CN101531971B - Bacillus subtilis and application thereof in prevention and treatment of continuous cropping obstacle of greenhouse vegetable - Google Patents
Bacillus subtilis and application thereof in prevention and treatment of continuous cropping obstacle of greenhouse vegetable Download PDFInfo
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- CN101531971B CN101531971B CN2008100524463A CN200810052446A CN101531971B CN 101531971 B CN101531971 B CN 101531971B CN 2008100524463 A CN2008100524463 A CN 2008100524463A CN 200810052446 A CN200810052446 A CN 200810052446A CN 101531971 B CN101531971 B CN 101531971B
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Abstract
The invention relates to Bacillus subtilis and the application thereof in prevention and treatment of continuous cropping obstacle of greenhouse vegetable. The Bacillus subtilis is M-2 strain with the preservation number of CGMCC No. 2271. The M-2 strain is plant growth-promoting rhizobacteria separated from oceanic mangrove plant, which can restrain the growth of various harmful fungi such as fusarium wilt of cucumber, root rot of tomato, the root rot of soybean and the like, and can effectively prevent and treat the continuous cropping obstacle of the greenhouse vegetable and increase the quality and output of the greenhouse vegetable.
Description
Technical field
The invention belongs to biotechnology and agricultural disease Prevention Technique field, specifically, it relates to a kind of subtilis Bacillus subtilis and the application in the facility for prevention and control obstruction of vegetable continuous cropping thereof.
Background technology
Show that according to Food and Argriculture OrganizationFAO's survey data the annual farm crop of being seized by disease and pest in the whole world account for 20%～30% of harvest, the financial loss that causes thus reaches 1,200 hundred million dollars.Mainly be to prevent and treat at present through spraying chemical reagent.But, even poisonous substance occurs and be accumulated in the phenomenon that causes poisoning in domestic animal and the human body gradually because a large amount of chemical pesticides that use have caused the pollution in various degree of air, water source, soil and food.The whole world has 2,000,000 people to poison because of chemical pesticide every year approximately, and wherein nearly 40,000 people are dead.And some chemical pesticide of life-time service also can make insect develop immunity to drugs, and the disclosed statistical information of a tree name shows to have existing 417 kinds at present of drug-fast insects.In recent years, in agriculture prodn, applying biological agricultural chemicals substituting chemical pesticide becomes a kind of trend gradually.Countries in the world are to all drawbacks of chemical pesticide, developed that a series of selectivity are strong, efficient is high, cost is low, pollution-free, to person poultry harmless's biotechnological formulation.China also will progressively use the biological pesticide substituting chemical pesticide, particularly in the production of vegetables, will use biological pesticide as early as possible.
China's facilities vegetable cultivated area had surpassed 2,000,000 hectares in 2008, had become the maximum country of world's facilities horticulture area at present.Facilities vegetable has become the most great-hearted new industry in China's agriculture prodn, for huge effect has been played in the construction of China's vegetable basket project.But facilities vegetable has highly intensification, multiple crop index height and the single characteristics of kind; Continuous cropping for many years; Occurred that edatope deterioration, vegetables disease and pest are serious, output reduces, quality of vegetable becomes of inferior quality a series of bad phenomenon; The Sustainable development that the serious threat facilities vegetable is produced, the obstacle that continuous cropping causes have become a big technical barrier that needs to be resolved hurrily in the vegetable growing production.
Causing that crop comprises that the reason of facilities vegetable continuous cropping obstacle is complicated, is the external manifestation of two internal system variant factors of crop-soil exercising result.The reason that Different Crop produces continuous cropping obstacle possibly be different.Nineteen eighty-three, Japan's reason that will produce continuous cropping obstacle reduced the five big factors: (1) soil nutrient wanes; (2) soil reaction unusual (toxin); (3) soil physical property worsens; (4) from the objectionable impurities of plant; (5) soil microorganisms changes.Stress simultaneously: in this five big factor, the variation of soil microflora is the main factor of continuous cropping obstacle, and other is a cofactor.The research of root secretion was become the focus of announcement continuous cropping obstacle mechanism in recent years both at home and abroad.
