CN114854600B - Penicillium oxalate and application thereof - Google Patents
Penicillium oxalate and application thereof Download PDFInfo
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- CN114854600B CN114854600B CN202210281596.1A CN202210281596A CN114854600B CN 114854600 B CN114854600 B CN 114854600B CN 202210281596 A CN202210281596 A CN 202210281596A CN 114854600 B CN114854600 B CN 114854600B
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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
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- 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
Penicillium oxalicum and application thereof, the strain is preserved in a preservation unit appointed by the national intellectual property agency, the preservation date is No. 11/30 of 2021, and the preservation unit name is as follows: china general microbiological culture Collection center, with the preservation number: CGMCC No.23827. The strain of penicillium oxalicum can be stably propagated in soil, so that the enzyme activity related to the turnover of nitrogen and phosphorus nutrients in the soil is respectively improved by 86.77% and 72.92%, the content of abscisic acid in the root system of crops is improved by 21.95%, and the soil nutrient conversion, crop salt tolerance and growth are promoted. The strain of penicillium oxalicum is derived from saline soil farmland soil, is easy to adapt to the environment, and has green environmental protection safety. The strain of penicillium oxalicum has the advantages of 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
Soil phosphorus is an important factor affecting crop yield, and about 74% of farmland in China lacks phosphorus, and the shortage of available phosphorus in a considerable amount of farmland soil becomes a main factor limiting crop high yield, and even in some soils with high available phosphorus content, the application of phosphorus fertilizer is indispensable (Zhang Zhimeng, etc.. The effect of phosphorus application on soil available phosphorus, phosphorus absorption characteristics and wheat yield [ J)]Studies of the effect of phosphorus on potatoes in controlled-release compound fertilizers [ J ] Hebei academy of agricultural technology, 1999, 13 (1): 1115. Hu Yingying, zhang Min, song Fupeng]Plant nutrition and fertilisation journal, 2003, (02): 174-177.). For a long time, the application of phosphate fertilizer is an important measure for improving the grain yield. Research shows that the mobility of phosphorus in soil is poor, so that the utilization rate of the phosphorus fertilizer in the season is only 10% -25% (Gao Shan, yang Jinsong, yao Rongjiang, cao Yifan, zhu Hai, sun Yunpeng, wang Xiangping)Xie Wenping influence of regulatory measures on the phosphorus morphology of coastal saline soil and on the phosphorus uptake by 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 which is not beneficial to the absorption of plants. The phosphorus in the saline soil has a problem of low effectiveness, and phosphorus applied to the soil exists in the form of insoluble phosphate having low effectiveness due to the influence of the salinity of the soil. The phosphorus-dissolving microorganisms in the soil can convert the indissoluble 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. The improvement of 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 phosphate solubilizing function exist in plant rhizosphere soil (SON HJ, PARK GT, CHA M S, et al, solutionization of insoluble inorganic phosphates by a novel salt-and pH tolerant Pantoea agglomerans R-42 isolated from soybean rhizosphere[J ]. Bioresource Technoloogy,2006, 97:204-210.) and can convert ineffective-state phosphorus which is difficult to absorb and utilize in the soil into effective-state phosphorus which can be absorbed and utilized. The phosphorus-dissolving fungi reported at present mainly comprise aspergillus, mortierella, trichoderma, rhizopus and the like.
At present, researches on the aspect of activating and releasing soil phosphorus and promoting salt tolerance and growth of crops on penicillium oxalicum in saline soil are freshly reported.
Disclosure of Invention
The technical problems to be solved are as follows: the invention provides penicillium oxalicum and application thereof. The penicillium oxalicum is obtained from typical saline soil of Shandong's base of China academy of sciences genetic and developmental biology research institute, and has the advantages of high growth speed, low cost and easy culture. The strain has remarkable effect in promoting soil nutrient activation and crop growth. Meanwhile, the strain can promote the direct influence of the abscisic acid secreted by crops on the crop growth stress resistance.
The technical scheme is as follows: penicillium oxalatePenicillium oxalicumHSJ 002) that has been assigned to the national intellectual property agencyCollection of the collection unit, the collection date is No. 2021, 11 and 30, and the name of the collection unit is: china general microbiological culture Collection center, with the preservation number: CGMCC No.23827.
