Disclosure of Invention
Aiming at the salinization and biological character degradation of facility soil at present, the invention provides a biological activity conditioner for improving the salinization of the facility soil, the conditioner is prepared by taking a low-salt high-quality organic fertilizer, biochar, a high-biological activity trichoderma fungicide and a bacillus fungicide as raw materials, has the advantages of sufficient raw materials, low cost, easy implementation, environmental protection and the like, can obviously reduce the salt content of the facility soil, improves the microbial activity and the microbial flora of the soil, and increases the crop yield.
In order to realize the purpose, the invention firstly provides a bioactive conditioner for improving facility soil salinization, and the soil conditioner comprises the following raw materials: according to the mass percentage, the biochar accounts for 10-20 percent, and the trichoderma viride TV41 microbial inoculum (the bacterial content is more than 2 multiplied by 10)9CFU/g) accounts for 0.5-1%, and Bacillus amyloliquefaciens BS211 microbial inoculum (the bacterial content is more than 4 multiplied by 10)9CFU/g) accounts for 0.5-1 percent, and the balance is low-salt organic fertilizer; the technical indexes of the low-salt organic fertilizer are as follows: the organic content (calculated on a dry basis) is more than or equal to 45 percent; the content (calculated on a dry basis) of nitrogen, phosphorus pentoxide and potassium oxide is more than or equal to 5 percent; the water content is less than or equal to 30 percent; the pH value is 6.5-8.5;
in the invention, the low-salt organic fertilizer can be a commercially available product such as a traditional Chinese medicine residue organic fertilizer, a straw organic fertilizer, a mushroom residue organic fertilizer and the like, or an organic fertilizer obtained by taking traditional Chinese medicine residue, mushroom residue and straw as raw materials and performing high-temperature fermentation and after-ripening fermentation decomposition (the decomposition method is shown in Shandong province local standard DB 37T 2248 and 2012), and the aim of the invention can be achieved as long as the low-salt organic fertilizer is ensured to meet the technical indexes.
Further, in the bioactive conditioner for improving facility soil salinization disclosed by the invention, the biochar is commercial biochar or a solid product obtained by carbonizing agricultural production organic waste at the temperature of 300-; the agricultural production organic waste comprises at least one of corn straw, corncob, wheat straw, rice hull and soybean straw.
Further, in the bioactive conditioner for improving facility soil salinization disclosed by the invention, the trichoderma viride TV41 microbial inoculum is obtained by the following method:
a) inoculating Trichoderma viride TV41 strain to potato glucose agar culture medium, and culturing at 28 deg.C for 3 days; then transferring the Trichoderma viride strain to a potato glucose liquid culture medium, wherein the liquid loading amount is 80m1/250m1, and then carrying out shake cultivation at 28 ℃ for 3-4 days at the rotation speed of 160rpm of a shaking table;
b) inoculating 10% (volume/mass ratio, ml/kg) of the sterilized bran and straw with water content of 60% after the culture is finished, and culturing at 28 +/-2 ℃ for 5-7 days until sporulation is produced;
c) naturally air drying Trichoderma viride and bran or oven drying at 30 deg.C, pulverizing, and sieving with 10 mesh sieve to obtain Trichoderma viride TV41 microbial inoculum with bacteria content of more than 2 × 109CFU/g。
Further, in the bioactive conditioner for improving facility soil salinization disclosed by the invention, the bacillus amyloliquefaciens BS211 microbial inoculum is obtained by the following steps:
a) inoculating bacillus amyloliquefaciens BS211 into a beef extract peptone agar culture medium, and culturing for 3 days at 32 ℃; then transferring the bacillus amyloliquefaciens strain into a beef extract peptone liquid culture medium, wherein the liquid loading is 80m1/250m1, and then carrying out shake cultivation at 32 ℃ for 3-5 days at the rotation speed of 160rpm of a shaker;
b) inoculating 10% (volume/mass ratio, ml/kg) of the sterilized bran and straw with water content of 60% after the culture is finished, and culturing at 32 +/-2 ℃ for 3-5 days until sporulation is produced;
c) will solveNaturally air drying the bacillus amyloliquefaciens and bran and straws or drying the straws at 40 ℃, crushing the straws and sieving the straws with a 10-mesh sieve to obtain the bacillus amyloliquefaciens BS211 microbial inoculum with the bacterial content of more than 4 multiplied by 109CFU/g。
The invention further provides an application of the soil conditioner in improving the soil of the salinization facility, and the application method comprises the following steps: uniformly spreading the soil conditioner in the facility soil with the dosage of 300-.
