CN102888376A - Bacillus subtilis BC-198, and selenium-rich microbial inoculum and application thereof - Google Patents
Bacillus subtilis BC-198, and selenium-rich microbial inoculum and application thereof Download PDFInfo
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Abstract
The invention relates to Bacillus subtilis BC-198, and a selenium-rich microbial inoculum and application thereof. The collection number of the Bacillus subtilis BC-198 is CCTCC No.M2012321. The invention also provides a composite microbial inoculum of which the effective ingredients are the Bacillus subtilis BC-198 and Azotobacter chroococcum AS1.212. The preparation method of the selenium-rich microbial inoculum provided by the invention can utilize the existing universal fermentation plant, is simple to operate and has the advantage of low manufacturing cost; the prepared product has multiple functions of nitrogen fixation, growth promotion, fungus disease control and the like for crops; and after being applied to the crops, the selenium-rich microbial inoculum can increase the yield, improve the quality of the agricultural product, and enhance the selenium content in the agricultural product by more than 20 times, so that the agricultural product reaches the selenium-rich product quality standard.
Description
Technical field
The present invention relates to microbial technology field, particularly a kind of subtilis BC-198 and its rich selenium microbial inoculum and application.
Background technology
Selenium is the necessary trace element of human and animal, and anti-cancer and antitumous effect are arranged, and lacks for a long time selenium, can cause various diseases, is one of cause of disease that causes Keshan disease.People recognize and mend the selenium necessity now, mend selenium by food and are considered to the safest, the means of most convenient.
The same with world's most countries, China lacks the selenium area.China lacks selenium provinces and cities and reaches more than 20, the about 0.03-0.1mg/kg of Selenium in Soils content, and mending selenium to soil is a kind of effective way that obtains Se-rich farm products (food).For many years, people are with rich selenium ore, the raw materials such as rich selenium coal gangue, through pulverizing, alkaline purification obtains small part (20%) solubility inorganic selenium, with nitrogen, phosphorus, potassium, copper, zinc, iron, magnesium, the inorganic salt such as boron or organic compound become selenium-enriched fertilizer, or selenium is prepared into sodium selenate, Sodium Selenite, and be mixed into selenium-enriched fertilizer with inorganic or organic fertilizer again with its solution, spray application to crop leaf or directly execute in the soil, although these selenium-enriched fertilizers can be produced Se-rich farm products and selenium-enriched food, but to the 10%-15% only of inorganic selenium crop utilization rate in the fertilizer, still very low, using organoselenium may be to improve crop to the approach of se use efficiency.
The agricultural microorganism microbial inoculum is the functional microbial fertilizer that a class is different from inorganic composite fertilizer, organic fertilizer.Utilize beneficial microorganism to transform selenium, inorganic selenium is changed into organoselenium, to improve farm crop to se use efficiency, improve the agricultural-food selenium content.Selenium-enriched microbe comprises at present: yeast rich in selenium, selenium-enriched edible mushroom, photosynthetic bacterium etc.A kind of selenium-rich combination azotobacter agent of patent CN200710010871 invention, in the preparation microbial inoculum, add solubility selenium, utilize vinelandii that inorganic selenium is transformed, but only adopt vinelandii in its preparation, only have the selenium-enriched microbe fertilizer of single fixed nitrogen physiological function.Patent CN 102618469A, denomination of invention: the red bacterium of a kind of pod membrane and application thereof, the red bacterium AYGHJ-16 of a kind of pod membrane is disclosed, belong to photosynthetic bacterium, utilize luminous energy that material is carried out metabolism, promote conversion and the absorption of inorganic selenium, and fixed nitrogen and carbon solidification effect are arranged.Patent 100512666C, denomination of invention: a kind of selenium microorganism feed addictive and preparation method thereof that contains, disclose and utilized microorganism live bacteria to carry out liquid fermenting, solid fermentation for Rhodopseudomonas, Bacillus natto genus bacillus and yeast, realized the microbial transformation of inorganic selenium.Compare with the present invention, present microbial inoculum is to the conversion of selenium or adopt single bacterial strain or similar bacterial strain, and is still lower to the utilization of the conversion of selenium and Selenium By The Plants.And the functional microbial fertilizer of promoting the use of such as fixed nitrogen, potassium decomposing, phosphorus decomposing, to urge corruption, growth promotion, the fertilizer such as disease-resistant be the common micro-organisms microbial inoculum of function singleness mostly.
