CN103468592A - Fungus with straw degradation capability and application thereof in composting - Google Patents
Fungus with straw degradation capability and application thereof in composting Download PDFInfo
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- CN103468592A CN103468592A CN2013104432226A CN201310443222A CN103468592A CN 103468592 A CN103468592 A CN 103468592A CN 2013104432226 A CN2013104432226 A CN 2013104432226A CN 201310443222 A CN201310443222 A CN 201310443222A CN 103468592 A CN103468592 A CN 103468592A
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- penicillium oxalicum
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- 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
Abstract
The invention discloses a fungus with straw degradation capability and application thereof in composting and relates to the technical field of biological environmental protection. The straw degradation fungus JSD-17 is penicillium oxalicum (Penicillium oxalicum) and has a preservation number of CGMCC No.7743; the straw degradation fungus is further used for fermenting and composting agricultural waste straw. The penicillium oxalicum disclosed by the invention not only has strong cellulose enzyme synthesis capability, but also has the capability of decomposing hemicellulose and pectin components in the straw. By adopting the fungus disclosed by the invention, laccase (Lac), lignin peroxidase (Lip) and manganese peroxidase (Mnp) can be synthesized; the degradation speed of lignin components in the straw can be greatly increased.
Description
Technical field
What the present invention relates to is a kind of method of biological environmental production technical field, and specifically a kind of have the fungi of straw degradative ability and an application in compost thereof.
Background technology
China is large agricultural country, and annual all kinds of agricultural crop straw output is 700,000,000 tons of left and right.Since the later stage nineties 20th century, along with crop yield improves, the stalk total amount increases sharply, and vast farmers is for catching up with farming season, rob sowing, following the line of least resistance, and most areas just start to occur the crop straw burning phenomenon.The big area crop straw burning has not only caused great environmental pollution, and has wasted valuable resource.
The stalk main ingredient is that Mierocrystalline cellulose, xylogen, pectin, hemicellulose, crude protein and ash grade.Mierocrystalline cellulose in stalk, pectin, especially the lignin component structure is very stable, very slow in the occurring in nature degraded.Microorganism in environment has the characteristics such as rate of propagation is fast, feature richness is various, strong adaptability, and microbial method decomposition Treating straw has the irreplaceable advantage of many other physico-chemical processes, has a extensive future.Fungi generally has relatively complete enzyme system, more thorough to the degraded of stalk, therefore, screens efficient straw degradation bacteria, particularly fungi from nature just particularly important.
Through the retrieval to prior art, find, Chinese patent literature CN102559506, open day 2012-07-11 open day, discloses a kind of Penicillium fungal bacterial strain---penicillium oxalicum (Penicillium oxalicum) M.This bacterium can be turned out in a large number at short notice, the cellulose degrading enzyme of homogeneous produces bacteria culture fluid.This technology also provides the method for utilizing this bacterial strain fermentation culture production endoglucanase and exoglucanase in the liquid culture medium, and utilizes this strain fermentation to produce the method for enzyme glycolysis stalk cellulose.But this technology can't realize the degraded to pectin in stalk and hemicellulose component.
Summary of the invention
The present invention is directed to the prior art above shortcomings, a kind of the have fungi of straw degradative ability and the application in compost thereof are proposed, this bacterium not only has stronger cellulase synthesis capability, also has the capacity of decomposition of hemicellulose and pectin fraction in stalk simultaneously; Bacterial strain of the present invention can synthesize laccase (Lac), lignin peroxidase (Lip) and manganese peroxidase (Mnp), has greatly accelerated the degradation speed to lignin component in stalk.
The present invention is achieved by the following technical solutions:
The present invention relates to a strain straw degradative fungi JSD-17 is penicillium oxalicum (Penicillium oxalicum), and deposit number is CGMCC No.7743.
Described straw degradative fungi is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), and address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Chinese science institute of microbiology; Date saved is on June 19th, 2013, and deposit number is CGMCC No.7743; Taxonomic species is called penicillium oxalicum (Penicillium oxalicum).
