A strain of Pteris grisea F27, a fungal agent and its application in agriculture

Abstract

The present invention belongs to the technical field of microbial agents, and specifically relates to a strain of Emericellopsispallida F27, a microbial agent and its application in the field of agriculture. The present invention provides a strain of Emericellopsispallida F27, with a deposit number of CGMCC No. 40123, and a microbial agent is prepared using Emericellopsispallida F27. The application results show that the strain of Emericellopsispallida F27 can increase the degradation rate of corn stalks by 30.55%, and the reducing sugar concentration in the culture solution reaches 25.3 mg / L. Emericellopsispallida F27 can significantly increase the proportion of large aggregates in the soil and significantly inhibit the growth of various sickle fungi. The application of the Emericellopsispallida F27 microbial agent can promote the growth of corn and wheat, improve the stress resistance of corn, and increase soil microbial biomass carbon, microbial nitrogen, soil soluble nitrogen, soil oxygen content and straw degradation rate.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial agents, and specifically relates to a gray-white winged spore shell F27, a microbial agent and applications thereof in the agricultural field. Background Art

[0002] Emericellopsis pallida was first isolated in Xinjiang, China in 2012 (Su Jun, Ma Xiangru, et al., Emericellopsis pallida, a new record species in China, Journal of Xinjiang University (Natural Science Edition), 20120815), and its ability to produce exopolysaccharides with straw extract as raw material in the presence of sucrose and ammonium chloride was further verified (Su Jun, Feng Xinzhong, et al., Optimization of exopolysaccharide production by fermentation of Emericellopsis pallida by response surface methodology, Food Industry Science and Technology, 20080825). Currently, there are very few studies on Emericellopsis pallida, and there are no reports on the application of Emericellopsis pallida in the agricultural field. Summary of the invention

[0003] The purpose of the present invention is to provide a strain of Emericellopsis pallida F27. The Emericellopsis pallida F27 and the microbial agent prepared by the Emericellopsis pallida F27 provided by the present invention can improve the growth of wheat or corn, increase the straw degradation rate in the soil environment, promote the formation of soil aggregates, increase the organic carbon and soluble carbon and nitrogen content in the soil, increase the soil microbial biomass, and resist pathogenic bacteria to promote the improvement of soil fertility.

[0004] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0005] The invention provides a strain of Emericellopsis pallida F27, whose preservation number is CGMCC No.40123.

[0006] The present invention provides a bacterial agent containing the gray-white wing spore shell described in the above technical solution, wherein the number of active hyphae of gray-white wing spore shell F27 in the bacterial agent is ≥10 6 cfu / g.

[0007] The present invention provides a method for preparing the bacterial agent described in the above technical solution, comprising the following steps: a first fermentation culture of Pterospora grisea F27 is performed to obtain a first fermentation mixture, and the first fermentation mixture is subjected to a second fermentation culture to obtain a bacterial agent; the first fermentation culture and the second fermentation culture are both performed under dark conditions; the first fermentation culture time is 7 to 10 days, and the second fermentation culture time is 3 to 6 days.

[0008] Preferably, the culture medium used in the second fermentation culture comprises the following components in parts by weight: 65-75 parts of wheat bran, 20-30 parts of soybean cake powder, 2-4 parts of sucrose, 0.1-0.2 parts of poly-gamma-glutamic acid and 90-110 parts of water.

[0009] Preferably, the molecular weight of the poly-gamma-glutamic acid is 600,000 to 800,000.

[0010] Preferably, the medium used in the first fermentation culture is PD liquid medium, the temperature of the first fermentation culture is 25-30° C., and the rotation speed of the first fermentation culture is 120-180 rpm.

[0011] Preferably, the temperature of the second fermentation culture is 25-30° C., and the volume ratio of the first fermentation mixture to the second fermentation culture medium is (15-25):100.

[0012] The present invention provides the use of the gray white wing spore shell F27 described in the above technical solution or the bacterial agent described in the above technical solution or the bacterial agent prepared by the preparation method described in the above technical solution in one or more of the following (1) to (7);

[0013] (1) Improve the degradation rate of straw in the soil environment;

[0014] (2) Promote the formation of soil aggregates;

[0015] (3) Increase the soluble carbon and nitrogen content in the soil;

[0016] (4) Increase soil microbial biomass;

[0017] (5) Inhibit the growth of Fusarium;

[0018] (6) Promote plant growth;

[0019] (7) Improve plant resistance to stress.

[0020] Preferably, the plant comprises wheat or corn.

[0021] Preferably, the Hoary White Pterospora F27 promotes plant growth by secreting gibberellins and / or zeatin riboside; the Hoary White Pterospora F27 promotes plant stress resistance by secreting abscisic acid.

[0022] Beneficial effects of the present invention: The present invention provides a strain of Emericellopsis pallida F27, with a preservation number of CGMCC No. 40123. The Emericellopsis pallida of the present invention has ecological environmental safety. The strain of Emericellopsis pallida is derived from farmland soil where straw has been returned to the field for a long time, and can achieve green ecological agriculture to ensure the quality and safety of food. The Emericellopsis pallida of the present invention has long-lasting effect, can grow and reproduce on its own in the soil, and reach an ecologically stable balance with the indigenous microbial community without repeated addition. The Emericellopsis pallida of the present invention is operable, and the expansion culture process of the strain of Emericellopsis pallida is simple to operate and low in cost. It can be prepared into a microbial agent and promoted and applied in the agricultural field. Emericellopsis pallida F27 mainly degrades straw through endo-β-1,4-glucanase and β-sucrasidase, and the enzyme activities are 130 mg reducing sugar / h / g mycelium and 370 mg reducing sugar / h / g mycelium, respectively. The application results showed that the degradation rate of corn straw by the strain of Pteris glaucosa F27 was 30.55% within 7 days, and the reducing sugar concentration in the culture solution reached 25.3 mg / L. The results of the soil culture experiment showed that Pteris glaucosa could significantly increase the composition ratio of large aggregates in the soil. The results of the plate confrontation experiment showed that Pteris glaucosa could significantly inhibit the growth of various sickle fungi. The biomass of corn inoculated with Pteris glaucosa increased by 13%, and the SPDA value of corn chlorophyll index increased by 14%. The soil microbial biomass carbon treated with Pteris glaucosa increased by 15%, the microbial nitrogen content increased by 75%, the soil soluble nitrogen increased by 23%, and the straw degradation rate increased by 10%.

[0023] Biological Deposit Description

[0024] Emericellopsis pallida F27 was deposited in the China General Microbiological Culture Collection Center (CGMCC) on March 31, 2022, with the deposit number CGMCC No.40123. The address of the deposit unit is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.

