CN110218657A - One plant of long shoot trichoderma MD30 and its biological organic fertilizer of development - Google Patents
One plant of long shoot trichoderma MD30 and its biological organic fertilizer of development Download PDFInfo
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- CN110218657A CN110218657A CN201910545893.0A CN201910545893A CN110218657A CN 110218657 A CN110218657 A CN 110218657A CN 201910545893 A CN201910545893 A CN 201910545893A CN 110218657 A CN110218657 A CN 110218657A
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- trichoderma
- organic fertilizer
- long shoot
- fermentation
- biological organic
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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
-
- 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
-
- 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 one plant of long shoot trichoderma MD30 and its biological organic fertilizers of development.A kind of long shoot trichoderma MD30, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC NO.17467, which normal growth and can generate a large amount of spores under 40 DEG C of hot conditions.The biological organic fertilizer is added obtained by function stem MD30 in common organic fertilizer, and viable count is preferably >=2.0 × 107CFU/g organic fertilizer dry weight.It is good to the growth-promoting effect of cucumber at seedling stage using the product, growth and yield after kind of transplantation of seedlings can be effectively facilitated.
Description
Technical field
The invention belongs to field of agricultural microorganism, and in particular to one plant of long shoot trichoderma MD30 and its biological organic of development
Fertilizer.
Background technique
Plant growth-promoting rhizobacteria (Plant Growth-promoting Rhizobacteria, PGPR), which refers to, is present in plant
In object rhizosphere surface and root system surrounding soil microenvironment, micro- life of plant growth can be remarkably promoted by direct or indirect mode
Object (Ahmad et al, 2008).The one kind of trichoderma as PGPR, can be effectively facilitated plant growth, while have at low cost
It is honest and clean, using safe, lasting effect is good, volume increase is stable, to environment and food safety the features such as.Trichoderma is explored to crop yield and life
The influence of long character can be Trichoderma preparation reasonable using scientific basis is provided in the production of green non-pollution vegetables, to saving
Fertilizer resources, Development of Organic Agriculture are of great significance.But common trichoderma strain is generally more sensitive to temperature on the market, no
It can be grown under the conditions of being higher than 37 DEG C of temperature.Therefore, the quasi- screening from the soil of China hot-zone of the present invention adapts to higher soil
Temperature and the trichoderma strain that plant growth can be effectively facilitated, and its biological organic fertilizer of preparation, to promote trichoderma in hot-zone
Application effect.
The numerous patents document report such as CN107828670A, CN103451112A Changzhi trichoderma to phytopathogen
Biological and ecological methods to prevent plant disease, pests, and erosion effect, but there is not yet the growth-promoting function of Changzhi trichoderma is disclosed in the prior art.
Summary of the invention
It is an object of the invention to for the practical problem and demand in production practices, provide one plant adapt to higher soil temperature and
Effectively facilitate the long shoot trichoderma MD30 of plant growth.
It is a further object of the present invention to provide the applications of the fungi.
It is yet another object of the invention to provide the bio-feritlizer of fungi preparation and its applications.
The purpose of the present invention can be achieved through the following technical solutions:
One plant, for promoting the growth-promoting bacterial strain MD30 of plant growth, is isolated from Hainan Mango soil, classification naming is length
Branch trichoderma Trichoderma longibrachiatum, is preserved in China Committee for Culture Collection of Microorganisms's commonly micro- life
Object center, the deposit date is on April 12nd, 2019, deposit number was CGMCC No.17467.
Bacterial strain MD30 is grown comparatively fast on PDA plate, and fluffy white mycelia largely generates green spores, has white miscellaneous
Will forms concentric wheel stripe, generates yellow diffusivity pigment.The phylogenetic tree analytical table of bacterial strain ITS rDNA sequence construct
Bright, bacterial strain MD30 is located in Trichoderma longibrachiatum branch, is accredited as MD30 in conjunction with physiological and biochemical test
Long shoot trichoderma.