Show according to disclosed research data: no matter booth or open country, the kind of mikrobe all is various in the soil.In each monoid mikrobe, generally all be at most with the bacterial number, secondly be actinomycetes, fungi quantity occupies the 3rd; Mentioned microorganism quantity and microorganism active all are higher than open country in the booth.Be main with penicillium spp, aspergillus tubigensis, little Ke Yinhan mould in the fungi, facultative parasite Helminthosporium sp. bacterium, powder spore are mould etc., and pathogenic fungi all has existence in booth, open country soil.Therefore, these fungus-caused diseases are not facility greenhouse, the distinctive obstruction factor of plastic shed soil.See that from the plantation time limit bacterial number does not have considerable change, and in plantation year limit for length's the booth fungi, actinomycetes quantity plant in the short booth of the time limit less, the generation of this situation maybe with higher humidity in the booth and relevant to its inhibition than supersalinity.
Disclosed research data also shows: the soil-borne disease insect pest is to cause the topmost factor in the continuous cropping obstacle factor; Relevant continuous cropping obstacle causal investigation result shows, causes that about 70% plot of obstruction of vegetable continuous cropping is because the insect pest of soil infection venereal disease is caused.Continuous cropping provides the host that root diseases depends on for existence and the place of breeding; Make the pathogenic bacteria quantity in the soil constantly increase; Particularly cause of disease antimicrobial short of money reduces in the soil because too much use chemical fertilizer brings in recent years, has more increased the weight of the generation of soil-borne disease insect pest.
Rhizospheric microorganism is of a great variety and active, and the mikrobe that wherein can promote plant-growth, prevent and treat disease, increase crop yield is called as short livings bacterium (Plant Growth-Promoting Rhizobacteria, abbreviation PGPR).Between they and plant is intergrowth relation, interacts, mutually promotes with root system of plant.Mikrobe accumulates in around the root system in a large number, changes organism into inorganics, for plant provides effective nutriment; Simultaneously, mikrobe can also secrete VITAMINs, and growth stimulant etc. promote plant-growth.In growing process, the dead root system and the cast (root hair, epidermic cell, root cap etc.) of root, and root system is important source of nutrition of mikrobe and energy derive to exocrine inorganics of root and organism; Because interting of root system is superior to outside the rhizosphere aeration condition of rhizosphere and water regime, thereby form the ecotope that is beneficial to mikrobe.
PGPR can suppress the breeding of harmful pathogenic micro-organism in the soil; And can produce the meta-bolites of useful plants growth; Thereby can be used for preventing and treating continuous cropping obstacle; Promote growth and development of plant, improve the quality of vegetables, its research haves a bright future with industrialization, inspiring, fact proved that the PGPR industrialization is the key areas and the direction that develop of agricultural biological technical field both at home and abroad from now on.
Summary of the invention
The objective of the invention is to the serious day by day technical problem of industrialized agriculture continuous cropping obstacle, and a kind of subtilis and the application in the facility for prevention and control obstruction of vegetable continuous cropping thereof are provided.
The subtilis that the present invention relates to (Bacillus subtilis) M-2 bacterial strain is preserved on November 29th, 2007 that " China Committee for Culture Collection of Microorganisms's common micro-organisms " center ", its preserving number are CGMCC No.2271.
Said subtilis (Bacillus subtilis) M-2 CGMCC No.2271 is from the mangrove of ocean, to separate to obtain, and after the substratum enrichment, obtains through the inhibition zone experiment sieving.
The cell morphological characteristic of said subtilis (Bacillus subtilis) M-2 CGMCC No.2271: gram-positive microorganism, shaft-like, size is 0.5～1.0 μ m * 3.5～6.0 μ m, and the gemma end is given birth to or inferior end is given birth to, and peritrichous can move.
The physiological and biochemical property of said subtilis (Bacillus subtilis) M-2 CGMCC No.2271: 27 ℃～45 ℃ of growth temperatures, pH4～9, NaCL tolerance 2%～6%, catalase test, H
2S test, starch hydrolysis experiment, cellulose hydrolysis test and MR test are all positive, and the test of phenylalanine(Phe) deamination, urease test, oxidase test, gelatin liquification test, tyrosine degraded test and V-P test are all negative, produce acid-utilising N.F,USP MANNITOL; Can not utilize pectinose; Semi-lactosi, wood sugar, rhamnosyl; Sucrose, inose and Citrate trianion.
The Nucleotide of the 16S rRNA of said subtilis (Bacillus subtilis) M-2 CGMCC No.2271 has sequence shown in sequence table.
The application of said subtilis Bacillus subtilis M-2 CGMCC No.2271 in the facility for prevention and control obstruction of vegetable continuous cropping.
The application method of said subtilis Bacillus subtilis M-2 CGMCC No.2271 bacterial strain is fermentation culture → dilution → sprinkling → effect measuring.
