CN112159761A - Preparation method of penicillium oxalicum and application of penicillium oxalicum in phosphate solubilizing, growth promoting and fusarium graminearum antagonism - Google Patents
Preparation method of penicillium oxalicum and application of penicillium oxalicum in phosphate solubilizing, growth promoting and fusarium graminearum antagonism Download PDFInfo
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Abstract
The invention discloses a preparation method of penicillium oxalicum and application of the penicillium oxalicum in phosphate solubilizing, growth promoting and fusarium graminearum antagonism, and belongs to the technical field of environmental biology. The penicillium oxalicum MEM02 can inhibit germination of fusarium graminearum spores in soil, infection of the fusarium graminearum spores to wheat is remarkably reduced, and meanwhile, the strain MEM02 can efficiently dissolve phosphorus and promote plant growth. Therefore, the penicillium oxalicum MEM02 has a good application prospect in the aspect of developing new biological pesticides or new biological control microbial agents.
Description
Technical Field
The invention belongs to the technical field of environmental biology, and particularly relates to a preparation method and application of a penicillium oxalicum strain MEM 02.
Background
Phosphorus is one of three essential mineral nutrient elements for plant growth, phosphate fertilizer is an important substance for grain yield increase and grain safety guarantee, and improving the utilization efficiency of phosphorus and phosphate fertilizer in soil is an important measure for saving phosphate fertilizer and sustainable development of agriculture. About 74% of cultivated land soil in China is lack of phosphorus, most of phosphorus is solidified phosphorus which is difficult to absorb and utilize by plants, and the problem of phosphorus deficiency of soil is improved by applying phosphate fertilizer in agricultural production. However, the utilization rate of the crops in the season of the phosphate fertilizer applied to the soil is low, so that the phosphorus in the soil is continuously accumulated, the soil is hardened, the quality is degraded, and the ecological environment is polluted. Improving the utilization rate of phosphorus has important significance for improving the soil structure and protecting the ecological environment.
Research shows that a large number of microorganisms in the soil of the root system of the plant have strong phosphate solubilizing function and can convert the phosphorus which is difficult to be absorbed and utilized by the plant and is in an invalid state into an absorbable and utilizable form, and the microorganisms with the capability are called phosphate solubilizing bacteria and have reported that aspergillus (Aspergillus (A) (phosphorus solubilizing bacteria)Aspergillus) Rhizopus (A) and (B)Rhizopus) Fusarium (F.), (Fusarium) Microsclerotium bacterium (C.) (Sclerotium) And the like.
Wheat scab is a global wheat disease and is one of the main diseases harmful to wheat production in China, particularly fusarium graminearum in winter wheat areas in the middle and lower reaches of Yangtze river. Its occurrence and development are related to the accumulation amount of local bacteria sources, meteorological factors and agricultural measures. After the wheat is diseased, the yield is generally reduced by about 20 percent, the ear of the seriously diseased land can exceed 90 percent, the wheat is basically harvested absolutely, and huge economic loss is brought to agricultural production. It not only causes large-area yield reduction, influences the seed quality and reduces the rate of emergence, but also is easy to be poisoned after people and livestock eat sick wheat, and leaves huge hidden troubles for food safety.
The strain MEM02 is a penicillium oxalicum strain, can efficiently dissolve phosphorus and promote growth, can inhibit germination of fusarium graminearum spores, particularly can obviously inhibit germination of the fusarium graminearum spores in soil containing rice straws, can control the quantity of the fusarium graminearum from the source and reduce the possibility of wheat infection. The invention also provides a simple and efficient fermentation culture method for improving the number of spores of penicillium oxalicum. At present, the use of penicillium oxalicum spore fungicide for preventing and treating wheat scab in rice and wheat crop rotation areas is rarely reported.
Disclosure of Invention
The penicillium oxalicum MEM02 can inhibit germination of fusarium graminearum spores in soil, infection of the fusarium graminearum spores to wheat is remarkably reduced, meanwhile, the strain MEM02 can efficiently dissolve phosphorus and promote plant growth, and the fungus MEM02 has a good application prospect in the aspect of developing new biological pesticides or new biological control inoculants.
After a lot of experiments and continuous efforts of the inventor, a strain of penicillium oxalicum is finally obtained, wherein the penicillium oxalicum is penicillium (II)Penicillium oxalicum) MEM02, deposited at the chinese type culture collection with accession number: CCTCC NO, M20191002, preservation date: 12/3/2019, address: chinese Wuhan dynastyWuhan university.
