CN109836190A - A method of biocontrol bacterial fertilizer is prepared using biocontrol bacteria and eliminates muskmelon continuous cropping obstacle - Google Patents
A method of biocontrol bacterial fertilizer is prepared using biocontrol bacteria and eliminates muskmelon continuous cropping obstacle Download PDFInfo
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Abstract
A method of biocontrol bacterial fertilizer being prepared using biocontrol bacteria, using the separation, the identification that, as bacterial strain, carry out strain in the isolated bacillus subtilis of kiwi fruit tree garden；It is added to the soy meal of sterilizing, ammonium nitrate, sucrose, magnesium sulfate, manganese sulfate in the carrier matrix of sterilizing, and the seed liquor of sterile water and biocontrol bacteria is added, stir, obtains hair fermentation raw material；Fermentation raw material is fermented, ensures that biocontrol bacteria number reaches 10 in medium carrier10CFU/g DW carrier matrix, the previous moon is transplanted in Muskmelon Seedlings, the biocontrol bacterial fertilizer of preparation even is spread in melonry according to 135kg/ is per mu, it can inhibit soil fungi growth, balanced soil microbiologic population balance, the content for reducing muskmelon root exudates main component coumaric acid in soil, to eliminate the continuous cropping obstacle in muskmelon production, it realizes agricultural sustainable development, safeguards ecological environment.
Description
Technical field
The invention belongs to planting industry technical fields, are related to Muskmelon Planting, in particular to a kind of to utilize biocontrol bacteria preparation life
Anti- bacterial manure and the method for eliminating muskmelon continuous cropping obstacle.
Background technique
Continuous cropping obstacle refers to after of the same race or equal crop is planted in same plot for years, even if using normal cultivation
The phenomenon that management measure can also make plant growing way die down, pest and disease damage aggravates, yield and quality decline.Ancient Times in China just has
About after continuous cropping disease aggravate record, be referred to as continuous cropping disease, American-European countries be referred to as soil-borne disease, continuous cropping obstacle,
Replant disease.Japan is referred to as to avoid ground phenomenon.Continuous cropping obstacle shows themselves in that seedling (strain) growth is weak, and root system development is bad, susceptible
Disease, plant resistance reduction etc..
In recent years, with the quickening that rural laborer's population shifts, in addition cultivated area is limited, especially because facility melon
The appearance of the production models such as industrialization, Focus, the scale of fruit vegetable production, more exacerbates gourd, fruit and vegetable continuous cropping obstacle phenomenon
Generation, less serious case causes the underproduction reduction of income, and severe one causes to have no harvest, and seriously constrains the sustainable development of facility gourd, fruit and vegetable production.
Muskmelon Planting mostly uses greenhouse to produce at present.Greenhouse production can be effectively controlled investment risk, planting industry investment repayment
Rate is higher, and benefit is stablized.But continuous cropping obstacle easily occurs for greenhouse production.The cause that continuous cropping obstacle occurs is extremely complex, is to plant
Many factors are interactional as a result, still generally believing at present causes the principal element of continuous cropping obstacle to be native biography in object and soil
Disease, soil physico-chemical property and Auto toxicity.
Although soil-borne disease can eliminate soil fungi by measures such as pouring roots, going out in applying pesticides except soil is true
While bacterium, other microorganisms in soil have also been killed；Long-term one crop varieties of production list, eventually lead to soil physical chemistry
The deterioration of matter is unfavorable for the growth of crop；And soil cannot be eliminated though can promote plant growth by applying agricultural and chemical fertilizer
The suppression harmful bacteria that root system of plant generates in earth.
Summary of the invention
In order to overcome the disadvantages of the above prior art, biocontrol bacteria preparation life is utilized the purpose of the present invention is to provide a kind of
Anti- bacterial manure and the method for eliminating muskmelon continuous cropping obstacle ferment the biocontrol bacteria screened the organic of generation using wheat bran as carrier
Bio-bacterial manure is manured into soil, to inhibit soil fungi growth, balanced soil microbiologic population balance, reduce muskmelon root system in soil
The content of secretion main component coumaric acid realizes agricultural sustainable development, dimension to eliminate the continuous cropping obstacle in muskmelon production
Protect ecological environment.
