CN117683644A - Plant endophytic fungus and application thereof in plant growth promotion - Google Patents
Plant endophytic fungus and application thereof in plant growth promotion Download PDFInfo
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- 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
- C12N1/145—Fungal isolates
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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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- A—HUMAN NECESSITIES
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Abstract
The invention discloses a plant endophytic fungusParaphaeosphaeria sardoa) And the application of the strain in plant growth promotion can effectively promote the growth of arabidopsis thaliana and tomatoes; after co-culturing with the strain, the fresh weight of tomato seedlings is increased by 175.97% compared with that of a control group; after the strain fermentation liquor filtrate is applied, the dry weight of the overground part of tomato seedlings in a potting growth promotion test is increased by 36.70 percent compared with that of a control group, the dry weight of the underground part is increased by 76.74 percent, the content of soluble protein is increased by 56.65 percent, and the content of soluble sugar is increased by 156.96 percent. The strain and the filtrate of the fermentation liquor thereof provided by the invention can exert obvious growth promoting effect and have wide development and utilization in the field of agricultural microbial fertilizersThe method has a prospect and is worth to be widely popularized.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a plant endophytic fungus and application thereof in plant growth promotion.
Background
Endophytic fungi (endophytic fungi) are fungi which are parasitic in plants, participate in the whole life history of host plants and have no obvious harm to the host plants. Endophytes are a large group of fungi, including mycorrhizal fungi, surface-growing saprophytes, latent pathogenic bacteria that are temporarily harmless to the host, and the like. Endophytic fungi are found in almost all living plants and can be isolated from surface-sterilized plant tissue.
Endophytic fungi can establish a reciprocal symbiotic relationship with most plants in nature. Plants provide habitat and nutrition for endophytes, while secondary metabolites produced by endophytes can regulate hormone levels in plants. After the endophytic fungi and the host plants are symbiotic, the growth of the host plants can be promoted, and the negative influence of the severe environment on the growth of the host plants can be reduced. By utilizing the symbiotic relationship of the plant and the endophytic fungi, the growth promotion effect of the endophytic fungi on the plant can be realized, and the nutrition absorption of the plant can be improved. Thus, isolation and utilization of endophytic fungus strains having growth promoting ability to crops helps to reduce the use of fertilizers in agricultural ecosystems.
The fungus Paraphaosphaeria is a newly discovered genus in recent years, and has no accurate Chinese name. Studies have shown that Paraphaophaeria fungi can improve plant disease resistance, and no study has been reported on whether they can promote plant growth.
Disclosure of Invention
The invention aims to: in order to solve the defects in the prior art, the invention aims to solve the technical problem that a plant endophyte (Paraphaeosphaeria sardoa) capable of promoting plant growth is separated from the root system of apocarya, and research on the plant growth promoting effect is carried out, so that a foundation is provided for subsequent deep development of microbial fertilizers and reduction of chemical fertilizers.
The invention also solves the technical problem of providing a fermentation liquor filtrate capable of promoting plant growth and a preparation method thereof.
The invention also solves the technical problem of providing a microbial preparation capable of promoting plant growth.
The invention finally solves the technical problem of providing the application of endophytic fungi CNBG-PGPF-2, fermentation liquor filtrate and/or preparation thereof in promoting plant growth.
The technical scheme is as follows: in order to solve the technical problems, the invention provides a plant endophyte (Paraphaeosphaeria sardoa) CNBG-PGPF-2, wherein the plant endophyte (Paraphaeosphaeria sardoa) CNBG-PGPF-2 is preserved in China general microbiological culture Collection center of China general microbiological culture Collection center, with the preservation number of 2023, 5 months and 22 days: CGMCC No.40636.
The invention also comprises a fermentation liquor filtrate which contains the endophyte (Paraphaeosphaeria sardoa) CNBG-PGPF-2.
The invention also discloses a preparation method of the fermentation liquor filtrate, which comprises the following steps: obtaining fresh cultured endophytic fungus mycelium blocks by using a puncher, putting the mycelium blocks into a PDB (PDB) culture medium, culturing in the dark for 7-10 days, collecting fermentation liquor, filtering the fermentation liquor by using gauze, and diluting the fermentation liquor by using sterile distilled water to obtain fermentation liquor filtrate.
Wherein, the formula of the PDB culture medium is as follows: 3g of potato soaked powder, 20g of glucose, 1000mL of deionized water and high-pressure steam sterilization at 121 ℃ for 20min.
The invention also comprises a microbial preparation which contains the endophyte (Paraphaeosphaeria sardoa) CNBG-PGPF-2 or the fermentation broth filtrate.
The invention also comprises the application of the endophyte (Paraphaeosphaeria sardoa) CNBG-PGPF-2, the fermentation broth filtrate or the microbial preparation in promoting plant growth.
Wherein the concentration of plant endophytic fungi (Paraphaeosphaeria sardoa) CNBG-PGPF-2 spore in the fermentation broth filtrate is 5×10 4 Per mL-2×10 5 And each mL.
