CN107325821B - Biomass charcoal-based microbial inoculant for preventing and treating watermelon fusarium wilt and preparation method and application thereof - Google Patents
Biomass charcoal-based microbial inoculant for preventing and treating watermelon fusarium wilt and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a biomass charcoal-based microbial inoculant for preventing and treating watermelon fusarium wilt, and a preparation method and application thereof, wherein the inoculant comprises the following components: biomass charcoal, attapulgite, watermelon wilt biocontrol bacteria, calcium superphosphate or nitrohumic acid and sterile water; the preparation method of the microbial inoculum comprises the following steps: adsorbing a biocontrol bacterium liquid on biomass charcoal, adding attapulgite and calcium superphosphate or nitrohumic acid, mixing uniformly, adding a proper amount of sterile water, blending, granulating, drying and ventilating under natural conditions, and drying to obtain the microbial inoculum, wherein the viable count in the microbial inoculum is more than or equal to 50 hundred million cfu/g, the particle diameter is 3-6 mm, and the pH value is 6.5-7.5; the microbial inoculum of the invention has the functions of improving the physical and chemical properties of soil, improving the activity of soil enzyme, promoting the growth of crops, increasing the yield of the crops and the like, and has better control effect on watermelon wilt.
Description
Technical Field
The invention relates to a microbial inoculum carried by biomass charcoal in the technical field of agriculture, in particular to a preparation method of a solid microbial inoculum suitable for various facility crops and application thereof in preventing and treating watermelon wilt.
Background
In recent years, facility vegetables in China are rapidly developed, the planting area reaches 5832.6 ten thousand mu, the original ecological environment of soil is changed by covering and cultivating the ground surface for a long time under the condition of facility cultivation, the soil is often in a closed circulating metabolic environment with high temperature, high humidity, high evaporation and no rain leaching, the physical and chemical properties and the biological properties of the soil are changed, the soil is hardened, the porosity is reduced, the phenomenon of soil salt content accumulation is obvious, and the soil quality is continuously worsened. The added pesticide and chemical fertilizer are excessively applied and deposited, toxic and harmful substances such as heavy metal, antibiotics and the like in soil are gradually accumulated, the microbial population quantity and the structure are changed, and the soil biotoxicity is enhanced, so that the continuous cropping production obstacle contradiction of soil acidification, secondary salinization and soil-borne disease aggravation is prominent, and the continuous cropping production obstacle contradiction becomes a main factor for restricting the continuous and efficient development of the facility-cultivated vegetables.
At present, the agricultural measures for preventing and treating continuous cropping obstacles mainly comprise: (1) and (5) performing rotation. Crop rotation and intercropping crop rotation are the simplest and most effective methods for solving continuous cropping obstacles. The number of pathogenic bacteria in the soil can be obviously reduced by crop rotation with pathogenic bacteria non-host plants or by error planting in a period of easy occurrence of high-temperature continuous cropping diseases, but in many cases, continuous cropping of certain vegetables is forced to be carried out in order to improve economic benefit, and the method cannot be practically implemented. (2) And (5) disinfecting soil. Soil disinfection includes chemical disinfection and physical disinfection. The chemical method mainly comprises mixing formalin, carbendazim or quintozene with soil or sterilizing with sulfur powder, or spraying or mixing with medicinal liquid for trench application or hole application to treat soil; or fumigating the soil by adopting methyl bromide. The physical method mainly adopts high temperature or low temperature to reduce the number of harmful bacteria. In high temperature season in summer, the greenhouse is sealed at high temperature after irrigation, and the soil can be subjected to disinsection and sterilization by high temperature steam, so that the number of soil-borne diseases and insect pests can be greatly reduced. The low-temperature freezing method is used for protecting land which is cold in winter in the north and cannot be utilized, and can be used for removing films and then deeply ploughing