To achieve the goals above, the technical solution adopted by the present invention is that:
A method of biocontrol bacterial fertilizer being prepared using biocontrol bacteria, is included the following steps:
(1) in the isolated bacillus subtilis of kiwi fruit tree garden (Bacillus subtilis, preservation date: 2018
In on December 3, in, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), preservation are compiled
Number: CGMCC NO.16850) it is bacterial strain, carries out separation, the identification of strain；
(2) carrier matrix sterilizes；
(3) by the soy meal of sterilizing, ammonium nitrate, sucrose, magnesium sulfate, manganese sulfate, it is added to the carrier matrix of step (2)
In, and the seed liquor of sterile water and biocontrol bacteria is added, it stirs, obtains hair fermentation raw material；
(4) fermentation raw material for obtaining step (3) ferments, and ensures that biocontrol bacteria number reaches 10 in medium carrier10CFU/g
DW carrier matrix.
In the step (2), carrier matrix is wheat bran or corn material.
In the step (2), the single size of carrier matrix is greater than 2mm.
In the step (2), under the conditions of 100Kpa, 120 DEG C, moist heat sterilization.
In the step (3), soy meal, ammonium nitrate, sucrose, magnesium sulfate, manganese sulfate dosage be followed successively by fermentation substrate weight
2%, 2%, 0.5%, 0.1%, the 0.05% of amount, sterile water consumption are the 60% of carrier matrix weight, the seed liquor of biocontrol microorganisms
Dosage is the 1% of carrier matrix weight, total plate count 4 × 109CFU/mL。
In the step (3), the sterile water of addition will guarantee that carrier matrix moisture content is no more than 60%, the biocontrol microorganisms
Seed liquor is the fresh bacterium solution for the resting period being no more than 48h.
In the step (4), to ferment in Full-automatic heat preservation device, initial stage temperature is set as 35 DEG C, it ferments for 24 hours, the megathermal period
Ventilation quantity is adjusted to 0.15m3/(m3Min), guarantee that carrier matrix temperature is maintained to 45 DEG C of fermentation 48h.
The present invention also provides a kind of methods for eliminating muskmelon continuous cropping obstacle using biocontrol bacteria, to separate in kiwi fruit tree garden
Obtained bacillus subtilis is bacterial strain, and manner described above prepares biocontrol bacterial fertilizer, transplants the previous moon in Muskmelon Seedlings, will prepare
Biocontrol bacterial fertilizer even be spread in melonry according to 135kg/ is per mu.
In this method, it can match and apply dry sheep manure or 2000kg/ mus of dry cow dung, rotary tillage.
Compared with prior art, the beneficial effects of the present invention are:
(1) biocontrol microorganisms that the present invention uses have eccrine fiber element, chitinase and phosphoric acid enzyme viability, can effectively inhibit
Soil fungi growth promotes soil bacteria growth, balanced soil microbiologic population balance.
(2) biocontrol microorganisms that the present invention uses have the function of decomposing muskmelon root exudates main component coumaric acid.Application
After this bacterial manure, the phenolic acid even being eliminated in continuous cropping soil can be reduced, the Auto toxicity of muskmelon is eliminated.
(3) the biocontrol bacteria bacterial manure produced using wheat bran or corn broken particle, is applied in muskmelon continuous cropping soil, can be brought into
Biocontrol microorganisms can also bring a large amount of organic matters into soil.The physicochemical property of soil can be improved, can also improve the health of continuous cropping soil
State.
(4) present invention eliminates the suppression harmful bacteria that plant generates in soil using soil native country biocontrol bacteria, produces biological and ecological methods to prevent plant disease, pests, and erosion
The simple process of bacterial manure, raw material are easy to get, are cheap, is easy to operate, and bacterial fertilizer products stablize safety.
Detailed description of the invention
Fig. 1 is antagonism effect diagram of the bacterial strain of the present invention to pathogen.
Fig. 2 is that the present invention is based on the chadograms of bacterial strain 16SrDNA sequence.
Fig. 3 is the decomposition diagram of SBRh5 p-Coumaric Acid of the present invention, wherein figure a is LB culture medium, it is unique that figure b, which is CA,
The minimal medium of carbon source.
Fig. 4 is SBRh5 of the present invention to the decomposition amount of CA in inorganic salt liquid culture medium and its in minimal medium
Quantity changes schematic diagram.