Long shoot trichoderma MD30 of the present invention is promoting the application in plant growth.
Application of the long shoot trichoderma MD30 of the present invention in the bio-feritlizer that preparation promotes plant growth.
The bio-feritlizer prepared by long shoot trichoderma MD30 of the present invention.
The bio-feritlizer preferably mainly passes to lower section method preparation: by the long shoot wood that deposit number is CGMCC No.17467
Mould MD30 is inoculated into PDA liquid medium, is carried out liquid fermentation, and fermentation temperature is 28 DEG C, and mixing speed is 170 revs/min,
Fermentation time is 96h, is fermented under illumination condition more excellent, and bacteria containing amount >=1 × 10 in fermentation liquid are made6A/mL.Every dry weight 100g stalk
It is mixed with 200ml water, is added to 6.6ml Liquid amino acid compound (ZL201410042218.3) and mixes thoroughly.115 DEG C of sterilizing 1h.
After sterilization and cooling, 20% trichoderma fermentation liquid is accessed, mixes thoroughly, uniformly spreads out in turnover box, after 28 DEG C are cultivated 4-5 days, production
At trichoderma strain, it is put in 4 DEG C of refrigerators and saves.By the trichoderma strain of addition 5%~15% in organic fertilizer, it is raw that production becomes trichoderma
Organic fertilizer material.PDA culture liquid making method used is, for preparing 1L culture medium: being cut into after being peeled with 200g potato small
Block is put into water and boils, and 20min is boiled after boiling, after filtering in filtrate plus 20g common glucose sugar, be settled to 1L, pH value naturally,
115 DEG C of sterilizing 30min.
Bio-feritlizer of the present invention is promoting the application in plant growth.
The utility model has the advantages that
The present invention mainly using the application for separating the long shoot trichoderma that rhizosphere is effectively colonized, significantly improves crops seedling stage
The speed of growth, promote crop nutrient in soil is absorbed and utilized, to promote plant growth.Promoting bacteria MD30 of the present invention exists
The stabilization growth-promoting ability shown in more season pot experiments.Bio-feritlizer of the present invention is obvious using rear growth-promoting effect, is suitble to vegetables
Crop producing region large area uses.
Detailed description of the invention
The phylogenetic tree that Fig. 1 uses adjacent method to establish based on the ITS rDNA gene order of bacterial strain MD30
The upgrowth situation comparison diagram of Fig. 2 trichoderma at different temperatures
Fig. 3 first season cucumber pot test effect figure
Fig. 4 second season cucumber pot test effect figure
Biomaterial preservation information
Bacterial strain MD30, classification naming are long shoot trichoderma Trichoderma longibrachiatum, are preserved in the micro- life of China
Object culture presevation administration committee common micro-organisms center, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, in
Institute of microbiology, the academy of sciences, state, the deposit date is on April 12nd, 2019, deposit number was CGMCC No.17467.
Specific embodiment
The separation and identification of embodiment 1, function stem
5g soil sample is placed in the triangular flask for filling 45ml sterile water, soil is made in 28 DEG C of oscillation 30min (170r/min)
Suspension.It draws 0.1mL soil supension to be placed in trichoderma selectivity plate and be coated with, is placed in 28 DEG C of incubators and cultivates 48h, observe
Bacterium colony growing state on plate selects the purifying of difference bacterium colony, 4 DEG C will be placed in strain inoculated to PDA culture medium after purification
Refrigerator saves stand-by.
The trichoderma strain selected is inoculated in PDA culture medium, is respectively placed in 28 DEG C, 37 DEG C and 40 DEG C incubators,
It is observed after culture 5d, filtering out can be in the high temperature trichoderma of 40 DEG C of normal growths.
