CN101050425A - Trichodermin capable of decomposing and fermenting organic composting material in animality - Google Patents
Trichodermin capable of decomposing and fermenting organic composting material in animality Download PDFInfo
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- CN101050425A CN101050425A CN 200610034868 CN200610034868A CN101050425A CN 101050425 A CN101050425 A CN 101050425A CN 200610034868 CN200610034868 CN 200610034868 CN 200610034868 A CN200610034868 A CN 200610034868A CN 101050425 A CN101050425 A CN 101050425A
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Abstract
This invention discloses a trichoderma strain that can decompose and ferment organic compost (such as cattle manure) rich in cellulose. The conidiophore of the trichoderma strain has multiple lateral ramifications, each of which can be diverged to form small branches in pairs. Each small branch has 2-3 whorled or opposite generator cells. The generator cells are bottle shape, and most are vertical with a small part bending. The sizes of the generator cells are 6.9-11.8X1.5-3.1 mu.m. The conidiophore is quasi-sphere or ellipse shape, and is green. The size of conidiophore is 3.1-4.6X2.4-3.7 mu.m. The surface of mature conidiophore is smooth or rough, and has unobvious spur-shaped objects, which are cone or prismoid shape and spread on the whole surface of conidiophore under SEM observation.
Description
Technical field
The invention relates to and a kind ofly can decompose the organic Trichoderma bacterial strain of cellulosic (Trichoderma sp.) fast.
Background technology
Use biotechnological formulation for cultivating employed new technology now in the arviculture process, wherein the kind of biotechnological formulation comprises that (1) promotes plant-growth person, as mycorhiza bacterium, vinelandii and root circle beneficial microorganism group; (2) disease and pest biological control person is as Su Li bacterium, Trichoderma and Bacillus subtilus; (3) person that improves the crop growth environment is as phosphorus-solubilizing bacteria.The function of biotechnological formulation is mainly promoting plant-growth, increasing output and is reducing disease and pest, and other function still comprises the generation of promotion plant hormone, inducing plant disease resistance response, delays soil acidification, lowers the accumulation of soil salt and lures that other beneficial microorganism produces into.The existing in recent years considerable research input of these biotechnological formulations.
The definition of biotechnological formulation except that comprising farm crop itself, still comprises with farm crop the various biologies that react/act on as with biological species differentiation widely.With microbial preparation, then biotechnological formulation only comprises microbe itself or its associated metabolic product as only.But the benefit rough segmentation that microbial preparation is applied on the crop growth is direct benefit and indirect benefit.The microorganism of tool direct benefit comprises subfamily symbiosis root nodule bacterium (Rhizobium), with the symbiotic actinomycetes of forest crop (Frankia), promote nitrogen-fixing bacteria, capsule arbuscular mycorrhizal fungus (vesicualar-arbuscularmycorrhizal fungi) and the ectotrophic mycorrhiza (ectomycorrhizal fungi) of plant-growth, it all can directly promote the dietetic alimentation of plant.And the microorganism of tool indirect benefit comprises the biological control actor to phytopathogen, plant insect and weed competition, and it has the indirect mechanism of the poor environment factor effect of the crop growth that weakens the effect.No matter should mechanism why, these microorganisms all can manifest in experimentation and promote to replace mineral manure and chemical insecticide by crop growth potential, reduce production costs and reduce environmental pollution, and can guarantee crop yield.
General agricultural wastes all have contaminative and resource concurrently.Utilize microorganism can deal carefully with agricultural wastes, it suitably is converted into required nutrient source of agriculture production (as nitrogen, phosphorus and potassium) and energy source (as carbon).If agricultural wastes can be returned in the farmland, then not only conform with the natural law of resource reutilization, and also be the important directions of handling a large amount of organic wastes now.