The repair of muskmelon continuous cropping soil is shown in the influence and SBRh5 that Fig. 5, which is muskmelon continuous cropping soil, grows Muskmelon Seedlings
It is intended to.
Fig. 6 is the influence schematic diagram that external source CA and SBRh5 grows Muskmelon Seedlings.
Fig. 7 is influence schematic diagram of the SBRh5 to muskmelon growing way under field condition.
Bacillus subtilis Bacillus subtilis of the present invention has been preserved in State Intellectual production on December 3rd, 2018
Specified China Committee for Culture Collection of Microorganisms's common micro-organisms center, depositary institution, power office, deposit number CGMCC
NO.16850, preservation address BeiJing, China.
Specific embodiment
Illustrate a specific embodiment of the invention below in conjunction with drawings and examples.
The present invention is a kind of method for preparing biocontrol bacterial fertilizer using biocontrol bacteria and eliminating muskmelon continuous cropping obstacle, process packet
It includes:
1, strain isolation, identification and culture
The screening of 1.1 high-temperature resistant strains
15 parts of rhizosphere soils back will be adopted after 90 borders handle 20min from the healthy Chinese gooseberry garden in Shaanxi Mei County area,
2g is weighed, is put into the 50mL conical flask bottle for filling 18mL sterile water, it is to dilute 10 times that 180r/min, which vibrates 30min, under room temperature
Bacteria suspension.It is coated on LB solid medium after above-mentioned bacteria suspension is diluted 10 times again uniform.Observe the form of single colonie
Feature and progress Gram's staining, primarily determine the bacterial strain of 5 plants of doubtful bacillus subtilises.
The screening of 1.2 pathogenic bacteria antagonistic strains
By above-mentioned 5 finally obtained plant bacterial strain, plate dual test is carried out with soil soil-borne disease bacterium sickle-like bacteria respectively, therefrom
One plant of antagonistic effect bacterial strain the most apparent is chosen to expand culture and Molecular Identification (antagonistic effect such as Fig. 1).
The Molecular Identification (the hot Gadamer Biotechnology Co., Ltd in Xi'an) of 1.3 antagonistic strains
The strain genomic DNA, PCR amplification (PFU PCR mix, Tiangeng) bacterium are extracted by gene rapid fractionation method
The area 16SrDNA, primer sequence are as follows:
Lactic acid bacteria-F AgAgTTTgATCCTggCTCAg
Lactic acid bacteria-R CTACggCTACCTTgTTACgA
When 16S amplified production length is to 1500bp or so, sequence information is obtained.With DNA gel purification kit
(AXYGEN) glue purification recycling is cut, NL4 carries out DNA sequencing (3730xl, ABI) to recycling DNA fragmentation.
The sequence that the bacterial strain PCR amplification comes out is compared with sequence existing in gene pool, finds its 16S rDNA
Sequence and the similitude of the bacillus subtilis SBRh5 (Sequence ID:HQ443229.1) published are higher, reach
99%, it is accredited as bacillus subtilis (Fig. 2).
The culture of 1.4 SBRh5 seed liquors
The complete single colonie of oese picking is used in superclean bench, is transferred into 100mL LB liquid medium,
180r/min shaken cultivation about 12h (30 DEG C), the seed liquor as prepared.
2.SBRh5 eliminating the correlation test of muskmelon continuous cropping obstacle
The shadow of 2.1 SBRh5 and muskmelon root exudates main component coumaric acid (CA) to bacterium in soil and fungi
It rings
Method: taking 4g wheat topsoil native, add 36mL sterile water, after being placed in 180r/min shaking table shake 30min, takes this soil
Thallus suspension liquid 2mL is separately added into following three kinds of fluid nutrient mediums: I minimal medium (NH4NO3—1.0g；K2HPO4—
1.5g；NaCl—0.5g；(NH4)2SO4—0.5g；MgSO4·7H2O—0.5g；KH2PO4- 0.5g) in, II contains 100 μ g/
The minimal medium of mLCA；SBRh5 seed liquor (is inoculated in by the supernatant of the SBRh5 of III minimal medium culture for 24 hours
In inorganic salt liquid culture medium, 30 DEG C, 180r/min shaking table culture for 24 hours, 10000rpm/min, is centrifuged 10min, by fermentation liquid mistake
(remove thallus) after 0.22 μm of miillpore filter, as SBRh5 sterile supernatant).Then by it is each processing be placed on 30 DEG C,
On 180r/min shaking table cultivate 18h after, in plate count method culture solution bacterium and fungi number.