Furthermore, the facility soil is preferably sunlight greenhouse or greenhouse soil with salt content of 2.0-6.0 g/kg.
The applicant obtains the biological activity conditioner for salinization of green improved facility soil through multiple formula improvement, pot culture experiments and field experiments, can improve a microflora and biological activity, balance soil nutrients, reduce the total salt content of the soil, relieve continuous cropping obstacles and improve the yield and quality of crops, and has the following beneficial effects compared with the prior art:
(1) meanwhile, the method has the functions of reducing facility soil salinization and improving soil biological activity.
(2) The invention has reasonable raw material compounding, and each raw material has specific functions: the low-salt decomposed organic fertilizer is not easy to increase the salt content of soil, effectively increases the organic matter content and active substances of the soil, and is a good living organic carrier of effective bacteria; the biochar can improve the organic carbon content of soil, improve the water and fertilizer retention performance of the soil, improve the buffering capacity of the soil on high salinity and acidification, has large specific surface area, and can provide living space for beneficial microorganisms; the TV41 microbial inoculum and the BS211 microbial inoculum have high effective microbial activity, can effectively improve soil microflora, improve soil bioactivity and inhibit pathogenic microorganisms, and in addition, a large number of metabolites in the microbial inoculum have the effects of promoting crop growth and inhibiting pathogenic microorganisms.
(3) The fertilizer is applied to soil as a base fertilizer, and the application amount of organic materials and fertilizers is effectively reduced.
(4) The invention has sufficient main raw materials and low price, and the field application method is simple, thus being an environment-friendly conditioner for preventing and controlling continuous cropping obstacles of greenhouse vegetables.
Detailed Description
The technology related to the present invention will be further described with reference to specific examples.
The examples relate to media and strains:
potato dextrose agar medium: peeling 200g potato, cutting into pieces, cooking for 30min, filtering with four layers of gauze to obtain filtrate, adding 20g glucose, diluting to 1L, adding 20g agar, and sterilizing at 121 deg.C for 30 min.
Potato dextrose liquid medium: peeling 200g potato, cutting into pieces, cooking for 30min, filtering with four layers of gauze to obtain filtrate, adding 20g glucose, diluting to 1L, and sterilizing at 121 deg.C for 30 min.
Beef extract peptone agar medium: 5g of beef extract, 10g of peptone, 5g of NaCl, 20g of agar and 1L of water, and the pH value is 7.2-7.4. Sterilizing at 121 deg.C for 30 min.
Beef extract peptone liquid medium: 5g of beef extract, 10g of peptone, 5g of NaCl, 1L of water and 7.2-7.4 of pH. Sterilizing at 121 deg.C for 30 min.
The preparation method of the solid culture medium of bran and rice straw comprises mixing bran and rice straw (length less than 4mm) at a mass ratio of 9:1, adding water to make water content in the mixture 60%, sterilizing at 126 deg.C for 60min, and cooling.
Trichoderma viride (Trichoderma harzianum) used in the examples was Trichoderma viride TV41 strain with a collection number of CGMCC No.9293, as disclosed in document 104982198A and provided by the academy of agricultural sciences of Jiangsu province.
The Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) BS211 strain used in the examples (see the literature, "antagonistic activity and potting control effect of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) BS211 in watermelon", Mayan et al, Jiangsu agricultural bulletin, 2006) was provided by the agricultural academy of Jiangsu province.
The examples relate to the starting materials:
the low-salt organic fertilizer is purchased from Chinese medicine residue organic fertilizer of mucelis fertilizer industry ltd in Jiangsu province; the organic content (calculated on a dry basis) is more than or equal to 45 percent; the content (calculated on a dry basis) of nitrogen, phosphorus pentoxide and potassium oxide is more than or equal to 5 percent; the water content is less than or equal to 30 percent; the pH value is 6.5-8.5.
Biochar: purchased from Li Yang Shang De Sheng active carbon factory, is rice straw and rice hull biochar.
Example 1 determination of acid and salt resistance of Trichoderma viride TV41 bacteria
Inoculating Trichoderma viride TV41 strain into potato glucose agar culture medium with pH of 3, 4, 5, 6, 7, 8, 9, 10, 11 in sequence, and culturing at 28 deg.C for 3 days, wherein the photograph of the culture dish is shown in FIG. 1; as shown in FIG. 3, the total number of spores on the medium was determined, and it can be seen that the Trichoderma reesei (TV 41) produced large amount of spores under acidic condition of pH4-5, which can reach 2.9 × 109Above CFU.