Summary of the invention
The object of the invention provides bacillus subtilis BC-198 and an application thereof.
Another purpose of the present invention provides the microbial inoculum that contains above-mentioned subtilis BC-198.
Still a further object of the present invention provides the composite fungus agent that a kind of effective constituent is subtilis BC-198 and blown-ball Azotobacter AS1.212.
Another object of the present invention provides the preparation method of described composite fungus agent.
Subtilis Bacillus Subitilis BC-198 provided by the invention is preserved in Chinese Typical Representative culture collection center on August 30th, 2012 and (is called for short CCTCC; Address: Wuhan, China, Wuhan University; Postcode: 430072), deposit number CCTCCNo.M2012321.
The present invention also provides the microbial inoculum that contains described subtilis BC-198.
The present invention further provides a kind of composite fungus agent, its effective constituent is subtilis BC-198 and blown-ball Azotobacter AS1.212.The viable count of described subtilis BC-198 is 10-25 hundred million/ml, and the viable count of described blown-ball Azotobacter AS1.212 is 10-25 hundred million/ml.
Preferably, the viable count of described subtilis BC-198 is 15-19 hundred million/ml, and the viable count of described blown-ball Azotobacter AS1.212 is 15-19 hundred million/ml.
Composite fungus agent of the present invention also contains sanitas, protective material;
Wherein, described sanitas is one or more in benzoic acid and sodium benzoate, Sorbic Acid and sodium salt thereof, propionic acid and the sodium salt thereof; Described protective material is polyhydroxylated polymer, and wherein said polyhydroxylated polymer is one or more in Lalgine and sodium salt, polyglutamic acid and sodium salt thereof, carboxymethyl cellulose and the sodium salt thereof.
The present invention also provides the preparation method of described composite fungus agent, may further comprise the steps:
1) preparation subtilis BC-198 seed liquor and blown-ball Azotobacter AS1.212 seed liquor;
2) subtilis BC-198 seed liquor and blown-ball Azotobacter AS1.212 seed liquor fermentation culture, the preparation fermented liquid.
Wherein, described seed culture condition is: seed culture medium is at the 500 milliliters of bottled 100-130 milliliter of triangle substratum, 28-35 ℃ constant-temperature shaking culture 12-16 hour seed liquor.
The seed culture medium of cultivating subtilis BC-198 forms: add glucose 0.5%, starch 0.3%, peptone 0.6%, yeast extract paste 0.25%, sodium-chlor 0.4%, dipotassium hydrogen phosphate 0.1%, PH7.2 in 100 milliliters of the Se solutions;
The seed culture medium of blown-ball Azotobacter AS1.212 forms: add 0.3% glucose, 0.8% sucrose, 0.2% peptone, 0.6% yeast extract paste, 0.4% sodium-chlor, saltpetre 0.2%, PH7.4 in 100 milliliters of the Se solutions.
Wherein, described fermentation culture conditions is: stocking volume is the 65-70% of fermentor tank volume.The seed access amount of subtilis and blown-ball Azotobacter is 1.0-2.0% and 1.5-2.5% with the nutrient solution volume ratio respectively, temperature 28-35 ℃, tank pressure 0.04-0.06MPa, stirring velocity are 190220 rev/mins in the tank, the nutrient solution volume with pass into volume of air than 1:0.2-0.4(v/v), mixed fermentation 14-19 hour fermented liquid.
Fermention medium consists of: add 0.3-0.5% glucose, 0.3-0.7% sucrose, 0.4-0.6% peptone, 0.3-0.5% yeast extract paste, 0.2-0.5% sodium-chlor, PH7.4 in Se solution.
Further add sanitas, protective material in the described fermented liquid.It is 0.10%-0.50% that described sanitas adds mass volume ratio by fermentating liquid volume.It is 0.15%-1.0% that described protective material adds mass volume ratio by fermentating liquid volume.
Described subtilis BC-198 or described microbial inoculum or described composite fungus agent can be applicable to produce rich selenium microbial inoculum, are used for improving absorption and the utilization of Selenium By The Plants.
Advantage of the present invention is: bacterial strain provided by the invention can be used for the preparation of rich selenium microbial inoculum. and the preparation method can utilize ready-made general fermentation equipment, simple to operate, cost of manufacture is low, the product that makes has the several functions such as fixed nitrogen, growth promotion, control fungal disease to crop, solubility selenium further activates and transforms through microorganism and more is conducive to crop to selenium absorption and utilization, can increase output behind the crop applying, improve quality of agricultural product, produce the agricultural-food selenium content and improve 20 times, reach rich selenium target level of product quality.