As a result: as shown in table 1.The results show that CA promotes the quantity of fungi in soil, bacterial number is influenced less, but
SBRh5 remarkably promotes bacterial number in soil, reduces fungi.
Influence of table 1 SBRh5 and CA to bacterium and fungi in wheat soil
Conclusion: CA can promote the growth of soil fungi；SBRh5 can remarkably promote bacterial growth, inhibit fungi growth.
Decomposition of 2.2 SBRh5 to muskmelon root exudates main component CA
2.2.1 SBRh5 decomposes muskmelon root exudates main component coumaric acid under condition of culture in room
Method: being connected to the inorganic salts using CA as sole carbon source for SBRh5 point and (form with 2.1) on solid medium, 30 trainings
8d is cultivated, growing state of the SBRh5 on culture medium is observed.
As a result: as Fig. 3 shows.The results show that SBRh5 being capable of normal growth on the culture medium using CA as sole carbon source.
Conclusion: SBRh5 can decompose CA, the carbon source as its growth.
2.2.2 decomposition amount of the SBRh5 to CA in efficient liquid phase measurement fluid nutrient medium
Method: SBRh5 is inoculated in the liquid inorganic salting liquid containing XX CA, after shaking table culture 18h, with efficient liquid
The variable quantity of CA content before and after the culture of phase chromatographic determination.And the quantity of SBRh5 in culture solution is counted with colony counting method.
Chromatographic condition: CA assay uses HPLC method (Waters 1525, the U.S.) in fermentation liquid.It is used before sample feeding
The filtering with microporous membrane of 0.22 μm of m, chromatography column are SB-C18 4.6mm × 250mm), flow velocity 1mLmin-1, sample volume
10 μ L, 25 DEG C of column temperature, Detection wavelength 280nm, mobile phase A: methanol (chromatographic grade) and Mobile phase B: the acetic acid aqueous solution of pH 2.8
(chromatographic grade), the separation of sample achieve the purpose that separation by the method for gradient elution and shorten phenolic acid retention time.
Elution requirement B:70% (0min) → 50% (15min) → 50% (7min) → end.
As a result: HPLC measurement result is shown, in minimal medium, SBRh5 can be in 18h, by CA by initial
61.318μg·mL-1It is degraded to 37.858 μ gmL-1, and 3.196log (cfumL of its own quantity by-1),18h
Inside increase 5.858log (cfumL-1)。
Conclusion: SBRh5 can decompose muskmelon root exudates main component CA.
The influence that 2.3 continuous cropping soil and CA grow Muskmelon Seedlings
2.3.1 repair of the influence and SBRh5 that muskmelon continuous cropping soil grows Muskmelon Seedlings to continuous cropping soil
Method: it is tested using miniature alms bowl.Muskmelon seeds are impregnated 15 minutes or so in 1% aqueous sodium hypochlorite solution,
After cleaning, it is put into 55 DEG C or so water, room temperature to be down to, seed is pulled out and is put into vernalization in 30 DEG C of insulating boxs.After seed shows money or valuables one carries unintentionally
Nursery, long to one heart stage an of leaf to seedling, selection grows consistent seedling and moves to miniature alms bowl, every one seedling of alms bowl.Processing are as follows: I muskmelon
Non- continuous cropping soil；II muskmelon continuous cropping soil；III muskmelon continuous cropping soil admixes 1gkg-1Soil SBRh5 pulvis；IV muskmelon continuous cropping soil is mixed
Enter 3gkg-1Soil SBRh5 pulvis.Every alms bowl fills soil 0.5kg.Being placed in culturing room, (23 DEG C of temperature, humidity 49-51%, illumination is strong
Degree is 2850Lx).Normal water and fertilizer management.
As a result: as shown in figure 5, seedling is small weak with regard to obvious more non-continuous cropping soil when muskmelon 2 generation of continuous cropping；
When admixing SBRh5 pulvis in continuous cropping soil, Muskmelon Seedlings growing way is similar to the growing way of seedling in non-continuous cropping soil.
Conclusion: (1) there are serious continuous cropping obstacle phenomenons for muskmelon；(2) SBRh5 can effectively overcome muskmelon continuous cropping obstacle.