The penicillium oxalicum MEM02 can inhibit germination of fusarium graminearum spores and is used for preventing and treating wheat scab.
The penicillium oxalicum MEM02 can efficiently dissolve phosphorus and promote plant growth, and is used for preparing a microbial preparation for promoting crop growth.
The method for producing the high-concentration spore preparation by the solid fermentation of the penicillium oxalicum comprises the following steps:
(1) taking a penicillium oxalicum block from a penicillium oxalicum flat plate by using a puncher with the diameter of 0.7 cm, inoculating the penicillium oxalicum block into a PDA solid culture medium (200 g of potato, 20 g of glucose, 15g of agar, 1.0L of deionized water, sterilizing at 115 ℃ for 30min, and naturally adjusting pH) to activate for 3 days, and washing with 10 mL of sterile water to obtain a penicillium oxalicum spore suspension;
(2) and (2) uniformly inoculating 5% of the spore suspension activated in the step (1) into a fermentation solid culture medium, fermenting for 5 days at 28 ℃, and uniformly mixing conical flasks once a day to obtain a solid fermentation product, namely the penicillium oxalicum MEM02 solid fermentation product for producing the high-concentration spore preparation.
Preferably, the solid fermentation method for producing the high-concentration spore preparation by the penicillium oxalicum strain comprises the following steps of: wheat bran, rice chaff, rice straw powder, K2HPO4、FeSO40.005、MgSO4 .7H2O、CaCO3And deionized water.
Preferably, in the method for producing a high-concentration spore preparation by solid fermentation of the penicillium oxalicum strain, the fermentation solid medium in the step (2) comprises the following components in parts by weight: 25 parts of wheat bran, 10 parts of rice chaff, 15 parts of rice straw powder and K2HPO41.0 part of FeSO40.005 part of MgSO 24 .7H20.05 part of O, CaCO3 0.05 part and 50 parts of deionized water.
Compared with the prior art, the invention has the following beneficial effects:
(1) penicillium oxalicum of the present inventionThe MEM02 has effect in inhibiting Fusarium graminearum with a bacteriostasis rate of 56.45%, and is 2 × 107CFU/mL Penicillium oxalicum MEM02 spores were significantly inhibited by 2X 105The CFU/mL fusarium graminearum spores germinate in the soil containing the rice straws and can promote the decomposition of the rice straws. The quantity of fusarium graminearum can be controlled from the source, and the infection of the fusarium graminearum on wheat is reduced.
(2) The phosphorus-dissolving concentration of the penicillium oxalicum MEM02 in the PKO culture medium containing calcium phosphate can reach 1250.68 mg/L.
(3) After the penicillium oxalicum MEM02 is fermented for 5 days by a solid fermentation culture medium, the concentration of the penicillium oxalicum spore preparation reaches 2.29 multiplied by 109 CFU/g。
(4) The growth promotion effect of the penicillium oxalicum MEM02 on the pakchoi is obviously higher than that of a control group, compared with a control group without inoculation, the growth rates of fresh weight, plant height, stem thickness and root length of a plant taking phosphate rock powder as a phosphorus source are 191.86%, 28.46%, 66.67% and 22.9% respectively, and the effective phosphorus in soil is improved by 561.72%.
(5) The penicillium oxalicum MEM02 provided by the invention is derived from soil in rice and wheat crop rotation areas of Zhongxiang phosphorite Hubei as a biological control and plant growth promoting material for wheat scab, and the fungus has good application prospect no matter in the aspect of developing new biological pesticides or new biological control microbial inoculum.
Drawings
FIG. 1 is a soluble phosphor ring of MEM02 on a PKO plate.
Fig. 2 is a micrograph of spores of MEM 02.
FIG. 3 is a phylogenetic tree of MEM 02.
FIG. 4 shows the phosphorus dissolving effect of MEM02 on calcium phosphate, magnesium phosphate, and powdered rock phosphate.
FIG. 5 is a graph of the effect of different temperatures, pH, carbon and nitrogen sources on the phosphorus solubilizing effect of MEM 02.
FIG. 6 shows MEM02 in opposition to a plate of Fusarium graminearum.
FIG. 7 is a photograph of an experiment on the resistance of MEM02 spores to Fusarium graminearum spores, 2X 10 on the left5CFU/mL Fusarium graminearum spores, 2X 10 in the right5CFU/mL fusarium graminearum spores+2×107CFU/mL Penicillium oxalicum spores.