Description of drawings
Fig. 1 is Bc-198 cell Electronic Speculum figure, magnification 10k;
Fig. 2 is the growing way design sketch that cucumber is used subtilis BC-198;
Fig. 3 is the growing way design sketch that cucumber is not used subtilis BC-198.
Embodiment
Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.Without departing from the spirit and substance of the case in the present invention, modification or replacement to the inventive method, step or condition are done all belong to scope of the present invention.
If specialize, the conventional means that used technique means is well known to those skilled in the art among the embodiment.
The percentage sign that relates among the present invention " % " is not if specify the forefinger mass percent; But the per-cent of solution except as otherwise herein provided, refers to contain among the solution 100ml the some grams of solute; Per-cent between the liquid, the ratio of capacity when referring to 20 ℃.
Embodiment 1 rich selenium multi-functional bacterial strain separates and identifies
From the selenium-rich tobacco waste of enshi city Baiyangping, by at 0.15%NaSeO
3, 0.6% glucose, 0.5% sucrose, 0.8% peptone, 0.3% yeast extract paste, 0.4% sodium-chlor, 100 milliliters of preparations of water substratum in the concentration and separation purifying.A strain bacterial strain BC-198 is screened to get in antagonistic effect and growth promotion test, and this bacterial strain fermentation liquor is to soybean seeds percentage of germination, long, the contrast raising 30-40% of root; Sickle-like bacteria, mould are suppressed circle all greater than 1.5 centimetres.Also have proteolytic enzyme, cellulase activity; This strain morphology or physiological and biochemical property are; Gram-positive, cell are shaft-like, diameter＜1um, as shown in Figure 1, form gemma, do not expand, without parasporal crystal; Catalase, oxidase positive, aerobic growth, VP tests positive, and VP＜PH6 tests positive, VP＞PH7 negative, methyl red test is negative, utilizes glucose, wood sugar, L-arabinose, N.F,USP MANNITOL to produce acid; Utilize not aerogenesis of glucose; Utilize Citrate trianion, reduction nitrate, hydrolyzed starch, liquefy gelatin, decomposition casein.In PH5.7,7%NaCl, 50 ℃ of all growths; This bacterial strain 16SrRNA gene sequencing result is shown in SEQ ID NO.1.
According to cellular form, biochemical test result, and the gene sequencing result, bacterial strain BC-198 is accredited as subtilis Bacillus Subitilis.This bacterial strain is preserved in Chinese Typical Representative culture collection center on August 30th, 2012 and (is called for short CCTCC; Address: Wuhan, China, Wuhan University; Postcode: 430072), deposit number CCTCCNo.M2012321.
Embodiment 2 subtilis BC-198 and the test of blown-ball Azotobacter compatibility
(1) test medium forms: 1) 0.6% glucose, 0.5% sucrose, 0.8% peptone, 0.3% yeast extract paste, 100 milliliters in 0.4% sodium-chlor water, PH7.4; (2) add 1.8% agar in the solid medium
(2) subtilis BC-198 and blown-ball Azotobacter are cultivated
250 milliliters of triangular flasks are loaded onto and are stated 50 milliliters of nutrient solutions, 0.1MPa sterilization 25 minutes, access respectively subtilis and blown-ball Azotobacter, place on the bottle swingging machine, 28-30 ℃ shaking culture 14-20 hour for subsequent use.
(3) antagonistic effect
By 100 milliliters of solid mediums of above-mentioned formulated, 0.1MPa sterilize 25 minutes, add 1 milliliter of blown-ball Azotobacter nutrient solution of cultivating in advance 14 hours when being cooled to 45 ℃, behind the mixing, draw 15 milliliters of culture dish that move into 9 centimetres of diameters, make the agar culture dish on the desktop of placement level, cool off after 40 minutes, place again the Oxford cup, in the cup of Oxford, drip 0.3 milliliter of centrifuged supernatant of cultivating in advance 20 hours withered grass gemma nutrient solution, put 28-30 ℃ of thermostat container and cultivate 24-36, circle does not appear suppressing in Oxford cup outer rim.Judge that subtilis BC-198 is to the growth unrestraint effect of blown-ball Azotobacter.The similar test method judges that blown-ball Azotobacter is to bacillus subtilis bacteria growing also unrestraint.