The present invention relates to the application of a kind of straw degradative fungi in straw degradative, be specially: described straw degradative fungi is gone forward side by side for the treatment of agricultural wastes straw, and windrow is fertile to ferment.
Prepared in the following manner by described straw compost: by the penicillium oxalicum spore inoculating in the PD substratum, 32 ℃, 150r/min shake flask fermentation; Collect bacterium liquid after 72h, every liter of bacterium liquid and about 1.0-1.5kg rice straw mix, and hatch 24h afterwards in 30 ℃ of incubators.
Described fermentation concrete steps are: rice straw is crushed to 5-10cm through disintegrating apparatus, forms stalk section, press stalk weight 1.5% and add compost bacterium, regulate heap body water-content in 65% left and right; Measure heap temperature every day, just started heap temperature and can rise to rapidly 70 ℃, and maintained 1-2 days, when temperature no longer rises and drops to gradually below 45 ℃, be described the one time fermentation cycle, correspondence is carried out turning, i.e. every 7-8 days turning 1 time, and turning is 3 times altogether; After turning for the third time, complete fermentation.
Described degraded is preferably the cellulosic component in stalk.
Described straw degradative fungi carries out preservation in the following manner: adopt 4 ℃ of inclined-plane freezing methods, make microorganism in lesser temps, thereby reach the purpose of preservation of bacteria strain to reduce its metabolism speed.
Technique effect
Compared with prior art, advantage of the present invention comprises:
1, the present invention not only has stronger cellulase synthesis capability, also has the capacity of decomposition of hemicellulose and pectin fraction in stalk simultaneously;
2, the present invention can synthesize laccase (Lac), lignin peroxidase (Lip) and manganese peroxidase (Mnp), has greatly accelerated the degradation speed to lignin component in stalk;
3, the present invention can improve the biological organic fertilizer organic content, has using value well.
The accompanying drawing explanation
Fig. 1 is penicillium oxalicum lithograph of the present invention.
Fig. 2 is penicillium oxalicum 18S rDNA electrophorogram of the present invention.
Embodiment
Below embodiments of the invention are elaborated, the present embodiment is implemented take technical solution of the present invention under prerequisite, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
Embodiment 1
The separation of bacterial strain, purifying and preservation
Getting rotten stalk and the soil of handing over large Pujiang base rice terrace in Minhang District, Shanghai Pujiang town is sample.Adopt the shaking flask enrichment culture, different samples are got 10g after grinding and being mixed, and are placed in the 100mL enrichment and support base, 30 ℃, 150rpm constant temperature culture.Every 5 days, access another enrichment with 10% inoculum size and support in base, so repeatedly tame 4 times.Enrichment medium adopts take the substratum that CMC-Na is sole carbon source.
Get 100 μ L pregnant solutions and be coated with on the Congo red culture medium flat plate of CMC-, be placed in 28 ℃ of fixed temperature and humidity incubators and be inverted cultivation, observed after 5 days.The larger thalline of picking hydrolytic circle, be inoculated on the PDA substratum, until obtain single bacterium colony.Single bacterium colony that will obtain again, passed for 4 generations continuously take on the inorganic salt solid medium that CMC-Na is sole carbon source, after colonial morphology is stable, obtains pure culture, as shown in Figure 1.
The formula of described inorganic salt solid medium is: peeling potato 200g, glucose 20g, agar 30-35g, distilled water 1000mL.
Embodiment 2
The evaluation of penicillium oxalicum (Penicillium oxalicum) bacterial strain
In conjunction with Physiology and biochemistry, identify and 18S rDNA ITS sequence alignment analysis (shown in Fig. 2), identify that this bacterium is mould, taxonomic species is called penicillium oxalicum (Penicillium oxalicum).
Naming this bacterial strain is JSD-17, on June 19th, 2013, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Chinese science institute of microbiology, deposit number is CGMCC No.7743.