It is named as MD30 by screening finally one plant of trichoderma of acquisition, bacterial strain MD30 is grown comparatively fast on PDA plate, villiform
White hypha largely generates green spores, there is white magazine, forms concentric wheel stripe, generates yellow diffusivity pigment.Bacterial strain
The phylogenetic tree of ITSrDNA sequence construct analysis shows, bacterial strain MD30 is located at Trichoderma longibrachiatum points
MD30 is classified as Trichoderma longibrachiatum by Zhi Shang, is preserved in Chinese microorganism strain preservation management
Committee's common micro-organisms center, deposit number are CGMCC No.17467.
Growth measurement under embodiment 2, different temperatures
Bacterial strain MD30 (CGMCC No.17467) is seeded in PDA culture medium at different temperatures, it is observed and grows shape
Condition.Set temperature is 37 DEG C, 40 DEG C and 42 DEG C, is compareed as Guizhou trichoderma NJAU4742 that deposit number is CGMCC NO.12166
Bacterial strain is disclosed in CN108675895A.
As a result, it has been found that bacterial strain MD30 remains to grow under 38 DEG C of hot conditions, 4742 bacterial strains then can not normal growth；40
Under DEG C hot conditions, bacterial strain MD30 remains to grow, and 4742 bacterial strains can not then be grown；Under 42 DEG C of hot conditions, bacterial strain MD30 and
4742 bacterial strains can not be grown.
1,40 DEG C of trichoderma mycelial growth rate of table
The generation of embodiment 2, function bacterium bio-feritlizer
Bacterial strain MD30 (CGMCC No.17467) is seeded to liquid fermentation in PDA culture solution, the condition of fermenting and producing are as follows:
Fermentation temperature is 28 DEG C, and mixing speed is 170 revs/min, and fermentation time 96h makes bacteria containing amount >=1 × 10 in fermentation liquid6A/
mL.Every dry weight 100g stalk and the mixing of 200ml water, are added 6.6ml Liquid amino acid compound and mix thoroughly.115 DEG C of sterilizing 1h.Sterilizing
After cooling, 20% trichoderma fermentation liquid is accessed, mixes thoroughly, uniformly spreads out in turnover box, after 28 DEG C are cultivated 4-5 days, is fabricated to wood
Mould species are put in 4 DEG C of refrigerators and save.By the trichoderma strain of addition 5%~15% in organic fertilizer, production has as trichoderma as biological
Machine fertilizer.
Wherein PDA used culture liquid making method is, for preparing 1L culture medium: being cut into after being peeled with 200g potato small
Block is put into water and boils, and 20min is boiled after boiling, after filtering in filtrate plus 20g common glucose sugar, be settled to 1L, pH value naturally,
115 DEG C of sterilizing 30min.
Embodiment 3, potting growth-promoting effect test
Application effect of the 3.1 growth-promoting bio-fertilizers on cucumber (first season, the second season)
Growth-promoting pot experiment carries out in national organic fertilizer Engineering Technical Research Centre (Yixing) Greenhouse, for examination soil
Earth is the healthy sandy soil without pathogen, is the rich cucumber of dew without pathogen for examination cucumber variety.Cucumber seeds are immersed in
1h in normal-temperature water is placed on wet gauze in 30 DEG C of incubators vernalization 12~for 24 hours, after the sprouting of 75% or more seed,
It chooses the seed to show money or valuables one carries unintentionally and moves to seedlings nursing plate nursery, selected after growing 2 true leaves in the consistent seedling replanting basin of growing way.Test setting
Two processing: 1) CK does not apply any fertilizer；2) OF is handled, common organic fertilizer is applied；3) MD30 is handled, MD30 trichoderma is applied
Biological organic fertilizer.2% common organic fertilizer is added in 6 basins of each processing, every basin dress 1.5kg soil, OF, and 2% MD30 is added in MD30
Trichoderma as biological organic fertilizer, Routine Management, the physical signs of the rear measurement plant of 30d or so.