[0026] Figure 1 The colony morphology of the gray-white winged spore shell F27 on the PDA plate in Example 1;

[0027] Figure 2 This is the plate confrontation experiment of gray white wing spore shell F27 and Fusarium in Example 2, Figure 2 The middle is a clear antagonistic line. Figure 2Y1 refers to Fusarium fujikura; Y7 refers to Fusarium laminarum; F33 refers to Fusarium oxysporum;

[0028] Figure 3 The colony morphology of the gray-white winged spore shell F27 cultured in soil in Example 3;

[0029] Figure 4 This is a comparison chart of wheat seedling growth in Example 4 and Comparative Example 3. Figure 4 The three wheat plants in the comparative example are comparative example 3, and the three wheat plants in the example are example 4;

[0030] Figure 5 This is the phylogenetic tree of Emericellopsis pallida F27. DETAILED DESCRIPTION

[0031] The present invention provides a strain of Emericellopsis pallida F27, with a deposit number of CGMCC No. 40123. The ITS1 gene sequence of the Emericellopsis pallida F27 of the present invention is shown in SEQ ID NO.1.

[0032] SEQ ID NO.1 sequence:.

[0033] The gray-white winged spore shell of the present invention is preferably isolated and cultured from the long-term straw return to field treatment plough layer soil in the national fluvo-aquic soil fertility and fertilizer efficiency test in Yuanyang County, Henan Province. The strain is isolated using Bengal red medium (peptone 5.0g, agar 15g, sucrose 10.0g, MgSO4·7H2O0.5g, KH2PO41.0g, Bengal red 0.03g and chloramphenicol 0.10g, pH7.0), and the bacterial solution is diluted by 10 3, and further purified on potato sucrose solid medium. The purified fungi were lysed and their DNA was extracted using Lysis Buffer for Microorganism to DirectPCR reagent produced by Takara. The ITS1 region of the extracted fungal DNA was amplified, and PCR amplification was performed using ITS1F / ITS4 primers. The amplification system and procedure are shown in Table 1-1. The amplified products were verified by 2% agarose gel electrophoresis and sequenced using Sanger sequencing technology. The specific identification method is as follows:

[0034] 1) Take 5 μL of Lysis Buffer for Microorganism to Direct PCR into a sterile centrifuge tube.

[0035] 2) Pick up a single colony with a sterilized toothpick or pipette tip, place it in a centrifuge tube, stir it a few times and then take it out.

[0036] 3) After heat denaturation at 80°C for 15 minutes, take out the tube and shake it quickly to ensure sufficient heat denaturation, and centrifuge it at 5000 rpm.

[0037] 4) Take the supernatant after lysis as a template for PCR reaction. The primers used are ITS1 / ITS4 primers for amplifying the ITS1 and ITS2 regions of fungi. The PCR amplification system and amplification program are shown in Table 1-1.

[0038] Table 1-1 PCR amplification system and amplification procedure

[0039]

[0040] 5) After amplification, the samples were tested by 1.2% agarose gel electrophoresis, and qualified samples were subjected to conventional sequencing.

[0041] 6) After the primer sequences were manually removed from the sequenced sequences, BLAST sequence alignment was performed using the National Center for Biotechnology Information database and a phylogenetic tree was constructed using the maximum likelihood approach.

[0042] The method for constructing the phylogenetic tree is:

[0043] 1. The ITS1 sequence of P. glaucum F27 obtained by sequencing was put into NCBIblast for comparison. The website is https: / / blast.ncbi.nlm.nih.gov / Blast.cgi?PROGRAM=blastn&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome;

[0044] 2. The data used was Internal transcribed spacer region (ITS) from Fungi type and reference. The alignment optimization condition was Highly similar sequences (megablast). The other conditions were kept at default conditions.

[0045] 3. Input the sequences with the highest similarity and use Mega7 to construct the phylogenetic tree. The method uses MaximumLikelihood and the model uses Tamura-Nei model. Figure 5 The obtained fungal species was determined to be Emericellopsis pallida and named Emericellopsis pallida F27.

[0046] The gray-white wing spore shell of the invention has ecological environmental safety. The gray-white wing spore shell is derived from farmland soil where straw is returned to the field for a long time, and can realize green ecological agriculture to ensure the high quality and safety of food.

[0047] The gray-white wing spore shell of the present invention has long-term effect, can grow and reproduce in the soil by itself, and reaches an ecologically stable balance with the indigenous microbial community without repeated addition. The gray-white wing spore shell of the present invention is operable, and the expansion culture process of the gray-white wing spore shell of the strain is simple to operate, low in cost, and can be prepared into a microbial agent for application in the agricultural field. The gray-white wing spore shell of the present invention promotes the improvement of soil fertility by promoting the effective degradation of straw, promoting the formation of soil aggregates, and resisting pathogens, which can promote the growth of corn or wheat and improve the stress resistance of corn.

[0048] The present invention provides a bacterial agent containing the gray-white wing spore shell described in the above technical solution, wherein the number of active hyphae of gray-white wing spore shell F27 in the bacterial agent is ≥10 6 cfu / g.

[0049] The present invention provides a method for preparing the bacterial agent described in the above technical solution, comprising the following steps: a first fermentation culture of Pterospora grisea F27 is performed to obtain a first fermentation mixture, and the first fermentation mixture is subjected to a second fermentation culture to obtain a bacterial agent; the first fermentation culture and the second fermentation culture are performed under dark conditions.

[0050] In the present invention, the Pteris grisea F27 is preferably stored in a PDA medium before being inoculated into a PD liquid medium.

[0051] The Pteris grisea F27 of the present invention is preferably inoculated into a PD liquid culture medium for a first fermentation culture to obtain a first fermentation mixture. The present invention has no particular limitation on the inoculation amount and inoculation method, and conventional inoculation methods and inoculation amounts can be used.

[0052] In the present invention, the PD liquid culture medium of the present invention preferably comprises 180-220 parts of peeled potatoes, 18-22 parts of sucrose and 900-1100 parts of water, more preferably 200 parts of peeled potatoes, 20 parts of sucrose and 1000 parts of water, and the pH is natural. The present invention has no particular limitation on the sterilization method of the PD liquid culture medium, and a conventional method can be used.

[0053] In the present invention, the time of the first fermentation culture is preferably 7 to 10 days, more preferably 7.5 to 9 days, and more preferably 8 days. The first fermentation temperature is preferably 25 to 30°C, more preferably 26 to 29°C, and more preferably 28°C. The first fermentation speed is preferably 120 to 180 rpm, more preferably 130 to 170 rpm, and more preferably 150 rpm. In the present invention, the time of the first fermentation is preferably based on the appearance of white soybean-sized mycelial balls during the culture of Hoary White Winged Sporangia F27. The purpose of the first fermentation culture of the present invention is to obtain a sufficient amount of active bacteria or spores of Hoary White Winged Sporangia F27.