Embodiment 3
Penicillium oxalicum (Penicillium oxalicum) inulinase-producing activity is measured
Adopt the DNS method, measure the activity of optimizing in secondary fermentation liquid two kinds of cellulase CMCase and FPase and be respectively 72.69U/mL and 40.55U/mL; The activity of measuring polygalacturonase and zytase is 5.86U/mL and 3.26U/mL.Wherein, an enzyme unit alive (U) is the enzyme amount that every milliliter of enzyme liquid catalyzed reaction thing of per minute generates 1 μ g monose product.
Respectively with ABTS, veratryl alcohol, Mn
2+for reaction substrate, adopt spectrophotometry, measure laccase (Lac), lignin peroxidase (Lip) and manganese peroxidase (Mnp) activity in fermented liquid.After measured, the activity of three kinds of enzymes is respectively 15.8U/L, 1.94U/L, 5.54U/L.Define per minute oxidation 1 μ mol substrate (ABTS, veratryl alcohol, Mn herein
2+) be enzyme unit (U) alive.
Visible, penicillium oxalicum of the present invention has stronger cellulase, hemicellulase, polygalacturonase is synthetic and secretion capacity, there is in addition certain lignin degradation ability, can not have a negative impact to the nature carbon cycle, can be to the environment embodiment 4 that works the mischief
The preparation of penicillium oxalicum (Penicillium oxalicum) compost bacterium
By the spore inoculating of described CGMCC No.7743 penicillium oxalicum in the PD substratum, 32 ℃, 150r/min shake flask fermentation.Collect bacterium liquid after 72h, every liter of bacterium liquid and about 1.0-1.5kg rice straw mix, and hatch 24h afterwards in 30 ℃ of incubators.
The formula of described PD substratum is: peeling potato 200g, glucose 20g, agar 15g, distilled water 1000mL.
Embodiment 5
Compost bacterium is applied in the rice straw compost
Rice straw is crushed to 5-10cm through disintegrating apparatus, forms stalk section, press stalk weight 1.5% and add compost bacterium, regulate heap body water-content in 65% left and right.Measure heap temperature every day, just started heap temperature and can rise to rapidly 70 ℃, and maintained 1-2 days, when temperature no longer rises and drops to gradually below 45 ℃, be described the one time fermentation cycle, carry out turning.Within approximately every 7-8 days, turning is 1 time, and turning is 3 times altogether.Treat that turning completes for the third time, just obtain the high-efficiency rice straw biologic organic fertilizer.
Embodiment 6
Biological organic fertilizer pH and organic index determining
Get the biological organic fertilizer sample, measure pH and organic content.Sample and water are measured the pH value with pH meter after pressing the 1:5 dilution; In sample, organic content is measured by potassium bichromate titrimetric method.After measured, the biological organic fertilizer sample pH value is 7.53, and the contrast of not adding microbial inoculum is 6.87; The contrast organic content is for being 15.71%, and in biological organic fertilizer, organic content, up to being 21.34%, exceeds 35.8% than contrast.To sum up, add the processing of penicillium oxalicum microbial inoculum, on pH and organic index, all be better than control treatment.
Claims (7)
1. a straw degradative fungi, is characterized in that, this straw degradative fungi JSD-17 is penicillium oxalicum (Penicillium oxalicum), and deposit number is CGMCC No.7743, and date saved is on June 19th, 2013.
2. the application of straw degradative fungi in straw degradative according to claim 1, is characterized in that, described straw degradative fungi is carried out to compost fermentation for the treatment of agricultural wastes straw.
3. application according to claim 2, is characterized in that, prepared in the following manner by described straw compost: by described CGMCC No.7743 penicillium oxalicum spore inoculating in the PD substratum, 32 ℃, 150r/min shake flask fermentation; Collect bacterium liquid after 72h, every liter of bacterium liquid and about 1.0-1.5kg rice straw mix, and hatch 24h afterwards in 30 ℃ of incubators.