2.3.2 the influence that external source CA grows Muskmelon Seedlings
Method: it is tested using miniature alms bowl.Muskmelon seeds are impregnated 15 minutes or so in 1% aqueous sodium hypochlorite solution,
After cleaning, it is put into 55 DEG C or so water, room temperature to be down to, seed is pulled out and is put into vernalization in 30 DEG C of insulating boxs.After seed shows money or valuables one carries unintentionally
Nursery, long to one heart stage an of leaf to seedling, selection grows consistent seedling and moves to miniature alms bowl, every one seedling of alms bowl.Processing are as follows: I muskmelon
Non- continuous cropping soil；The non-continuous cropping soil+CA of II (400 μ g/kg soil)；III muskmelon continuous cropping soil+CA (400ug/kg soil)+SBRh5 pulvis
4g·kg-1Soil；IV muskmelon continuous cropping soil+CA (400ug/kg soil)+SBRh5 pulvis 8gkg-1Soil.Every alms bowl fills soil 0.5kg.Culture
Condition and the same 2.3.1 of management.
As a result: as Fig. 6 shows.After external source CA is added in the non-continuous cropping soil of muskmelon, muskmelon growth is obviously inhibited；External source is added
While CA, when different amounts of SBRh5 pulvis is added, muskmelon growing way obviously takes a turn for the better, and compares indifference.
Plant agronomic traits data (table 2) display, the main component CA of muskmelon root exudates have seriously affected muskmelon children
The growth of seedling: plant height, the number of blade, root long and biomass are all significantly lower than control level；But after applied SBRh5, especially work as
SBRh5 amount of application is 8gkg-1When native, the economical character of Muskmelon Seedlings has obviously relatively compareed preferably: plant height, the number of blade, root long
It is higher by control 15%, 28% and 37% respectively, and ground fresh weight is higher by 87% than compareing, root system fresh weight is higher by control 245%.Root system
Vigor is the 177% of control, is the 417% of external source CA processing.
The influence that 2. external source coumaric acid (CA) of table and SBRh5 grow muskmelon
Conclusion: (1) CA is the principal element for leading to muskmelon continuous cropping obstacle.(2) SBRh5 can eliminate the CA in soil, promote
Into the growth of muskmelon.
SBRh5 acts on the elimination of continuous cropping obstacle in 2.4 cultivations of fields
Method: test is on July 2,18 days to 2017 January in 2017, (34 ° 16 ' of Yang Ling Zhong Lai crop cultivation speciality cooperative society
N, 108 ° of 0 ' E, the village Ling Wan, the township Yangling Area Rou Gu) it carries out.Test process are as follows: sheep dung 1200kg uniformly is spread fertilizer over the fields per acre, then
By the solid fungicide 75kg of SBRh5 in sheep dung, rotary tillage after being uniformly sprinkling upon thereon, ridging transplants muskmelon seedling after one month.
As a result 1: the application SBRH5 influence to soil bacteria fungi
Just in contrast, measurement result such as table 3, it can be seen that bacterial number is gradually to rise from February to April in soil
High, reach maximum value April, is then gradually reduced later；Fungi from 2 months to April, change it is smaller, but to May and
It is declined slightly June, July slightly rises.
Influence of the SBRH5 to the different month soil bacterias, fungi of muskmelon Growing season is applied under 3 field condition of table
Apply microbial inoculum after, whether the quantity variation tendency of bacterium or fungi with compare it is similar.But in quantity
It sees, application SBRh5 microbial inoculum processing, under identical month, bacterial number relatively control has to be increased by a relatively large margin, especially bacterium
The April of frequent activity.And fungi is then declined.
From the point of view of bacterial fungus ratio, the bacterial fungus ratio of control applies SBRH5 microbial inoculum in maximum in April, at any time
It elapses its variation and is presented wavy decline, 2 months are compared with being March height, and April was high compared with 2 months, and had reached peak；
Subsequent 5,6, July, value is compared with being April low.Totality is seen, under identical month, relatively control is high to application SBRh5,2 months
Part differs maximum with control, and April is minimum.
As a result 2: the application SBRh5 influence to muskmelon growing way
After application SBRh5 biocontrol microorganisms (Fig. 7), muskmelon blade color is green and thick, and drought tolerance, heat resistanceheat resistant are strong.