[0054] After obtaining the first fermentation mixture, the present invention performs a second fermentation culture on the first fermentation mixture to obtain a bacterial agent. The present invention preferably inoculates the first fermentation mixture into a second fermentation medium for the second fermentation culture.

[0055] In the present invention, the second fermentation medium preferably comprises the following components in parts by weight: 65-75 parts of wheat bran, 20-30 parts of soybean cake powder, 2-4 parts of sucrose, 0.1-0.2 parts of poly-gamma-glutamic acid and 90-110 parts of water, with a natural pH.

[0056] The molecular weight of the poly-gamma-glutamic acid of the present invention is preferably 600,000 to 800,000, more preferably 700,000. The wheat bran of the present invention is preferably dry wheat bran that is not moldy. The second fermentation medium of the present invention is preferably prepared by the following method: the wheat bran and bean cake powder are mixed in a dry state, sucrose and poly-gamma-glutamic acid are dissolved in water, and the mixture is mixed with the wheat bran and bean cake powder for 30 minutes to obtain a mixture, and sterilized at 121°C for 20 minutes to obtain the second fermentation medium. The role of the wheat bran of the present invention is to provide a slow-acting carbon source for the growth of white-winged spore shell F27, the role of the bean cake powder is to provide a nitrogen source for the growth of white-winged spore shell F27, the role of the sucrose is to provide a fast-acting carbon source for the growth of white-winged spore shell F27, the role of the poly-gamma-glutamic acid is to regulate the growth of gray-white winged spore shell F27, and the role of the second fermentation medium is to make the gray-white winged spore shell F27 hyphae colonize on the solid matrix, which is convenient for application and activity maintenance. The second fermentation culture of the present invention is carried out until the gray-white winged spore shell F27 hyphae completely covers the fermentation medium.

[0057] In the present invention, the inoculation amount of the first fermentation mixture inoculated into the second fermentation medium is preferably 15-25 mL of the first fermentation culture per 100 mL of the second fermentation medium, more preferably 18-23 mL of the first fermentation culture per 100 mL of the second fermentation medium, and more preferably 20 mL of the first fermentation culture per 100 mL of the second fermentation medium.

[0058] In the present invention, the time of the second fermentation culture is preferably 3 to 6 days, more preferably 4 to 6 days, and more preferably 4 to 5 days. The second fermentation culture temperature is preferably 25 to 30°C, more preferably 27 to 29°C, and more preferably 28°C. The second fermentation culture method of the present invention is preferably static culture. The present invention does not specifically limit the container for the second fermentation culture, and a conventional container can be used. In an embodiment of the present invention, the container for the second fermentation culture is preferably a polyethylene square culture box. In the present invention, the second fermentation culture is preferably cultured until the mycelium of the gray-white winged spore shell F27 completely covers the fermentation medium, and the spore and mycelium density is ≥10 6 The static culture of the present invention is to allow the mycelium of Aspergillus niger F27 to be tightly combined with the solid matrix, and the second fermentation time and temperature parameters of the present invention are set to obtain more active and dense Aspergillus niger F27 active bacteria in a short time.

[0059] The present invention provides the use of the gray white wing spore shell F27 described in the above technical solution or the bacterial agent described in the above technical solution or the bacterial agent prepared by the preparation method described in the above technical solution in one or more of the following (1) to (6);

[0060] (1) Improve the degradation rate of straw in the soil environment;

[0061] (2) Promote the formation of soil aggregates;

[0062] (3) Increase the soluble carbon and nitrogen content in the soil;

[0063] (4) Increase soil microbial biomass;

[0064] (5) Inhibit the growth of Fusarium;

[0065] (6) Promote plant growth;

[0066] (7) Improve plant resistance to stress.

[0067] In the present invention, the plant preferably includes wheat or corn.

[0068] In the present invention, P. hoaricum F27 preferably promotes plant growth by secreting gibberellins and / or zeatin riboside; P. hoaricum F27 promotes plant stress resistance by secreting abscisic acid.

[0069] The Emericellopsis pallida F27 microbial agent provided by the present invention is preferably applied to the soil, and the mass ratio of the microbial agent to the soil during the application is preferably 1: (200-500), further preferably 1: (300-400), and more preferably 1: 350. In an embodiment of the present invention, 20 g of the Emericellopsis pallida F27 microbial agent is preferably applied to 7 kg of soil. After application, the Emericellopsis pallida F27 microbial agent provided by the present invention can increase the degradation rate of straw, inhibit the growth of various Fusarium, promote the growth of wheat and corn, improve the stress resistance of corn, soil microbial biomass nitrogen, soil microbial biomass carbon, soil soluble carbon, soil soluble nitrogen, and soil oxygen content.

[0070] In order to further illustrate the present invention, the technical solution provided by the present invention is described in detail below in conjunction with the accompanying drawings and embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0071] Example 1 Plate confrontation experiment between gray white winged spore shell F27 and plant pathogenic bacteria

[0072] 1. Preparation of culture medium and bacterial blocks

[0073] PDA medium is potato sucrose agar medium, composed of 200 g potato, 20 g sucrose, 20 g agar and 1000 mL deionized water, natural pH.

[0074] Preparation of the bacterial tuber of Pteris palustris F27: The Pteris palustris F27 stored on the PDA slant was inoculated into a PDA medium with a diameter of 90 mm for activation, cultured at 25°C for 5 days, and agar tubers with a diameter of 0.5 cm were obtained by punching with a cork hole. The colony morphology of Pteris palustris F27 on the PDA plate is shown in Figure 1 The colony morphology of Pteris pallida F27 is: light yellow colonies, vigorous aerial hyphae, and light red pigment will appear in the late growth stage.

[0075] Preparation of pathogenic Fusarium fungus blocks: Fusarium fujikura, Fusarium lamina, and Fusarium oxysporum stored on PDA slopes were inoculated into PDA culture medium with a diameter of 90 mm for activation, cultured at 25°C for 5 days, and punched with a cork puncher to obtain fungal blocks with a diameter of 0.5 cm.

[0076] 2 Antagonism experiment

[0077] (1) The prepared Pteris cinerea F27 and Fusarium fujikura fungus blocks were inoculated at 1 / 4 and 3 / 4 of the diameter of a 90 mm PDA plate; 5 parallel experiments were set up.

[0078] (2) The prepared Pteris cinerea F27 and Fusarium solani fungi were inoculated at 1 / 4 and 3 / 4 of the diameter of a 90 mm PDA plate; five parallel experiments were set up.

[0079] (3) The prepared Pteris cinerea F27 and Fusarium oxysporum fungi were inoculated at 1 / 4 and 3 / 4 of the diameter of a 90 mm PDA plate; five parallel experiments were set up.

[0080] The inoculated PDA plates obtained in steps (1) to (3) were placed in a 25°C incubator for dark culture for 3 to 4 days, during which the growth rate of the colonies and whether there was an obvious antagonistic effect were observed. Figure 2 .