FIG. 8 is a graph of the effect of MEM02 spores on the germination rate of Fusarium graminearum spores in rice straw-containing soil.
Detailed Description
The following are specific embodiments of the present invention and further description of the technical solutions of the present invention, but the present invention is not limited to the scope of the embodiments, and all changes or equivalent substitutions that do not depart from the spirit of the present invention are included in the scope of the present invention.
The embodiment relates to the components of the culture medium and the reagent:
inorganic phosphorus medium (PKO medium): glucose 10.0 g, (NH)4）2SO40.5 g, yeast powder 0.5 g, NaCl 0.3 g, KCl 0.3 g, FeSO4·7H2O 0.03 g、MgSO4·7H2O 0.3 g、MnSO4·4H2O 0.03 g、Ca3(PO4)25.0 g, 1.0L deionized water and 20.0 g agar, sterilizing at 115 ℃ for 30min, and adjusting the pH value to 7.0-7.2.
Potato dextrose agar medium (PDA medium): 200 g of potato, 20 g of glucose, 15g of agar and 1.0L of deionized water, and sterilizing at 115 ℃ for 30min, wherein the pH value is natural.
Potato glucose medium (PDB medium) comprises potato 200 g, glucose 20 g, deionized water 1.0L, and sterilizing at 115 deg.C for 30min with natural pH.
Yeast extract peptone glucose agar medium (YPD medium): 1.0% yeast extract, 2.0% peptone, 2.0% glucose, 2.0% agar powder, sterilized at 115 ℃ for 30min, pH 7.0.
PBS culture medium: KH (Perkin Elmer)2PO4 0.24g，Na2HPO43.63 g, NaCl 8g, KCl 0.2 g, adding deionized water to 1L, sterilizing at 121 deg.C for 20min, and naturally adjusting pH.
CMC culture medium: 20 g of CMC-Na, 2.5 g of Na2HPO4，1.5g KH2PO42.5 g of peptone, deionized water was added to 1L, 121 ℃, 20min, pH 7.0-7.5.
The high-yield spore solid fermentation culture medium comprises the following components in parts by weight: 25 parts of wheat bran, 10 parts of rice chaff, 15 parts of rice straw and K2HPO4 1.0 part of FeSO4 0.005 part of MgSO 24 .7H20.05 part of O, CaCO3 0.05 part and 50 parts of deionized water, sterilizing at 121 ℃ for 20min, and naturally adjusting the pH value.
EXAMPLE 1 screening of the strains of the invention
10 g of field rhizosphere soil sample in the Zhongxiang phosphorite Zhendao rice and wheat rotation region in Hubei province is weighed and put into a conical flask filled with 90 mL of PKO culture medium, and cultured for 2 days at 28 ℃ and 150 rpm. And (3) taking 0.2 mL of soil suspension after enrichment culture, coating the soil suspension on an inorganic phosphorus solid culture medium, carrying out inverted culture at 28 ℃ for 7 days, and purifying the strain with a phosphorus-soluble ring. Inoculating the primarily screened fungus on a PDA plate, taking a piece of fungus block with a hole puncher with the diameter of 0.7 cm after the colony grows to be full, inoculating the fungus block into a conical flask filled with 200 mL of PKO culture medium, and carrying out shake culture at 28 ℃ and 150 rpm. The supernatant was taken every day to determine pH and available phosphorus content (Mo-Sb colorimetric resistance method), and the measurement was continued for 8 days. Through the screening, 1 strain which can take inorganic phosphorus as the only phosphorus source and generate obvious phosphorus dissolving rings on the screening plate is obtained as shown in figure 1, and the number is MEM02, namely the strain in the patent.
Example 2 morphological Observation and PCR identification of the strains
The strain MEM02 is inoculated on a PDA plate and is inversely cultured for 7 days at the temperature of 28 ℃, the colony of the strain MEM02 is grey, the reverse side is light yellow, a large number of conidia are generated, the edge is neat, the opacity is realized, and no exudate exists. The strain MEM02 plate was washed with sterile water to obtain Penicillium oxalicum MEM02 spores, and the shape of the spores was oval as shown in FIG. 2. Inoculating high-efficiency phosphorus-dissolving fungus MEM02 on YPD plate, and performing inverted culture at 28 deg.C for 3-4 days until large amount of hypha is produced. Adopts a fungus universal primer ITSl (5)，-TCCGTAGGTGAACCTGCGG-3，) And ITS4 (5)，-TCCTCCGCTTATTGATATGC-3，）
Amplification is performed.
