CN103719159B - A kind of solanaceous vegetables complex microbial inoculum - Google Patents
A kind of solanaceous vegetables complex microbial inoculum Download PDFInfo
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- CN103719159B CN103719159B CN201310659244.6A CN201310659244A CN103719159B CN 103719159 B CN103719159 B CN 103719159B CN 201310659244 A CN201310659244 A CN 201310659244A CN 103719159 B CN103719159 B CN 103719159B
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- zymotic fluid
- bacillus
- megaterium
- aztobacter
- bacillus megaterium
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Abstract
The present invention discloses a kind of solanaceous vegetables complex microbial inoculum, its preparation method and application; Does it adopt colloid bacillus cereus (Bacillus? mucilaginosus) zymotic fluid, bacillus megaterium (Bacillus? megaterium) zymotic fluid, aztobacter sp (Azotobacter? chroococcum) zymotic fluid, bacillus subtilis (Bacillus? subtilis) after zymotic fluid mixing as mix bacterium agent, obtain through fermentation again.This composite bacteria agent capable takes full advantage of the interaction between each bacterial classification, play stimulating plant growth, alleviate the hardening soil, improve the effect such as ecological environment of soil, disease resistance, fruit appearance quality can be improved and improve fruit quality, effect of increasing production is obvious, can economic benefit be significantly improved, reach the object of volume increase, high yield.
Description
Technical field
The invention belongs to agricultural and biological technical field, relate to a kind of solanaceous vegetables complex microbial inoculum, its preparation method and application.
Background technology
The birth of chemical fertilizer brings the hope of good harvest in the regular hour to peasant.But the drawback that Long-Time Service chemical fertilizer produces is following, the hardening soil, chemical element are residual, environmental pollution etc. series of problems emerges.Chemical fertilizer can not meet the needs of sustainable development, more can not become the pillar of green agriculture.
And solanaceous vegetables is the vegetables that a class is relatively grown based on edible fresh and tender fruit, growth cycle, mainly comprise cucumber, tomato, capsicum etc.These vegetables have similarity to nutrient demand, compare with leaf vegetables, relatively many to phosphorus, potassium demand in N P and K three elements, and nitrogen requirement is less, simultaneously because their vegetative period are long, so require fertilizer releasing period long, in order to avoid the later stage take off fertilizer.Existing production model adopts chemical fertilizer etc. to be main, and be generally fermented together by all raw materials, nutrient major part in organic matter is discharged, this later stage fruit growth for solanaceous vegetables is unfavorable.Solve appeal problem and will change existing production model, thus make nutrients release meet the requirement of solanaceous vegetables Nutrient Absorption.Along with the reach of science, the effect of microorganism in agricultural reveals gradually.Utilize microorganism formulation improve soil, degraded chemical residual, improve the ecological environment, crop yield, bacteriostatic and disease prevention etc., become the focus of research of agricultural science.Complex microbial inoculum is the guarantee of development " high yield, high-quality, efficient, ecological, safety " agricultural.
Summary of the invention
Problem to be solved by this invention adopts multiple bacterial classification to prepare composite bacteria agent capable, and utilize this composite bacteria agent capable to make plant biotechnology regulator to be applied in the plantation of crops.Make full use of the interaction between bacterial classification, play stimulating plant growth, alleviate the hardening soil, improve the effect such as ecological environment of soil, disease resistance, thus reach the object solving volume increase, high yield.
Solanaceous vegetables complex microbial inoculum of the present invention, it adopts colloid bacillus cereus (Bacillus mucilaginosus) CGMCC1.153 zymotic fluid, bacillus megaterium (Bacillus megaterium) ACCC10010 zymotic fluid, aztobacter sp (Azotobacter chroococcum) CGMCC1.142 zymotic fluid, as mix bacterium agent after the mixing of bacillus subtilis (Bacillus subtilis) ACCC01183 zymotic fluid, obtain through fermentation again.
For technical scheme mentioned above, in preferred situation: described colloid bacillus cereus (Bacillus mucilaginosus) CGMCC1.153 zymotic fluid, bacillus megaterium (Bacillus megaterium) ACCC10010 zymotic fluid, aztobacter sp (Azotobacter chroococcum) CGMCC1.142 zymotic fluid, the mass percent of bacillus subtilis (Bacillus subtilis) ACCC01183 zymotic fluid mixing is respectively 30 ~ 40%, 30 ~ 40%, 10 ~ 20%, 5 ~ 10%.