[0081] according to Figure 2It can be seen that there is a significant antagonistic effect between F27 and plant pathogenic bacteria, indicating that F27 can inhibit the activity of Fusarium fujikuroi, Fusarium serriola, and Fusarium oxysporum. Fusarium is a harmful fungus that can cause significant agricultural losses. Fusarium fujikuroi is the main cause of wild lettuce disease in Korea (Kim, BR, et al., 2021. Fusarium fujikuroi Causing Fusarium Wilt of Lactuca serriola in Korea. Plant Disease 105, 502-502.) At the same time, it has been verified that it is the main pathogenic microorganism of rice seedling disease in my country (Chen, et al, 2020. Invasion and Colonization Pattern of Fusarium fujikuroi in Rice. Phytopathology 110, 1934-1945.). Fusarium oxysporum can cause wilt disease and is difficult to control. Fusarium can produce fusarium toxins in the parasitic host. On the one hand, these toxins are closely related to the pathogenicity of Fusarium to host plants as pathogenic factors (Huo Yingying et al., 2022 Research progress on the potential of Fusarium oxysporum related pathogenic factors and volatile oils to inhibit Fusarium oxysporum. Pesticides. 61, 79-86). On the other hand, it can lead to a decline in livestock production performance and the occurrence of related diseases, thereby affecting the agricultural ecosystem and posing a threat to human health (Zhang Jiacheng et al., 2021 Types of toxins produced by plant pathogenic Fusarium and their hazards. Grassland Science, 38, 1513-1524.). Fusarium spp. infects rice, wheat, corn, asparagus, onion and other plants, causing wilt and rot. In addition to causing yield loss, the pathogen can also produce fumonisin to affect quality (Li Jiayang, et al. 2022. Comparison of biological characteristics, pathogenicity and fumonisin-producing B1 of Fusarium spp. causative agent of corn ear rot in Liaoning Province. Maize Science, 30(05):163-170.). The strain of the present invention, the gray-white winged spore shell F27, has obvious antagonism to Fusarium spp., Fusarium oxysporum and Fusarium fujikura, can prevent and control diseases caused by these fungi, has broad prospects for agricultural application, and has certain practical value.

[0082] Example 2 Determination of the straw degradation ability of the gray white winged spore shell

[0083] Prepare enough clean and uniform straw, cut it into 1 cubic centimeter size with scissors, and use it to prepare the medium for measuring straw degradation. The composition of the medium for measuring straw degradation is as follows: 0.50g yeast extract, 20.0g straw, 5.0g tricalcium phosphate, 0.50g ammonium sulfate, 0.20g potassium chloride, 0.10g magnesium sulfate heptahydrate, 0.0001g manganese sulfate monohydrate and 0.0001g ferrous sulfate heptahydrate per liter of medium, adjust the pH value to 7.0 with 2N hydrochloric acid and 0.2M NaOH, sterilize at 121℃ for 20min, and let it cool for use.

[0084] Two agar blocks of Pterospora grisea F27 strain with a diameter of 1 cm were inoculated into the culture medium for measuring straw degradation, and cultured in a shaking incubator at 25°C and 160-180 rpm for 7 days. This experiment was set up in triplicate.

[0085] Comparative Example 1

[0086] The same amount of blank agar blocks as in Example 2 were inoculated into the culture medium for measuring straw degradation prepared in Example 2. The culture was carried out in a shaking incubator at 25°C and 160-180 rpm for 7 days. This experiment was set up in 3 parallels.

[0087] After the culture was completed, the supernatant of the culture medium of Example 2 and Comparative Example 1 was taken out after standing to determine the concentration of reducing sugar.

[0088] The reducing sugar determination method is the DNS method (dinitrosalicylic acid method), and the steps are as follows:

[0089] Preparation method of DNS solution: Weigh 3.25g 3,5-dinitrosalicylic acid and dissolve it in a small amount of water. Transfer it into a 500mL volumetric flask, add 162.5mL 2M sodium hydroxide, then add 22.5g propylene glycol, shake well and adjust the volume to 500mL. Store in a 4℃ refrigerator away from light for later use.

[0090] After Example 2 and Comparative Example 1 were allowed to stand, the supernatant of the culture medium was taken into a 2 mL glass test tube, 1.5 mL of DNS was added, and the mixture was boiled in a water bath for 5 min, immediately cooled on ice, and color developed at room temperature for 20 min. Colorimetry was performed using an enzyme reader at 540 nm.

[0091] Preparation method of standard curve: Take 2g of sucrose and dry it at 98℃ for 2h to constant weight. Accurately weigh 1.0000g of dried sucrose and dilute to 1L. Set different gradients of 0, 0.1, 0.3, 0.5, 0.7, 0.9mg / mL. After boiling in water bath for 5min, immediately cool on ice for 10min, color develop at room temperature for 20min, and colorimetric at 540nm. The test tube number and the amount of each solution added during the determination of the standard curve are shown in Table 1-2. The standard curve is y=0.2713x, R 2 =0.9935, x represents the absorbance value, and y represents the reducing sugar concentration.

[0092] Table 1-2 Test tube numbers and the amount of each solution added when measuring the standard curve.

[0093]

[0094] After the culture was completed, the straw degradation rate in the culture medium of Example 2 and Comparative Example 1 was calculated by using the drying difference subtraction method: the straw in the culture bottle was dried to constant weight at 50°C for 12-24 hours, and weighed after drying to constant weight. The calculation formula (1) is as follows:

[0095] Straw degradation rate = (straw dry weight - straw dry weight after degradation) / straw dry weight * 100% (1)

[0096] Method for determination of available phosphorus: weigh 50mL of culture solution into a 250mL conical flask, add a spoonful of phosphorus-free activated carbon, plug the bottle tightly, and shake on a shaker for 30 minutes. Immediately filter with phosphorus-free filter paper, collect the filtrate in a 100mL conical flask, draw 1mL of filtrate into a 25mL volumetric flask, accurately add 15mL of distilled water with a burette, then add 2.5mL of molybdenum antimony reagent with a pipette, shake well and make up to volume, let stand for 30 minutes, shake well again and perform colorimetry at a wavelength of 800nm. The standard curve is y=1.3935x, R 2 =0.9959, y represents the concentration of phosphorus in the corresponding 25mL colorimetric tube, and x represents the absorbance value.

[0097] SPSS20.0 statistical software was used for analysis. The experimental data were expressed as "mean ± standard error". Tukeys-b (K) was used to test data homogeneity. The results are shown in Tables 1-3 and 1-4.

[0098] According to Table 1-3, the inoculation of Pteris palustris F27 in the culture medium containing straw significantly increased the straw degradation rate and the content of available phosphorus in the degradation solution, that is, accelerated the dissolution of tricalcium phosphate. The degradation rate of corn straw by this strain of Pteris palustris was 30.55% within 7 days, and the reducing sugar concentration in the culture solution reached 25.3 mg / L.