The preparation of embodiment 3 rich selenium microbial inoculums
(1) rich selenium agricultural microorganism microbial inoculum adopts bacterial classification: subtilis BC-198;
The seed culture medium of cultivating subtilis BC-198 forms: add glucose 0.5%, starch 0.3%, peptone 0.6%, yeast extract paste 0.25%, sodium-chlor 0.4%, dipotassium hydrogen phosphate 0.1%, PH7.2 in 100 milliliters of the Se solutions (0.249mg/ml);
By above-mentioned formulated seed culture medium, loading amount is 100 milliliters in 500 milliliters of triangular flasks, 0.11MPa sterilization is 25 minutes under the pressure, behind the cool to room temperature, with inoculation bamboo shovel picking withered grass gemma BC-198 bacterial classification primary and secondary kind, in the access triangular flask substratum, on the bottle swingging machine of 190 rev/mins of rotating speeds, 28 ℃ ± 1 ℃ constant-temperature shaking culture 14 hours makes the subtilis seed liquor;
(2) the tank top fermentation of rich selenium agricultural microorganism microbial inoculum is cultivated
Rich selenium agricultural microorganism microbial inoculum carries out at 5 liters of stainless steel automatically controlled fermentors.In 3 liters of Se solutions (0.3mg/ml), add 0.5% glucose, 0.6% sucrose, 0.6% peptone, 0.5% yeast extract paste, 0.4% sodium-chlor, PH7.4.Substratum 0.12MPa sterilization 25 minutes, substratum drop in temperature to 35 ℃ accesses 1.8% subtilis seed liquor, set tank pressure 0.04-0.06MPa, 30 ± 2 ℃ of tank temperature, ventilation flow rate 1:0.2(v/v), 190 rev/mins of stirring velocitys, fermenting got 2.86 liters of fermentation cultures in 18 hours.
(3) Sodium Benzoate of adding 0.19%, 0.9% sodium alginate, 0.09% polyglutamic acid stirs, and must contain selenium 0.284mg/ml, the selenium-enriched microbe microbial inoculum of bacterium viable count 34.8 hundred million/ml.
The preparation of embodiment 4 Se-enriched microbial inoculums
(1) seed culture of selenium-enriched microbe microbial inoculum
1) the rich selenium agricultural microorganism microbial inoculum of preparation adopts bacterial classification: subtilis BC-198, and blown-ball Azotobacter AS1.212.
2) rich selenium agricultural microorganism microbial inoculum seed culture medium forms:
A. contain the preparation of the selenium aqueous solution: selenium mineral powder is broken, crosses the 100-200 order, and ratio adding 3%-4% ammoniacal liquor and 1%-3% sodium hydroxide mixing solutions in selenium mineral powder mass volume ratio 1:2-3 stirred 24-36 hour, used H
3PO
4Transfer PH8.5-9.0 to filter to get supernatant liquor, measuring and adjusting selenium concentration is 0.25-1.0mg/ml, or directly uses NaSeO
3Preparation.
B. substratum forms
The seed culture medium of cultivating subtilis BC-198 forms: add glucose 0.5%, starch 0.3%, peptone 0.6%, yeast extract paste 0.25%, sodium-chlor 0.4%, dipotassium hydrogen phosphate 0.1%, PH7.2 in 100 milliliters of the Se solutions (0.25mg/ml).
The seed culture medium of blown-ball Azotobacter AS1.212 forms: add 0.3% glucose, 0.8% sucrose, 0.2% peptone, 0.6% yeast extract paste, 0.4% sodium-chlor, saltpetre 0.2%, PH7.4 in 100 milliliters of the Se solutions (0.25mg/ml).
C. seed culture condition
Prepare seed culture medium 100 milliliters of loading amounts in 500 milliliters of triangular flasks, 0.1MPa sterilize 25 minutes, access respectively subtilis BC-198 and blown-ball Azotobacter AS1.212, place on 200 rev/mins of rotary shaking bottle machines 30-32 ℃ of seed liquor that got the selenium-enriched microbe microbial inoculum in constant-temperature shaking culture 12-16 hour.