The fermention medium of said subtilis Bacillus subtilis M-2 CGMCC No.2271 bacterial strain consists of g/L: glucose 10, and yeast extract paste 10, Carnis Bovis seu Bubali cream 4, peptone 4, NaCl 25, pH7.2～7.4.
The bacterial strain that the present invention relates to; Content with reference to " Bergey ' s Mannual of Systematic Bacteriology " Vol.VIII; According to its morphological specificity and physiological and biochemical property; And 16S rRNA gene order result, identify it is a kind of subtilis (Bacillus subtilis) through polyphase sort, called after M-2.
M-2CGMCC No.2271 strain bacterium provided by the invention; It is the short living bacterium (PGPR) of a kind of good rhizosphere; Fungal disease in the facilities vegetable continuous cropping obstacle had very strong restraining effect; Can suppress the growth of multiple pathomycete, particularly prevent and treat the mould and mould Plant diseases that causes of epidemic disease of cucumber fusarium axysporum, cotton wilt, damping-off and various corruption.Growth for promoting plant has outstanding advantages such as efficient, pollution-free.
Description of drawings
Accompanying drawing 1 is the Photomicrograph of subtilis Bacillus subtilis M-2 CGMCC No.2271 bacterial strain.
Accompanying drawing 2 is that subtilis Bacillus subtilis M-2 CGMCC No.2271 bacterial strain is to fungi exercising result mensuration figure in the continuous cropping soil.
To be the different extension rates of subtilis Bacillus subtilis M-2 CGMCC No.2271 bacterial strain fermentation liquor compare comparison film to the seedling growth-promoting effect of taking root with accompanying drawing 3.
Accompanying drawing 4 is subtilis Bacillus subtilis M-2 CGMCC No.2271 bacterial strains to the restraining effect of cotton wilt germ than comparison film, wherein A for effect after, B for effect before.
Accompanying drawing 5 is to the photo of growth of mung beans influence, after wherein A is effect, before B is effect after the subtilis Bacillus subtilis M-2 CGMCC No.2271 inoculation.
Accompanying drawing 6 is to the photo of Chinese cabbage growth of seedling influence, after wherein A is effect, before B is effect after the subtilis Bacillus subtilis M-2 CGMCC No.2271 inoculation.
Embodiment
Further specify the present invention below in conjunction with embodiment and accompanying drawing thereof.
Embodiment 1:
The screening and the breeding of subtilis Bacillus subtilis M-2 bacterial strain provided by the invention.
The root of ocean mangrove is immersed in about 20min in a large amount of sterilized waters, and flush away sticks to the soil on the root, and then with the residual soil of root under the rinsed with sterile water, this part soil is the rhizosphere soil sample.Water sample is collected in that 100mL is clean, in the wide-mouth Plastic Bottle of sterilization, gets 15mL after the concussion evenly, and (form: peptone 5g by substratum in the 100mLZoBell substratum for aseptic inoculation; Yeast extract paste 1g, tertiary iron phosphate 0.1g, Chen Haishui 1000mL; PH7.6～7.8), 36 ℃ of concussions were cultivated 6 days.On the LB flat board, observe, cultivate colony diameter 3～5mm of one day, rounded, neat in edge, surface wettability.After will cultivating 6 days enrichment culture liquid mixings; Getting 0.1mL coats in the nutrient agar; 36 ℃ of constant temperature culture 2 days are numbered according to colonial morphology, are indicator with the myxococcus albus with all mikrobes that are separated to; Screen through inhibition zone method, finally obtain the best M-2 bacterial strain of fungistatic effect as shown in Figure 1.
Embodiment 2:
The morphological specificity of subtilis Bacillus subtilis M-2 bacterial strain provided by the invention and the evaluation of physiological and biochemical property.
TP with reference to " Bergey ' s Mannual of Systematic Bacteriology " Vol.VIII is carried out, and detects its gramstaining, and thalline size and form have atrichia and gemma, growth temperature, pH scope.The NaCL tolerance.Catalase, H
2S test, starch hydrolysis, cellulose hydrolysis, MR test, phenylalanine(Phe) deamination, urease test, oxydase, gelatine liquefication, tyrosine degraded, V-P test, and utilize N.F,USP MANNITOL; Pectinose, semi-lactosi, wood sugar; Rhamnosyl, sucrose, inose and citrate test.