For technical scheme mentioned above, in preferred situation: the inoculum concentration of described mix bacterium agent through again fermenting is that mix bacterium agent accounts for 1 ~ 5% of total fermentation medium quality.
For technical scheme mentioned above, in preferred situation: the described fermentation medium again fermented is: corn flour 2 ~ 20g/100mL, beans Hectometer powder 1 ~ 10g/100mL, yeast extract 0.05 ~ 0.2g/100mL, KH
2pO
40.2 ~ 2g/100mL, K
2hPO
40.5 ~ 2g/100mL, MgSO
47H
2o 0.001 ~ 0.02g/100mL, NaCl 0.5 ~ 0.8g/100mL, (NH
4)
2sO
40.001 ~ 0.01g/100mL, solvent is distilled water.
For technical scheme mentioned above, in preferred situation: described colloid bacillus cereus (Bacillus mucilaginosus) CGMCC1.153 zymotic fluid, bacillus megaterium (Bacillus megaterium) ACCC10010 zymotic fluid, the bacterial classification quantity of aztobacter sp (Azotobacter chroococcum) CGMCC1.142 zymotic fluid is all not less than 10
8individual/mL.
For technical scheme mentioned above, in preferred situation: the end-product total bacteria count amount 6 ~ 10 × 10 that described fermentation again obtains
8individual/mL.
For technical scheme mentioned above, in preferred situation: described condition of again fermenting is cultivate 4 ~ 7 days at 30 DEG C.
For technical scheme mentioned above, in preferred situation: the firsts and seconds fermentation broth formula preparing colloid bacillus cereus (B. mucilaginocus) (Bacillusmucilaginosus) CGMCC1.153 zymotic fluid used is: KH
2pO
40.06g, MgSO
47H
2o 0.05g, CaSO
45H
2o 0.004g, peptone 5g, corn steep liquor (50% dry weight) 2.5g, lactose 15g, NaCl 4g, distilled water 1000mL, pH7.1;
The firsts and seconds fermentation broth formula preparing bacillus megaterium (Bacillus megaterium) ACCC10010 zymotic fluid used is: beef extract 3g, peptone 5g, NaCl5g, distilled water 1000mL, pH7.0;
The firsts and seconds fermentation broth formula preparing aztobacter sp (Azotobacter chroococcum) CGMCC1.142 zymotic fluid used is: KH
2pO
40.2g, K
2hPO
40.8g, MgSO
47H
2o 0.2g, CaSO
42H
2o 0.1g, Na
2moO
42H
2o trace, yeast extract 0.5g, mannitol 20g, FeCl
3trace, distilled water 1000mL, pH7.2;
The firsts and seconds fermentation broth formula preparing bacillus subtilis (Bacillus subtilis) ACCC01183 zymotic fluid used is: beef extract 10g, peptone 10g, NaCl5g, glucose 10g, distilled water 1000mL, pH7.0,
The invention has the beneficial effects as follows: this solanaceous vegetables complex microbial inoculum adopts bacillus megaterium, the circular azotobacter of brown, colloid bacillus cereus (B. mucilaginocus), bacillus subtilis as preparing bacterial classification.Above bacterial classification is cultivated respectively through first class inoculum, and two, three grade fermemtation, make bacterium number in zymotic fluid reach 6 ~ 10 × 10
8individual/mL, obtains solanaceous vegetables complex microbial inoculum.This product is through verifying on the spot for a long time, and rationally, stable effect, can promote plant growth, improve fruit quality, urge precocious, increase yield, improve utilization rate of fertilizer formula, improvement ecological environment of soil, and has certain disease resistance effect.
Embodiment
The present invention adopts colloid bacillus cereus (B. mucilaginocus) (Bacillus mucilaginosus) CGMCC1.153, bacillus megaterium (Bacillus megaterium) ACCC10010, aztobacter sp (Azotobacter chroococcum) CGMCC1.142, bacillus subtilis (Bacillus subtilis) ACCC01183, all can be bought by commercial sources and obtain, hereinafter for convenience of describing, be abbreviated as colloid bacillus cereus, bacillus megaterium, aztobacter sp, bacillus subtilis respectively.