soil to freeze germs and worm eggs. Soil disinfection methods generally kill plant diseases and insect pests and also cause destructive damage to beneficial organisms, particularly chemical disinfection methods are expensive, short in duration, and not ideal in prevention and treatment effect, and are not beneficial to large-area popularization, and soil structure and environment pollution can be possibly damaged. (3) Application of resistant varieties. The aggravation of diseases and insect pests is an important factor causing continuous cropping obstacle of crops, new varieties with resistance are bred, and the crops which are difficult to breed into disease-resistant varieties are grafted and cultivated by adopting resistant stocks, so that various soil-borne diseases and the harm of nematodes can be effectively prevented. However, due to the problems of the limitation of disease-resistant variety resources, the breeding cycle and the like, the breeding work of the disease-resistant variety is far lagged behind the speed of disease occurrence; the grafting method effectively solves the problems that the quality and the taste of crop products are reduced while soil-borne diseases are solved. (4) And (5) reasonably applying fertilizer. The special fertilizer applied in the current production has obvious effect on reducing the continuous cropping obstacles of crops, and in addition, the additional application of organic fertilizer is an effective measure for reducing the continuous cropping obstacles. The continuous cropping obstacle is generated by a plurality of biological and non-biological complex factors such as crops, soil, environment and the like, and the mutual influence of the factors makes the problem to be solved more complicated, and the ideal effect is hardly achieved by any single measure or a few measures. Therefore, the key to solving the continuous cropping obstacle of the protected cultivation vegetable production is to improve the physical and chemical properties of the soil, increase the number of beneficial microbial populations in the soil and improve the soil fertility.
Disclosure of Invention
The invention aims to provide a microbial inoculum carried by biomass charcoal for preventing watermelon fusarium wilt, which does not contain harmful substances to the environment, has good fertilizer efficiency and strong stress resistance, has soil improvement property, improves the comprehensive effects of soil micro-ecological structure and the like, can effectively prevent watermelon fusarium wilt, and is realized by the following steps:
a biomass charcoal-based microbial agent for preventing and treating watermelon wilt comprises a solid-phase component and a liquid-phase component;
solid phase component: by mass percentage, 40-50% of biomass charcoal, 20-25% of attapulgite, and calcium superphosphate or nitrohumic acid for complementing the balance;
liquid phase components: comprises biocontrol bacteria fermentation liquor (the bacteria content is 1-9 multiplied by 10)12cfu/mL) and sterile water;
wherein, the volume mass ratio (L/Kg) of the biocontrol bacteria fermentation liquor to the solid phase component is 0.01-0.03:1, the mass of the sterile water is 30-50% of the mass of the solid phase component, and the viable count of the biocontrol bacteria in the microbial inoculum is more than or equal to 50 hundred million cfu/g.
Furthermore, the biocontrol bacteria in the invention are biocontrol bacteria of watermelon fusarium wilt, and specifically can be one or more of bacillus tequilensis, bacillus vallismortis, pseudomonas oryza sativa or bacillus subtilis.
Further, in the present invention, the biomass char is obtained by: agricultural wastes such as rice hulls, crop straws and the like are used as raw materials, heat preservation treatment is carried out for 1-2 hours in a high-temperature open-type tubular furnace under the anaerobic condition of 400-700 ℃, the raw materials are naturally cooled to room temperature, ground and sieved by a 40-mesh sieve, and sterilization is carried out for 1 hour at 121 ℃, so that the biomass carbon is obtained.
Further, in the invention, the attapulgite is an agricultural attapulgite slow-release granulating agent, preferably, by mass, the attapulgite contains 50.4% of silicon dioxide, 9.8% of aluminum oxide, 4% of ferric oxide, 0.04% of manganese oxide, 1.0% of potassium oxide, 1.8% of calcium oxide, 0.04% of magnesium oxide, 13.8% of caustic soda and the balance of soil components; the calcium superphosphate is a granular fertilizer of calcium superphosphate sold in the market, preferably 80-95% of the granular fertilizer is dissolved in water, and the content of available phosphorus is more than or equal to 12%.