[0099] Compared with Comparative Example 1, the reducing sugar concentration in the culture medium supernatant of Example 2 increased, mainly because the significantly increased straw degradation rate increased the reducing sugar concentration in the solution.

[0100] Table 1-3 Determination of the straw degradation rate of gray-white winged spore shell and the activity of straw degradation-related enzymes in Example 2 and Comparative Example 1

[0101]

[0102]

[0103] Note: N=3. * indicates significance at the p<0.05 level, ** indicates significance at the p<0.01 level.

[0104] After the culture was completed, 2 g of fresh mycelial pellets in the culture medium of Example 2 were taken, and the pellets were simply absorbed with sterile filter paper and weighed. After grinding, 5 mL of 0.05 M PBS solution was added to dissolve the pellets for 30 min, and the pellets were centrifuged at 8000 rpm for 10 min. The supernatant obtained was the crude enzyme solution, which was used to determine the activities of endo-β-1,4-glucanase, exo-β-1,4-glucanase, and β-sucrasidase.

[0105] Determination of endo-β-1,4-glucanase (endo-β-1,4-glucanase EC) enzyme activity: Take 1.8 mL of substrate 2% (w / v) sodium carboxymethyl cellulose (sodium carboxymethyl cellulose is obtained by dissolving 2.0 g sodium carboxymethyl cellulose in 100 ml 0.1 mol / L pH = 5 sodium acetate buffer), add 0.2 mL of crude enzyme solution, react in a 37°C water bath for 30 min, finally add 0.5 mL 0.5 mol / L pre-cooled sodium carbonate solution to terminate the reaction, and measure the absorbance at 373 nm.

[0106] Determination of exo-β-1,4-glucanase activity: Take 1.8 mL of 0.5% (w / v) microcrystalline cellulose solution as substrate, add 0.2 mL of crude enzyme solution, incubate in a 37°C water bath for 30 min, and finally add 0.5 mL of 0.5 mol / L cold sodium carbonate solution to terminate the reaction, and measure the absorbance at 373 nm.

[0107] Method for preparing microcrystalline cellulose solution: weigh 8.2g sodium acetate and dissolve it in 800mL water, adjust the pH to 6.5 with glacial acetic acid, make the volume to 1L, and prepare sodium acetate buffer; weigh 0.5g microcrystalline cellulose and dissolve it in 100mL 0.1mol / L sodium acetate buffer (pH=6.5) to obtain microcrystalline cellulose solution.

[0108] Determination of β-glucosidase activity: Take 1.8 mL of 5 mmol / L 4-nitrobenzene-β-D-pyranosylsugar (β-PNPG) substrate (preparation method: β-PNPG is prepared by dissolving 0.150625 g of 4-nitrobenzene-β-D-pyranosylsugar in 100 mL of 0.1 mol / LTris-HCl buffer (pH = 8)), add 0.2 mL of crude enzyme solution, place in a 37°C water bath for 30 min, add 0.5 mL of 0.5 mol / L cold sodium carbonate solution to terminate the reaction, and measure the absorbance at 373 nm.

[0109] The standard curve used for the determination of endo-β-1,4-glucanase, exo-β-1,4-glucanase, and β-sucrasidase activities is: y = 240.86x, R 2 =0.9999, x represents the absorbance value, and y represents the p-nitrophenol concentration in mg / L.

[0110] SPSS20.0 statistical software was used for analysis, and the experimental data were expressed as "mean ± standard error". Tukeys-b (K) was used to test the homogeneity of the data. The results are shown in Tables 1-4. According to Tables 1-4, Pterospora grisea F27 mainly degrades straw through endo-β-1,4-glucanase and β-sucrasidase.

[0111] Table 1-4 Determination of the activity of enzymes related to straw degradation in Example 2 and Comparative Example 1

[0112]

[0113]

[0114] Note: N=3. * indicates significance at the p<0.05 level, ** indicates significance at the p<0.01 level.

[0115] Example 3 Experiment on promoting soil aggregate formation by inoculation of gray white winged spore shell F27

[0116] 100g of dry soil through a 100 mesh sieve was selected. The soil was sandy tidal soil in Fengqiu, Henan Province, with a sand content of more than 70%, and there was basically no agglomerate greater than 2mm in the natural state. After adding straw powder through a 60 mesh sieve at a ratio of 5mg / kg to the dry soil and mixing (the straw powder was added in the experiment to evaluate the ability of the field of the application of straw to the field of the gray-white winged spore shell F27 under laboratory conditions), it was then placed in a triangular bottle and sealed with a breathable sealing film and sterilized at high temperature. The conditions for high temperature sterilization were 121°C, 20min, and the triangular bottle was sterilized at high temperature again after cooling, and sterilization was repeated 3-4 times. Under sterile conditions, the soil after high temperature sterilization was spread flat in a sterile culture dish with a diameter of 70mm, 15g of sterilized soil in each culture dish, 5mL of sterile water was added, and then the gray-white winged spore shell F27 stored on the PDA slant was inoculated into the culture dish, and 5 parallel experiments were set. After 20 days of incubation at 25°C in the dark, the soil culture morphology of the gray-white winged spore shell F27 is shown in Figure 3 .according to Figure 3 It can be seen that the hyphae of P. hoary white pterygosporium F27 can effectively promote the formation of soil aggregate structure, and large aggregates are visible to the naked eye.

[0117] Comparative Example 2

[0118] 100g of dry soil passed through a 100-mesh sieve was selected. The soil was sandy tidal soil in Fengqiu, Henan Province, with a sand content of more than 70%, and there was basically no aggregate larger than 2mm in the natural state. Straw powder passed through a 60-mesh sieve was added to the dry soil at a ratio of 5mg / kg, and then mixed and placed directly in a sterile culture dish without sterilization. 15g of sterilized soil was added to each culture dish, 5mL of sterile water was added, and then the gray-white winged spore shell F27 preserved on the PDA slant was inoculated into the culture dish and cultured in the dark at 25°C for 20 days. Five parallel experiments were set up.

[0119] The soil aggregates in the culture dishes of Example 3 and Comparative Example 2 were classified and the extracellular polymers of each aggregate were determined. Five parallel experiments were set for determination. The soil aggregates were classified by the soil wet sieving method: 50g of air-dried soil was soaked in water for 2min, and then vibrated with a sieve at 30 times / min, 60 times, and an amplitude of 5cm; the apertures of the sieves were 1mm, 0.25mm, and 0.053mm, respectively.