The cultivation method of the penicillium oxalicum comprises the following steps: selecting penicillium oxalicum hyphae, inoculating the penicillium oxalicum hyphae into a PD liquid culture medium, and culturing the penicillium oxalicum hyphae in a constant-temperature shaking table at 27 ℃ and 220 rpm for 5-7 days to obtain mycelium pellets; filtering under aseptic condition, adding 5 g mycelium pellets into 50 g wheat bran-bean cake powder-flour culture medium, and immediately shaking; standing at 27deg.C for culturing; after the mycelium solidifies 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; then placing in a 27 ℃ incubator until the mycelium is completely covered with wheat bran-bean cake powder-flour culture medium, and mashing.
The PD liquid culture medium comprises the following components: peeled potato 200 g, glucose 20 g, water 1000 mL; the bran-bean cake powder-flour culture medium comprises the following components: wheat bran 15 g, bean cake powder 20 g, flour 15 g and water 50 mL, wherein the water is added with the bean cake powder to moisten for 30 min, then the wheat bran and the flour are added to be uniformly mixed, and the mixture is sterilized at 121 ℃ for 20 min and then uniformly shaken to obtain the bean cake.
The penicillium oxalicum plays a role in promoting crop growth.
The application of the penicillium oxalicum in improving the activities of soil phosphatase and N-acetyl-glucosaminidase.
The application of the penicillium oxalicum in synthesizing abscisic acid from plant root systems.
The application of the penicillium oxalicum in increasing the soil phosphorus conversion is provided.
The composition of the inorganic phosphorus culture medium for increasing the phosphorus conversion of the soil is as follows: glucose 10.0. 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
The beneficial effects are that: (1) The strain of penicillium oxalicum can obviously improve the activity of alkaline phosphatase and N-acetyl-glucosaminidase in soil (figure 1:a), improve the content of quick-acting phosphorus (figure 1:c), and have certain phosphorus dissolving capacity (figure 2), which shows that the strain plays an obvious role in nitrogen-phosphorus conversion in soil. (2) The content of abscisic acid in the root system of the crops is improved by 21.95 percent (figure 1:f), which shows that the strain improves the capability of the crops to resist salt stress, and simultaneously, the biomass of the plants is improved by 70.74 percent (figure 1:d), so that the salt content is reduced by 30.17 percent (figure 1:e) to a certain extent, and the salt toxicity of the plants is relieved to a certain extent. (3) The strain is used as rhizosphere microorganism, and is inoculated into soil to improve the biomass carbon of the microorganism in the rhizosphere soil by 60.73% (shown in 1:b), so that the soil nutrient is promoted to be effective to a certain extent, and the microbial community structure in the rhizosphere soil area of crops is changed through root soil communication so as to regulate the growth of the crops (shown in fig. 3).
Drawings
FIG. 1 shows the effect of Penicillium oxalate inoculation on soil alkaline phosphatase activity, N-acetyl-glucosaminidase activity, soil microbial biomass carbon content, quick-acting phosphorus content, salinity content, wheat plant biomass and root system abscisic acid content. Wherein a is the relation between the activity of the oxalic acid penicillium and alkaline phosphatase and the activity of N-acetyl-glucosaminidase, b is the relation between the oxalic acid penicillium and the biomass carbon content of microorganisms, c is the relation between the oxalic acid penicillium and the content of quick-acting phosphorus, d is the relation between the oxalic acid penicillium and the biomass of wheat plants, e is the relation between the oxalic acid penicillium and the content of salt in soil, and f is the relation between the oxalic acid penicillium and the content of abscisic acid in root systems of crops.
FIG. 2 shows the growth of Penicillium oxalicum on inorganic phosphorus medium and its phosphorus dissolution.
FIG. 3 shows the effect of inoculation with Penicillium oxalicum on soil rhizosphere fungal community composition.