The invention discloses a preparation method of a biomass charcoal-based microbial inoculant for preventing and treating watermelon fusarium wilt, which comprises the following specific steps:
step one, fermenting biocontrol bacteria to prepare the biocontrol bacteria with the bacteria content of 1012cfu/mL fermentation liquor for standby;
and step two, uniformly mixing the biomass charcoal and the biocontrol bacterium fermentation liquor to ensure that the biocontrol bacteria are fully adsorbed by the biomass charcoal and uniformly mixed.
And step three, uniformly mixing the solid-phase components (biomass charcoal, attapulgite, calcium superphosphate or/and nitrohumic acid) according to a proportion under an aseptic condition, adding sterile water, fully stirring, transferring to a disc granulator to prepare granules with the particle diameter of 3-6 mm and the pH value of 6.5-7.5, taking out the granules, and ventilating and drying under natural conditions until the humidity of the granules is less than or equal to 20% (about 3-4 days), wherein the viable count of biocontrol bacteria in the microbial inoculum is more than or equal to 50 hundred million cfu/g, thus obtaining the biomass charcoal-based microbial inoculum.
Further, in the preparation method of the biomass charcoal-based microbial agent for preventing and treating watermelon fusarium wilt, the biocontrol bacterium fermentation broth in the step one is as follows: inoculating biocontrol bacteria into LB liquid culture medium, performing shake culture at 37 + -1 deg.C and 200rpm until the bacteria content is 1012cfu/mL (about 48 hours);
the biomass charcoal-based microbial agent disclosed by the invention is applied to prevention and treatment of watermelon fusarium wilt.
Further, the application of the biomass carbon-based microbial agent in preventing and treating watermelon fusarium wilt provided by the invention is as follows: the microbial inoculum is applied in holes when 2 true leaves of watermelon seedlings are transplanted, and the dosage of each plant is 100 g.
In the invention, the LB liquid culture medium is: 10g of tryptone, 5g of yeast powder and 10g of sodium chloride, and adding water to 1L, wherein the pH value is 7.0-7.5.
The invention utilizes the characteristics of the biochar of improving the soil performance and effectively adsorbing soil microorganisms, utilizes the advantages of rich carbon source, proper pore structure, water retention and the like, carries plant endophytic biocontrol bacteria by the biochar, and prepares an excellent facility cultivation soil conditioner, thereby achieving the purposes of improving the soil performance and introducing beneficial microorganisms at the rhizosphere of crops.
Compared with the prior art, the invention takes the wastes such as rice hulls, crop straws and the like as raw materials, the materials are easy to obtain, the preparation method is simple, and the use is convenient; the survival time of the biocontrol bacteria can be effectively prolonged; the quantity of beneficial bacteria in soil is increased, the soil enzyme activity is enhanced, the soil fertility is improved, and the soil conditioner has stronger prevention and treatment effects on various soil-borne diseases such as watermelon fusarium wilt and the like.
Drawings
FIG. 1 is a picture of biomass charcoal-based microbial inoculum particles for controlling watermelon fusarium wilt according to the present invention;
FIG. 2 is a schematic diagram showing the detection result of the biomass carbon-based microbial agent for preventing and treating watermelon fusarium wilt in shelf life according to the present invention;
FIG. 3 is a schematic diagram showing the survival amount of biocontrol bacteria in soil after the biomass charcoal-based microbial inoculant for controlling watermelon fusarium wilt is applied;
FIG. 4 is a schematic diagram showing the effect of the biomass charcoal-based microbial inoculum for controlling watermelon fusarium wilt on the activity of soil enzymes;
FIG. 5 is a schematic diagram showing the effect of the biomass charcoal-based microbial inoculum for controlling watermelon fusarium wilt on the growth of watermelon in seedling stage and the control effect of the biomass charcoal-based microbial inoculum for controlling watermelon fusarium wilt.
Detailed Description
To help better understand the essence of the idea of the present invention, the following embodiments of the present invention are described in detail by specific examples, which are not intended to limit the scope of the present invention.