[0120] The extracellular polymers were measured as follows:

[0121] The determination method is

[0122] Select 4 boards with better growth and mix them gently to form a mixed soil sample. There are 5 mixed soil samples in total. (1) to (5) are all carried out in a clean bench:

[0123] (1) The mixed soil sample was placed on a 250 μm sieve and flooded for 5 min;

[0124] (2) vibration for 2 min, amplitude of 3 cm, frequency of 30 times per minute;

[0125] (3) The soil on the sieve was transferred to a sterile petri dish, with aggregates >250 μm;

[0126] (4) The water-soil mixture was passed through a 53 μm sieve, and the soil on the sieve was transferred to a sterile petri dish, with aggregates of >53 μm and <250 μm;

[0127] (5) The remaining water-soil mixture was kept in a fresh-keeping box and placed in a clean bench overnight to absorb excess moisture. The water-soil mixture was dried in a clean bench until the water content was 20%, then collected in a fresh-keeping bag, divided into two parts, and stored at -80°C for the determination of EPS content.

[0128] The EPS extraction method is as follows:

[0129] (1) Add 30 mL of 0.1 M CaCl2 solution to a 50 mL polypropylene centrifuge tube to remove soluble organic matter, and add 3 g of a water-soil mixture with a water content of 20%. Place the centrifuge tube in ice and vibrate it back and forth at 120 rpm for 1 h. Centrifuge at 3200 g for 30 min, discard the supernatant, and extract EPS from the remaining material.

[0130] (2) 15.98 g CER and 25 ml buffer were added to the precipitate. The mixture was shaken at 120 rpm for 2 h and then centrifuged at 4000 g for 30 min.

[0131] (3) The buffer composition was 2 mM Na3PO4·12H2O (0.760 g / L), 4 mM NaH2PO4·H2O (0.552 g / L), 9 mM NaCl (0.526 g / L), 1 mM KCl (0.0746 g / L), and finally 1 M HCl was used to adjust the pH to 7 and precooled at 4°C. CER (ion exchange resin 'Marathon C' sodium form, strongly acidic, 20-50 mesh) was prewashed twice in the above buffer. The CER content was determined based on the 178 mg CER mg-1 SOC required for the highest organic carbon soil.

[0132] (4) The supernatant containing EPC was transferred to a new test tube, quickly frozen in liquid nitrogen, and stored at -80°C for the determination of EPS uronic acid, EPS protein, and EPS humic acid.

[0133] EPS uronic acid determination method:

[0134] (1) Select 200 μL of supernatant containing EPC and add 1.2 mL of sodium tertraborate (0.0125 M, concentrated sulfuric acid as solvent) solution. Vortex for 45 seconds, place in a 100°C water bath for 5 minutes, cool on ice for 3 minutes, then add 20 μL of m-hydroxybiphenyl solution (CAS#: 580-51-8) and mix well. Let stand for 4 minutes and measure the absorbance at 520 nm.

[0135] (2) Preparation method of standard curve: D-glucuronic acid (CAS#: 6556-12-3) curve Prepare 25μg / mL D-glucuronic acid stock solution. Take 0, 4, 20, 60, 100 and 200ul of the stock solution respectively, and add deionized water to 200ul. Then repeat step (1) to obtain the absorbance values ​​of different stock solutions and prepare the standard curve. The standard curve is: y = 0.0053x + 0.0812, R 2 =0.9978, where y represents absorbance and x represents uronic acid concentration.

[0136] The contents of EPS protein and EPS humic acid were determined by Lowry method.

[0137] Solution A: Dissolve 3.5 g of copper sulfate pentahydrate in 100 mL of water to obtain solution (1); dissolve 7.0 g of sodium potassium tartrate in 100 mL of water to obtain solution (2); dissolve 70 g of sodium carbonate in 1 L of 0.35 N sodium hydroxide solution (3). Solution (1), solution (2) and solution (3) are mixed in a volume ratio of 1:1:100 to obtain solution A.

[0138] Solution B: deionized water, solution (2) and solution (3) are mixed in a volume ratio of 1:1:100 to obtain solution B.

[0139] 1. Repeat each sample 3 times, add 50 μL of supernatant sample containing EPC to each well of plate A and plate B, and add PBS to 100 μL.

[0140] 2. Add 100 μL of A solution and mix by pipetting. For the control, add 100 μL of B solution.

[0141] 3. Place the two plates in the dark at room temperature for 10 minutes.

[0142] 4. Immediately add 100 μL of Folin-Phenol reagent to terminate the reaction.

[0143] 5. After 30 minutes, measure the absorbance at 750 nm.

[0144] Folin-Phenol reagent: 2N Folinphenol was diluted 10 times with water.

[0145] The calculation formula of EPS protein and EPS humic acid is as follows: Abs A Represents the absorbance of the plate with solution A added, Abs B Represents the absorbance of the plate with solution B added.

[0146] Abs 蛋白 =1.25*(Abs A -Abs B );

[0147] Abs 蛋白 The standard curve is: y = 0.0009x + 0.0089, R 2 =0.9965; x is the EPS protein concentration, y is 1.25*(Abs A -Abs B );

[0148] Abs腐殖酸 =AbS B -0.2Abs 蛋白 ;

[0149] Abs 腐殖酸 =AbS B -0.2*1.25*(Abs A -Abs B );

[0150] The standard curve of Abs humic acid is: y = 0.0016x + 0.0732, R 2 =0.9943; x is the concentration of EPS humic acid, y is 0.2*1.25*(Abs A -Abs B ).

[0151] SPSS 20.0 statistical software was used for analysis. The experimental data were expressed as “mean ± standard error” and Tukeys-b (K) was used to test data homogeneity.

[0152] Table 2 Soil aggregate classification results of Example 3 and Comparative Example 2

[0153]

[0154] Note: N=5. * indicates significance at the p<0.05 level, ** indicates significance at the p<0.01 level.

[0155] According to Table 2, the inoculation of gray-white winged spore shells in Example 3 significantly increased the content of 0.25-1mm aggregates in sandy tidal soil, significantly reduced the percentage of <0.053mm aggregates, and significantly increased the average weight diameter of aggregates. From the results of extracellular polymers EPS, the inoculation of F27 significantly increased the content of EPS protein, EPS uronic acid, and EPS humic acid in large aggregates, and also increased the content of EPS components in other aggregates to varying degrees, indicating that the inoculation of gray-white winged spore shells F27 in the soil has a significant promoting effect on the soil aggregation process, and can promote soil aggregation by secreting EPS substances, which helps to improve soil structure and promote soil fertility. However, it has limited effect on promoting the formation of large aggregates. The EPS content in each aggregate size in the soil of Example 3 and Comparative Example 2 is shown in Table 3-1 to Table 3-4.

[0156] Table 3-1 EPS uronic acid content in each particle size aggregate in the soil of Example 3 and Comparative Example 2

[0157]

[0158] Note: Independent sample T test was used to determine the significant difference between the comparative example and the example. Capital letters represent significant differences at the 0.01 level, and lowercase letters represent significant differences at the 0.05 level. No letter marks indicate that no significant difference was reached. The same applies below.

