CN110229757B - Trichoderma citrinoviride JS84 capable of effectively promoting crop growth and bio-organic fertilizer developed by trichoderma citrinoviride JS84 - Google Patents
Trichoderma citrinoviride JS84 capable of effectively promoting crop growth and bio-organic fertilizer developed by trichoderma citrinoviride JS84 Download PDFInfo
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- CN110229757B CN110229757B CN201910545892.6A CN201910545892A CN110229757B CN 110229757 B CN110229757 B CN 110229757B CN 201910545892 A CN201910545892 A CN 201910545892A CN 110229757 B CN110229757 B CN 110229757B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/885—Trichoderma
Abstract
The invention discloses trichoderma citrinoviride JS84 for effectively promoting crop growth and a biological organic fertilizer developed by the trichoderma citrinoviride JS 84. Trichoderma citrinoviride JS84 is preserved in China general microbiological culture Collection center with preservation number of CGMCC NO.17466, and can antagonize various fusarium oxysporum such as banana, cucumber and the like. The biological organic fertilizer is obtained by adding a functional strain JS84 into a common organic fertilizer, and the number of viable bacteria is preferably more than or equal to 2.0 multiplied by 107CFU/g organic fertilizer dry weight. The product has good growth promoting effect on cucumber seedling stage, can cultivate high-quality seedlings with a large amount of active functional microorganisms at the rhizosphere, and promotes the growth and yield of the transplanted seedlings.
Description
Technical Field
The invention belongs to the field of agricultural microorganisms, and particularly relates to trichoderma citrinoviride JS84 capable of effectively promoting crop growth and application thereof.
Background
Plant Growth-promoting Rhizobacteria (PGPR) refers to a microorganism which is present in the microenvironment of the soil around the surface and root system of a Plant rhizosphere and can significantly promote the Growth of the Plant by direct or indirect means (Ahmad et al, Microbiological Research,2008,163(2): 173-181). Trichoderma, as one kind of PGPR, can promote the growth of plant effectively, and has the features of low cost, high safety, high lasting effect, stable yield increasing, environment friendship, etc. Therefore, the influence of trichoderma on crop yield and growth traits is explored, scientific basis can be provided for reasonable utilization of trichoderma preparations in green pollution-free vegetable production, and the method has important significance for saving fertilizer resources and developing organic agriculture.
Trichoderma citrinoviride is disclosed in various documents (CN101637187A, CN105219650A) to be capable of controlling plant fungal diseases, but no report is found about its growth promoting effect.
Disclosure of Invention
The invention aims to provide a Trichoderma citrinoviride JS84 which can effectively promote the growth of crops aiming at the practical problems and needs in production practice.
Another object of the invention is to provide the use of the fungus.
The invention also aims to provide a biological fertilizer prepared by the fungus and application thereof.
The purpose of the invention can be realized by the following technical scheme:
a growth-promoting strain JS84 for promoting crop growth is separated from Jiangsu farmland soil, is classified and named as Trichoderma citrinoviride, is preserved in China general microbiological culture Collection center, has the preservation date of 2019, 4 months and 12 days, and has the preservation number of CGMCC No. 17466.
The strain JS84 grows faster on the PDA plate, and hypha grows radially, a large amount of spores and green spores are produced, and spore-forming clusters are white to green. Phylogenetic tree analysis of the strain ITS rDNA sequence shows that the strain JS84 is located on the Trichoderma citrinovride branch, and JS84 is identified as Trichoderma citrinoviride by combining physiological and biochemical tests.
The invention discloses application of Trichoderma citrinoviride JS84 in promoting plant growth.
The invention discloses application of Trichoderma citrinoviride JS84 in preparation of a biological fertilizer for promoting plant growth.
The biological fertilizer prepared by the Trichoderma citrinoviride JS84 is provided.