Detailed Description
The following examples are given solely for the purpose of illustrating the general principles and features of the present invention and are not meant to limit the scope of the invention in any way, as will be apparent to those skilled in the art. All equivalent changes or modifications made according to the spirit of the present invention should be included in the scope of the present invention. The strain of penicillium oxalicum can be stably propagated in soil, so that the enzyme activity related to the turnover of nitrogen and phosphorus nutrients in the soil is respectively improved by 86.77% and 72.92%, the content of abscisic acid in the root system of crops is improved by 21.95%, and the soil nutrient conversion, crop salt tolerance and growth are promoted. The composition of the inorganic phosphorus culture medium for increasing the soil phosphorus conversion of the penicillium oxalicum is as follows: glucose 10.0 g, (NH 4) 2SO4 0.1 g, mgCl2.6H2O 5.0 g, mgSO4.7H2O 0.25 g, KCl 0.2 g, ca3 (PO 4) 2.0 g. The strain of penicillium oxalicum is derived from saline soil farmland soil, is easy to adapt to the environment, and has green environmental protection safety. The strain of penicillium oxalicum has the advantages of simple culture and preparation process and low cost, and is favorable for popularization.
Example 1
Separating and culturing a strain of penicillium oxalicum from Shandong-base typical salty soil of China academy of sciences of developmental biologyPenicillium oxalicumHSJ 002). The strain shows under a microscope that the sporangium at the top of the upright hypha is broom-shaped, and the spore forms strings which can float everywhere, and can develop into a new individual under proper environmental conditions, and obvious phosphate dissolving transparent rings can be generated on inorganic phosphorus culture medium. The strain is preserved in a preservation unit appointed by the national intellectual property agency, the preservation date is No. 2021, 11 and 30, and the preservation unit name is as follows: china general microbiological culture Collection center, address: beijing, chaoyang area, north Chen Xi Lu 1, 3, china academy of sciences microbiological institute, deposit number: CGMCC No.23827. The nutrient indexes of the surface soil are as follows: pH 8.59, conductivity 4.66 ds/m, organic carbon content of soil 6.53 g/kg, total nitrogen content of soil 0.53 g/kg, total phosphorus content of soil 0.71 g/kg.
Implementation steps
Step 1: good growth of penicillium oxalicumPenicillium oxalicumHSJ 002) strain is inoculated into PD liquid culture medium, and shake culture is carried out for 5-7 days at the temperature of 27 ℃ and the rpm of 220 rpm.
Step 2: liquid culturing Penicillium oxalicumPenicillium oxalicumHSJ 002) was filtered under sterile conditions and the filtered mycelia were rinsed clean with sterile deionized water.
Step 3: 3 g clean mycelium is dissolved in 50 mL sterile deionized water, bacterial liquid is added into 1.5 kg soil, and the mixture is evenly mixed and then is filled into a basin.
Step 4: and (5) planting crops after fully watering.
Comparative example 1
Implementation steps
Step 1: accurately measuring 50 mL sterile deionized water without mycelium, adding 1.5 kg soil, and then filling into a basin.
Step 2: and (5) planting crops after fully watering.
Example 2
Implementation steps
Step 1: good growth of penicillium oxalicumPenicillium oxalicumHSJ 002) is inoculated into PD liquid culture medium, and shake culture is carried out for 5-7 days at the temperature of 27 ℃ and the rpm of 220 rpm until the liquid culture medium is turbid and a large amount of mycelium pellets grow.
Step 2: liquid culturing Penicillium oxalicumPenicillium oxalicumHSJ 002) was filtered under sterile conditions and the filtered mycelia were rinsed clean with sterile deionized water.
Step 3: inoculating clean mycelium into sterilized wheat bran-bean cake powder-flour culture medium, standing at 27deg.C, and culturing in dark until mycelium completely covers the culture medium.
Step 4: and (3) mashing wheat bran, bean cake powder and flour culture medium of hyphae in different triangular flasks, taking out, and uniformly mixing to obtain the microbial inoculum.
Step 5: accurately weighing 3 g microbial inoculum, adding into 1.5 kg soil, fully mixing, and filling into a basin.
Step 6: and (5) planting crops after fully watering.
Comparative example 2
Implementation steps
Step 1: accurately weighing wheat bran-bean flour cake-flour culture medium 3 g which is sterilized and does not add hypha, adding into 1.5 kg soil, fully mixing, and filling into a basin.
Step 2: and (5) planting crops after fully watering.