Microorganism is served as the role of organism decomposition and compost stabilization in composting fermentation process.Inoculate suitable microorganism and can quicken compost fermentation.For reaching efficient composting effect, except that adding suitable microorganism, in piling up environment, keep the condition that is fit to microorganism growth and make its fully activity and breeding, more can quicken the fermentation and the decomposition of compost.The breeding of the microorganism that compost fermentation is used can utilize the nature composting process, promptly according to the composting condition, in wood chip, dregs of beans composting material, adjust suitable material size, moisture content and air permeability etc., pile up again and become thoroughly decomposed, make the occurring in nature microbial growth in this composting material, promptly contain the microbial strains of multiple energy decomposing organic matter at last in this matured compost.These microbial strainss can be used as the microbial strains that the next batch compost is made.But,, add suitable microbial strains and will be the important step in the compost making processes at the organic characteristic of difference in order to promote the decomposition efficiency of composting material fermentation.Wherein relevantly utilize the key mechanism of microbial strains should comprise the suitable microbial strains of screening, set up efficient spawn culture and propagation method and be applied to inoculation method etc. in the composting material.
Found Trichoderma (Trichoderma sp.) but except that antagonism certain plants pathogenic bacteria (such as da mping-off fungi (Pythium sp) and dry thread Pyrenomycetes (Rhizoctonia solani) etc.) to reduce the farm crop death; its part bacterial strain still can with the symbiosis of farm crop root system to overcome environmental stress, reach its root system of protection, promote nutrient and absorb, promote growth and improve effects such as output.Utilizing gene to change plantation technology can import disease-resistant gene in the plant materials and reach disease-resistant function.Because Trichoderma has all advantages on multi-functional, existing at present this bacterium is developed to the biotechnological formulation that biological control is used.Trichoderma commonly used comprises T.virens, T.harzianum, T.atroviride and T.asperellum, and the farm crop of using comprise vegetables, fruit tree, flowers and turf.
Summary of the invention
The objective of the invention is to be to provide a kind of trichoderma strain that the organic compost material breakdown of animality can be fermented.
Being characterized as of trichoderma strain of the present invention: conidinphore has the adnation branch of plural number, and each branch can diverge, and often forms living branchlet; Can produce 2～3 verticillate or conidiogenous cells on each sprig to give birth to arranging; Conidiogenous cell is a bottle stalk shape, and its kenel mostly is uprightly, the part slight curvature, and the wide middle shape is like Pear-Shaped, and size is 6.9～11.8 * 1.5～3.11 μ m; Conidium is subsphaeroidal to wide ellipse, and part is avette a little, and size is 3.1～4.6 * 2.4～3.7 μ m, is green; Ripe spore surface is smooth to coarse, the not obvious thorn-like jewelry of tool; Conidial cell wall surface jewelry is steeple shape or trapezoid platform shape under scanning electronic microscope, be distributed in whole spore surface.
The strong trichoderma strain of a kind of fiber capacity of decomposition provided by the invention, this bacterial strain can quickly diffuse to whole composting material in the initial stage of compost reaction, so that the decomposition rapidly in composting fermentation process of organism such as composting material, and can be used as the use of organic compost material acid ferment additive.
The trichoderma strain that the present invention filters out has high definition to organic compost, and can quicken compost maturity, reduces the composting time-histories, reduces the compost cost of manufacture.
Description of drawings
Fig. 1 be for trichoderma strain of the present invention at 25 ℃ of colonial morphology figure of seven days of growth regulation in the substratum that contains 2% Fructus Hordei Germinatus extract;
Fig. 2 be for trichoderma strain of the present invention 25 ℃ the time in potato glucose-agar medium seven days colonial morphology figure of growth regulation;
Fig. 3 is the mode of appearance figure that presents luxuriant shape projection for the product spore zone of trichoderma strain of the present invention under 63 times of enlargement ratios;
Fig. 4 is the conidinphore of trichoderma strain of the present invention and the branch mode of appearance figure at 200 times of enlargement ratios;
Fig. 5 is the conidinphore of trichoderma strain of the present invention and branch another mode of appearance figure at 200 times of enlargement ratios;
Fig. 6 is the conidinphore of trichoderma strain of the present invention and the branch mode of appearance figure at 1000 times of enlargement ratios; And
Fig. 7 is the conidinphore of trichoderma strain of the present invention mode of appearance figure at 2000 times of enlargement ratios.