(2) fermentation culture: the preparation of agricultural microorganism microbial inoculum is carried out in the 10L fermentor tank, and its substratum forms: add 0.5% glucose, 0.6% sucrose, 0.6% peptone, 0.5% yeast extract paste, 0.4% sodium-chlor, PH7.4 in 6.5L Se solution (0.25mg/ml).The substratum stocking volume is 65% of fermentor tank volume, 0.12MPa sterilize 25 minutes, substratum drop in temperature to 32 ℃, access simultaneously 1.4% subtilis BC-198 seed liquor and 1.5% blown-ball Azotobacter AS1.212 seed liquor, temperature is 30 ℃ in the control tank, tank internal pressure 0.06MPa, 190 rev/mins of stirring velocitys, substratum with pass into volume of air and compare 1:0.4, the access seed liquor rises, pH value and bacterial cell concentration every sampling and measuring fermentation in 2 hours, find fermentation 18 hours and 19 hours, bacterial cell concentration does not rise appreciably, and fermentation culture fermented 18-19 hour to get.
(3) emit in the tank 5.8 liters of nutrient solutions, add 0.27% Sodium Benzoate, 0.34% polyglutamic acid, the selenium-enriched microbe microbial inoculum of selenium content 0.24mg/ml stirs to get.Subtilis BC-198 viable count 18.3 hundred million/ml, blown-ball Azotobacter AS1.212 viable count 16.7 hundred million/ml.
The preparation of embodiment 5 Se-enriched microbial inoculums
(1) seed culture of selenium-enriched microbe microbial inoculum
1) the rich selenium agricultural microorganism microbial inoculum of preparation adopts bacterial classification: subtilis BC-198, and blown-ball Azotobacter AS1.212
2) rich selenium agricultural microorganism microbial inoculum seed culture medium forms:
A. contain the preparation of the selenium aqueous solution: the selenium that contains of preparation substratum leaches the water preparation: pulverize through ball mill in the selenium ore deposit, crosses 100 orders, adds 3% ammoniacal liquor, the 2% sodium hydroxide mixing solutions of 2 times of volumes of selenium mineral powder quality, stirred 30 hours, uses H
3PO
4Transfer PH8.7 centrifuging to get the rich selenium aqueous solution, the regulator solution selenium content is 0.3mg/ml.
B. substratum forms
The seed culture medium of cultivating subtilis BC-198 forms: add glucose 0.5%, starch 0.3%, peptone 0.6%, yeast extract paste 0.25%, sodium-chlor 0.4%, dipotassium hydrogen phosphate 0.1%, PH7.2 in 100 milliliters of the Se solutions.
The seed culture medium of blown-ball Azotobacter AS1.212 forms: add 0.3% glucose, 0.8% sucrose, 0.2% peptone, 0.6% yeast extract paste, 0.4% sodium-chlor, saltpetre 0.2%, PH7.4 in 100 milliliters of the Se solutions.
C. spawn culture
The seed culture medium that forms preparation by above-mentioned substratum, 120 milliliters of substratum of loading amount in 500 milliliters of triangular flasks, 0.12MPa sterilize 25 minutes, move on to respectively in the triangular flask nutrient solution with inoculation shovel picking subtilis BC-198 and female kind of blown-ball Azotobacter AS1.212, place on 200 rev/mins of rotary shaking bottle machines 30-32 ℃ of constant temperature culture 15 hours.
(2) fermentation culture: the fermentation of rich selenium agricultural microorganism microbial inoculum is carried out in 50 liters of stainless steel automatically controlled fermentors.In 35 liters of Se solutions, add 0.5% glucose, 0.6% sucrose, 0.6% peptone, 0.5% yeast extract paste, 0.4% sodium-chlor, PH7.4.The substratum stocking volume be the fermentor tank volume 70%, 0.12MPa sterilization 25 minutes, the substratum drop in temperature is to 30-35 ℃, the subtilis seed liquor of access 1.14% and 2.43% blown-ball Azotobacter seed liquor, set 195 rev/mins of fermentation stirring velocitys, temperature is 28 ℃ in the tank, air flow quantity 1:0.3(v/v in the tank internal pressure 0.05MPa, tank).Behind the access seed, find every 3 hours sampling and measuring fermented liquid PH and bacterial cell concentration that the bacterial cell concentration of ferment 18 hours the time is close with 15 hours, the unobvious increase of cell concn.