Trichoderma viride TV41 strain was inoculated into potato dextrose agar medium with salt concentration (salinity adjusted by NaCl reagent) of 0%, 0.6%, 1.2%, 1.8%, 2.4%, 3%, 3.6%, 4.2%, 4.8%, and cultured at 28 deg.C for 2 days, and the photograph of the dish is shown in FIG. 2; the results of detecting the total number of spores on the culture medium and the amount of spores are shown in FIG. 4, and it can be seen that the amount of spores produced by Trichoderma TV41 at 1.2% salt concentration is relatively large, and is substantially consistent with the amount of spores produced at 0% salt concentration.
Example 2 determination of acid and salt resistance of Bacillus amyloliquefaciens BS211 bacteria
The number of viable bacteria detected after the bacillus amyloliquefaciens BS211 strain is inoculated into beef extract peptone culture media with different pH values and cultured for 3 days at 30 ℃ is shown in figure 5, the number of viable bacteria cultured by the bacillus amyloliquefaciens BS211 strain under the acidic condition of pH4-5 can still be larger and can reach 5.1 multiplied by 108Above CFU.
In addition, the BS211 strain is inoculated into beef extract peptone culture media with different salt concentrations (the salt concentration is adjusted by NaCl reagent) and cultured for 3 days at 30 ℃, and then the viable count is detected, and the detection result is shown in figure 6, so that the viable count of the BS211 bacillus cultured under the salt concentration of 3% is still high and is basically consistent with the viable count under the salt concentration of 0%.
Example 3 preparation of Trichoderma viride TV41 inoculum
a) Inoculating the Trichoderma viride TV41 strain preserved on the inclined plane into a potato glucose agar culture medium, and culturing at 28 ℃ for 3 days to generate green conidia; and inoculating the trichoderma viride mycelia with conidia into a potato glucose liquid culture medium, wherein the liquid filling amount of the liquid culture medium is 80m1/250m1, and 3 trichoderma viride mycelia with the diameter of 8mm are inoculated into each bottle. Then culturing at 28 deg.C with shaking table rotation speed of 160rpm for 3-4 days to obtain mycelium solution.
b) Inoculating 80ml of trichoderma mycelium liquid into 1.5kg of sterilized bran and straw solid culture medium, uniformly stirring under aseptic condition, spreading the solid culture in a shallow tray (the shallow tray is soaked in formalin solution overnight for airing), wherein the thickness is 3-5cm, spreading filter cloth subjected to high-pressure sterilization treatment below the shallow tray, covering double layers of gauze subjected to high-pressure sterilization treatment above the shallow tray, and culturing at 28 +/-2 ℃ for 5-7 days until spore production is complete.
c) Naturally air drying Trichoderma viride and wheat bran or oven drying at 30 deg.C until water content is less than 10%, pulverizing, and sieving with 10 mesh sieve to obtain Trichoderma viride TV41 microbial inoculum with bacteria content of more than 2 × 109CFU/g。
Example 4 preparation of Bacillus amyloliquefaciens BS211 inoculant
a) Streaking and inoculating the bacillus amyloliquefaciens BS211 strain stored on the inclined plane into a beef extract peptone agar culture medium, and culturing for 2 days at 32 ℃; then, the bacterium blocks with the BS211 colonies are inoculated into a beef extract peptone liquid medium, the liquid loading capacity of the liquid medium is 80m1/250m1, and 3 bacterium blocks are inoculated in each bottle. Then culturing for 3-5 days at 32 ℃ and 180rpm of a shaking table to obtain a bacterial liquid.
b) Inoculating 80ml of cultured bacillus liquid into 1.5kg of sterilized bran and straw solid culture medium, uniformly stirring under aseptic condition, spreading the solid culture in a shallow tray (the shallow tray is soaked in formalin solution overnight for airing) with the thickness of 3-5cm, spreading filter cloth subjected to high-pressure sterilization treatment below the shallow tray, covering double layers of gauze subjected to high-pressure sterilization treatment above the shallow tray, and culturing at 32 +/-2 ℃ for 4-6 days until spore production is complete.
c) Naturally air drying Bacillus together with bran and straw or oven drying at 40 deg.C until water content is less than 10%, pulverizing, and sieving with 10 mesh sieve to obtain Bacillus amyloliquefaciens BS211 microbial inoculum with bacterial content of more than 4 × 109CFU/g。
Example 5 growth promoting effects of Trichoderma viride TV41 microbial inoculum on tomatoes in saline soil
NaCl is added into the plough layer soil until the NaCl concentration in the soil is 0 percent (CK), 0.2 percent, 0.4 percent, 0.6 percent and 0.8 percent, the soil with the salt concentration is divided into two groups, 0 percent (control group) and 1 percent (experimental group) of the trichoderma viride TV41 microbial inoculum prepared in the example 3 are respectively added in mass percent, and the potted landscape after 30 days of planting the tomatoes is shown in figure 7.