Embodiment
Trichoderma strain of the present invention is separable from natural origin, for example soil, the homemade compost of farm crop root system and farmers', and (Wateragar, W.A.) the agar flat band method is carried out microorganism and is separated wherein to utilize 2% water agar.Through the soil segregator, take by weighing 1g soil and place the 10c.c. sterilized water, after vibrating, take out filtrate and carry out the plate streaking separation.The segregator adopts the root segment partition method to the farm crop root system, cuts 0.5 centimeter segment after root segment is cleaned, and wherein 5 segments place water agar; After each composting material is subdivided into 0.5 centimeter earlier, getting wherein, 5 segments place water agar, more than every kind of material prepare 5 culture dish, after irradiation was not cultivated three days in 28 ℃ of incubators, picking fragment mycelia tip moved to it on potato glucose substratum, after cultivating 7 days, observe each mycelial growth state, by the bacterial strain of picking Trichoderma mycelia form wherein, after the secondary pure lines were separated, the mycelia piece that cuts 0.5cm2 placed the spiral test tube of 5c.c. sterilized water to preserve; Each isolated strains is after potato glucose agar medium (PDA) is cultivated seven days, cut the mycelia piece, place the 10g substratum to cultivate, the mycelia of observing each bacterial strain is twined and the growth situation, and observe it and produce the spore situation, select the bacterial strain that can twine the paddy substratum fast and can produce spore according to this.
The selected bacterial strain that goes out applicable to cow dung compost is to be Trichodermaasperellum (TCT301) after BCRC identifies, it is characterized by: conidinphore has the adnation branch of plural number, and each branch can diverge, and often forms living branchlet; Can produce 2～3 verticillate or conidiogenous cells (extremely shown in Figure 5 as Fig. 3) to giving birth to arrangement on each sprig, conidiogenous cell is a bottle stalk shape, and its kenel mostly is uprightly, the part slight curvature, and the wide middle shape is like Pear-Shaped, and size is 6.9～11.8 * 1.5～3.1 μ m; Conidium is subsphaeroidal to wide ellipse, and part is avette a little, and size is 3.1～4.6 * 2.4～3.7 μ m, is green; Ripe spore surface is smooth to coarse, the not obvious thorn-like jewelry of tool; Conidial cell wall surface jewelry is steeple shape or trapezoid platform shape under the scan-type electricity shows, be distributed in whole spore surface (as Figure 6 and Figure 7).
Below be the strain characteristic of test gained Trichoderma under different growth conditionss.
In MA2 substratum (substratum that contains 2% Fructus Hordei Germinatus extract), growth is quick down to place 25 ℃ with aforementioned inoculation, and mycelia can extend to whole 9 centimeters culture dish (as shown in Figure 1) after 3～4 days; Conidiogenous cell is powdery or particulate state in the peripheral pencil Xiao Cong that forms of culture dish; Ripe spore ball presents deep green; The bacterium colony back side is white in color.
In Fructus Hordei Germinatus extract agar medium (MEA), growth is quick down to place 25 ℃ with aforementioned inoculation, and mycelia can extend to whole 9 centimeters culture dish (as shown in Figure 1) after 3 days, bacterium colony is open and flat, the product spore is dense, and conidiogenous cell is scattered to be distributed in whole culture dish, is powdery or particulate state; Ripe spore ball presents sap green; The bacterium colony back side is white in color to milk yellow.
In potato glucose agar medium (PDA), growth is quick down to place 25 ℃ with aforementioned inoculation, and mycelia can extend to whole 9 centimeters culture dish (as shown in Figure 2) after 3 days, bacterium colony is open and flat, the product spore is dense, and conidiogenous cell is scattered to be distributed in whole culture dish, is powdery or particulate state; Ripe spore ball presents olive-green to deep green; The bacterium colony back side is white in color to milk yellow.
Trichoderma strain of the present invention can be the varient that separates from the trichoderma strain of natural origin, for example handle resulting varient through ultraviolet ray (UV) irradiation, the processing of methylsulfonic acid second fat or nitrous acid, can also be to utilize cytogamy or recombinant DNA (deoxyribonucleicacid, DNA) technology is through the genetic engineering modified recombinant bacterial strain that produces.
