CN102888376B - 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, has anti-cancer and antitumous effect, lacks for a long time selenium, can cause various diseases, is one of cause of disease causing Keshan disease.People recognize and mend selenium necessity now, mend selenium and are considered to the safest, the means of most convenient by food.
The same with world's most countries, China lacks selenium area.Que Xi provinces and cities of China reach 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 become selenium-enriched fertilizer with inorganic or organic fertilizer mixture again with its solution, spray application to crop leaf or directly execute in soil, although these selenium-enriched fertilizers can be produced Se-rich farm products and selenium-enriched food, but to only 10%-15% of inorganic selenium crop utilization rate in fertilizer, still very low, using organoselenium may be to improve the approach of crop to se use efficiency.
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 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 microbial inoculum, add solubility selenium preparing, utilize vinelandii to transform inorganic selenium, but only adopt vinelandii in its preparation, only there is 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 to carry out metabolism to material, promote conversion and the absorption of inorganic selenium, and have fixed nitrogen and carbon solidification effect.Patent 100512666C, denomination of invention: a kind of containing selenium microorganism feed addictive and preparation method thereof, 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.Compared with the present invention, the conversion of current microbial inoculum to selenium or adopt single bacterial strain or similar bacterial strain, the conversion to selenium and the utilization of Selenium By The Plants are still lower.And the functional microbial fertilizer of promoting the use of 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 is to provide bacillus subtilis BC-198 and an application thereof.
Another object of the present invention is to provide the microbial inoculum that contains above-mentioned subtilis BC-198.
Still a further object of the present invention is to provide 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 is to provide 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 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 sodium salt thereof.
The present invention also provides the preparation method of described composite fungus agent, comprises the following steps:
1) prepare 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, prepare fermented liquid.
Wherein, described seed culture condition is: seed culture medium is at the 500 milliliters of bottled 100-130 milliliter of triangle substratum, and 28-35 DEG C obtains seed liquor for constant-temperature shaking culture 12-16 hour.
Cultivate the seed culture medium composition of subtilis BC-198: in 100 milliliters of Se solutions, 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;
The seed culture medium composition of blown-ball Azotobacter AS1.212: 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 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 nutrient solution volume ratio respectively, 190220 revs/min of temperature 28-35 DEG C, tank pressure 0.04-0.06MPa, stirring velocity in tank, 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: in Se solution, 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 described fermented liquid, further add sanitas, protective material.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, 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. preparation method can utilize ready-made general fermentation equipment, simple to operate, cost of manufacture is low, the product making has the several functions such as fixed nitrogen, growth promotion, control fungal disease to crop, solubility selenium further activates and transforms through microorganism and is more conducive to crop to selenium absorption and utilizes, after crop applying, can increase output, improve quality of agricultural product, produce agricultural-food selenium content and improve 20 times, reach rich selenium target level of product quality.
Brief description of the 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 illustrating 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, the amendment that the inventive method, step or condition are done or replacement, all belong to scope of the present invention.
If specialize, the conventional means that in embodiment, technique means used is well known to those skilled in the art.
The percentage sign " % " relating in the present invention, if not specified, forefinger mass percent; But the per-cent of solution, unless otherwise specified, refers to and in solution 100ml, contains some grams of solutes; Per-cent between liquid, the ratio of capacity while referring to 20 DEG C.
The rich selenium multi-functional bacterial strain of embodiment 1 separates and qualification
From the selenium-rich tobacco waste of enshi city Baiyangping, by 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 concentration and separation purifying.Antagonistic effect and growth promotion test, screen to obtain a strain bacterial strain BC-198, and this bacterial strain fermentation liquor improves 30-40% to soybean seeds percentage of germination, root length, ratio contrast; Sickle-like bacteria, mould are suppressed to circle and be all greater than 1.5 centimetres.Also there is 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 feminine gender, 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 DEG C of all growths; This bacterial strain 16SrRNA gene sequencing result is as shown in SEQ ID NO.1.