(3) emit 33 liters of the interior zymocyte liquids of tank, the Sodium Benzoate of adding 0.2% and 0.8% polyglutamic acid stir, and get the rich selenium microbial inoculum of the rich selenium multifunctional agricultural of selenium content 0.304mg/ml microorganism.Subtilis BC-198 viable count 15.3 hundred million/ml, blown-ball Azotobacter AS1.212 viable count 18.7 hundred million/ml.
The preparation of embodiment 6 Se-enriched microbial inoculums
(1) seed culture of selenium-enriched microbe microbial inoculum
1) the rich selenium agricultural microorganism microbial inoculum of preparation adopts bacterial classification: subtilis BC-198, and blown-ball Azotobacter AS1.212
2) rich selenium agricultural microorganism microbial inoculum seed culture medium forms:
A. contain the preparation of the selenium aqueous solution:
B. substratum forms
The seed culture medium of cultivating subtilis BC-198 forms: add glucose 0.5%, starch 0.3%, peptone 0.6%, yeast extract paste 0.25%, sodium-chlor 0.4%, dipotassium hydrogen phosphate 0.1%, PH7.2 in 100 milliliters of the Se solutions (0.249mg/ml).
The seed culture medium of blown-ball Azotobacter AS1.212 forms: add 0.3% glucose, 0.8% sucrose, 0.2% peptone, 0.6% yeast extract paste, 0.4% sodium-chlor, saltpetre 0.2%, PH7.4 in 100 milliliters of the Se solutions (0.249mg/ml)
C. seed culture
By above-mentioned formulated seed culture medium, loading amount is 100 milliliters in 500 milliliters of triangular flasks, 0.11MPa sterilization is 25 minutes under the pressure, behind the cool to room temperature, with inoculation bamboo shovel difference picking withered grass gemma BC-198 and blown-ball Azotobacter AS1.212 bacterial classification primary and secondary kind, access respectively in the triangular flask substratum, on the bottle swingging machine of 190 rev/mins of rotating speeds, 28 ℃ ± 1 ℃ constant-temperature shaking culture 14 hours makes subtilis seed liquor and blown-ball Azotobacter seed liquor.
(2) the tank top fermentation of rich selenium agricultural microorganism microbial inoculum is cultivated
Rich selenium agricultural microorganism microbial inoculum carries out at 200 liters of stainless steel automatically controlled fermentors.In 140 liters of Se solutions (0.3mg/ml), add 0.5% glucose, 0.6% sucrose, 0.6% peptone, 0.5% yeast extract paste, 0.4% sodium-chlor, PH7.4.The substratum stocking volume be the fermentor tank volume 70%, 0.12MPa sterilization 25 minutes, substratum drop in temperature to 35 ℃, access 1.2% rise the subtilis seed liquor, 1.8% blown-ball Azotobacter seed liquor, set tank pressure 0.04-0.06MPa, 30 ± 2 ℃ of tank temperature, ventilation flow rate 1:0.2(v/v), 190 rev/mins of stirring velocitys, fermenting got 132 liters of fermentation cultures in 18 hours.
(3) Sodium Benzoate of adding 0.3%, 0.5% sodium alginate, 0.15% polyglutamic acid stirs, and must contain the selenium-enriched microbe microbial inoculum of selenium 0.284mg/ml.Subtilis BC-198 viable count 15.1 hundred million/ml, blown-ball Azotobacter AS1.212 viable count 14.9 hundred million/ml.
The effect test of embodiment 7 microbial inoculums
Winter in 2008 was used the selenium-enriched microbe microbial inoculum of embodiment 5 preparations potato: every strain is used and is contained the rich selenium agricultural microorganism of selenium 0.304mg/ml microbial inoculum 0.065kg, the result shows: use selenium-enriched microbe microbial inoculum fertilizer group potato mean yield 3.51kg/ strain, yampi ratio 70.98%, potato on average contains selenium 2.11mg/kg, does not use on average to contain selenium 0.102mg/kg; Do not use the potato mean yield 2.86kg/ strain of this fertilizer, yampi ratio 60.39%; Use the late blight of potato diseased plant rate 4.51% of this fertilizer; Do not use the late blight of potato diseased plant rate 79.12% of this fertilizer; Use the agricultural-food selenium content behind this fertilizer to improve 20.68 times, reach rich selenium standard.