As a result 3: the application SBRh5 influence to muskmelon quality
Muskmelon quality results such as table 4.After applying SBRh5, single fruit weight relatively compares high by 10%；Soluble sugar is higher by than control
20%；Soluble protein content is higher by 25%；Vc content is higher by 10%, and nitrate nitrogen content declines 50%.It can be seen that application
After SBRh5 microbial inoculum, the continuous cropping effect of muskmelon can be significantly overcome, improve melon fruit quality.
Influence after 4 SBRh5 of table application to Melon fruit quality
Conclusion: 75kg SBRh5 microbial inoculum is applied under field condition per acre with organic manure is applied, Melon can be substantially eliminated
Continuous cropping effect.
Claims (10)
1. a kind of method for preparing biocontrol bacterial fertilizer using biocontrol bacteria, which comprises the steps of:
(1), as bacterial strain, to carry out the separation of strain in the isolated bacillus subtilis of kiwi fruit tree garden, identify；
(2) carrier matrix sterilizes；
(3) it by the soy meal of sterilizing, ammonium nitrate, sucrose, magnesium sulfate, manganese sulfate, is added in the carrier matrix of step (2), and
The seed liquor of sterile water and biocontrol bacteria is added, stirs, obtains hair fermentation raw material；
(4) fermentation raw material for obtaining step (3) ferments, and ensures that biocontrol bacteria number reaches 10 in medium carrier10CFU/g DW is carried
Body matrix.
2. the method for preparing biocontrol bacterial fertilizer using biocontrol bacteria according to claim 1, which is characterized in that the step (2)
In, carrier matrix is wheat bran or corn material.
3. the method according to claim 1 or claim 2 for preparing biocontrol bacterial fertilizer using biocontrol bacteria, which is characterized in that the step
(2) in, the single size of carrier matrix is greater than 2mm.
4. the method for preparing biocontrol bacterial fertilizer using biocontrol bacteria according to claim 1, which is characterized in that the step (2)
In, under the conditions of 100Kpa, 120 DEG C, moist heat sterilization.
5. the method for preparing biocontrol bacterial fertilizer using biocontrol bacteria according to claim 1, which is characterized in that the step (3)
In, soy meal, ammonium nitrate, sucrose, magnesium sulfate, manganese sulfate dosage be followed successively by fermentation substrate weight 2%, 2%, 0.5%,
0.1%, 0.05%, sterile water consumption is the 60% of carrier matrix weight, and the seed liquor dosage of biocontrol microorganisms is carrier matrix weight
1%, total plate count 4 × 109CFU/mL。
6. the method for preparing biocontrol bacterial fertilizer using biocontrol bacteria according to claim 1, which is characterized in that the step (3)
In, the sterile water of addition to guarantee carrier matrix moisture content be no more than 60%, the seed liquor of the biocontrol microorganisms be the resting period not
Fresh bacterium solution more than 48h.
7. the method for preparing biocontrol bacterial fertilizer using biocontrol bacteria according to claim 1, which is characterized in that the step (4)
In, it ferments in Full-automatic heat preservation device, initial stage temperature is set as 35 DEG C, and for 24 hours, megathermal period ventilation quantity is adjusted to 0.15m for fermentation3/
(m3Min), guarantee that carrier matrix temperature is maintained to 45 DEG C of fermentation 48h.
8. a kind of method for eliminating muskmelon continuous cropping obstacle using biocontrol bacteria, which is characterized in that with isolated in kiwi fruit tree garden
Bacillus subtilis be bacterial strain, prepare biocontrol bacterial fertilizer, transplant the previous moon in Muskmelon Seedlings, by the biocontrol bacterial fertilizer of preparation according to
135kg/ is per mu even to be spread in melonry.
9. the method for eliminating muskmelon continuous cropping obstacle using biocontrol bacteria according to claim 8, which is characterized in that match Shi Ganyang
Excrement or 2000kg/ mus of dry cow dung, rotary tillage.
10. the method for eliminating muskmelon continuous cropping obstacle using biocontrol bacteria according to claim 8, which is characterized in that the life
The preparation method of anti-bacterial manure such as claim 1.
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| CN113336601A (en) * | 2021-07-21 | 2021-09-03 | 广西壮族自治区农业科学院 | Method for preparing biocontrol bacterial manure by bacteria and method for eliminating continuous cropping obstacles of melons |
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