According to cellular form, biochemical test result, and 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 composition: 1) 0.6% glucose, 0.5% sucrose, 0.8% peptone, 0.3% yeast extract paste, 100 milliliters, 0.4% sodium-chlor water, PH7.4; (2) in solid medium, add 1.8% agar
(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 sterilizing 25 minutes, access respectively subtilis and blown-ball Azotobacter, place on bottle swingging machine, 28-30 DEG C shaking culture 14-20 hour for subsequent use.
(3) antagonistic effect
By 100 milliliters of solid mediums of above-mentioned formulated, 0.1MPa sterilizing 25 minutes, while being cooled to 45 DEG C, add 1 milliliter to cultivate in advance the blown-ball Azotobacter nutrient solution of 14 hours, after mixing, draw 15 milliliters of culture dish that move into 9 centimetres of diameters, on the desktop of placement level, make agar culture dish, after cooling 40 minutes, place again Oxford cup, in the cup of Oxford, drip 0.3 milliliter of centrifuged supernatant of cultivating in advance the withered grass gemma nutrient solution of 20 hours, putting 28-30 DEG C of thermostat container and cultivate 24-36, there is not suppressing circle in Oxford cup outer rim.Judge the growth unrestraint effect of subtilis BC-198 to blown-ball Azotobacter.Similar test method judges that blown-ball Azotobacter is to bacillus subtilis bacteria growing also unrestraint.
The preparation of the rich selenium microbial inoculum of embodiment 3
(1) rich selenium agricultural microorganism microbial inoculum adopts bacterial classification: subtilis BC-198;
Cultivate the seed culture medium composition of subtilis BC-198: in 100 milliliters of Se solutions (0.249mg/ml), 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;
By above-mentioned formulated seed culture medium, 100 milliliters of loading amounts in 500 milliliters of triangular flasks, sterilizing 25 minutes under 0.11MPa pressure, after cool to room temperature, by inoculation bamboo shovel picking withered grass gemma BC-198 bacterial classification primary and secondary kind, in access triangular flask substratum, on the bottle swingging machine of 190 revs/min of rotating speeds, 28 DEG C ± 1 DEG C constant-temperature shaking culture 14 hours, makes 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 on 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 sterilizing 25 minutes, substratum temperature drops to 35 DEG C, access 1.8% subtilis seed liquor, set tank pressure 0.04-0.06MPa, 30 ± 2 DEG C of tank temperature, ventilation flow rate 1:0.2(v/v), 190 revs/min of stirring velocitys, ferment and within 18 hours, obtain 2.86 liters of fermentation cultures.
(3) add 0.19% Sodium Benzoate, 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) prepare rich selenium agricultural microorganism microbial inoculum and adopt bacterial classification: subtilis BC-198, and blown-ball Azotobacter AS1.212.
2) rich selenium agricultural microorganism microbial inoculum seed culture medium composition:
A. containing the preparation of the selenium aqueous solution: selenium mineral powder is broken, crosses 100-200 order, adds 3%-4% ammoniacal liquor and 1%-3% sodium hydroxide mixing solutions in the ratio of selenium mineral powder mass volume ratio 1:2-3, stirs 24-36 hour, uses H
3pO
4adjust PH8.5-9.0 to filter to obtain supernatant liquor, measuring and adjust selenium concentration is 0.25-1.0mg/ml, or directly uses NaSeO
3preparation.
B. substratum composition
Cultivate the seed culture medium composition of subtilis BC-198: in 100 milliliters of Se solutions (0.25mg/ml), 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.
The seed culture medium composition of blown-ball Azotobacter AS1.212: 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 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 sterilizing 25 minutes, access respectively subtilis BC-198 and blown-ball Azotobacter AS1.212, place on 200 revs/min of rotary shaking bottle machines, 30-32 DEG C obtains the seed liquor of selenium-enriched microbe microbial inoculum for constant-temperature shaking culture 12-16 hour.