The PCR amplification system is 50 μ L: 1. mu.L each of the upstream and downstream primers (10. mu. mol/L) ITSl and ITS4, 2. mu.L of DNA template, Easy Taq DNA Polymerase (5U)/μL) 0.5 μL，10×Easy Taq buffer 5 μL，dNTP (2.5 mM) 6.5 μL，ddH2O34. mu.L. 5 min at 95 ℃; 30 cycles at 95 ℃ for 30 s, 51 ℃ for 1 min, 72 ℃ for 45 s; and (5) 10 min at 72 ℃.
The PCR amplification product is sent to Biotech company of Wuhan department for sequence analysis, the sequencing result (SEQ ID NO: 1) is subjected to Blast comparison analysis on NCBI, MEGA4.1 is utilized to construct phylogenetic tree as shown in figure 3, and MEM02 is identified as penicillium oxalicum (Merll) (Merlol & ltm & gt, Merlol & ltm & gt, MerlPenicillium oxalicum）。
Example 3 preparation of Penicillium oxalicum MEM02 high spore productivity solid fermentation inoculum
A puncher with the diameter of 0.7 cm is used for taking a penicillium oxalicum block from a penicillium oxalicum MEM02 flat plate, inoculating the penicillium oxalicum block into a PDA solid culture medium for activation for 3 days, and 10 mL of sterile water is used for washing to obtain a penicillium oxalicum spore suspension. The activated spore suspension is uniformly inoculated into the MEM02 high-efficiency spore-producing solid fermentation medium designed by the patent according to the inoculation amount of 5 percent (5 percent of the mass of the fermentation medium) for fermentation for 5 days at 28 ℃, and the Erlenmeyer flasks are uniformly mixed once a day. The obtained solid fermentation product is the penicillium oxalicum MEM02 solid fermentation product to produce the high-concentration spore preparation. The concentration of Penicillium fragrans spores is 2.29 multiplied by 10 after counting by a blood cell counting plate9 CFU/g。
Example 4 phosphate solubilizing Effect of Penicillium oxalicum MEM02
The MEM02 was inoculated on a PDA plate, after the colony growth was completed, the colony blocks were taken out with a punch having a diameter of 0.7 cm, and inoculated in conical flasks containing 200 mL and 5 g/L of insoluble phosphates (calcium phosphate, magnesium phosphate, powdered rock phosphate), respectively, and cultured with shaking at 28 ℃ and 150 rpm, and the supernatant was taken every day to measure the pH and the available phosphorus content (MoSb colorimetric method), and continuously measured for 8 days. The phosphorus dissolving effect of MEM02 on calcium phosphate, magnesium phosphate and ground phosphate rock is shown in FIG. 4. After fungi MEM02 is inoculated, the content of available phosphorus in the culture medium taking calcium phosphate, magnesium phosphate and ground phosphate rock as phosphorus sources can reach 1250.68 mg/L, 1461.78 mg/L and 103.42 mg/L at most.
The content of soluble phosphorus was measured after culturing for 6 days with PKO as a basal medium and calcium phosphate as a phosphorus source under conditions of different temperatures (20 ℃, 23 ℃, 25 ℃, 28 ℃, 31 ℃, 34 ℃, 37 ℃ and 40 ℃), different pHs (4, 5, 6, 7, 8 and 9), different carbon sources (soluble starch, maltose, lactose, sucrose and glucose) and different nitrogen sources (potassium nitrate, urea, ammonium sulfate and ammonium chloride). As shown in FIG. 5, the optimum phosphorus dissolution temperature is 28 ℃ and the high phosphorus dissolution effect can be maintained at 28-31 ℃. The optimum pH value is 7, and the phosphorus dissolving amount is 5-9, so that the high phosphorus dissolving effect can be kept. The most suitable carbon source is sucrose. The urea can obviously promote the phosphorus dissolving effect of the MEM02 to reach 1327.78 mg/L.
Example 5 growth promotion experiment of Penicillium oxalicum MEM02
Experimental materials: the test plant is original seed Shanghai Qing (pakchoi). The soil tested was from the flower bed of the integrated building of the university in Hubei.