First season cucumber growth-promoting effect such as Fig. 2, biomass situation are as shown in table 1, after potting culture 30d, application MD30 wood
Mould biological organic fertilizer processing (MD30), application of organic fertilizers (OF) processing and do not apply fertilizer (CK) processing cucumber plant plant height,
There is significant difference in terms of top fresh weight and overground part dry weight.Compared with CK, the processing of MD30 trichoderma as biological organic fertilizer is applied
In terms of plant height, stem thickness, chlorophyll, overground part fresh weight and overground part dry weight, 15%, 7%, 3%, 22% and has been increased separately
39%.Compared with OF, the processing of MD30 trichoderma as biological organic fertilizer is applied in plant height, stem thickness, overground part fresh weight and overground part dry weight
Aspect has increased separately 4%, 4%, 14% and 20%.
Second season cucumber growth-promoting effect such as Fig. 3, biomass situation are as shown in table 2, after potting culture 30d, application MD30 wood
Plant height in cucumber plant of mould biological organic fertilizer processing (MD30), application of organic fertilizers (OF) processing and (CK) processing of not applying fertilizer, stem
Slightly, there is significant difference in terms of chlorophyll, overground part fresh weight and overground part dry weight.Compared with CK, MD30 trichoderma as biological is applied
The processing of organic fertilizer in terms of plant height, stem thickness, chlorophyll, overground part fresh weight and overground part dry weight, increased separately 53%,
32%, 9%, 52% and 33%.Compared with OF, apply MD30 trichoderma as biological organic fertilizer processing plant height, stem thickness, chlorophyll,
In terms of overground part fresh weight and overground part dry weight, 18%, 13%, 1%, 13% and 7% has been increased separately.
To sum up the potting of two seasons the result shows that, with do not apply fertilizer (CK) processing and organic fertilizer (OF) handle cucumber in plant height, stem
Slightly, difference is obvious between chlorophyll, overground part fresh weight and overground part dry weight, illustrates that the growth-promoting effect of MD30 bacterial strain is obvious.To sum up
The potting of two seasons the result shows that, application MD30 trichoderma as biological organic fertilizer (MD30) processing cucumber plant growing way is not better than applying fertilizer at (CK)
Reason and common organic fertilizer (OF) processing, and reach aobvious in terms of plant height, stem thickness, chlorophyll, overground part fresh weight and overground part dry weight
Sex differernce is write, shows that MD30 bacterial strain has apparent growth-promoting effect.
Table 2, first season cucumber growth-promoting pot experiment biomass situation
Table 3, second season cucumber potting growth-promoting test organism amount situation
Claims (9)
1. a kind of long shoot trichoderma MD30 for adapting to higher soil temperature, it is commonly micro- to be preserved in China Committee for Culture Collection of Microorganisms
Bio-Centers, preservation address are Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, preservation day
Phase is on April 12nd, 2019, and deposit number is CGMCC NO.17467；The higher soil temperature is the soil temperature no more than 40 DEG C.
2. long shoot trichoderma MD30 described in claim 1 answering in the functional biological organic fertilizer that preparation promotes plant growth
With.
3. a kind of functional biological organic fertilizer for adapting to higher soil temperature, it is characterised in that add claim in common organic fertilizer
Long shoot trichoderma MD30 strain described in 1.
4. the functional biological organic fertilizer of long shoot trichoderma MD30 according to claim 3, it is characterised in that the function
Viable count >=2.0 × 10 long shoot trichoderma MD30 described in type vegetables nursery bio-matrix7CFU/g organic fertilizer dry weight, organic matter
>=45%, total nutrient (N+P2O5+K2O) > 5%, moisture≤30%.
5. functional biological organic fertilizer according to claim 3, it is characterised in that the long shoot trichoderma MD30 strain is logical
Cross following fermentation process preparation:
The long shoot trichoderma MD30 that deposit number is CGMCC NO.17467 is inoculated into PDA culture solution, production is carried out liquid fermentation,
28 DEG C of fermentation temperature range, mixing speed is 170 revs/min, and fermentation time 96h makes bacteria containing amount >=1 × 10 in fermentation liquid8
A/ml；Every dry weight 100g stalk and the mixing of 200ml water, are added 6.6ml Liquid amino acid compound and mix thoroughly；115 DEG C of sterilizing 1h.