Embodiment 1:
Described solanaceous vegetables complex microbial inoculum preparation process is as follows:
1. inoculate: described each bacterial classification (colloid bacillus cereus, bacillus megaterium, aztobacter sp, bacillus subtilis) is connected on slant medium respectively; Bacillus megaterium is 27 ~ 37 DEG C, and pH is 6.5 ~ 7.5, and incubation time is 4 days; Aztobacter sp cultivation temperature 25 ~ 30 DEG C, pH is 7.0 ~ 8.0, and incubation time is 5 days; Bacillus subtilis cultivation temperature 27 ~ 37 DEG C, pH is 6.5 ~ 7.5, and incubation time is 4 days; Colloid bacillus cereus (B. mucilaginocus) cultivation temperature 25 ~ 35 DEG C, pH is 7.5 ~ 8.0, and incubation time is 5 days;
2. first class inoculum is cultivated: the bacterial classification described step 1 prepared is inoculated in by the amount of oese one ring respectively and fills in the 50mL conical flask of respective liquid medium, and medium pH is 6.5 ~ 7.5, and conical flask puts into shaking table vibration, and temperature is 30 DEG C, vibration 24 ~ 48h; Employing blood counting chamber counts, and in bacterium liquid, bacterial classification quantity reaches 10 respectively
8individual/mL;
3. second order fermentation: the bacterial classification described step 2 prepared is inoculated in by 5% respectively and fills in the 1000mL conical flask of liquid nutrient medium, medium pH 6.5 ~ 7.5, put into shaking table vibration, temperature is 30 DEG C, vibration 48 ~ 72h; Bacterial classification quantity is made to reach 10 respectively
8individual/more than mL, thus obtained colloid bacillus cereus (B. mucilaginocus) zymotic fluid, bacillus megaterium zymotic fluid, aztobacter sp zymotic fluid and fermentation of bacillus subtilis liquid.
Again ferment after each bacterial classification is mixed by mass percentage: colloid bacillus cereus (B. mucilaginocus) zymotic fluid 30 ~ 40%, bacillus megaterium zymotic fluid 30 ~ 40%, aztobacter sp zymotic fluid 10 ~ 20%, fermentation of bacillus subtilis liquid 5 ~ 10%, as table 1 common prescription table.
Table 1. common prescription table
| Strain fermentating liquid title | Formula 1 | Formula 2 | Formula 3 | Formula 4 | Formula 5 | Formula 6 |
| Colloid bacillus cereus | 35% | 40% | 30% | 40% | 40% | 40% |
| The circular azotobacter of brown | 20% | 20% | 20% | 15% | 16% | 10% |
| Bacillus megaterium | 35% | 30% | 40% | 40% | 39% | 40% |
| Bacillus subtilis | 10% | 10% | 10% | 5% | 5% | 10% |
4. three grade fermemtation: the mixed bacterium liquid described step 3 prepared is inoculated in the fermentation tank filling composite bacteria agent capable medium, composite bacteria agent capable medium pH 6.5 ~ 7.5, inoculum concentration is cultivate 4 ~ 7 days at 5%, 30 DEG C of composite bacteria agent capable medium gross mass, makes bacterium number in zymotic fluid reach 6 ~ 10 × 10
8individual/mL, namely obtains solanaceous vegetables complex microbial inoculum, and pH is 6.8, and bacterium number content all reaches 9 × 10
8individual/mL.
Described slant medium formula is:
1. the slant medium formula that bacillus subtilis adopts is: beef extract 10g, peptone 10g, NaCl 5g, glucose 10g, agar 15g, distilled water 1000mL, pH7.0;
2. the slant medium formula that aztobacter sp adopts is: KH
2pO
40.2g, K
2hPO
40.8g, MgSO
47H
2o0.2g, CaSO
42H
2o 0.1g, Na
2moO
42H
2o trace, yeast extract 0.5g, mannitol 20g, FeCl
3trace, agar 15g, distilled water 1000mL, pH7.2;
3. the slant medium formula that bacillus megaterium adopts is: beef extract 3g, peptone 5g, agar 15g, NaCl 5g, distilled water 1000mL, pH7.0;
4. the slant medium formula that colloid bacillus cereus adopts is: KH
2pO
40.06g, MgSO
47H
2o 0.05g, CaSO
45H
2o 0.004g, peptone 5g, corn steep liquor (50% dry weight) 2.5g, lactose 15g, NaCl 4g, distilled water 1000mL, agar 25g, pH7.1;
Described liquid culture based formulas is:
1. the firsts and seconds fermentation broth formula that bacillus subtilis adopts is: beef extract 10g, peptone 10g, NaCl 5g, glucose 10g, distilled water 1000mL, pH7.0;
2. the firsts and seconds fermentation broth formula that aztobacter sp adopts is: KH
2pO
40.2g, K
2hPO
40.8g, MgSO
47H
2o 0.2g, CaSO
42H
2o 0.1g, Na
2moO
42H
2o trace, yeast extract 0.5g, mannitol 20g, FeCl
3trace, distilled water 1000mL, pH7.2;
3. the firsts and seconds fermentation broth formula that bacillus megaterium adopts is: beef extract 3g, peptone 5g, NaCl5g, distilled water 1000mL, pH7.0;
4. the firsts and seconds fermentation broth formula that colloid bacillus cereus adopts is: KH
2pO
40.06g, MgSO
47H
2o0.05g, CaSO
45H
2o 0.004g, peptone 5g, corn steep liquor (50% dry weight) 2.5g, lactose 15g, NaCl 4g, distilled water 1000mL, pH7.1;
Described composite bacteria agent capable culture medium prescription is: corn flour 2 ~ 20g/100mL, beans Hectometer powder 1 ~ 10g/100mL, yeast extract 0.05 ~ 0.2g/100mL, KH
2pO
40.2 ~ 2g/100mL, K
2hPO
40.5 ~ 2g/100mL, MgSO
47H
2o 0.001 ~ 0.02g/100mL, NaCl 0.5 ~ 0.8g/100mL, (NH
4)
2sO
40.001 ~ 0.01g/100mL, all the other are distilled water.