The result shows that this bacterium is Gram-positive, and is shaft-like, and size is 0.5 μ m～1.0 μ m * 3.5 μ m～6.0 μ m, and the gemma end is given birth to or inferior end is given birth to, and peritrichous can the motion (see figure 1).27～45 ℃ of growth temperatures, pH4～9, NaCL tolerance 0～2%.Catalase, H
2S test, starch hydrolysis, cellulose hydrolysis, MR test are all positive, phenylalanine(Phe) deamination, urease test, oxydase, gelatine liquefication, tyrosine degraded,, the V-P test is all negative, can utilize N.F,USP MANNITOL; Can not utilize pectinose; Semi-lactosi, wood sugar, rhamnosyl; Sucrose, inose and Citrate trianion.
Embodiment 3:
The pcr amplification and the sequencing of the 16S rRNA gene of subtilis Bacillus subtilis M-2 bacterial strain provided by the invention.
With this inoculation in the LB substratum, 120rpm, 37 ℃ of concussions were cultivated 10 hours; Centrifugal collection thalline; The back that suspends adds N,O-Diacetylmuramidase and SDS broken wall, adopts phenol-chloroform method extraction genomic dna, with positive primer 2 7F (5 '-GAGAGTTTGATCCTGGCTCAG-3 ') and reverse primer 1541R (5 '-AAGGAGGTGATCCAGCC-3 '); Its 16S rRNA gene is carried out pcr amplification, the product of amplification is won companies by Beijing three check order.The PCR condition is: 94 ℃, and 10min; 94 ℃, 45s; 55 ℃ of 45s; 72 ℃, 90s; 30 circulations, 4 ℃ of preservations.16S rRNA gene order length is 1543bp, and the number of landing in GenBank is EU333948, with Bacillus subtilis (AB110598) similarity be 99%.Its result is shown in sequence table.
Embodiment 4:
The fermentation culture of subtilis Bacillus subtilis M-2 bacterial strain provided by the invention.
On the LB substratum, 36 ℃ of following activation 24 hours are transferred to activatory M-2 bacterial strain then and carry out shake flask fermentation in the basic medium with the M-2 inoculation, and temperature is 30～36 ℃, and shaking speed is 200rpm, and fermentation time is 72 hours.Wherein fermention medium consists of g/L: glucose 10, and yeast extract paste 10, Carnis Bovis seu Bubali cream 4, peptone 4, NaCl 25, pH7.2～7.4.
Embodiment 5:
Adopt dilution-plate method, respectively the fungi quantity of soil behind continuous cropping soil and the inoculation M-2 bacterial strain is measured, substratum is the Ma Dingshi substratum, and every ware adds 1 paraxin and suppresses the bacterium control.Place 28 ℃ thermostat container to cultivate counting 3 days.The result shows that marine bacteria M-2 can effectively suppress to cause harmful bacteria growing in the continuous cropping soil, and the exercising result fungi quantity of inoculating after 20 weeks as shown in Figure 2 is reduced to 30% of control soil.
Embodiment 6:
Subtilis Bacillus subtilis M-2 bacterial strain provided by the invention is to the restraining effect test of various hypha,hyphaes.
Employing contains toxic medium method; The fermented liquid that in the 20mLPDA substratum, adds 2mL M-2, the flat board of falling PDA uses the pathogenic fungi mycelia piece of punch tool cut-off footpath as 5mm; Move into dull and stereotyped center; Cultivate 24h, 48h, 72h and 96h for 28 ℃ and measure mycelial growth diameter (mm), repeat 3 times, calculate the bacteriostasis rate under each concentration of treatment.
Bacteriostasis rate=[(the mycelial growth diameter is handled in contrast mycelial growth diameter-test)/contrast mycelial growth diameter] * 100%.The result shows; Its inhibiting rate to the canker of apple fruit germ is 99.5%, is 99.1% to the inhibiting rate of hami melon root rot germ, is 98.5% to the inhibiting rate of cotton wilt germ; Inhibiting rate to the corn southern leaf blight germ is 97.5%; The inhibiting rate of rice seedling blight germ is 98.3%, is 96.5% to the inhibiting rate of root rotof flax germ, is 97.5% to the inhibiting rate of Alternaria alternate germ.As shown in Figure 4, this bacterium can effectively suppress the cotton wilt pathogen growth.
Embodiment 7:
Subtilis Bacillus subtilis M-2 bacterial strain provided by the invention is to the promoter action test of mung bean seedlings root growth.
