CN114854600A - Penicillium oxalicum and application thereof - Google Patents
Penicillium oxalicum and application thereof Download PDFInfo
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- CN114854600A CN114854600A CN202210281596.1A CN202210281596A CN114854600A CN 114854600 A CN114854600 A CN 114854600A CN 202210281596 A CN202210281596 A CN 202210281596A CN 114854600 A CN114854600 A CN 114854600A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
- A01N63/36—Penicillium
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
Abstract
The strain is preserved in a preservation unit appointed by the national intellectual property office, the preservation date is No. 11/30 in 2021, the name of the preservation unit is as follows: the China general microbiological culture Collection center has the following preservation numbers: CGMCC No. 23827. The penicillium oxalicum can be stably propagated in soil, so that the enzyme activities related to turnover of nitrogen and phosphorus nutrients of the soil are respectively improved by 86.77% and 72.92%, the content of abscisic acid of crop roots is improved by 21.95%, and the soil nutrient conversion, salt tolerance and growth of crops are promoted. The penicillium oxalicum is derived from saline soil farmland soil, is easy to adapt to the environment, and has green, environment-friendly and safe properties. The penicillium oxalicum preparation has simple culture and preparation process and low cost, and is favorable for popularization.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to functional penicillium oxalicum screened from saline soil and application thereof.
Background
The soil phosphorus element is an important factor influencing crop yield, according to statistics, about 74 percent of farmland soil in China is lack of phosphorus, the insufficient utilization of available phosphorus in considerable farmland soil becomes a main factor limiting the high yield of crops, and the application of phosphate fertilizer is indispensable (Zhang Chi Chong, etc. the influence of phosphorus application on the available phosphorus, phosphorus absorption property and wheat yield of soil [ J]Proceedings of the institute of agricultural and technical sciences, Hebei, 1999, 13 (1): 1115, lip-ying, Zhang Min, Song Fu Peng, Effect study of phosphorus in controlled release compound fertilizer on potato [ J]Plant Nutrition and Fertilizer bulletin, 2003, (02): 174-. The application of phosphate fertilizers has long been an important measure for improving the yield of food. Research shows that the mobility of phosphorus in soil is poor, so that the utilization rate of the current-season phosphate fertilizer is only 10% -25% (Gaoshan, Populon, Yaoangjiang, Caoaofan, Zhuhai, Sunjiapeng, Wangzhenping, Shexin. the regulation and control measures influence the phosphorus form of the coastal saline soil and the phosphorus absorption of crops [ J]Soil 2020,52(04): 691-698), most of the phosphorus is associated with Ca in the soil 2+ 、Fe 3+ 、Al 3+ 、Fe 2+ The plasma is combined and converted into insoluble phosphorus, so that the insoluble phosphorus is not beneficial to the absorption of plants. Phosphorus in saline soil has a problem of low availability, and phosphorus applied to soil exists in the form of poorly soluble phosphate with low availability due to the influence of soil salinity. And the phosphorus-dissolving microorganisms in the soil can convert insoluble phosphorus in the soil into soluble phosphorus which can be absorbed and utilized by plants, so that the utilization rate of the phosphate fertilizer is improved, the phosphorus absorption amount of crops is increased, and the improvement of the crop yield is facilitated. 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 with the function of phosphate solubilizing exist in the rhizosphere soil of plants (SON HJ, PARK GT, CHA M S, et al. solution of insoluble inorganic phosphates by a novel salt and pH tolerant Pantoea agglomerans R-42 isolatoted from soybean rhizosphere and rhizosphere [ J ]. Bioresource Technolook, 2006,97: 204. sup. 210.), and can convert ineffective phosphorus which is difficult to absorb and utilize in the soil into available phosphorus which can be absorbed and utilized. The phosphorus-solubilizing fungi reported at present mainly comprise aspergillus, mortierella, trichoderma, rhizopus and the like.
At present, researches on the aspects of soil phosphorus activation and release and crop salt tolerance growth promotion of penicillium oxalicum in saline soil are rarely reported.
Disclosure of Invention
The technical problem to be solved is as follows: the invention provides penicillium oxalicum and application thereof. The penicillium oxalicum is from typical saline soil of Shandong base of institute of genetics and developmental biology of Chinese academy of sciences, and has the advantages of high growth speed, low cost and easy culture. The strain has obvious effect on promoting soil nutrient activation and crop growth. Meanwhile, the strain can promote crops to secrete abscisic acid to directly influence the growth stress resistance of the crops.
