |
| Ab | 1.67 | 14.00 | 93.50 |
| Bb | 3.00 | 9.27 | 93.00 |
| Cb | 1.00 | 5.67 | 76.00 |
| Ac | 1.67 | 4.88 | 87.00 |
| Bc | 2.67 | 5.22 | 46.00 |
| Cc | 2.50 | 3.63 | 52.00 |
| Farm manure | 1.80 | 10.33 | 41.33 |
| Organic fertilizer | 1.25 | 7.56 | 31.00 |
| Blank control | 0.00 | 0.00 | 27.00 |
TABLE 20 average fruit weight and length and width of passion fruit treated by each group
| Numbering | Average fruit weight (g) | Average fruit length (cm) | Average fruit width (cm) |
| Aa | 70.25 | 6.65 | 5.90 |
| Ba | 67.20 | 6.60 | 6.15 |
| Ca | 52.03 | 6.43 | 5.60 |
| Ab | 62.65 | 7.00 | 6.20 |
| Bb | 69.30 | 6.93 | 6.06 |
| Cb | 49.50 | 6.25 | 5.60 |
| Ac | 51.45 | 5.85 | 5.25 |
| Bc | 76.40 | 6.00 | 5.80 |
| Cc | 52.90 | 6.50 | 5.60 |
| Farm manure | 48.35 | 7.05 | 5.65 |
| Organic fertilizer | 59.40 | 6.10 | 5.65 |
| Blank control | 54.50 | 6.77 | 5.83 |
As can be seen from tables 15-20, the average plant diameter of the formulations Ac (20.59mm), Cb (20.04mm), farmyard manure (19.45mm) and Aa (19.91mm) was higher than that of the other treatments at 150 days; the average plant height after farmyard manure treatment at 150d was highest (284.3cm), followed by Bc (263.3cm), Bb (262.8cm) and Ac (262.43 cm); the average number of leaves treated with Bb at 150d was the highest (284 leaves), followed by Ca (260 leaves) and Cb (255 leaves); at 150d, the average number of flowers after Ab treatment was the highest (14), followed by Aa (12.55) and farmyard manure (10.33), and Bb treatment was 9.27; the number of average strains after Ba treatment at 150d was the largest (97), followed by Ab (93.5), Bb (93) and Ac (87); as can be seen from the above table, at 150d, the difference in average fruit length and width was not significant for each treatment, with the average fruit weight of Bc being the heaviest (76.4g), followed by Aa (70.3g), Bb (69.3g) and Ba (67.2 g); since the passion fruit blooms and fruits for many times a year, the fertilizer effect of the passion fruit cannot be accurately explained by using yield data of one time alone, therefore, the research integrates multiple factors such as the growth state of the plant, the number of blossoms and fruits and the like, the obtained Bb, Ac and Cb formulas have strong growth promoting effect on the plant, the Aa, Ab, Ba and Bb have great influence on the blossoming and fruits of the plant, and the combination is superior to that of a farmyard manure and organic fertilizer treatment group.
Example 7
The materials obtained from the formulas in Table 14 in example 6 are extracted according to a water-fertilizer ratio of 2:1, after filtration, filtrate is taken and centrifuged for 20min, supernatant is absorbed and distilled water with the concentration of 100%, 75% and 50% in corresponding proportion is mixed, 2mL of the supernatant is respectively absorbed and placed in a culture dish paved with filter paper, 10 radish seeds are placed in the culture dish, and the radish seeds are cultured in a constant temperature incubator at 25 ℃ for 72h in the dark, and meanwhile, distilled water control is carried out. Then calculating the seed germination potential of each concentration by using the following formula, and finally taking the average of 3 concentrations as the seed germination potential of the sample;
TABLE 21 average germination potential of the treated radish seeds of each group
The germination vigor measurement results of radish seeds of each group are shown in table 21; therefore, the following steps are carried out: after 72 hours of treatment, the germination vigor of the formulas Ba, Bc and Ca is higher and is more than 100%, the germination rates of Ca and Cb are 100%, the average bud length of Ba is the longest and is 1.79cm, and Bc and Ca are 1.44 cm and 1.43cm respectively and are more than the bud length of CK control; therefore, Ba, Bc and Ca formulations have a promoting effect on the germination of seeds.
Claims (10)
1. A walnut green husk biological organic fertilizer is characterized by being prepared by mixing 4-8 parts by weight of rice bran, 5-10 parts by weight of sawdust, 6-12 parts by weight of mushroom sticks, 20-35 parts by weight of walnut green husks, 10-30 parts by weight of oil cakes and 5-10 parts by weight of macadamia nut peels, adding a microbial suspension liquid with cellulose and lignin degrading functions into raw materials according to the proportion of 8-10 ml of the suspension liquid per gram of the walnut green husks, naturally fermenting for 60-65 days, and turning over piles once a week in the natural fermentation process.