In the following examples, Bacillus vallismortis, Bacillus tequilensis and Bacillus subtilis were isolated into healthy tomato plants, purified, identified and stored by the institute of habitat environmental of the academy of agricultural sciences of Jiangsu province. The watermelon wilt pathogen is separated into diseased watermelon plants, and the diseased watermelon plants are separated, purified and stored by an environmental research laboratory of quality safety institute of Jiangsu academy of agricultural sciences.
Raw materials and media referred to in the examples:
LB liquid medium (1L): 10g of tryptone, 5g of yeast powder and 10g of sodium chloride, and adding water to 1L, wherein the pH value is 7.0-7.5;
LB solid medium: 10g of tryptone, 5g of yeast powder, 10g of sodium chloride and 20g of agar powder, adding water to 1L, adjusting the pH value to 7.0-7.5, sterilizing at 121 ℃ for 20min, and cooling for later use;
LB plate: melting LB solid culture medium in a microwave oven, cooling to about 50 ℃, pouring the melted LB solid culture medium into a sterilized culture dish in a sterile operating platform, and cooling to obtain an LB flat plate;
attapulgite clay: agricultural attapulgite sustained-release granulating agent purchased from Changzhou Dingbang mineral products science and technology Limited; wherein the content of silicon dioxide is 50.4 percent, the content of aluminum oxide is 9.8 percent, the content of ferric oxide is 4 percent, the content of manganese oxide is 0.04 percent, the content of potassium oxide is 1.0 percent, the content of calcium oxide is 1.8 percent, the content of magnesium oxide is 0.04 percent, the content of caustic soda is 13.8 percent, and the balance is soil;
the calcium superphosphate is a commercially available calcium superphosphate granular fertilizer: is purchased from chemical fertilizer Limited company, wherein 80 to 95 percent of the fertilizer is dissolved in water, and the content of available phosphorus is more than or equal to 12 percent.
Experimental equipment:
the high-temperature open-type tube furnace is purchased from Shanghai optical precision machinery research institute of Chinese academy of sciences, model SG-GL 1100K;
example 1 preparation of Biomass charcoal-based microbial inoculum for controlling watermelon wilt
Step one, taking purified biocontrol bacteria (Bacillus tequilensis and Bacillus vallismortis), inoculating the biocontrol bacteria into an LB liquid culture medium, and culturing for 48 hours in a shaker (150 rpm) at 37 +/-1 ℃ to obtain bacteria with the bacteria content of 1012cfu/mL biocontrol bacteria fermentation liquor for later use.
And step two, using rice hulls as raw materials, preparing the biomass carbon under the anaerobic condition of adopting a high-temperature open-type tubular furnace at 500 ℃ (in the specific implementation process, the temperature of the tubular furnace can be controlled within the range of 400-.
Step three, taking 20Kg of rice hull biomass carbon, attapulgite and calcium superphosphate (the rice hull biomass carbon, the attapulgite and the calcium superphosphate are mixed according to the mass ratio of 4:2: 1), taking 0.2L of biocontrol bacteria fermentation liquor in the step one, and uniformly mixing the biocontrol bacteria fermentation liquor with the biomass carbon to enable the biomass carbon to fully adsorb biocontrol bacteria; then adding the rest components, mixing uniformly, adding sterile water accounting for 50% of the total amount of the solid phase components, stirring uniformly, transferring to an organic fertilizer disc granulator, preparing granules with the particle size of 3-6 mm at 220v and 45 r/min for 2 hours, and then carrying out ventilation drying at the indoor temperature of 25 ℃ for 3-4 days with the humidity of less than or equal to 20%. Thus obtaining the biomass charcoal-based microbial agent (as shown in figure 1), wherein the number of the biocontrol bacteria in the microbial agent is about 56 hundred million cfu/g.
In the specific implementation process, one or more of bacillus tequilensis, bacillus vallismortis, pseudomonas oryzae or bacillus subtilis can be selected to prepare the biocontrol bacteria fermentation liquor as long as the biocontrol bacteria content in the microbial inoculum is ensured to be 1012cfu/mL, the purpose of the invention can be realized; the amount of sterile water added can be selected within the range of 30-50% by mass of the solid phase components.