Penicillium oxalicum MEM02 growth promotion experiment on Chinese cabbage potting: the fertilizer dosage N is 0.15g/kg, K: 0.15g/kg, phosphorus control was fertilized at P:0.1g/kg (N: P: K = 3: 2: 3). The fertilizer source is as follows in sequence: urea, potassium sulphate and calcium phosphate. The application method comprises the following steps: applying the base fertilizer once and mixing evenly. Each pot is filled with 2.5kg of soil. The experiment totaled 5 treatments, each treatment was repeated 3 times. (flowerpot height 15cm, pot mouth diameter 20cm, soil 2.5 kg)
TABLE 1 design of the experiments
Taking a plurality of pakchoi seeds, disinfecting and accelerating germination, and selecting pakchoi seedlings with almost the same growth vigor when the seedlings begin to cross, wherein each pot is used for fixing 3 seedlings. After the seedlings are settled, the roots are irrigated for inoculation and are randomly placed. Growing under natural conditions, and performing daily management such as watering. Periodically collecting the root soil of the pakchoi, and measuring the content of available phosphorus in the soil by using sodium bicarbonate (OLSEN colorimetric method). Harvesting after 30 days, and measuring the fresh weight, the plant height, the stem thickness and the root length of the pakchoi.
The influence of the penicillium oxalicum MEM02 on the available phosphorus content of soil in the potting test is shown in the table 1, and the MEM02 can obviously improve the available phosphorus content of soil. Under the condition of taking the ground phosphate rock as a phosphorus source, the content of available phosphorus in the soil reaches 36.99mg/kg at the maximum at 24 days, and is improved by 561.72 percent compared with a non-inoculated contrast.
TABLE 1 Effect of phosphorus solubilizing bacteria on soil available phosphorus content in potting experiments
The influence of the penicillium oxalicum MEM02 on the pakchoi biomass in the pot culture test is shown in the table 2, and the pakchoi biomass is obviously improved in the pot culture inoculated with the penicillium oxalicum MEM02 bacterial solution compared with the pot culture of a control group. Compared with a non-inoculated control, the growth rates of fresh weight, plant height, stem thickness and root length of a plant of a pot plant inoculated with the penicillium oxalicum MEM02 bacterial liquid are 191.86%, 28.46%, 66.67% and 22.9% respectively under the condition that the phosphate rock powder is used as a phosphorus source.
TABLE 2 influence of phosphorus solubilizing bacteria on the biomass of pakchoi in potting experiments
EXAMPLE 6 Penicillium oxalicum MEM02 antagonism of Fusarium graminearum experiments
The penicillium oxalicum MEM02 is inoculated on the center of a PDA culture medium plate, after 5 days, a puncher with the diameter of 7mm is used for punching bacterial blocks at the edge of a penicillium oxalicum bacterial colony, then the bacterial blocks are inoculated on one side of the center of the PDA plate by 20 mm, Fg bacterial blocks are symmetrically spotted on the other side of the center of the plate by 20 mm after 3 days, and the distance between the penicillium oxalicum MEM02 bacterial blocks and fusarium graminearum bacterial blocks is 40 mm. After 4d, the control plate is full, the width of the antibacterial zone (the distance between the edge of the antagonistic bacterium colony and the edge of the fusarium graminearum colony) is measured, and the non-inoculated penicillium oxalicum mass is used as a control. Calculating the bacteriostasis rate: bacteriostatic rate = (control colony diameter-treated colony diameter)/control colony diameter × 100%. Fig. 6 shows photographs of the penicillium oxalicum MEM02 against a plate of fusarium graminearum, and the penicillium oxalicum MEM02 has a significant effect of inhibiting fusarium graminearum, with a bacterial inhibition rate of 56.45%.
Example 7 Penicillium oxalicum MEM02 spore antagonism to Fusarium graminearum spores experiment
The penicillium oxalicum MEM02 and the sickle graminisKnife bacteria are inoculated on the center of a PDA culture medium plate, penicillium oxalicum MEM02 is inoculated on the center of the PDA culture medium plate, 10 mL of sterile water is used for washing the penicillium oxalicum plate and the fusarium graminearum plate respectively after 7 days, and penicillium oxalicum spores and fusarium graminearum spores with certain concentrations are obtained. Then 1 mL of 2X 107Penicillium oxalicum MEM02 spores in 1 mL of 2X 105Fusarium graminearum Fg spores were mixed and placed on a shaker at 180rpm for 4 h. After 4h, a 7mm piece of filter paper was placed in the center of the PDA plate, and then 10. mu.L of the mixture was spotted onto the piece of filter paper. And (6) recording the photographed image. The results are shown in FIG. 7, 2X 107Penicillium oxalicum MEM02 spores were able to completely inhibit 2X 10 spores5Fusarium graminearum Fg spores (only the Penicillium oxalicum spores germinated and grew).