Get the M-2 fermented liquid, centrifugal, get supernatant, it is diluted 5,10,100,1000 and 10000 times respectively; Zero(ppm) water is set does contrast (Control), each diluent is joined in the small test tube of 10mL, with kraft paper with seal film and seal; A hole is made a call in the centre therein, and green bean-leaf is planted into, one of every pipe; Cultivated 7 days, four repetitions are set, the result sees table 1 and Fig. 3.
Table 1 M-2 is to the promoter action of mung bean seedlings root growth
Table 1 explanation, M-2 bacterial strain can effectively promote the seedling growth of taking root, and effect is best when wherein diluting 100 times.Among Fig. 3 CK, a, b, c, d, e be respectively zero(ppm) water contrast, during with 5,10,100,1000 and 10000 times of fermented liquid dilutions to the influence of green bean-leaf root growth, can find out that therefrom the M-2 bacterial strain has good promoter action to the seedling growth of taking root.
Embodiment 8:
After the subtilis Bacillus subtilis M-2 CGMCC No.2271 inoculation provided by the invention to the influence of growth of mung beans.
Mung bean is planted in the soil after the continuous cropping, a winding kind M-2 bacterial strain, another group is not inoculated as blank (Control), cultivates the growing state of observing mung bean after 8 days.As can be seen from Figure 5 inoculate the growth that can effectively promote green bean-leaf behind the M-2 bacterial strain.
Embodiment 9:
After the subtilis Bacillus subtilis M-2 CGMCC No.2271 inoculation provided by the invention to the influence of growth of mung beans.
Mung bean is planted in the soil after the continuous cropping, a winding kind M-2 bacterial strain, another group is not inoculated as blank (Control), cultivates the growing state of observing mung bean after 8 days.As can be seen from Figure 6 inoculate the growth that can effectively promote green bean-leaf behind the M-2 bacterial strain.
Embodiment 10:
Subtilis Bacillus subtilis M-2 bacterial strain provided by the invention is at cucumber test Tanaka's application test.
Can suppress cucumber fusarium axysporum through fermented liquid, detailed process is: M-2 ferments to bacterial strain, 1000 times of sprayings of fermented liquid dilution, and the cucumber seedling transplant survival begins to spray medicine after one week, and each 20-25 days at interval, 3-4 time continuously.Its biocontrol effect can reach 73%.Fermentation culture conditions is: on the LB substratum, 36 ℃ of following activation 24 hours are transferred to activatory M-2 bacterial strain then and carry out fermentation culture in the basic medium with the M-2 inoculation, and temperature is 30～36 ℃, and shaking speed is 200rpm, and fermentation time is 72 hours.Wherein fermention medium consists of g/L: glucose 10, and yeast extract paste 10, Carnis Bovis seu Bubali cream 4, peptone 4, NaCl 25, pH7.2.
Embodiment 11:
The application of subtilis Bacillus subtilis M-2 bacterial strain provided by the invention in the tobacco experimental plot.
Can suppress Alternaria alternate through fermented liquid; Detailed process is: M-2 ferments to bacterial strain, 1000 times of sprayings of fermented liquid dilution, the 1st the foliar spray dispenser of seedbed cross phase; Execute medicine behind the transplant survival after 5-7 days the 2nd time; Interval dispenser in 20 days later on 1 time, totally 3 times, whole growing reagent 4 times.Its biocontrol effect can reach 71%.Fermentation culture conditions is: on the LB substratum, 36 ℃ of following activation 24 hours are transferred to activatory M-2 bacterial strain then and carry out fermentation culture in the basic medium with the M-2 inoculation, and temperature is 30～36 ℃, and shaking speed is 200rpm, and fermentation time is 72 hours.Wherein fermention medium consists of g/L: glucose 10, and yeast extract paste 10, Carnis Bovis seu Bubali cream 4, peptone 4, NaCl 25, pH7.2.
Embodiment 12:
The application of Bacillus subtilis M-2 bacterial strain provided by the invention in the tobacco experimental plot.