The biological fertilizer is preferably prepared mainly by the following method: inoculating Trichoderma citrinoviride JS84 with the preservation number of CGMCC No.17466 into a PDA liquid culture medium for liquid fermentation, wherein the fermentation temperature is 28 ℃, the stirring speed is 170r/min, the fermentation time is 96h, the fermentation is better under the illumination condition, and the bacteria content in the fermentation liquid is more than or equal to 1 x 106one/mL. Mixing 100g straw and 200ml water per dry weight, adding 6.6ml amino acid, and stirring. Sterilizing at 115 deg.C for 1 h. Sterilizing, cooling, inoculating 20% Trichoderma fermentation broth, stirring, spreading in a turnover box, culturing at 28 deg.C for 4-5 days, making into Trichoderma strain, and storing in 4 deg.C refrigerator. 5 to 15 percent of trichoderma is added into the organic fertilizer to prepare the trichoderma bio-organic fertilizer. The preparation method of the PDA culture solution is as follows, taking 1L of culture medium as an example: peeling 200g potato, cutting into small pieces, decocting in water, boiling, decocting for 20min, filtering, adding 20g common glucose into the filtrate, diluting to 1000ml, adjusting pH to natural, and sterilizing at 115 deg.C for 30 min.
The biological fertilizer is applied to promoting the growth of plants.
Has the advantages that:
the method mainly utilizes the application of trichoderma citrinoviride which is effectively planted and planted in the rhizosphere and is separated and screened, so that the growth speed of the seedling stage of the crops is obviously improved, and the absorption and utilization of nutrients in the soil by the crops are promoted, thereby promoting the growth of the crops. The growth-promoting bacterium JS84 shows stable growth-promoting capability in a multi-season potting test. The biological fertilizer has obvious growth promoting effect after being applied and is suitable for being used in large area of vegetable crop producing area.
Drawings
FIG. 1 phylogenetic Tree based on ITS rDNA Gene sequence of Strain JS84 established by adjacency method
FIG. 2 is a diagram of the effect of first-season cucumber potting
FIG. 3 is a diagram of the effect of second-season cucumber potting
Biological material preservation information
The strain JS84 is classified and named as Trichoderma citrinoviride, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, has the preservation address of No. 3 of Xilu-Tai-Chen-Yang district of Beijing, and the microorganism research institute of Chinese academy of sciences, has the preservation date of 2019, 4 and 12 days, and the preservation number of CGMCC No. 17466.
Detailed Description
Example 1 isolation and identification of functional strains
Placing 5g of soil sample in a triangular flask containing 45ml of sterile water, and shaking at 28 deg.C for 30min (170r/min) to obtain soil suspension. Sucking 0.1mL of soil suspension, placing the soil suspension on a trichoderma selective plate, coating the plate, placing the plate in an incubator at 28 ℃ for culturing for 48h, observing the growth condition of bacterial colonies on the plate, selecting differential bacterial colonies for purification, inoculating the purified bacterial strains on a PDA culture medium, and placing the PDA culture medium in a refrigerator at 4 ℃ for storage for later use.
Inoculating the selected trichoderma strains on a PDA culture medium, respectively placing in incubators at 28 ℃, 37 ℃ and 40 ℃, culturing for 5 days, observing, and screening out high-temperature trichoderma capable of normally growing at 40 ℃.
The trichoderma strain is finally obtained by screening and named as JS84, the strain JS84 grows faster on a PDA plate on a strain JS84 of the PDA plate, hypha grows radially, a large amount of spores are produced, green spores are produced, and spore producing clusters are white to green. Phylogenetic tree analysis constructed by the strain ITSrDNA sequence shows that the strain JS84 is positioned on the branch of Trichoderma citrinovride, JS84 is classified as Trichoderma citrinovride, and the strain JS84 is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No. 17466.
Example 2 production of functional bacterial biofertilizer
Inoculating strain JS84(CGMCC No.17466) into PDA culture solution for liquid fermentation, wherein the conditions of fermentation production are as follows: the fermentation temperature is 28 ℃, the stirring speed is 170r/min, the fermentation time is 96h, and the bacteria content in the fermentation liquor is more than or equal to 1 multiplied by 106one/mL. 100g of straw per dry weight was mixed with 200ml of water, added to 6.6ml of acid-hydrolyzed amino acid solution (liquid amino acid complex disclosed in ZL 201410042218.3) and stirred. Sterilization at 115 ℃ 1h. Sterilizing, cooling, inoculating 20% Trichoderma fermentation broth, stirring, spreading in a turnover box, culturing at 28 deg.C for 4-5 days, making into Trichoderma strain, and storing in 4 deg.C refrigerator. 5 to 15 percent of trichoderma is added into the organic fertilizer to prepare the trichoderma bio-organic fertilizer.