The soil to be tested is yellow river delta eastern nutrient farmland saline soil, and the soil nutrient indexes are as follows: the salt content of the soil is 3.50 g/kg, the organic carbon of the soil is 11.46 g/kg, the total nitrogen of the soil is 1.15 g/kg, the total phosphorus of the soil is 1.07 g/kg, and the quick-acting phosphorus of the soil is 32.07 mg/kg. The planted crop is wheat (variety Jinan 177), and three plants are reserved in each pot in the three-leaf period. Wheat is cultivated in a greenhouse environment, during which the moisture content is kept at 60%. And (5) watering and weeding are carried out according to the need in the period. Culturing to flowering period, sampling, and measuring soil enzyme activity, wheat biomass, and wheat root hormone content.
As shown in FIG. 1, the test results were inoculated with Penicillium oxalatePenicillium oxalicumHSJ 002) the biomass of the treated wheat increased by 70.74%, the wheat root system abscisic acid content increased by 21.95%, the alkaline phosphatase activity increased by 72.92% and the N-acetyl-glucosaminidase activity increased by 86.77%. Thus, inoculating penicillium oxalicumPenicillium oxalicum HSJ 002) remarkably improves the activity of alkaline phosphatase and N-acetyl-glucosaminidase in saline soil, the content of available phosphorus and plant biomass, promotes the activation of nitrogen and phosphorus nutrients in saline soil, improves the absorption and utilization of nitrogen and phosphorus nutrients by crops, and further promotes the growth of crops. The strain of penicillium oxalicumPenicillium oxalicumHSJ 002) stimulates the secretory production of abscisic acid in crops, thereby improving salt tolerance and stress resistance of crops to a certain extent.
Claims (7)
1. Penicillium oxalatePenicillium oxalicum) HSJ002, the strain has been deposited in the accession number specified by the national intellectual property agency, accession number 2021, 11, 30, accession number: china general microbiological culture Collection center, with the preservation number: CGMCC No.23827.
2. The method for culturing penicillium oxalicum HSJ002 according to claim 1, wherein the steps are as follows: selecting penicillium oxalicum hyphae, inoculating the penicillium oxalicum hyphae into a PD liquid culture medium, and culturing the penicillium oxalicum hyphae in a constant-temperature shaking table at 27 ℃ and 220 rpm for 5-7 days to obtain mycelium pellets; filtering under aseptic condition, adding 5 g mycelium pellets into 50 g wheat bran-bean cake powder-flour culture medium, and immediately shaking; standing at 27deg.C for culturing; after the mycelium solidifies 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; then placing in a 27 ℃ incubator until the mycelium is completely covered with wheat bran-bean cake powder-flour culture medium, and mashing.
3. The method for culturing penicillium oxalicum HSJ002 according to claim 2, wherein the PD liquid medium composition is: peeled potato 200 g, glucose 20 g, water 1000 mL; the bran-bean cake powder-flour culture medium comprises the following components: wheat bran 15 g, bean cake powder 20 g, flour 15 g and water 50 mL, wherein the water is added with the bean cake powder to moisten for 30 min, then the wheat bran and the flour are added to be uniformly mixed, and the mixture is sterilized at 121 ℃ for 20 min and then uniformly shaken to obtain the bean cake.
4. The use of penicillium oxalicum HSJ002 according to claim 1 to promote wheat growth.
5. Use of penicillium oxalicum HSJ002 according to claim 1 for increasing soil phosphatase and N-acetyl-glucosaminidase activity.
6. The use of penicillium oxalicum HSJ002 according to claim 1 in wheat root synthesis of abscisic acid.
7. Use of penicillium oxalicum HSJ002 according to claim 1 for increasing soil phosphorus conversion.
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| US7176027B2 (en) * | 2001-12-20 | 2007-02-13 | Pioneer Hi-Bred International, Inc. | Genes and regulatory DNA sequences associated with stress-related gene expression in plants and methods of using the same |
| CN105296363B (en) * | 2015-10-28 | 2019-06-04 | 南京农业大学 | A kind of penicillium oxalicum NJDL-03 bacterial strain and its application |
| CN110129208B (en) * | 2019-04-28 | 2021-05-04 | 湘潭大学 | Penicillium oxalicum with broad-spectrum acid production characteristic |
| CN112159761B (en) * | 2020-04-28 | 2021-10-22 | 湖北大学 | Preparation method of penicillium oxalicum and application of penicillium oxalicum in phosphate solubilizing, growth promoting and fusarium graminearum antagonism |
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