The technical scheme is as follows: penicillium oxalicum (B)Penicillium oxalicumHSJ 002), which has been deposited in a depository designated by the national intellectual property office, with a date of deposit No. 11/30/2021, and the name of the depository: the China general microbiological culture Collection center has the following preservation numbers: CGMCC number 23827.
The method for culturing the penicillium oxalicum comprises the following steps: selecting penicillium oxalicum mycelium, inoculating the penicillium oxalicum mycelium into a PD liquid culture medium, and placing the penicillium oxalicum mycelium in a constant-temperature shaking table at 27 ℃ and 220 rpm for culturing for 5-7 days to form mycelium pellets; filtering under aseptic condition, adding 5 g of mycelium pellet into 50 g of wheat bran-bean cake powder-flour culture medium, and shaking up immediately; standing at 27 deg.C for culture; when the hyphae solidify the wheat bran-bean cake powder-flour culture medium into blocks, the two sides of the fungus cake are contacted with air for continuous culture; placing in 27 deg.C incubator until the mycelium completely covers the culture medium of testa Tritici-bean cake powder-flour, and mashing.
The PD liquid culture medium comprises the following components: peeled potato 200 g, glucose 20 g and water 1000 mL; the bran-bean cake powder-flour culture medium comprises the following components: 15 g of wheat bran, 20 g of bean cake powder, 15 g of flour and 50 mL of water, wherein the bean cake powder is added into the water firstly for moistening for 30 min, then the wheat bran and the flour are added for stirring, and the mixture is sterilized at 121 ℃ for 20 min and then shaken uniformly to prepare the wheat bran-soybean cake flour.
The penicillium oxalicum has the function of promoting the growth of crops.
The use of penicillium oxalicum for increasing the activity of soil phosphatases and N-acetyl-glucosaminidase.
The application of the penicillium oxalicum in the synthesis of abscisic acid by a plant root system is disclosed.
The application of the penicillium oxalicum in increasing the phosphorus conversion of soil is disclosed.
The inorganic phosphorus culture medium for increasing the phosphorus conversion of soil comprises the following components: glucose 10.0 g, (NH) 4 ) 2 SO 4 0.1 g、MgCl 2 ·6H 2 O 5.0 g、MgSO 4 ·7H 2 O 0.25 g、KCl 0.2 g、Ca 3 (PO 4 ) 2 5.0 g
Has the advantages that: (1) the penicillium oxalicum can obviously improve the alkaline phosphatase activity and the N-acetyl-glucosaminidase activity of soil (figure 1: a), improve the content of available phosphorus (figure 1: c), and has certain phosphorus dissolving capacity (figure 2), which shows that the strain plays a significant role in the nitrogen-phosphorus conversion of soil. (2) The content of abscisic acid in the root system of the crop is increased by 21.95 percent (figure 1: f), which shows that the strain improves the salt stress tolerance of the crop, simultaneously the biomass of the plant is also increased by 70.74 percent (figure 1: d), the salt content is also reduced by 30.17 percent (figure 1: e) to a certain extent, and the salt toxicity of the plant is relieved to a certain extent. (3) The strain is used as rhizosphere microorganism, is inoculated into soil to improve the biomass carbon of the rhizosphere soil microorganism by 60.73 percent (figure 1: b), promotes the soil nutrient validation to a certain extent, and changes the microbial community structure of the rhizosphere soil area of the crop through the root-soil communication so as to regulate the crop growth (figure 3).
Drawings
FIG. 1 shows the effect of penicillium oxalicum inoculation on soil alkaline phosphatase activity, N-acetylglucosaminidase activity, soil microbial biomass carbon content, available phosphorus content, salt content, wheat plant biomass, and root abscisic acid content. Wherein a is the relationship between the activity of the penicillium oxalicum and alkaline phosphatase and the activity of the N-acetyl-glucosaminidase, b is the relationship between the penicillium oxalicum and the biomass carbon content of the microorganism, c is the relationship between the penicillium oxalicum and the content of the available phosphorus, d is the relationship between the penicillium oxalicum and the biomass of the wheat plant, e is the relationship between the penicillium oxalicum and the content of soil salt, and f is the relationship between the penicillium oxalicum and the content of abscisic acid in the root system of the crop.
FIG. 2 shows the growth of Penicillium oxalicum in inorganic phosphorus medium and its phosphorus dissolution.
FIG. 3 shows the effect of the inoculation of Penicillium oxalicum on the composition of the rhizosphere fungal community of the soil.