4. application according to claim 2, is characterized in that, described fermentation concrete steps are: rice straw is crushed to 5-10cm through disintegrating apparatus, forms stalk section, press stalk weight 1.5% and add compost bacterium, regulate heap body water-content 65%; Measure heap temperature every day, just started heap temperature and can rise to rapidly 70 ℃, and maintained 1-2 days, when temperature no longer rises and drops to gradually below 45 ℃, be described the one time fermentation cycle, correspondence is carried out turning, i.e. every 7-8 days turning 1 time, and turning is 3 times altogether; After turning for the third time, complete fermentation.
5. application according to claim 2, is characterized in that, described degraded is mainly the cellulosic component in stalk.
6. application according to claim 2, it is characterized in that, described straw degradative fungi carries out preservation in the following manner: adopt 4 ℃ of inclined-plane freezing methods, make microorganism in lesser temps, thereby reach the purpose of preservation of bacteria strain to reduce its metabolism speed.
7. application according to claim 3, is characterized in that, the formula of described PD substratum is: peeling potato 200g, glucose 20g, agar 15g, distilled water 1000mL.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105801210A (en) * | 2016-03-09 | 2016-07-27 | 南京农业大学 | Method for directly preparing fertilizer by using straws and prepared straw organic fertilizer |
| CN109054843A (en) * | 2018-08-03 | 2018-12-21 | 北京农业生物技术研究中心 | A kind of soil conditioner and preparation method and its application as pesticide synergistic agent |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101580853A (en) * | 2008-03-06 | 2009-11-18 | 王宝维 | Straw fermenting technique with goose source penicillium oxalicum |
| CN101838613A (en) * | 2009-09-30 | 2010-09-22 | 河南省恒隆态生物工程股份有限公司 | Agricultural straw fast-degradation decomposition agent strain composition |
| CN102559506A (en) * | 2010-12-07 | 2012-07-11 | 中国农业科学院作物科学研究所 | Penicillium oxalicum and application thereof |
| CN103160441A (en) * | 2011-12-14 | 2013-06-19 | 河北农业大学 | Preparation process for Penicillium oxalicum fermentation broth and application thereof in agriculture |
-
2013
- 2013-09-25 CN CN2013104432226A patent/CN103468592A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101580853A (en) * | 2008-03-06 | 2009-11-18 | 王宝维 | Straw fermenting technique with goose source penicillium oxalicum |
| CN101838613A (en) * | 2009-09-30 | 2010-09-22 | 河南省恒隆态生物工程股份有限公司 | Agricultural straw fast-degradation decomposition agent strain composition |
| CN102559506A (en) * | 2010-12-07 | 2012-07-11 | 中国农业科学院作物科学研究所 | Penicillium oxalicum and application thereof |
| CN103160441A (en) * | 2011-12-14 | 2013-06-19 | 河北农业大学 | Preparation process for Penicillium oxalicum fermentation broth and application thereof in agriculture |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105801210A (en) * | 2016-03-09 | 2016-07-27 | 南京农业大学 | Method for directly preparing fertilizer by using straws and prepared straw organic fertilizer |
| CN105801210B (en) * | 2016-03-09 | 2016-12-28 | 南京农业大学 | A kind of method utilizing the direct composting of straw and the Straw manures of preparation |
| US10287216B2 (en) | 2016-03-09 | 2019-05-14 | Nanjing Agricultural University | Method for directly preparing a fertilizer by using straws and prepared straw organic fertilizer |
| CN109054843A (en) * | 2018-08-03 | 2018-12-21 | 北京农业生物技术研究中心 | A kind of soil conditioner and preparation method and its application as pesticide synergistic agent |
| CN109054843B (en) * | 2018-08-03 | 2020-09-15 | 北京农业生物技术研究中心 | Soil conditioner, preparation method and application of soil conditioner as pesticide synergist |
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Application publication date: 20131225 |