The results of the above-ground fresh weight and the underground fresh weight of the tomato are shown in fig. 8 and fig. 9, the trichoderma can obviously promote the above-ground and underground parts of tomato plants in the soil with 0-0.8% of NaCl concentration, and the higher the NaCl concentration is, the more obvious the promotion effect is.
Example 6 growth promoting action of Trichoderma viride TV41 microbial inoculum and Bacillus amyloliquefaciens BS211 microbial inoculum on pepper in salinized soil
Taking plough layer soil, adding NaCl until the NaCl concentration in the soil is 0.8%, and dividing the treatment into four treatments: the method comprises the steps of (1) adding no fungicide (CK), preparing 1% of bacillus amyloliquefaciens BS211 fungicide by mass percent in the addition example 4 (control group 1), preparing 1% of trichoderma viride TV41 fungicide by mass percent in the addition example 3 (control group 2), simultaneously adding 1% of BS211 fungicide by mass percent and 1% of TV41 fungicide by mass percent (experimental group), and planting pepper for 30 days, wherein the pot culture scene is shown in figure 10.
The results of the aboveground fresh weight and underground fresh weight of the pepper are shown in fig. 11, and the pepper plants treated by the aseptic agent have extremely poor growth and high mortality rate. The BS211 microbial inoculum and the TV41 microbial inoculum which are independently applied have better growth promotion effect on pepper plants, the BS211 microbial inoculum and the TV41 microbial inoculum are applied in a compounding way, the effect is best, the growth promotion effect on the pepper plants is very obvious, and the death rate is low.
Example 7 application in greenhouse amaranth soil
Uniformly mixing 90kg of traditional Chinese medicine residue organic fertilizer, 10kg of charcoal, 0.5kg of trichoderma viride TV41 microbial inoculum and 0.5kg of bacillus amyloliquefaciens BS211 microbial inoculum to prepare the soil conditioner.
Selecting a greenhouse with green vegetables in a sky, blue and earth green vegetable production base in a tin-free urban area, wherein the service life of the greenhouse is more than 8 years. The experiment set up 2 treatments: conventional treatments and soil conditioner treatments. Each processing 3 repeated cells with a cell area of 6m2And are randomly arranged. The conventional treatment of 15-15-15 compound fertilizer is 30 kg/mu, and the soil conditioner treatment conditioner is 300 kg/mu. And (5) uniformly spreading the amaranth seeds on the soil surface, fully ploughing, watering until the maximum water holding capacity of the field is reached, and planting the amaranth seeds after keeping moisture for 10 days. The results are shown in tables 1 and 2.
Table 1 shows that the soil conditioner has the effect on the yield of amaranth and the salt content of soil
The results in table 1 show that, compared with the conventional control, the soil conditioner can significantly reduce the content of soluble salt in soil by 17.1% and increase the yield of amaranth by 17.5%.
TABLE 2 influence of soil conditioners on the number of bacteria, number of fungi, number of beneficial bacteria, microbial activity and microbial diversity in facility amaranth soil
The results in Table 2 show that the number of bacteria increased by 18.9% and the number of fungi decreased by 15.6% for the soil conditioner treatment, indicating that the soil conditioner inhibited soil mycosis and increased the bacteria fraction. In addition, the conditioner treated trichoderma increased by 4.75 × 102CFU/g, the number of bacillus increased by 0.77X 106CFU/g, which indicates that the application of conditioner is beneficial for increasing the number of soil-beneficial microorganisms. Biolog carbon source utilization detection of AWCD values can be a sensitive indicator of microbial activity. Therefore, the conditioner also has a remarkable promoting effect on the activity and the microbial diversity of soil microorganisms. The increase of beneficial microorganisms in the soil is beneficial to promoting plants to absorb nutrients, so that the salt content of the soil is reduced, and healthy disease-inhibiting soil is formed.
Example 8 application in greenhouse Chinese little greens soil
90kg of straw organic fertilizer, 10kg of charcoal, 0.5kg of trichoderma viride TV41 microbial inoculum and 0.5kg of bacillus amyloliquefaciens BS211 microbial inoculum are mixed uniformly to prepare the soil conditioner.