The preparation method of the PDA culture solution comprises the following steps of preparing 1L of culture medium: peeling 200g potato, cutting into small pieces, decocting in water, boiling, decocting for 20min, filtering, adding 20g common glucose into the filtrate, diluting to 1000ml, adjusting pH to natural, and sterilizing at 115 deg.C for 30 min.
Example 3 potted plant growth promoting Effect test
3.1 application effect of growth-promoting biological fertilizer on cucumber (first season, second season)
The growth-promoting pot experiment is carried out in a warm-net room of the national center for organic fertilizer engineering technology research (Yixing), the soil to be tested is healthy sandy soil without pathogenic bacteria, and the cucumber variety to be tested is Lufeng cucumber without pathogenic bacteria. Soaking cucumber seeds in normal-temperature water for 1h, placing the cucumber seeds on moist gauze, accelerating germination for 12-24 h in an incubator at the temperature of 30 ℃, selecting the seeds with exposed white after more than 75% of the seeds germinate, transferring the seeds to a seedling tray for seedling culture, and selecting seedlings with consistent growth vigor to be transplanted into a pot after 2 true leaves grow out. The experiment set up two treatments: 1) CK, no fertilizer application; 2) treating OF, applying common organic fertilizer; 3) treating JS84, and applying a trichoderma bio-organic fertilizer. And (3) filling 1.5kg OF soil in each treatment pot, adding 2% OF common organic fertilizer into OF, adding 2% OF trichoderma bio-organic fertilizer into JS84, performing conventional management, and measuring physiological indexes OF the plants after about 30 d.
The growth promoting effect OF the cucumber in the first season is shown in fig. 2, and the biomass condition is shown in table 1, after 30 days OF pot culture, the cucumber plant is treated by JS84 trichoderma bioorganic fertilizer (JS84), Organic Fertilizer (OF) and non-fertilizer (CK), wherein the cucumber plant has significant differences in plant height, stem thickness, fresh weight OF the overground part and dry weight OF the overground part. Compared with treatment without fertilization (CK), the plant height, stem thickness, chlorophyll, fresh weight of the overground part and dry weight of the overground part of the cucumber plant are respectively increased by 17%, 8%, 5%, 18% and 36% by applying JS84 trichoderma bio-organic fertilizer treatment (JS 84). Compared with OF, the treatment with JS84 trichoderma bioorganic fertilizer increases 5%, 6%, 2%, 9% and 16% OF plant height, stem thickness, chlorophyll, fresh weight OF overground part and dry weight OF overground part, respectively.
The growth promoting effect OF the cucumbers in the second season is shown in figure 3, and the biomass condition is shown in table 2, after 30 days OF pot culture, the cucumber plants are treated by JS84 trichoderma bioorganic fertilizer (JS84), Organic Fertilizer (OF) and non-fertilizer (CK), wherein the cucumber plants have significant differences in plant height, stem thickness, chlorophyll, fresh weight OF the overground part and dry weight OF the overground part. Compared with CK, the treatment by applying JS84 trichoderma bio-organic fertilizer increases 60%, 30%, 11%, 58% and 41% of plant height, stem thickness, chlorophyll, fresh weight of overground part and dry weight of overground part respectively. Compared with OF, the treatment with JS84 trichoderma bioorganic fertilizer increases 30%, 11%, 3%, 23% and 18% OF plant height, stem thickness, chlorophyll, fresh weight OF overground part and dry weight OF overground part, respectively.
In conclusion, the two-season potting results show that the cucumber has obvious differences between the plant height, the stem thickness, the chlorophyll, the fresh weight OF the overground part and the dry weight OF the overground part compared with the cucumber treated by no fertilization (CK) and Organic Fertilizer (OF), and the JS84 strain has obvious growth promotion effect. In conclusion, the two-season potting results show that the cucumber plant growth vigor treated by the JS84 trichoderma bio-organic fertilizer (JS84) is superior to that treated by the non-fertilization (CK) and the common Organic Fertilizer (OF), and the significant differences in plant height, stem thickness, chlorophyll, fresh weight OF the overground part and dry weight OF the overground part are achieved, so that the JS84 strain has obvious growth promoting effect.