Following examples are to be used to further specify the present invention, but scope of the present invention is not limited to following embodiment:
Embodiment 1. uses Trichoderma to make the temperature effective of compost
Each composition dry matter usage ratio of cow dung organic compost material is (cow dung 1000kg, rice husk 500kg, rice bran 100kg) in regular turn; Take an amount of Trichoderma of the present invention (bacterium number about 1 * 1012spore/ml) again after composting material mixed, elder generation's thin up becomes bacteria suspension for 200 times, again the bacterium diluent is sneaked in the composting material, at last the composting material moisture content is adjusted to 60%, piling height is kept about 1.5～2.0m, thereafter pile up making immediately, utilized shovel loader turning every day second inferior in about 5～7 days during the compost corruption, continue to till the compost maturity.
Cow dung compost material the inoculation Trichoderma is made the 0-5 day of piling up, compost body temperature degree rises to more than 60 ℃, can reach 67 ℃, this pliotherm period (＞60 ℃) is maintained until about the 15th day, thereafter temperature reduces gradually, and about the 31st～35 day, compost temperature was reduced to about 44 ℃, about the 36th～45 day, compost temperature can be reduced to about 41 ℃;
Do not inoculate the cow dung compost of Trichoderma, piling up making the 6th～10 day, compost body temperature degree rises to more than 60 ℃, reach as high as 61 ℃, about 36-35 day, about 48 ℃ of compost temperature, about the 36th～45 day, compost temperature just is reduced to about 44 ℃, shows that the cow dung compost that the inoculation Trichoderma is handled can promote compost high temperature (＞60 ℃), reaches the stage of becoming thoroughly decomposed at last ahead of time on the 4th～6.
Use temperature effective that Trichoderma makes compost as shown in Table 1:
Table one, inoculation Trichoderma are to the influence of cow dung compost material temperature
| Number of days is handled | 0～5 | 6～10 | 11～15 | 16～20 | 21～25 | 26～30 | 31～35 | 36～45 |
| The inoculation Trichoderma | 63 | 67 | 62 | 57 | 55 | 51 | 44 | 41 |
| Not inoculation | 57 | 60 | 61 | 60 | 58 | 54 | 48 | 44 |
Unit (℃)
Embodiment 2. uses the influence of Trichoderma to the compost nutrient content
Compost is made with embodiment 1, nutrient content analytical results by cow dung compost shows below table two, utilizes that the cow dung compost nitrogen content of Trichoderma inoculation is about 2.51%, phosphorus content is about 1.27%, potassium content is about 2.70%, calcium contents is about 1.59%, Mg content is about 1.67%, organic content is about 60.7%, the about 85ppm of zinc content, the about 32ppm of copper content, Trichoderma several 6.2 * 10
5Spore (spore)/g.
Do not inoculate that the cow dung compost nitrogen content of Trichoderma is about 2.44%, phosphorus content is about 1.25%, potassium content is about 2.62%, calcium contents is about 1.39%, Mg content is about 1.61%, organic content is about 64.7%, the about 84ppm of zinc content, the about 30ppm of copper content, Trichoderma several 1.1 * 10
2Spore (spore)/g.
Obviously have nitrogen, phosphorus, potassium, calcium, magnesium, zinc and the copper equal size of the cow dung compost of inoculation Trichoderma all to handle, and its nutrient content is more stable a little more than not inoculating.Inoculation Trichoderma bacterial classification is arranged in compost making processes, and in the finished product such as cow dung compost, all separable bacterial classification that goes out to be added, separation rate is about 1 * 10
5To 1 * 10
6Spore/g, the Trichoderma bacterial classification that demonstration is added can be survived in composting process and compost finished product.
Table two, inoculation Trichoderma are to the cow dung compost Temperature Influence
| Nutrient is handled | Nitrogen | Phosphorus | Potassium (%) | Calcium (%) | Magnesium | Organic | Zinc (ppm) | Copper (ppm) | Trichoderma spore/g |
| The inoculation Trichoderma | 2.51 | 1.27 | 2.70 | 1.59 | 1.67 | 60.7 | 85 | 32 | 6.2×10 2 |
| Not inoculation | 2.44 | 1.25 | 2.62 | 1.39 | 1.61 | 64.7 | 84 | 30 | 1.1×10 2 |
Implementation method of the present invention has been specified in the previous embodiment, and this embodiment is not in order to limiting claim of the present invention, does not allly break away from the equivalence that skill spirit of the present invention does and implements or change, all should be contained in the claim of this case.