In the embodiment, the biomass charcoal prepared from the rice hulls is used, and in the specific operation, the biomass charcoal prepared from other plant straws can be used.
EXAMPLE 2 shelf-Life testing of microbial Agents
(1) Diluting the biocontrol bacteria fermentation liquor obtained in the first step of example 1 by using normal saline to ensure that the biocontrol bacteria fermentation liquor has the same initial average amount as the biomass charcoal-based microbial inoculum obtained in the third step and the bacterial content is about 56 hundred million cfu/g, and hermetically storing the biocontrol bacteria fermentation liquor at room temperature.
(2) And (3) measuring the quantity of biocontrol bacteria in biocontrol bacteria fermentation liquor and biomass carbon-based microbial inoculant: taking 50 mL of biocontrol bacteria fermentation broth (or 50g of biomass charcoal-based microbial inoculum), adding sterile water to a constant volume of 1000 mL, and placing the mixture into a shaking table to vibrate at a temperature of 200r/min for 30 min; taking 10 mL of solution into a 100 mL volumetric flask, diluting to 100 mL with constant volume, diluting for 5 times by 10 times, taking 100uL of solution, coating a plate on an LB (Langmuir-Blodgett) flat plate, carrying out inverted culture at 30 ℃ for 12 h, counting the number of colonies, and setting for 3 times of repetition; per ml of fermentation broth (Per gram of microbial inoculum) = (a × n × v)1)/(v2M), formula a: average plate colony count, n: dilution multiple, v1: initial dilution volume of product, v2: diluted bacterial liquid volume of the plating plate, m: the volume was sampled.
(3) The sampling time for shelf life testing lasted 12 months.
(4) As shown in FIG. 2, the experimental results show in FIG. 2 that the number of viable bacteria in the fermentation broth in the 7 th month test was 1.0X 107cfu/mL, and the viable count of the biomass charcoal-based microbial agent is 1.4 multiplied by 10 at the detection of 12 th month8cfu/g, the shelf life is obviously prolonged after the biomass charcoal is used for carrying microorganisms to prepare the biomass charcoal-based microbial agent.
Example 3 Effect of Biomass charcoal-based microbial Agents on soil enzyme Activity after application
(1) In order to eliminate the interference of mixed bacteria in soil, firstly, bacillus in the biomass charcoal-based microbial agent is subjected to antibiotic marking: inoculating bacillus on an LB (lysogeny broth) plate containing 1 mu g/mL of rifampicin, gradually increasing the concentration of rifampicin to 300 mu g/mL, screening an antibiotic-resistant strain which can stably grow and has physiological characteristics consistent with those of an original strain, and preparing a biomass charcoal-based microbial agent according to the steps of example 1;
(2) 2 treatments are set:
A. taking 10 kg of field soil, adding the biomass carbon-based microbial agent obtained in the step (1) according to the proportion of 2% by mass, uniformly mixing, placing at room temperature, and sealing for storage;
B. the biocontrol bacteria fermentation liquor is prepared by the resistant strain according to the method in the step 1, the biocontrol bacteria fermentation liquor is inoculated into 10 kg of field soil according to the total amount of the bacteria in the treatment A, and the biocontrol bacteria fermentation liquor is uniformly mixed and is sealed and stored at room temperature;
(3) determination of survival amount of biocontrol bacteria in soil: the measurement method was the same as in step (2) of example 2;
(4) and (3) soil enzyme activity determination: after 30 days of the above treatment, 10g of soil samples were taken for each treatment, and the activities of cellulase, neutral protease, sucrase, urease, catalase and dehydrogenase activities in the soil were measured, and the experimental procedures were described in the specification of soil enzyme measurement kit (http:// www.cominbio.com/a/chanpInzyhanshi/ziyoupaishijihe/turanxie /) manufactured by Suzhou Kemingming Biotechnology, Ltd.).