Used the selenium-enriched microbe microbial inoculum of embodiment 6 preparations in 2010-2012 continuous 3 years cucumber: every strain is used and is contained the rich selenium agricultural microorganism of selenium 0.284mg/ml microbial inoculum 0.062kg.Fresh cucumbers on average contains selenium 2.226mg/kg, and untapped fresh cucumbers on average contains selenium 0.11mg/kg, improves 20.23 times of selenium contents; Use the cucumber mean yield 3.36kg/ strain of this fertilizer, do not use the cucumber mean yield 1.94kg/ strain of this fertilizer, increase production extremely remarkable.Use the contained heavy metal of cucumber product of this fertilizer to reach the green organic product standard, and extremely significant effect of increasing production is arranged.Use the cucumber of this fertilizer disease-free, untapped powdery mildew of cucumber diseased plant rate 30.4%.Use the cucumber growing way effect of subtilis BC-198 as shown in Figure 2, plant is tall and big, and is with luxuriant foliage and spreading branches in leafy profusion, and leaf is large, and the institute cucumber that connects is large and many; Do not use the cucumber growing way effect of subtilis BC-198 as shown in Figure 3, plant is little, and branches and leaves are not in great numbers, and leaf is little, and the institute cucumber that connects is little and few.
Used the selenium-enriched microbe microbial inoculum of embodiment 6 preparations in 2010-2012 continuous 3 years potato, every strain is used and is contained the rich selenium agricultural microorganism of selenium 0.284mg/ml microbial inoculum 0.0625kg, potato on average contains selenium 1.935mg/kg, do not use on average to contain selenium 0.0941mg/kg, improve 20.56 times of selenium contents; Use the potato mean yield 3.56kg/ strain of this fertilizer, yampi ratio 70.82%; Do not use the potato mean yield 2.86kg/ strain of this fertilizer, yampi ratio 60.68%; Use the late blight of potato diseased plant rate 3.94% of this fertilizer; Do not use the late blight of potato diseased plant rate 82.3% of this fertilizer.
Conclusion: can increase output after the crop applying microbial inoculum of the present invention, reduce morbidity, improve quality of agricultural product, produce the agricultural-food selenium content and improve more than 20 times, reach rich selenium target level of product quality.
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (10)
1. subtilis (Bacillus Subitilis) BC-198, its preserving number is CCTCCNo.M2012321.
2. the microbial inoculum that contains the described subtilis BC-198 of claim 1.
3. composite fungus agent, its effective constituent is subtilis BC-198 and blown-ball Azotobacter AS1.212.
4. composite fungus agent according to claim 3 is characterized in that, the viable count of described subtilis BC-198 is 10-25 hundred million/ml, and the viable count of described blown-ball Azotobacter AS1.212 is 10-25 hundred million/ml.
5. composite fungus agent according to claim 4 is characterized in that, the viable count of described subtilis BC-198 is 15-19 hundred million/ml, and the viable count of described blown-ball Azotobacter AS1.212 is 15-19 hundred million/ml.
6. each described composite fungus agent is characterized in that according to claim 3-5, also contains sanitas, protective material;
Wherein, described sanitas is one or more in benzoic acid and sodium benzoate, Sorbic Acid and sodium salt thereof, propionic acid and the sodium salt thereof;
Described protective material is polyhydroxylated polymer, and wherein said polyhydroxylated polymer is one or more in Lalgine and sodium salt, polyglutamic acid and sodium salt thereof, carboxymethyl cellulose and the sodium salt thereof.
7. each described composite fungus agent is characterized in that according to claim 3-6, is prepared as follows:
1) preparation subtilis BC-198 seed liquor and blown-ball Azotobacter AS1.212 seed liquor;
2) subtilis BC-198 seed liquor and blown-ball Azotobacter AS1.212 seed liquor complex ferment are cultivated, the preparation fermented liquid.
8. composite fungus agent according to claim 7 is characterized in that, it is that access amount with subtilis BC-198 seed liquor, blown-ball Azotobacter AS1.212 seed liquor is respectively 1-2% and 1.5-2.5% that described complex ferment is cultivated.
9. according to claim 7 or 8 described composite fungus agents, it is characterized in that, further add sanitas, protective material.