Detailed Description
The following examples are only for illustrating the technical concept and features of the present invention, and are intended to enable those skilled in the art to understand the contents of the present invention and implement the present invention, and not to limit the protection scope of the present invention. All equivalent changes and modifications made according to the spirit of the present invention should be covered within the protection scope of the present invention. The penicillium oxalicum can be stably propagated in soil, so that the enzyme activities related to turnover of nitrogen and phosphorus nutrients of the soil are respectively improved by 86.77% and 72.92%, the content of abscisic acid of crop roots is improved by 21.95%, and the soil nutrient conversion, salt tolerance and growth of crops are promoted. The composition of the penicillium oxalicum in an inorganic phosphorus culture medium for increasing soil phosphorus conversion is as follows: 10.0 g of glucose, (NH4)2SO 40.1 g, MgCl 2.6H2O 5.0.0 g, MgSO 4.7H2O 0.25.25 g, KCl 0.2 g and Ca3(PO4) 25.0 g. The penicillium oxalicum is derived from saline soil farmland soil, is easy to adapt to the environment, and has green, environment-friendly and safe properties. The penicillium oxalicum preparation has simple culture and preparation process and low cost, and is favorable for popularization.
Example 1
A strain of penicillium oxalicum is separated and cultured from typical saline soil of Shandong base of institute of genetics and developmental biology, Chinese academy of sciencesPenicillium oxalicumHSJ 002). The strain shows that the tip of the vertical hypha is broom-shaped, the strung spores are grown on the brooms, the spores can float everywhere, and can develop into a new individual under the appropriate environmental condition, and the strain is inorganic phosphorusObvious phosphate-dissolving transparent rings are generated on the culture medium. The strain is preserved in a preservation unit appointed by the national intellectual property office, the preservation date is No. 11/30 in 2021, the name of the preservation unit is as follows: china general microbiological culture Collection center, address: west road No. 1, north west of the morning area, beijing, 3, institute for microbiology, china academy of sciences, accession number: CGMCC number 23827. The surface soil nutrient indexes are as follows: pH 8.59, conductivity 4.66 ds/m, soil organic carbon content 6.53 g/kg, soil total nitrogen content 0.53 g/kg, soil total phosphorus content 0.71 g/kg.
Carrying out the step
Step 1: a well-grown penicillium oxalicum (A)Penicillium oxalicumHSJ 002) strain is inoculated into PD liquid culture medium and is subjected to shake cultivation for 5-7 days at 27 ℃ and 220 rpm.
Step 2: subjecting liquid culture of Penicillium oxalicum (A)Penicillium oxalicumHSJ 002) was aseptically filtered, and the mycelia after filtration were rinsed with sterile deionized water.
And step 3: 3 g of clean mycelium is dissolved in 50 mL of sterile deionized water, and the bacterial liquid is added into 1.5 kg of soil, uniformly mixed and potted.
And 4, step 4: and (5) planting crops after fully watering.
Comparative example 1
Carrying out the step
Step 1: 50 mL of sterile deionized water containing no mycelia was accurately measured, and 1.5 kg of soil was added to the water and potted.
Step 2: and (5) planting crops after fully watering.
Example 2
Carrying out the step
Step 1: a well-grown penicillium oxalicum (A)Penicillium oxalicumHSJ 002) strains are inoculated into PD liquid culture medium and cultured for 5-7 days at 27 ℃ and 220 rpm in a shaking way until the liquid culture medium is turbid and full of a large number of mycelium pellets.
Step 2: subjecting liquid culture of Penicillium oxalicum (A)Penicillium oxalicumHSJ 002) was aseptically filtered, and the mycelia after filtration were rinsed with sterile deionized water.
And step 3: inoculating the clean mycelium into sterilized wheat bran-bean cake powder-flour culture medium, standing at 27 deg.C in dark for culturing until the mycelium completely covers the culture medium.
And 4, step 4: smashing wheat bran-bean cake powder-flour culture medium of hyphae in different triangular flasks, taking out, and uniformly mixing to obtain the microbial inoculum.
And 5: 3 g of microbial inoculum is accurately weighed, added into 1.5 kg of soil, fully and uniformly mixed and potted.
Step 6: and (5) planting crops after fully watering.
Comparative example 2
Carrying out the step
Step 1: 3 g of wheat bran-soybean cake-flour culture medium which is sterilized and is not added with hypha is accurately weighed, added into 1.5 kg of soil, fully and uniformly mixed and potted.