(2) fermentation culture: the preparation of agricultural microorganism microbial inoculum is carried out in 10L fermentor tank, its substratum composition: 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).Substratum stocking volume is 65% of fermentor tank volume, 0.12MPa sterilizing 25 minutes, substratum temperature drops to 32 DEG C, access 1.4% subtilis BC-198 seed liquor and 1.5% blown-ball Azotobacter AS1.212 seed liquor simultaneously, control 30 DEG C of the interior temperature of tank, tank internal pressure 0.06MPa, 190 revs/min of stirring velocitys, substratum with pass into volume of air and compare 1:0.4, access seed liquor rises, every pH value and the bacterial cell concentration of sampling and measuring fermentation in 2 hours, find fermentation 18 hours and 19 hours, bacterial cell concentration does not rise appreciably, fermentation culture ferments 18-19 hour to obtain.
(3) emit in 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 obtain.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) prepare rich selenium agricultural microorganism microbial inoculum and adopt bacterial classification: subtilis BC-198, and blown-ball Azotobacter AS1.212
2) rich selenium agricultural microorganism microbial inoculum seed culture medium composition:
A. containing the preparation of the selenium aqueous solution: the selenium that contains of preparation substratum leaches water preparation: pulverize through ball mill in 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, stirs 30 hours, uses H
3pO
4adjust PH8.7 centrifuging to obtain the rich selenium aqueous solution, regulator solution selenium content is 0.3mg/ml.
B. substratum composition
Cultivate the seed culture medium composition of subtilis BC-198: in 100 milliliters of Se solutions, 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.
The seed culture medium composition of blown-ball Azotobacter AS1.212: 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 Se solutions.
C. spawn culture
By the seed culture medium of above-mentioned substratum composition preparation, 120 milliliters of substratum of loading amount in 500 milliliters of triangular flasks, 0.12MPa sterilizing 25 minutes, plant and move on to respectively in triangular flask nutrient solution with inoculation shovel picking subtilis BC-198 and blown-ball Azotobacter AS1.212 mother, place on 200 revs/min of rotary shaking bottle machines 30-32 DEG C 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.Substratum stocking volume be fermentor tank volume 70%, 0.12MPa sterilizing 25 minutes, substratum temperature drops to 30-35 DEG C, the subtilis seed liquor of access 1.14% and 2.43% blown-ball Azotobacter seed liquor, set 195 revs/min of fermentation stirring velocitys, 28 DEG C of temperature in tank, tank internal pressure 0.05MPa, air flow quantity 1:0.3(v/v in tank).After access seed, find every 3 hours sampling and measuring fermented liquid PH and bacterial cell concentration, the bacterial cell concentration of fermenting 18 hours time is close with 15 hours, and cell concn is not significantly increased.
(3) emit in tank 33 liters of zymocyte liquids, add 0.2% Sodium Benzoate and 0.8% polyglutamic acid, stir, obtain 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) prepare rich selenium agricultural microorganism microbial inoculum and adopt bacterial classification: subtilis BC-198, and blown-ball Azotobacter AS1.212
2) rich selenium agricultural microorganism microbial inoculum seed culture medium composition:
A. containing the preparation of the selenium aqueous solution:
B. substratum composition
Cultivate the seed culture medium composition of subtilis BC-198: in 100 milliliters of Se solutions (0.249mg/ml), 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.
The seed culture medium composition of blown-ball Azotobacter AS1.212: 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 Se solutions (0.249mg/ml)
C. seed culture
By above-mentioned formulated seed culture medium, 100 milliliters of loading amounts in 500 milliliters of triangular flasks, sterilizing 25 minutes under 0.11MPa pressure, after cool to room temperature, by inoculation bamboo shovel picking withered grass gemma BC-198 and blown-ball Azotobacter AS1.212 bacterial classification primary and secondary kind respectively, access respectively in triangular flask substratum, on the bottle swingging machine of 190 revs/min of rotating speeds, 28 DEG C ± 1 DEG C 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 on 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.Substratum stocking volume be fermentor tank volume 70%, 0.12MPa sterilizing 25 minutes, substratum temperature drops to 35 DEG C, access 1.2% rise subtilis seed liquor, 1.8% blown-ball Azotobacter seed liquor, set tank pressure 0.04-0.06MPa, 30 ± 2 DEG C of tank temperature, ventilation flow rate 1:0.2(v/v), 190 revs/min of stirring velocitys, ferment and within 18 hours, obtain 132 liters of fermentation cultures.