TABLE 1 Biomass of first season cucumber growth promoting pot culture test
TABLE 2 growth promotion of cucumber in second season potted plants for Biomass test
Claims (7)
1. Trichoderma citrinoviride JS84 for promoting crop growth effectively is deposited in China general microbiological culture Collection center with the deposit address of No. 3 Xilu-Chen-Xilu-Shi-3 of the sunward area in Beijing and the deposit date of 12 days in 2019 and 4 months in China academy of sciences and with the deposit number of CGMCC NO. 17466.
2. The use of Trichoderma citrinoviride JS84 in preparing functional biological organic fertilizer for promoting crop growth as recited in claim 1.
3. A functional bio-organic fertilizer for effectively promoting the growth of crops is characterized in that the common organic fertilizer is added with the trichoderma citrinoviride JS84 solid strain of claim 1.
4. The functional bio-organic fertilizer of Trichoderma citrinoviride JS84 of claim 3, wherein the viable count of Trichoderma citrinoviride JS84 in the functional bio-organic fertilizer is not less than 2.0 x 107CFU/g organic fertilizer dry weight, organic matter content is more than or equal to 45 percent, total nutrient (N + P)2O5+K2O)>5 percent and the water content is less than or equal to 30 percent.
5. The functional bio-organic fertilizer of claim 3, wherein the Trichoderma citrinoviride JS84 solid strain is prepared by the following fermentation method:
inoculating Trichoderma citrinoviride JS84 with the preservation number of CGMCC NO.17466 into PDA culture solution for liquid fermentation production, wherein the fermentation temperature range is 28 ℃, the stirring speed is 170r/min, and the fermentation time is 96h, so that the bacteria content in the fermentation solution is more than or equal to 1.0 x 108Per ml; mixing 100g straws per dry weight with 200ml water, adding 6.6ml amino acid solution, stirring, and sterilizing at 115 deg.C for 1 h; sterilizing, cooling, inoculating 20% Trichoderma fermentation broth, stirring, spreading in a turnover box, culturing at 28 deg.C for 4-5 days, making into Trichoderma strain, and storing in 4 deg.C refrigerator.
6. The functional bio-organic fertilizer as claimed in claim 3, wherein said crop is cucumber.
7. The preparation method of the functional bio-organic fertilizer of claim 3, characterized by comprising the following steps:
(1) preparing Trichoderma citrinoviride JS84 Trichoderma citrinoviride solid strain with the preservation number of CGMCC NO. 17466;
(2) adding the trichoderma strains prepared in the step (1) into a common organic fertilizer according to 5-15% of the dry weight of the organic fertilizer, and uniformly mixing to obtain the functional biological organic fertilizer, wherein the viable count is more than or equal to 2.0 multiplied by 107CFU/g organic fertilizer dry weight.
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| CN114574372B (en) * | 2022-03-28 | 2023-06-20 | 山东省林业科学研究院 | Trichoderma citrinoviride and application thereof in degradation of fish protein |
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| CN101637187A (en) * | 2008-10-23 | 2010-02-03 | 中国农业科学院农业资源与农业区划研究所 | Microbial inoculum for preventing and curing fungal diseases of plants |
| CN107142211A (en) * | 2017-05-11 | 2017-09-08 | 沈阳农业大学 | The tangerine green trichoderma Snef1910 and metabolite of a kind of killing root-knot nematode and application |
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| WO2014104998A1 (en) * | 2012-12-28 | 2014-07-03 | Eltem Rengin | Production of trichoderma citrinoviride micropropagules as a biocontrol agent by means of an economical process |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101637187A (en) * | 2008-10-23 | 2010-02-03 | 中国农业科学院农业资源与农业区划研究所 | Microbial inoculum for preventing and curing fungal diseases of plants |
| CN107142211A (en) * | 2017-05-11 | 2017-09-08 | 沈阳农业大学 | The tangerine green trichoderma Snef1910 and metabolite of a kind of killing root-knot nematode and application |
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