(5) The results of the survival amount of the biocontrol bacteria in the soil are shown in FIG. 3, which indicates that the number of the viable bacteria detected by inoculating the biocontrol bacteria fermentation liquor for 7 days is 6.72 multiplied by 109cfu/g, but the viable count rapidly decreased with time, and was only 2.5X 10 by 120 days8cfu/g; the effective viable count of the biomass charcoal-based microbial agent is a process from slow release to rapid increase in number and slow decrease, and the viable count is only 7.03 multiplied by 10 when the biomass charcoal-based microbial agent is treated for 7 days9cfu/g, but the viable count reached a peak of 9.83X 10 at 21 days of treatment9cfu/g, when the viable count slowly decreases to 120 days after treatment, the effective viable count is still 2.98 multiplied by 109cfu/g shows that the microbial inoculum prepared by carrying bacillus by using biomass charcoal can obviously prolong the survival time of the biocontrol bacteria in soil.
(6) The results of soil enzyme activity are shown in FIG. 4, NS is field soil; b is a bacterial liquid (treatment B); BC is a microbial inoculum (treatment A);
as can be seen from FIG. 4, the activity of cellulase, sucrase, urease, catalase and dehydrogenase active enzymes in soil is significantly enhanced after the biomass charcoal-based microbial inoculant is used for treatment, but the activity of neutral protease is reduced, which may not be beneficial to the degradation of macromolecular proteins in soil into products such as amino acid.
Example 4 potted plant test of the effects of watermelon seedling growth and prevention and treatment of blight
The potting test was carried out in a sunlight greenhouse of the academy of agricultural sciences of Jiangsu province. Let 4 treatments:
A. sterilizing soil and inoculating watermelon fusarium wilt;
B. sterilized soil, inoculated watermelon wilt bacteria and biomass charcoal-based microbial inoculant auxiliary materials (sterile, specifically, the auxiliary materials for preparing the biomass charcoal-based microbial inoculant in the embodiment 1 do not contain biocontrol bacteria, the content of other components is the same as that of the biomass charcoal-based microbial inoculant obtained in the embodiment 1, and the addition amount of the auxiliary materials is 0.5 percent of the mass of the sterilized soil);
C. sterilized soil, watermelon wilt bacteria and 0.5 percent of biomass charcoal-based microbial inoculum (the biomass charcoal-based microbial inoculum prepared in the example 1 is added in an amount of 0.5 percent of the mass of the sterilized soil);
D. sterilized soil + watermelon wilt pathogen +2% biomass charcoal-based microbial agent (biomass charcoal-based microbial agent prepared in example 1, added in an amount of 2% by mass of sterilized soil).
The watermelon variety to be tested is Suchuang No. 1.
The test comprises the following specific steps:
(1) sterilizing the surface of watermelon seed, soaking in sterile water overnight, and accelerating germination at 30 deg.C for 2 days. After the white seeds are exposed, the seeds are sowed in a plug tray, and when the seedlings grow to 2 true leaves, the seedlings are transplanted into a plastic cup with the inner diameter of 10 cm and the height of 15 cm and filled with sterilized soil.
(2) The soil of the four treatment groups is 300 g of sterilized soil, and 0.5 percent and 2 percent of biomass charcoal-based microbial inoculum obtained in the third step of the example 1 are respectively added into the sterilized soil of the treatment groups C and D (accounting for the mass percent of the sterilized soil).
(3) While transplanting watermelon seedlings (2-leaf stage), root irrigation of 20 ml 1X 10 is carried out in the treatment groups A, B, C and D7cfu/mL watermelon wilt germ spore suspension is inoculated with watermelon wilt germ.
(4) After disease fungi are inoculated, the disease condition of the plants is investigated, after 60 percent of seedlings which are only inoculated with the pathogenic fungi (treated by A) have disease symptoms, the test is ended, and the growth condition, disease index and disease incidence of the watermelon seedlings are counted.