Step 2: and (5) planting crops after fully watering.
The soil to be tested is saline soil of Dongyang farmland in yellow river delta, and the soil nutrient indexes are as follows: the salt content of the soil is 3.50 g/kg, the organic carbon content of the soil is 11.46 g/kg, the total nitrogen content of the soil is 1.15 g/kg, the total phosphorus content of the soil is 1.07 g/kg, and the quick-acting phosphorus content of the soil is 32.07 mg/kg. The crop is wheat (variety: Jinan 177), and seedlings are set in the three-leaf period, and three plants are remained in each pot. The wheat was cultivated in a greenhouse environment, during which the water content was maintained at 60%. During the period, the water is watered and the weed is removed as required. Culturing to the flowering period, sampling, and measuring the soil enzyme activity, the wheat biomass, the wheat root system hormone content and the like.
The results of the measurement are shown in FIG. 1, and Penicillium oxalicum (A), (B)Penicillium oxalicumHSJ 002) the biomass of the treated wheat is increased by 70.74 percent, the content of abscisic acid in the root system of the wheat is increased by 21.95 percent, the activity of alkaline phosphatase is increased by 72.92 percent, and the activity of N-acetyl-glucosaminidase is increased by 86.77 percent. Thus, inoculating with Penicillium oxalicum (B) ((B))Penicillium oxalicum HSJ 002) can obviously improve the activities of alkaline phosphatase and N-acetyl-glucosaminidase in saline soil, the content of available phosphorus and plant biomass, promote the activation of nitrogen and phosphorus nutrients in saline soil, improve the absorption and utilization of nitrogen and phosphorus nutrients by crops, and further promote the growth of the crops. The strain of Penicillium oxalicum (Penicillium oxalicum HSJ002）Stimulates the secretion and generation of abscisic acid of crops, thereby improving the salt resistance and stress resistance of the crops to a certain extent.
Claims (8)
1. Penicillium oxalicum (B)Penicillium oxalicumHSJ 002), which has been deposited in a depository designated by the national intellectual property office, with a date of deposit No. 11/30/2021, and the name of the depository: the China general microbiological culture Collection center has the following preservation numbers: CGMCC number 23827.
2. The method for culturing penicillium oxalicum of claim 1, comprising the steps of: selecting penicillium oxalicum mycelium, inoculating the penicillium oxalicum mycelium into a PD liquid culture medium, and placing the penicillium oxalicum mycelium in a constant-temperature shaking table at 27 ℃ and 220 rpm for culturing for 5-7 days to form mycelium pellets; filtering under aseptic condition, adding 5 g of mycelium pellet into 50 g of wheat bran-bean cake powder-flour culture medium, and shaking up immediately; standing at 27 deg.C for culture; when the hyphae solidify the wheat bran-bean cake powder-flour culture medium into blocks, the two sides of the fungus cake are contacted with air for continuous culture; placing in 27 deg.C incubator until the mycelium completely covers the culture medium of testa Tritici-bean cake powder-flour, and mashing.
3. The method for culturing penicillium oxalicum as claimed in claim 2, wherein said PD liquid medium consists of: peeled potato 200 g, glucose 20 g and water 1000 mL; the bran-bean cake powder-flour culture medium comprises the following components: 15 g of wheat bran, 20 g of bean cake powder, 15 g of flour and 50 mL of water, wherein the bean cake powder is added into the water firstly for moistening for 30 min, then the wheat bran and the flour are added for stirring, and the mixture is sterilized at 121 ℃ for 20 min and then shaken uniformly to prepare the wheat bran-soybean cake flour.
4. The use of penicillium oxalicum of claim 1 for promoting crop growth.
5. Use of penicillium oxalicum as claimed in claim 1 for increasing the activity of soil phosphatases and N-acetyl-glucosaminidase.
6. Use of penicillium oxalicum according to claim 1 for the synthesis of abscisic acid from plant roots.
7. Use of the penicillium oxalicum of claim 1 for increasing phosphorus conversion in soil.
8. The use according to claim 7, wherein the composition of the inorganic phosphorus medium for increasing the phosphorus conversion in soil is: glucose 10.0 g, (NH) 4 ) 2 SO 4 0.1 g、MgCl 2 ·6H 2 O 5.0 g、MgSO 4 ·7H 2 O 0.25 g、KCl 0.2 g、Ca 3 (PO 4 ) 2 5.0 g。
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