Can suppress the tomato root rot through fermented liquid, detailed process is: M-2 is fermented 1000 times of sprayings of fermented liquid dilution; Medicine is executed in the 1st the foliar spray dispenser of seedbed cross phase the 2nd time after 5-7 days behind the transplant survival, interval dispenser in 20 days later on 1 time; Totally 3 times, whole growing reagent 4 times.Its biocontrol effect can reach 70%.Fermentation culture conditions is: on the LB substratum, 36 ℃ of following activation 24 hours are transferred to activatory M-2 bacterial strain then and carry out fermentation culture in the basic medium with the M-2 inoculation, and temperature is 30～36 ℃, and shaking speed is 200rpm, and fermentation time is 72 hours.Wherein fermention medium consists of g/L: glucose 10, and yeast extract paste 10, Carnis Bovis seu Bubali cream 4, peptone 4, NaCl 25, pH7.4.
Sequence table
Claims (6)
1. a subtilis (Bacillus subtilis) is that preserving number is the M-2 bacterial strain of CGMCC No.2271.
2. subtilis as claimed in claim 1, it is characterized in that: gram's staining is positive, and is shaft-like, and size is 0.5 μ m～1.0 μ m * 3.5 μ m～6.0 μ m, and the gemma end is given birth to or inferior end is given birth to, and peritrichous can move; 27 ℃～45 ℃ of growth temperatures, pH4～9, NaCl tolerance 0～2%; Catalase test, H2S test, starch hydrolysis experiment, cellulose hydrolysis test and MR test are all positive; The test of phenylalanine(Phe) deamination, urease test, oxidase test, gelatin liquification test, tyrosine degraded test and V-P test are all negative, produce acid-utilising N.F,USP MANNITOL.
3. subtilis as claimed in claim 1, the Nucleotide of its 16S rRNA is sequence shown in SEQ ID NO:1.
4. the application of subtilis Bacillus subtilis M-2CGMCC No.2271 as claimed in claim 1 in the facility for prevention and control obstruction of vegetable continuous cropping.
5. the application of subtilis Bacillus subtilis M-2CGMCC No.2271 as claimed in claim 4 in the facility for prevention and control obstruction of vegetable continuous cropping is characterized in that M-2 bacterial strain application method is fermentation culture → dilution → sprinkling → effect measuring.
6. the application of subtilis Bacillus subtilis M-2CGMCC No.2271 as claimed in claim 5 in the facility for prevention and control obstruction of vegetable continuous cropping; It is characterized in that fermention medium consists of: glucose 10g/L; Yeast extract paste 10g/L, Carnis Bovis seu Bubali cream 4g/L, peptone 4g/L; NaCl 25g/L, pH7.2～7.4.
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| CN102465096B (en) * | 2010-11-15 | 2015-04-15 | 中国科学院沈阳应用生态研究所 | Microbial agent as well as preparation method and application thereof |
| CN102845614B (en) * | 2012-09-11 | 2014-10-22 | 重庆市畜牧科学院 | Method for producing lactic acid-producing bacillus subtilis symbiotic feed additive bacteria solution |
| CN104611252B (en) * | 2014-12-13 | 2017-06-30 | 苏新宏 | A kind of tobacco rhizosphere Promoting bacteria TC6 and its application |
| CN104961538B (en) * | 2015-05-08 | 2017-11-14 | 淮阴工学院 | Recessed soil matrix immobilization complex micro organism fungicide for improveing capsicum continuous cropping obstacle and preparation method thereof |
| CN106222121B (en) * | 2016-09-23 | 2019-07-02 | 南京农业大学 | A kind of bacillus megaterium bacterial strain, biocontrol agent and the preparation method and application thereof |
| CN107937585A (en) * | 2018-01-18 | 2018-04-20 | 山东出入境检验检疫局检验检疫技术中心 | Specific primer, kit and detection method for Rapid identification life cancer enterobacteria |
| CN108934685B (en) * | 2018-06-26 | 2020-11-13 | 石阡县苔玺茶业有限公司 | Planting method for high-yield tea trees |
| CN109957535B (en) * | 2019-05-08 | 2020-09-11 | 河北省农林科学院遗传生理研究所（河北省农林科学院农产品质量安全研究中心） | Bacillus simplex, microbial agent prepared by using bacillus simplex, biological fertilizer and application |
| CN112167267A (en) * | 2020-11-04 | 2021-01-05 | 河南柏裕植物免疫科技有限公司 | Compound biological agent for preventing and treating tomato root rot and preparation method thereof |
| CN113139461A (en) * | 2021-04-23 | 2021-07-20 | 塔里木大学 | Wheat leaf disease and insect pest detection system for agricultural planting and management method thereof |
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| CN1621514A (en) * | 2003-11-28 | 2005-06-01 | 中国农业科学院生物防治研究所 | Lichen bacillus strain (B-0A12) and preparation thereof |
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