(3) add 0.3% Sodium Benzoate, 0.5% sodium alginate, 0.15% polyglutamic acid stirs, must be containing 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
The selenium-enriched microbe microbial inoculum that winter in 2008 is used embodiment 5 to prepare on potato: every strain is used containing the rich selenium agricultural microorganism of selenium 0.304mg/ml microbial inoculum 0.065kg, result shows: use selenium-enriched microbe microbial inoculum fertilizer group potato mean yield 3.51kg/ strain, yampi ratio 70.98%, potato, on average containing selenium 2.11mg/kg, is not used average containing 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 after this fertilizer to improve 20.68 times, reach rich selenium standard.
The selenium-enriched microbe microbial inoculum that uses embodiment 6 to prepare on cucumber in 2010-2012 continuous 3 years: every strain is used containing the rich selenium agricultural microorganism of selenium 0.284mg/ml microbial inoculum 0.062kg.Fresh cucumbers is on average containing selenium 2.226mg/kg, and untapped fresh cucumbers, on average containing 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 green organic product standard, and have extremely significant effect of increasing production.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, 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.
The selenium-enriched microbe microbial inoculum that uses embodiment 6 to prepare on potato in 2010-2012 continuous 3 years, every strain is used containing the rich selenium agricultural microorganism of selenium 0.284mg/ml microbial inoculum 0.0625kg, potato is on average containing selenium 1.935mg/kg, do not use the average selenium 0.0941mg/kg of containing, 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 crop applying microbial inoculum of the present invention, reduce morbidity, improve quality of agricultural product, produce 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 without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (8)
1. a composite fungus agent, its effective constituent is subtilis (Bacillus Subitilis) BC-198 and blown-ball Azotobacter AS1.212,
Wherein, subtilis BC-198, its preserving number is CCTCC No.M2012321.
2. composite fungus agent according to claim 1, 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.
3. composite fungus agent according to claim 2, 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.
4. according to the composite fungus agent described in claim 1-3 any one, it is characterized in that, also contain sanitas and protective material;
Wherein, described sanitas is one or more in benzoic acid and sodium benzoate, Sorbic Acid and sodium salt thereof, propionic acid and 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 sodium salt thereof.
5. according to the composite fungus agent described in claim 1-3 any one, it is characterized in that, be prepared as follows:
1) prepare 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, and prepare fermented liquid.
6. composite fungus agent according to claim 5, is characterized in that, in described complex ferment cultivation, the access amount of subtilis BC-198 seed liquor, blown-ball Azotobacter AS1.212 seed liquor is respectively 1-2% and 1.5-2.5%.
7. composite fungus agent according to claim 6, is characterized in that, further adds sanitas and protective material;
Wherein, described sanitas is one or more in benzoic acid and sodium benzoate, Sorbic Acid and sodium salt thereof, propionic acid and 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 sodium salt thereof.
8. the application of composite fungus agent in improving the absorption of Selenium By The Plants and utilizing described in claim 1.
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| 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 |
| CN105950503B (en) * | 2016-05-19 | 2019-04-16 | 湖北民族学院 | Bacillus subtilis SE201412 and organic selenium-rich microbial inoculum |
| CN105994937A (en) * | 2016-05-19 | 2016-10-12 | 湖北民族学院 | Green organic selenium-rich feed and preparation method thereof |
| 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 |
| CN110643526B (en) * | 2019-08-31 | 2022-06-14 | 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 |
| CN115261257B (en) * | 2022-05-25 | 2023-04-25 | 武汉轻工大学 | Selenium-enriched bacillus subtilis L11 from silage and application thereof |
| 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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Address after: 445000 Enshi city in Hubei Province Economic Development Zone Hongmiao Ping Liquidambar date dam Patentee after: Hubei and Connaught biological engineering Limited by Share Ltd Patentee after: Chen Jiaren Patentee after: Liu Jinlong Address before: 445000 Hubei Province, Enshi Tujia and Miao Autonomous Prefecture of Enshi Economic Development Zone Hongmiao Liquidambar Ping dam date Patentee before: Enshi Heno Biologic Engineering Co., Ltd. Patentee before: Chen Jiaren Patentee before: Liu Jinlong |