After sterilization and cooling, 20% trichoderma fermentation liquid is accessed, mixes thoroughly, uniformly spreads out in turnover box, after 28 DEG C are cultivated 4-5 days, production
At trichoderma strain, it is put in 4 DEG C of refrigerators and saves.
6. functional biological organic fertilizer according to claim 3, it is characterised in that the crop is cucumber.
7. the preparation method of functional biological organic fertilizer as claimed in claim 3, it is characterised in that comprise the steps of:
(1) the long shoot trichoderma MD30 trichoderma strain that deposit number is CGMCC NO.17467 is prepared；
(2) common organic fertilizer is added by the 5%~15% of organic fertilizer dry weight in the trichoderma strain of step (1) preparation to be uniformly mixed
Functional biological organic fertilizer makes wherein viable count >=1.0 × 106CFU/g organic fertilizer dry weight.
8. preparation method according to claim 7, it is characterised in that the long shoot trichoderma MD30 trichoderma strain preparation side
Method is as follows: the long shoot trichoderma MD30 that deposit number is CGMCC NO.17467 being inoculated into PDA culture solution, life is carried out liquid fermentation
Produce, 28 DEG C of fermentation temperature range, mixing speed be 170 revs/min, fermentation time 96h, make bacteria containing amount >=1 in fermentation liquid ×
108A/ml；Every dry weight 100g stalk and the mixing of 200ml water, are added 6.6ml Liquid amino acid compound and mix thoroughly；115 DEG C of sterilizings
1h.After sterilization and cooling, 20% trichoderma fermentation liquid is accessed, mixes thoroughly, uniformly spreads out in turnover box, after 28 DEG C are cultivated 4-5 days, system
It is made trichoderma strain, 4 DEG C of refrigerators is put in and saves.
9. deposit number is that the long shoot trichoderma MD30 trichoderma strain of CGMCC NO.17467 promotes the functional form of plant growth in preparation
Application in biological organic fertilizer.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115181675A (en) * | 2022-05-06 | 2022-10-14 | 南京思农生物有机肥研究院有限公司 | Trichoderma guizhou growth promoting companion aspergillus flavus and application thereof |
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| CN107142213A (en) * | 2017-05-25 | 2017-09-08 | 金正大生态工程集团股份有限公司 | One plant of trichoderma asperellum and its cultural method and application with growth-promoting functions |
| CN108675895A (en) * | 2018-06-22 | 2018-10-19 | 南京农业大学 | A kind of trichoderma as biological organic fertilizer and its preparation method and application dedicated for improvement salt-soda soil |
| CN108901755A (en) * | 2018-05-28 | 2018-11-30 | 南京军辉生物科技有限公司 | A kind of watermelon seedling culturing growth-promoting matrix and preparation method thereof |
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2019
- 2019-06-23 CN CN201910545893.0A patent/CN110218657B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107142213A (en) * | 2017-05-25 | 2017-09-08 | 金正大生态工程集团股份有限公司 | One plant of trichoderma asperellum and its cultural method and application with growth-promoting functions |
| CN108901755A (en) * | 2018-05-28 | 2018-11-30 | 南京军辉生物科技有限公司 | A kind of watermelon seedling culturing growth-promoting matrix and preparation method thereof |
| CN108675895A (en) * | 2018-06-22 | 2018-10-19 | 南京农业大学 | A kind of trichoderma as biological organic fertilizer and its preparation method and application dedicated for improvement salt-soda soil |
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| CN115181675A (en) * | 2022-05-06 | 2022-10-14 | 南京思农生物有机肥研究院有限公司 | Trichoderma guizhou growth promoting companion aspergillus flavus and application thereof |
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