Example 8 inhibition of Fusarium graminearum spores by Penicillium oxalicum MEM02 spores in straw-containing soil
50 g of soil powder is firstly filled in a 500 mL conical flask, then 5g of chopped straw is added into the upper layer of the soil, and 2 multiplied by 10 is added in equal proportion7CFU/mL Penicillium oxalicum spore preparation and 2X 105Fusarium graminearum spore solution, to which only 2X 10 is added5CFU/mL fusarium graminearum spores were used as a control, and germination of fusarium graminearum spores was observed after 5 days. As shown in FIG. 8, the germination rate of the fusarium graminearum spores in the control group is 67.17%, the straws are rotted, the germination rate of the fusarium graminearum spores in the experimental group is only 18.28%, and the germination of the fusarium graminearum spores can be obviously inhibited by applying MEM02 spores. At present, soil caused by straw returning in rice and wheat crop rotation areas is rich in organic matters, propagation of plant pathogenic fungi is facilitated, spore fungicide is applied, germination of fusarium graminearum spores growing in field soil with rice straws as organic matters is effectively reduced, and the quantity of fusarium graminearum is reduced from the source. The purpose of preventing and treating wheat scab is achieved.
Sequence listing
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Claims (6)
1. A strain of penicillium oxalicum is characterized in that: the penicillium oxalicum is penicillium oxalicum (A)Penicillium oxalicum) MEM02, deposited at the chinese type culture collection on 12/3 of 2019 with the deposit number: CCTCC NO: M20191002.
2. The use of a strain of penicillium oxalicum according to claim 1, wherein: the penicillium oxalicum MEM02 can inhibit germination of fusarium graminearum spores and is used for preventing and treating wheat scab.
3. The use of a strain of penicillium oxalicum according to claim 2, wherein: the penicillium oxalicum MEM02 can efficiently dissolve phosphorus and promote plant growth, and is used for preparing a microbial preparation for promoting crop growth.
4. A process for the production of highly concentrated spore preparations from the solid fermentation of Penicillium oxalicum according to claim 1, wherein: the method comprises the following steps:
(1) inoculating the penicillium oxalicum blocks into a PDA solid culture medium, activating for 3 days, and washing with sterile water to obtain penicillium oxalicum spore suspension;
(2) and (2) uniformly inoculating 5% of the spore suspension activated in the step (1) into a fermentation solid culture medium, fermenting for 5 days at 28 ℃, and uniformly mixing the fermentation solid culture medium once a day during fermentation to obtain a solid fermentation product, namely the high-concentration spore preparation generated by solid fermentation of the penicillium oxalicum MEM 02.
5. The method for producing a highly concentrated spore preparation by solid fermentation of penicillium oxalicum according to claim 4Characterized in that: the fermentation solid culture medium in the step (2) consists of the following components: wheat bran, rice chaff, rice straw powder, K2HPO4、FeSO40.005、MgSO4 .7H2O、CaCO3And deionized water.
6. The method for producing a high-concentration spore preparation by solid fermentation of penicillium oxalicum according to claim 5, wherein: the fermentation solid culture medium comprises the following components in parts by weight: 25 parts of wheat bran, 10 parts of rice chaff, 15 parts of rice straw powder and K2HPO41.0 part of FeSO4 0.005 part of MgSO 24 .7H20.05 part of O, CaCO3 0.05 part and 50 parts of deionized water.
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| CN115536446A (en) * | 2021-11-29 | 2022-12-30 | 云南大学 | Fertilizer for promoting plant growth and application thereof |
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| CN113333459A (en) * | 2021-06-08 | 2021-09-03 | 湘潭大学 | Solid fermentation of penicillium oxalicum and method for efficiently dissolving phosphorus and removing lead by using same |
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| CN114480246A (en) * | 2022-02-14 | 2022-05-13 | 山东省农业科学院作物研究所 | Method for inducing fusarium graminearum to produce spores in large quantity and preparing spore liquid |
| CN114854600A (en) * | 2022-03-21 | 2022-08-05 | 中国科学院南京土壤研究所 | Penicillium oxalicum and application thereof |
| CN114990000A (en) * | 2022-03-29 | 2022-09-02 | 华中农业大学 | Compound microbial agent for degrading DEHP and application of compound microbial agent in preparation of organic fertilizer |
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