[0159] Table 3-2 OD values ​​for measuring EPS uronic acid content in soil of each particle size in Example 3 and Comparative Example 2

[0160]

[0161] Table 3-3 EPS protein and EPS humic acid content in each particle size aggregate in the soil of Example 3 and Comparative Example 2

[0162]

[0163] Table 3-4 OD values ​​of the EPS protein and EPS humic acid content in each particle size aggregate in the soil of Example 3 and Comparative Example 2

[0164]

[0165] Example 4 Experiment on the growth of wheat root system by using gray-white winged spore shell F27

[0166] 80g of commercially available nutrient soil was placed in a 1L triangular flask, adjusted to a water content of 20%, sealed with a breathable sealing film, and sterilized at 121°C for 20min. After cooling, wheat seeds were sown in the nutrient soil under aseptic conditions, with a sowing depth of 1cm. At the same time, 3 agar blocks of the strain F27 of the gray-white winged spore shell with a diameter of 15mm prepared in Example 1 were inoculated into the triangular flask, with an inoculation depth of 1cm. After the inoculation was completed, the triangular flask was sealed and cultured in a biochemical incubator. After 2 days of emergence, the seedlings were removed under aseptic conditions, and 1 seedling was left in each bottle. The light was 25°C for 16h and the dark was 15°C for 8h. The experiment was terminated after culturing for 15 days. This experiment was set up in 3 parallels.

[0167] Comparative Example 3

[0168] Preparation method of blank agar block: Pour the prepared PDA culture medium into the culture dish, and after condensation, a blank agar block with a thickness of 0.3 cm can be obtained.

[0169] Comparative Example 3: Three blank agar blocks with a diameter of 15 mm were inoculated, the inoculation depth was 1 cm, and the other conditions were the same as those of Example 4. This experiment was set up in 3 parallels.

[0170] Example 4 and Comparative Example 3 The root system was scanned by a root scanner and analyzed using SPSS 20.0 statistical software. The experimental data were expressed as "mean ± standard error". Tukeys-b (K) test was used to test the homogeneity of the data. The results are shown in Table 4 and Figure 4 .according to Figure 4 As shown in Table 4, the soil of Example 4 was inoculated with Hoary Wing Sporangia F27, and the root crown number, number of intersections, total root length, total root area and total root volume of the wheat root system of Example 4 were significantly higher than those of Comparative Example 3. It can be seen that the inoculation of Hoary Wing Sporangia F27 can significantly promote the growth of the wheat root system and aboveground part.

[0171] Table 4 Growth parameter results of wheat root system in seedling stage of Example 4 and Comparative Example 3

[0172]

[0173]

[0174] Note: N=3. * indicates significance at the p<0.05 level, ** indicates significance at the p<0.01 level.

[0175] Example 5 Preparation of Gray White Wing Sporangia Agent

[0176] (1) A mycelium block of Pteris sphaerocephala F27 with a diameter of 15 mm that grows vigorously on PDA medium is inoculated into PD liquid medium and cultured in a constant temperature shaker at 28°C and 220 rpm for 4 to 5 days, and a large number of white bean-shaped mycelium balls appear to obtain a fermentation mixture; wherein the PD liquid medium is composed of: 200 g of peeled potatoes, 20 g of sucrose and 1000 mL of water.

[0177] (2) Aseptically taking 10 mL of the fermentation mixture obtained in step (1) and adding it to 20 g of the fermentation medium, immediately shaking it and placing it in a dark place at 28° C. to culture it; when the hyphae of Aspergillus niger F27 cover the surface and the inside of the medium, smash the hyphae with a glass rod to obtain a bacterial agent, and the number of active hyphae of Aspergillus niger F27 in the bacterial agent is 2×10 6 cfu / g.

[0178] The fermentation medium (wheat bran-sucrose medium) is: 70g wheat bran, 25g bean cake powder, 3g sucrose, 0.15g poly-gamma glutamic acid and 100g water. The preparation method of the fermentation medium is to mix the wheat bran and bean cake powder in a dry state, dissolve sucrose and poly-gamma glutamic acid in water, mix for 30 minutes to obtain a mixture, the pH is natural, put into a 200mL fresh-keeping box, seal with a breathable sealing film, sterilize at 121°C for 20 minutes, and shake it while hot.

[0179] Example 6

[0180] Accurately weigh 20g of the F27 bacterial agent prepared in Example 5, the water content of the bacterial agent is 15%, add it to 7kg of soil, mix it thoroughly and put it into the planting pot. Bury 10g of accurately weighed straw after drying at 15cm from the pot mouth, and put the straw into a 150-mesh double-layer nylon bag. In order to ensure the survival of F27, water it immediately after the pot is filled.

[0181] After watering, corn Denghai 605 was planted, with 5 corns planted in each planting pot. The corn in each planting pot was fixed at the three-leaf stage, and 1 corn seedling was left in each planting pot. Corn was cultured in an artificial climate chamber, with 14h light, 28℃ temperature under light conditions, 10h dark, 20℃ temperature under dark conditions, and 70% relative humidity. Watering and weeding were performed as needed during the period. This experiment was set up with 4 parallels.

[0182] Comparative Example 4

[0183] Accurately weigh 20g of sterilized wheat bran-sucrose medium, add it to 7kg of soil, mix it thoroughly and put it into the planting pot. After the planting pot is fully watered, plant corn Denghai 605, plant 5 corns in each planting pot, and the corn in each planting pot is fixed at the three-leaf stage, leaving 1 corn seedling in each planting pot. Corn is cultured in an artificial climate chamber, and the culture conditions are 14h of light, 28℃ under light conditions, 10h of darkness, 20℃ under dark conditions, and 70% relative humidity. Water and weed as needed during the period. This experiment is set up 4 parallels.

[0184] In the tasseling period of corn in Example 6 and Comparative Example 4, soil microbial biomass nitrogen, soil microbial biomass carbon, soil soluble carbon, soil soluble nitrogen, soil oxygen content, and straw degradation rate were measured, and the corn plant biomass, leaf chlorophyll, and corn root hormone content were also measured.

[0185] The soil microbial biomass carbon and nitrogen were measured using the chloroform fumigation-potassium sulfate extraction method: 5 g of fresh soil was weighed into two portions, one was fumigated with chloroform in the dark for 24 hours, and the other was not fumigated. 0.5 M K2SO4 was added and shaken at 25°C for 30 minutes. Then, mulit3100 was used to measure the water-soluble carbon and nitrogen, and the microbial biomass carbon and nitrogen were measured by subtraction.

[0186] Leaf chlorophyll was measured using the SPAD value method, and leaf SPAD was measured using a SPAD instrument (Aopgee MC-100).