10. the described subtilis BC-198 of claim 1 or the described microbial inoculum of claim 2 or the described composite fungus agent of claim 3 application in the absorption that improves Selenium By The Plants and utilization.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105441487A (en) * | 2014-09-03 | 2016-03-30 | 赵小勇 | Bioorganic selenium, preparation method and use thereof, and feed or fertilizer prepared from bioorganic selenium |
| CN105950503A (en) * | 2016-05-19 | 2016-09-21 | 湖北民族学院 | Bacillus subtilis SE201412 and organic-selenium-rich bactericide |
| CN105994937A (en) * | 2016-05-19 | 2016-10-12 | 湖北民族学院 | Green organic selenium-rich feed and preparation method thereof |
| CN108721337A (en) * | 2017-04-17 | 2018-11-02 | 江苏德禧生物科技有限公司 | A kind of microbial bacterial agent preventing chemotherapy of tumors gastrointestinal toxicity |
| CN110643526A (en) * | 2019-08-31 | 2020-01-03 | 暨南大学 | Method for promoting colonization of azotobacter chroococcum in plant rhizosphere |
| CN114133290A (en) * | 2021-12-21 | 2022-03-04 | 山东润银生物化工股份有限公司 | Biological organic fertilizer prepared from bacillus subtilis and used for increasing yield and improving quality of crops and preparation method |
| CN114230414A (en) * | 2022-01-20 | 2022-03-25 | 桂林融和农业开发有限公司 | Method for producing selenium-rich sugar oranges |
| CN115261257A (en) * | 2022-05-25 | 2022-11-01 | 武汉轻工大学 | Selenium-rich bacillus subtilis L11 derived from silage forage and application thereof |
| CN115851544A (en) * | 2022-12-16 | 2023-03-28 | 陕西省微生物研究所 | Bacillus subtilis and method for preparing selenium-rich natto by using same |
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Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105441487A (en) * | 2014-09-03 | 2016-03-30 | 赵小勇 | Bioorganic selenium, preparation method and use thereof, and feed or fertilizer prepared from bioorganic selenium |
| CN105950503A (en) * | 2016-05-19 | 2016-09-21 | 湖北民族学院 | Bacillus subtilis SE201412 and organic-selenium-rich bactericide |
| CN105994937A (en) * | 2016-05-19 | 2016-10-12 | 湖北民族学院 | Green organic selenium-rich feed and preparation method thereof |
| CN105950503B (en) * | 2016-05-19 | 2019-04-16 | 湖北民族学院 | Bacillus subtilis SE201412 and organic selenium-rich microbial inoculum |
| CN108721337B (en) * | 2017-04-17 | 2022-05-10 | 江苏德禧生物科技有限公司 | Microbial agent for preventing tumor chemotherapy intestinal toxicity |
| CN108721337A (en) * | 2017-04-17 | 2018-11-02 | 江苏德禧生物科技有限公司 | A kind of microbial bacterial agent preventing chemotherapy of tumors gastrointestinal toxicity |
| CN110643526B (en) * | 2019-08-31 | 2022-06-14 | 暨南大学 | Method for promoting colonization of azotobacter chroococcum in plant rhizosphere |
| CN110643526A (en) * | 2019-08-31 | 2020-01-03 | 暨南大学 | Method for promoting colonization of azotobacter chroococcum in plant rhizosphere |
| CN114133290A (en) * | 2021-12-21 | 2022-03-04 | 山东润银生物化工股份有限公司 | Biological organic fertilizer prepared from bacillus subtilis and used for increasing yield and improving quality of crops and preparation method |
| CN114230414A (en) * | 2022-01-20 | 2022-03-25 | 桂林融和农业开发有限公司 | Method for producing selenium-rich sugar oranges |
| CN115261257A (en) * | 2022-05-25 | 2022-11-01 | 武汉轻工大学 | Selenium-rich bacillus subtilis L11 derived from silage forage and application thereof |
| CN115261257B (en) * | 2022-05-25 | 2023-04-25 | 武汉轻工大学 | Selenium-enriched bacillus subtilis L11 from silage and application thereof |
| CN115851544A (en) * | 2022-12-16 | 2023-03-28 | 陕西省微生物研究所 | Bacillus subtilis and method for preparing selenium-rich natto by using same |
| CN115851544B (en) * | 2022-12-16 | 2024-02-09 | 陕西省微生物研究所 | Bacillus subtilis and method for preparing selenium-enriched natto by using same |
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Application publication date: 20130123 Assignee: Enshi Heno Biologic Engineering Co., Ltd. Assignor: Liu Jinlong|Chen Jiaren Contract record no.: 2014420000148 Denomination of invention: Bacillus subtilis BC-198, and selenium-rich microbial inoculum and application thereof Granted publication date: 20140709 License type: Exclusive License Record date: 20140717 |
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