Embodiment 2:
The solanaceous vegetables complex microbial inoculum adopting the formula 1 in table 1 of the present invention to prepare is effect test in tomato planting:
One, seed soaking trial
1. test material and method
Experimental cultivar: gold kind No. (Hailin City tomato)
Test method: four process are established in test, adopts randomized arrangement, repeats for three times, plot area 66.7 square metres.
4 described process: 1. spraying agricultural microbial agent is product of the present invention (diluting 500 times with water); 2. spraying inactivating microbial microbial inoculum is that this product is placed in 121 DEG C, 20min deactivation (diluting 500 times with water); 3. spray biological organic fertilizer, buy (diluting 500 times with water) by commercial sources; 4. CK(sprays equivalent clear water).
Investigation content:
(1) test is implemented: gold kind No., and field crops managerial skills are higher, and phase of bearing fruit, fruit expanding period, painted phase that test fertility is grown in tomato production respectively spray once.
(2) field management: test tomato sprays medicine prevention and elimination of disease and pests 8 times in breeding time, pours water 5 times.
(3) precipitation: average 425.6mm comparatively over the years increased 32.2mm precipitation and concentrated on 7,8,9 three months, accounts for total rainfall 85% at tomato growth stage co-falling water 457.8mm 3-9 month in 2013.
(4) temperature: 3-9 month in this year, at each monthly mean temperature of tomato growth stage, and lasts that average monthly difference is little.
(5) illumination: 3-9 month in this year is 1450.5 hours at tomato growth stage lighting delay number in this year, comparatively reduces 49.6 hours former years 1500.1.
In a word, within 2013, tomato natural conditions breeding time are better, are conducive to the growth of tomato.
2. test results and analysis
Described four process are that 1. to spray agricultural microbial agent be that 2. to spray inactivating microbial microbial inoculum be this product is placed in 121 DEG C, 20min deactivation to product of the present invention respectively.3. spray biological organic fertilizer 4. CK(spray equivalent clear water) be control group.
(1) on the impact of tomato yield
From table 2, contrast per mu yield is 3938 kilograms, and spraying agricultural microbial agent per mu yield is 4590 kilograms, to according to volume increase 16.56%, sprays deactivation artifact microbial inoculum per mu yield 4207.3 according to increase by 6.84%.Spray biological organic fertilizer per mu yield 4234.5, than contrast increase by 7.53%, from data result, spraying agricultural biological bacterium has extremely significant effect to tomato production promoting.
Table 2
(2) on the impact of tomato biological proterties
From table 3,4: by our the main plant proterties investigation to the fertility of test tomato: a, tomato spray agricultural microbial agent and spray inactivating microbial microbial inoculum, spray biological organic fertilizer, spray clear water and compare plant strain growth stalwartness, dark green leaf color, the incidences of disease such as blade is relatively many, stem stalk is sturdy, plant wither is few, bacterial wilt late blight are lower.B, plant height comparatively contrast increases 2cm, and stem slightly comparatively contrasts increases 0.5cm, and blade is dark green, and the degree of aging of bottom blade is low, and fruit-setting rate comparatively contrasts raising 11%, and large fruit rate improves 10.3%.C, single fruit weight comparatively contrast increases 27.3g, can promote that young fruit expands, and increases single fruit weight, improves output.