[0187] Maize biomass was measured by oven-drying weighing method.

[0188] The O2 and CO2 contents in the soil were measured using soil oxygen sensor SO-110 and soil CO2 sensor produced by eosGP.

[0189] The calculation of straw degradation rate is the same as formula (1).

[0190] The contents of maize root hormones indoleacetic acid, abscisic acid, gibberellin, and zeatin nucleoside were measured using enzyme-linked immunosorbent assay kits (Shanghai ELISA Biotechnology Co., Ltd.)

[0191] For the determination method of soil soluble carbon and nitrogen, please refer to the paper (Wu, J., Joergensen, RG, Pommerening, B., Chaussod, R., Brookes, PC, 1990. Measurement of soil microbial biomass C by fumigation--extraction--an automated procedure. Soil Biology & Biochemistry 22, 1167-1169.).

[0192] SPSS20.0 statistical software was used for analysis, and the experimental data were expressed as "mean ± standard error". Tukeys-b (K) was used to test the homogeneity of the data. The results of the determination of Example 6 and Comparative Example 4 are shown in Tables 5 and 6. According to Tables 5 and 6, the biomass of the corn in Example 6 applied with the F27 bacterial agent of the gray-white wing spore shell increased by 13%, and the SPDA value of the corn chlorophyll index increased by 14%. The application of the F27 bacterial agent of the gray-white wing spore shell significantly increased the concentration of gibberellins and zeatin nucleoside in the corn root system, indicating that the inoculation of the F27 gray-white wing spore shell has a strong biological stimulation effect on corn growth. In addition, the abscisic acid concentration of the corn root system applied with the F27 bacterial agent of the gray-white wing spore shell increased significantly, with an increase of up to 15%, indicating that the F27 gray-white wing spore shell has a good effect on improving the stress resistance of corn.

[0193] Example 6 The soil microbial biomass carbon of the soil treated with the F27 bacterial agent of the gray-white winged spore shell increased by 15%, the microbial nitrogen content increased by 75%, the soil soluble nitrogen increased by 23%, and the straw degradation rate increased by 10%. The soil microbial biomass carbon and nitrogen and the soil soluble carbon and nitrogen content of the soil treated with the F27 bacterial agent of the gray-white winged spore shell activated the soil ecosystem and promoted the activity of soil microorganisms. The application of the F27 bacterial agent of the gray-white winged spore shell significantly reduced the oxygen content of the cultivated layer soil by 0.3 percentage points. The reduction in oxygen content can effectively reduce the soil respiration intensity and promote the accumulation of soil organic matter.

[0194] Table 5 Measurement results of soil indicators of Example 6 and Comparative Example 4

[0195]

[0196] Note: N=4. * indicates significance at the p<0.05 level, ** indicates significance at the p<0.01 level.

[0197] Table 6 Determination results of hormone content in the aboveground part and root of corn in Example 6 and Comparative Example 4

[0198] Example 6 Comparative Example 4 <![CDATA[Aboveground biomass (gdwstain -1 )]]> 43.63±3.28* 38.57±2.50 <![CDATA[Leaf chlorophyll index (ngg -1 fw)]]> 40.46±4.35** 35.75±2.4 <![CDATA[Indole acetic acid content in maize (ng g -1 fw)]]> 54.12±1.67 51.03±2.84 <![CDATA[Abscisic acid content in maize (ng g -1 fw)]]> 575.64±34.32** 500.49±31.58 <![CDATA[Gibberellin content in maize roots (ng g -1 fw)]]> 231.33±14.45* 193.74±16.82 <![CDATA[Zeatin riboside content in maize roots (ng g -1 fw)]]> 39.62±2.18* 31.97±3.22

[0199] Note: N=4. * indicates significance at the p<0.05 level, ** indicates significance at the p<0.01 level.

[0200] In summary, the Emericellopsis pallida F27 provided by the present invention has a deposit number of CGMCC No. 40123, can increase the degradation rate of straw, inhibit the growth of various Fusarium, promote the growth of wheat and corn, improve the stress resistance of corn, soil microbial biomass nitrogen, soil microbial biomass carbon, soil soluble carbon, soil soluble nitrogen, and soil oxygen content. The Emericellopsis pallida F27 provided by the present invention and the microbial agent prepared therefrom have broad application prospects in the field of agriculture.

[0201] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A gray-white winged spore shell ( Emericellopsis pallida )F27, characterized in that The deposit number is CGMCC No.40123.

2. A fungal agent containing the gray-white winged spore shell according to claim 1, characterized in that The number of active hyphae of the gray-white winged spore shell F27 in the bacterial agent is ≥10 6 cfu / g.

3. The method for preparing the bacterial agent according to claim 2, characterized in that: The steps include: performing a first fermentation culture of Pterospora grisea F27 to obtain a first fermentation mixture, and performing a second fermentation culture of the first fermentation mixture to obtain a bacterial agent; the first fermentation culture and the second fermentation culture are both performed under dark conditions; The first fermentation culture time is 7 to 10 days, and the second fermentation culture time is 3 to 6 days.

4. The preparation method according to claim 3, characterized in that: The culture medium used in the second fermentation culture comprises the following components in parts by weight: 65-75 parts of wheat bran, 20-30 parts of soybean cake powder, 2-4 parts of sucrose, 0.1-0.2 parts of poly-gamma-glutamic acid and 90-110 parts of water.

5. The preparation method according to claim 4, characterized in that: The molecular weight of the poly-gamma-glutamic acid is 600,000 to 800,000.

6. The preparation method according to claim 3, characterized in that: The medium used in the first fermentation culture is PD liquid medium, the temperature of the first fermentation culture is 25-30° C., and the rotation speed of the first fermentation culture is 120-180 rpm.

7. The preparation method according to claim 3, characterized in that: The temperature of the second fermentation culture is 25-30° C., and the volume ratio of the first fermentation mixture to the second fermentation culture medium is (15-25):

100.

8. Use of the gray white wing spore shell F27 according to claim 1 or the bacterial agent according to claim 2 or the bacterial agent prepared by the preparation method according to any one of claims 3 to 7 in one or more of the following (1) to (6); (1) Improve the degradation rate of straw in the soil environment; (2) Promote the formation of soil aggregates; (3) Increase soil microbial biomass; (4) inhibiting the growth of Fusarium; the Fusarium is one or more of Fusarium fusarifolia, Fusarium laminarum and Fusarium oxysporum; (5) Promote plant growth; (6) Improve plant stress resistance.

9. The use according to claim 8, characterized in that: The plant includes wheat or corn.

10. The use according to claim 8 or 9, characterized in that: The gray-white wing spore shell F27 promotes plant growth by secreting gibberellins and / or zeatin riboside; the gray-white wing spore shell F27 promotes plant stress resistance by secreting abscisic acid.

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