2. The walnut green seedcase bio-organic fertilizer as claimed in claim 1, wherein the microorganism having the function of degrading cellulose and lignin is any one or more of Leuconobater sp, Stenotrophomonas rhizophila, Fusarium solani, Penicillium rolfsli, and Fungal endophytic sp.
3. The walnut green husk bio-organic fertilizer as claimed in claim 1 or 2, wherein the microorganism having the function of degrading cellulose and lignin is any one of the following compositions:
(1) mixing the leucobacter sp and the Fungal endophytic sp in any ratio;
(2) a mixture of Stenotrophomonas rhizophila and Fungal endophyte sp in any ratio;
(3) a mixture of the white bacillus Leucobater sp, Stenotrophomonas rhizophila Stenotrophora rhizophila and Fungal endophyte sp in any ratio.
4. The walnut green seedcase bio-organic fertilizer of claim 1, which is characterized in that the suspension liquid is prepared by the following steps: inoculating the microbial strain with the function of degrading cellulose and lignin into a liquid culture medium, and performing shake culture at 30 ℃ and 130rpm for 5 d.
5. The walnut green seedcase bio-organic fertilizer as claimed in claim 4, wherein the liquid culture medium is determined according to whether the microbial strains with the functions of degrading cellulose and lignin are fungi and bacteria, and is a Martin liquid culture medium when the strains are fungi and is a beef peptone culture medium when the strains are bacteria.
6. The walnut green seedcase bio-organic fertilizer as claimed in claim 5, wherein the Martinus culture medium: k2HPO41.0g，MgSO4·7H20.5g of O, 5g of peptone, 10g of glucose and 1000mL of water, and sterilizing at 121 ℃ for 20 min.
7. The walnut green seedcase bio-organic fertilizer as claimed in claim 5, wherein the beef extract peptone medium: 3.0g of beef extract, 10g of peptone, 5.0g of NaCl, 15-25g of agar and 1000mL of water, wherein the pH value is 7.4-7.6, the volume is adjusted to 1000mL by using distilled water, and the beef extract is sterilized at 121 ℃ for 20 min.
8. The application of the walnut green husk bio-organic fertilizer as claimed in any one of claims 1 to 7 in promoting radish seed germination, which comprises the following steps: taking a walnut green husk biological organic fertilizer, adding a functional microbial agent according to the amount of 0.57-1% of the mass of the walnut green husk, uniformly mixing to obtain a combined fertilizer, leaching according to the water-fertilizer ratio of 2:1, filtering, centrifuging the filtrate for 20min, absorbing the supernatant, adding distilled water to prepare a leaching solution with the volume concentration of 50-100%, absorbing 2mL of the leaching solution, placing the leaching solution in a culture dish paved with filter paper, placing 10 radish seeds in a constant-temperature incubator at 25 ℃ and culturing in the dark for 72 h.
9. The application of the walnut green husk bio-organic fertilizer as claimed in any one of claims 1 to 7 in passion fruit planting, which comprises the following steps: taking walnut green husk biological organic fertilizer, adding functional microbial agent according to the amount of 0.57-1% of the mass of the walnut green husk, uniformly mixing, using the obtained product as base fertilizer, applying the base fertilizer into soil, turning over, and standing for 7 days to plant passion fruit seedlings.
10. The use according to claim 8 or 9, wherein the functional microbial agent is any one or more of trichoderma harzianum, bacillus subtilis and bacillus megaterium.
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| CN102976803A (en) * | 2012-12-12 | 2013-03-20 | 中国科学院南京土壤研究所 | Additive for accelerating crop straws to efficiently maturate and application thereof |
| CN105347918A (en) * | 2015-11-03 | 2016-02-24 | 梁淑娟 | Method for preparing bio-organic fertilizer from walnut green seedcase |
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| CN102976803A (en) * | 2012-12-12 | 2013-03-20 | 中国科学院南京土壤研究所 | Additive for accelerating crop straws to efficiently maturate and application thereof |
| CN105347918A (en) * | 2015-11-03 | 2016-02-24 | 梁淑娟 | Method for preparing bio-organic fertilizer from walnut green seedcase |
| CN106495879A (en) * | 2016-10-26 | 2017-03-15 | 山东省葡萄研究院 | A kind of preparation method of Exocarpium Juglandis Immaturus residue organic fertilizer |
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