Table 3
Table 4
3. conclusion
After tomato sprays agricultural microbial agent, plant strain growth is healthy and strong, and blade is dark green, and leaf area increases, photosynthetic capacity strengthens, and tomato fruit is individual large, and fruit surface is bright and clean, painted bright-coloured, can improve fruit appearance quality and improve fruit quality, effect of increasing production is obvious, can significantly improve economic benefit.
The test effect of agricultural microbial agent on tomato proves, this product can expand demonstration area in tomato production, and carries out spread.
Claims (6)
1. a solanaceous vegetables complex microbial inoculum, it is characterized in that: adopt colloid bacillus cereus (Bacillusmucilaginosus) CGMCC1.153 zymotic fluid, bacillus megaterium (Bacillus megaterium) ACCC10010 zymotic fluid, aztobacter sp (Azotobacter chroococcum) CGMCC1.142 zymotic fluid, as mix bacterium agent after the mixing of bacillus subtilis (Bacillus subtilis) ACCC01183 zymotic fluid, obtain through fermentation again;
Described colloid bacillus cereus (Bacillusmucilaginosus) CGMCC1.153 zymotic fluid, bacillus megaterium (Bacillus megaterium) ACCC10010 zymotic fluid, aztobacter sp (Azotobacter chroococcum) CGMCC1.142 zymotic fluid, the mass percent of bacillus subtilis (Bacillus subtilis) ACCC01183 zymotic fluid mixing is respectively 30 ~ 40%, 30 ~ 40%, 10 ~ 20%, 5 ~ 10%.
2. according to the microbial bacterial agent described in claim 1, it is characterized in that: the inoculum concentration of described mix bacterium agent through again fermenting is that mix bacterium agent accounts for 1 ~ 5% of total fermentation medium quality.
3. according to the microbial bacterial agent described in claim 1, it is characterized in that: the described fermentation medium again fermented is: corn flour 2 ~ 20g/100mL, beans Hectometer powder 1 ~ 10g/100mL, yeast extract 0.05 ~ 0.2g/100mL, KH
2pO
40.2 ~ 2g/100mL, K
2hPO
40.5 ~ 2g/100mL, MgSO
47H
2o 0.001 ~ 0.02g/100mL, NaCl 0.5 ~ 0.8g/100mL, (NH
4)
2sO
40.001 ~ 0.01g/100mL, solvent is distilled water.
4. according to the microbial bacterial agent described in claim 1, it is characterized in that: described colloid bacillus cereus (Bacillusmucilaginosus) CGMCC1.153 zymotic fluid, bacillus megaterium (Bacillus megaterium) ACCC10010 zymotic fluid, the bacterial classification quantity of aztobacter sp (Azotobacter chroococcum) CGMCC1.142 zymotic fluid is all not less than 10
8individual/mL.
5. according to the microbial bacterial agent described in claim 1, it is characterized in that: the end-product total bacteria count amount 6 ~ 10 × 10 that described fermentation again obtains
8individual/mL.
6. according to the microbial bacterial agent described in claim 1, it is characterized in that: described condition of again fermenting is cultivate 4 ~ 7 days at 30 DEG C.
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| CN105859480A (en) * | 2016-04-13 | 2016-08-17 | 大连三科生物工程有限公司 | Preparation method of special bioorganic fertilizer for solanaceous fruit vegetables |
| CN112226391A (en) * | 2020-11-04 | 2021-01-15 | 河南柏裕植物免疫科技有限公司 | Microbial agent containing bacillus and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101774856A (en) * | 2010-01-26 | 2010-07-14 | 凯迪阳光生物能源投资有限公司 | Preparation method of microbial fertilizer |
| CN102531776A (en) * | 2011-12-23 | 2012-07-04 | 于永波 | Compound microbial fertilizer and method for producing same |
| CN102746064A (en) * | 2012-07-11 | 2012-10-24 | 德州市元和农业科技开发有限责任公司 | Microbial foliage fertilizer, and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101774856A (en) * | 2010-01-26 | 2010-07-14 | 凯迪阳光生物能源投资有限公司 | Preparation method of microbial fertilizer |
| CN102531776A (en) * | 2011-12-23 | 2012-07-04 | 于永波 | Compound microbial fertilizer and method for producing same |
| CN102746064A (en) * | 2012-07-11 | 2012-10-24 | 德州市元和农业科技开发有限责任公司 | Microbial foliage fertilizer, and preparation method and application thereof |
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