In sum, the trichoderma strain that the present invention filters out is to cow dung compost material prescription tool high definition, and can quicken compost maturity, reduces the composting time-histories, reduces the compost cost of manufacture.
Claims (10)
1. an energy is the trichoderma strain of the organic compost material breakdown fermentation of animality, being characterized as of this bacterial strain: conidinphore has the adnation branch of plural number, and each branch can diverge, and often forms living branchlet; Can produce 2～3 verticillate or conidiogenous cells to give birth to arranging on each sprig, conidiogenous cell be a bottle stalk shape, and its kenel mostly is uprightly, the part slight curvature, and the wide middle shape is Pear-Shaped seemingly, and size is 6.9～11.8 * 1.5～3.1 μ m; Conidium is subsphaeroidal to wide ellipse, and part is avette a little, and size is 3.1～4.6 * 2.4～3.7 μ m, is green; Ripe spore surface is smooth to coarse, the not obvious thorn-like jewelry of tool; Conidial cell wall surface jewelry is steeple shape or trapezoid platform shape under scanning electronic microscope, be distributed in whole spore surface.
2. by the described a kind of trichoderma strain that the organic compost material breakdown of animality can be fermented of claim 1, it is characterized in that: can grow when described trichoderma strain is inoculated in the substratum that contains 2% Fructus Hordei Germinatus extract and places 25 ℃ fast, mycelia can extend to whole 9 centimeters culture dish in 3～4 days; Produce spore and be configured in the peripheral pencil Xiao Cong that forms of culture dish, be powdery or particulate state; Ripe spore ball presents deep green; The bacterium colony back side is white in color.
3. by the described a kind of trichoderma strain that the organic compost material breakdown of animality can be fermented of claim 1, it is characterized in that: can grow when described trichoderma strain is inoculated in Fructus Hordei Germinatus extract agar medium substratum and places 25 ℃ fast, mycelia can extend to whole 9 centimeters culture dish in 3 days, bacterium colony is open and flat, the product spore is dense, produce the spore structure and intersperse among whole culture dish, be powdery or particulate state; Ripe spore ball presents sap green; The bacterium colony back side is white in color to milk yellow.
4. by the described a kind of trichoderma strain that the organic compost material breakdown of animality can be fermented of claim 1, it is characterized in that: described trichoderma strain is inoculated in the potato glucose agar medium, place 25 ℃, growth fast, mycelia can extend to whole 9 centimeters culture dish after 3 days, bacterium colony is open and flat, and it is dense to produce spore, produces the spore structure and intersperses among whole culture dish.
5. by the described a kind of trichoderma strain that the organic compost material breakdown of animality can be fermented of claim 1, it is characterized in that: described product spore is configured to powdery or particulate state.
6. by the described a kind of trichoderma strain that the organic compost material breakdown of animality can be fermented of claim 1, it is characterized in that: described ripe spore ball presents dark olive-green to deep green.
7. by the described a kind of trichoderma strain that the organic compost material breakdown of animality can be fermented of claim 1, it is characterized in that: the described bacterium colony back side is white in color to milk yellow.
8. by the described a kind of trichoderma strain that the organic compost material breakdown of animality can be fermented of claim 1, it is characterized in that: described trichoderma strain is the varient that separates from the trichoderma strain of natural origin.
9. by the described a kind of trichoderma strain that the organic compost material breakdown of animality can be fermented of claim 1, it is characterized in that: described trichoderma strain is to handle resulting varient through ultraviolet ray (UV) irradiation, the processing of methylsulfonic acid second fat or nitrous acid.
10. by the described a kind of trichoderma strain that the organic compost material breakdown of animality can be fermented of claim 1, it is characterized in that: described trichoderma strain is to utilize cytogamy or recombinant DNA technology through the genetic engineering modified recombinant bacterial strain that produces.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109717050A (en) * | 2019-02-22 | 2019-05-07 | 海盐森植生物科技有限公司 | A kind of culture substrate of cinnamomum camphora |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109717050A (en) * | 2019-02-22 | 2019-05-07 | 海盐森植生物科技有限公司 | A kind of culture substrate of cinnamomum camphora |
| CN109717050B (en) * | 2019-02-22 | 2021-04-06 | 海盐森植生物科技有限公司 | Culture medium for cinnamomum camphora |
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