(5) The results of the control effect experiment of watermelon wilt disease in seedling stage are shown in table 1 and fig. 5:
TABLE 1 prevention and treatment effect of biomass charcoal-based microbial inoculum on watermelon seedling blight
As can be seen from Table 1 and FIG. 5 (A-D in FIG. 5 correspond to treatments A-D respectively), the watermelon plants inoculated with the fungicide group (C, D treatment)) obtained in the inoculation example can effectively prevent the watermelon fusarium wilt, and compared with the treatment A, the incidence rate is remarkably reduced by about 70-85%, and the highest prevention effect reaches 74.6%.
(6) The results of the effect of watermelon seedling growth are shown in Table 2:
TABLE 2 influence of Biomass charcoal-based microbial inoculum on growth of watermelon seedling stage
Note: a, inoculating germs; b, inoculating germs and microbial inoculum auxiliary materials; c: inoculating bacteria and 0.5% of microbial inoculum; d: inoculating bacteria and 2% of microbial inoculum. Lower case letters in the table represent 0.05 level difference significance
Test results show that the microbial inoculum can remarkably inhibit the damage of watermelon fusarium oxysporum to the growth of watermelon seedlings and promote the growth of the watermelon seedlings, and after 2 percent of the solid bacterial manure is added into soil, the plant height and the root length of the watermelon seedlings are remarkably promoted to be increased by about 75-90 percent, and the biomass such as fresh weight, dry weight and the like is promoted to be increased by about 55 percent.
Claims (5)
1. A biomass charcoal-based microbial agent for preventing and treating watermelon fusarium wilt comprises the following solid-phase components and liquid-phase components;
solid phase component: by mass percentage, 40-50% of biomass charcoal, 20-25% of attapulgite, and calcium superphosphate or nitrohumic acid for complementing the balance;
liquid phase components: comprises biocontrol bacteria fermentation liquor and sterile water;
wherein the volume mass ratio of the biocontrol bacteria fermentation liquor to the solid phase component is 0.01-0.03:1; the mass of the sterile water is 30-50% of that of the solid phase components, and the viable count of the biocontrol bacteria in the microbial inoculum is more than or equal to 50 hundred million cfu/g;
the biomass charcoal is obtained by: taking agricultural wastes as raw materials, preparing the biomass charcoal under the anaerobic condition of 400-700 ℃, cooling, grinding, sieving with a 40-mesh sieve, and sterilizing at 121 ℃ for 1-2 hours to obtain the biomass charcoal;
the agricultural waste is one or two of rice hulls or straws;
the biocontrol bacteria are one or two of bacillus tequilensis and bacillus vallismortis.
2. The preparation method of the biomass charcoal-based microbial agent for preventing and treating watermelon fusarium wilt as claimed in claim 1, which comprises the following steps:
step one, fermenting biocontrol bacteria to prepare the biocontrol bacteria with the bacteria content of 1012cfu/mL biocontrol bacteria fermentation liquor for later use;
and step two, uniformly mixing the biocontrol bacteria fermentation liquor and the biomass charcoal, then adding the rest components, uniformly mixing, granulating until the particle diameter is 3-6 mm and the pH value is 6.5-7.5, and then drying until the particle humidity is less than or equal to 20% to obtain the biomass charcoal-based microbial inoculum for preventing and treating watermelon fusarium wilt.
3. The method for preparing the biomass charcoal-based microbial agent for preventing and treating watermelon fusarium wilt according to claim 2, wherein the biocontrol bacteria fermentation broth in the first step is: inoculating biocontrol bacteria into liquid LB culture medium, and culturing in a shaker at 37 + -1 deg.C and 200rpm until the bacteria content is 1012cfu/mL。
4. The use of the biomass charcoal-based microbial agent for controlling watermelon fusarium wilt as claimed in claim 1 for controlling watermelon fusarium wilt.
5. The use as claimed in claim 4, wherein the biomass charcoal based microbial agent for controlling watermelon fusarium wilt is applied in holes at the time of transplanting watermelon seedlings, and the dosage of each plant is 100g, so as to control watermelon fusarium wilt.
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