Selecting a greenhouse with the service life of more than 10 years in the vegetable production base of the longevity river in the Wuxi city mountainous area. The experiment set up 2 treatments: conventional treatments and soil conditioner treatments. Each processing 3 repeated cells with a cell area of 6m2And are randomly arranged. The conventional treatment of 15-15-15 compound fertilizer is 30 kg/mu, and the soil conditioner treatment conditioner is 300 kg/mu. And (3) uniformly spreading the water on the soil surface, fully ploughing, watering until the maximum water holding capacity of the field is reached, and planting the Chinese little greens after keeping moisture for 10 days. The results are shown in tables 3 and 4.
TABLE 3 soil conditioner on Chinese little greens yield and soil salinity
The results in table 3 show that, compared with the conventional control, the soil conditioner can significantly reduce the content of soluble salt in soil by 13.3% and increase the yield of the Chinese little greens by 24.5%.
TABLE 4 influence of soil conditioners on the number of bacteria, number of fungi, number of beneficial bacteria, microbial activity and microbial diversity in the soil of greenhouse salsola gallica
The results in table 4 show that the soil biological properties can be significantly improved by applying the soil conditioner. The soil conditioner treatment significantly increased the number of bacteria, trichoderma and bacillus, and inhibited soil mycosis compared to the conventional control. In addition, the soil conditioner treatment microbial activity and microbial diversity were also higher than the conventional controls.
Example 9 application in solar greenhouse tomato soil
80kg of traditional Chinese medicine residue organic fertilizer, 20kg of charcoal, 1kg of trichoderma viride TV41 microbial inoculum and 1kg of bacillus amyloliquefaciens BS211 microbial inoculum are uniformly mixed to prepare the soil conditioner.
Selecting a facility tomato greenhouse with the service life of more than 8 years in the vegetable production bases of Shuzhou suining county, Shuichouzhen village. The experiment set up 2 treatments: conventional treatments and soil conditioner treatments. Each treated 3 repeat cells, with a cell area of 0.25 mu, randomly arranged. Spreading 15-15-15 compound fertilizer on the ridges in the conventional treatment mode, wherein the using amount is 50 kg/mu. The conditioner is spread on the soil conditioner treatment ridges, and the dosage is 500 kg/mu. And uniformly spreading on the upper surface of the ridge, fully ploughing, watering until the maximum water holding capacity of the field is reached, and planting the tomatoes after keeping moisture for 10 days. The results are shown in tables 5 and 6.
TABLE 5 soil conditioner on tomato yield, tomato disease and soil salinity
The results in Table 5 show that compared with the conventional control, the soil conditioner can obviously reduce the content of soluble salt in the soil by 28.1 percent, reduce the incidence rate of leaf mold by 75.1 percent and increase the tomato yield by 51.3 percent.
TABLE 6 influence of soil conditioners on the number of bacteria, number of fungi, number of beneficial bacteria, microbial activity and microbial diversity in tomato soil of a facility
The results in Table 6 show that the soil conditioner treatment increased the number of bacteria and Trichoderma by 38.5% and 80.4%, respectively, and the fungus decreased by 22.9%, as compared to the conventional control. In addition, conditioner treatments may also increase soil microbial activity (FDAH enzyme) and microbial diversity.
In specific implementation, the low-salt organic fertilizer can be a commercial product of Chinese medicine residue organic fertilizer (come fertilizer industry ltd. good in Jiangsu province), mushroom residue organic fertilizer (Nanjing Ning grain bio-fertilizer company) and straw organic fertilizer (Nanjing Mingzhu fertilizer ltd. company), or an organic fertilizer which is prepared by taking Chinese medicine residue, mushroom residue and straw as raw materials and fully decomposed through high-temperature fermentation and after-ripening fermentation, as long as the low-salt organic fertilizer meets the following technical indexes: the organic content (calculated on a dry basis) is more than or equal to 45 percent; the content (calculated on a dry basis) of nitrogen, phosphorus pentoxide and potassium oxide is more than or equal to 5 percent; the water content is less than or equal to 30 percent; the pH value is 6.5-8.5, and the aim of the invention can be achieved.
The biochar can be selected from a commercially available product, and can also be prepared by crushing a solid product obtained by carbonizing agricultural production organic waste (such as at least one of corn straw, corncob, wheat straw, rice hull and soybean straw) at 600 ℃ and sieving the crushed solid product with a 40-mesh sieve.