Wherein the plant comprises, but is not limited to, one or more of Arabidopsis thaliana and Lycopersicon esculentum, and is also suitable for promoting growth of other plants.
Wherein, the application specifically includes: plant and plant endophytic fungi (Paraphaeosphaeria sardoa) CNBG-PGPF-2 were co-cultured on solid medium.
Wherein the solid medium includes, but is not limited to, MS medium, preferably 1/2MS solid medium.
The beneficial effects are that: compared with the prior art, the invention has the following advantages: the strain can effectively promote the growth of arabidopsis thaliana and tomatoes; after co-culturing with the strain, the fresh weight of tomato seedlings is increased by 175.97% compared with that of a control group; after the strain fermentation liquor filtrate is applied, the dry weight of the overground part of tomato seedlings in a potting growth promotion test is increased by 36.70 percent compared with that of a control group, the dry weight of the underground part is increased by 76.74 percent, the content of soluble protein is increased by 56.65 percent, and the content of soluble sugar is increased by 156.96 percent. The strain and the fermentation liquor filtrate thereof provided by the invention can exert obvious growth promoting effect, have wide development and utilization prospects in the field of agricultural microbial fertilizers, and are worth popularizing.
Drawings
FIG. 1 shows colony morphology of CNBG-PGPF-2 strain on corn meal medium (COM), wherein the left side is front side, and the right side is back side.
FIG. 2 shows the mycelium morphology of the CNBG-PGPF-2 strain on corn meal medium (COM).
FIG. 3 shows spore morphology of CNBG-PGPF-2 strain in corn meal medium (COM).
FIG. 4 is a phylogenetic tree of CNBG-PGPF-2 strain.
FIG. 5 shows the co-cultured pro-active effect of CNBG-PGPF-2 strain 1/2MS medium and Arabidopsis thaliana, with the upper panel being front side view and the lower panel being back side view (Control being Control group, CNBG-PGPF-2 being treatment group).
FIG. 6 shows the co-cultured pro-active effect of CNBG-PGPF-2 strain 1/2MS medium and tomato (Control as Control group, CNBG-PGPF-2 as treatment group).
FIG. 7 shows the effect of the 1/2MS culture medium of the strain CNBG-PGPF-2 on the growth index of tomato seedlings by co-cultivation of tomato, wherein the left side of the figure is fresh weight data of tomato seedlings, and the right side is root length data of tomato seedlings (Control is Control group, CNBG-PGPF-2 is treatment group).
FIG. 8 shows the colonization of the tomato root system by the strain CNBG-PGPF-2 (Control is Control group, CNBG-PGPF-2 is treatment group).
FIG. 9 shows the effect of the pot growth promoting test of the strain CNBG-PGPF-2 on the growth index of tomato seedlings, wherein the upper left part of the graph shows the growth rate data of tomato seedlings, the upper right part of the graph shows the dry weight data of the aerial parts of tomato seedlings, the lower left part of the graph shows the root length data of tomato seedlings, and the lower right part of the graph shows the dry weight data of underground parts of tomato seedlings (Control is a Control group, CNBG-PGPF-2 10%, CNBG-PGPF-2 20%, and CNBG-PGPF-2 50% is a treatment group).
FIG. 10 shows the effect of the potted plant growth promoting test of the strain CNBG-PGPF-2 on the physiological index of tomato seedlings, wherein the left side of the figure shows the soluble protein content data of tomato seedlings, and the right side shows the soluble sugar content data of tomato seedlings (Control is Control group, CNBG-PGPF-2 10%, CNBG-PGPF-2 20% and CNBG-PGPF-2% is treatment group).
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings, examples and experiments, but the present invention is not limited to the following technical schemes. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art. Those skilled in the art can make reference to various general specifications, technical and scientific literature or related specifications, manuals, etc. before the filing date of this invention.
EXAMPLE 1 isolation, purification and morphological observations of the endophytic fungus CNBG-PGPF-2
Taking root system of strong plant at planting base (period, jiangsu) of Carya illinoensis, washing root system surface with tap water, cutting sample into pieces, and placing on filter paper. Firstly, placing a cleaned sample into a 10mL centrifuge tube, repeatedly blowing with sterile water, and cleaning by vibration; secondly, adding 75% ethanol solution for disinfection, repeatedly blowing, oscillating and cleaning, naturally settling, and removing supernatant; repeatedly sucking sterile water, blowing, oscillating, washing off ethanol, and repeating for 3 times; then adding 10% sodium hypochlorite solution for disinfection, repeatedly blowing, oscillating and cleaning, naturally settling and removing supernatant; finally, absorbing sterile water, repeatedly blowing, oscillating and washing sodium hypochlorite, and repeating for 3 times.
After the sample sterilization is completed, the sample is placed in an ultra-clean workbench, split by a sterilized knife, placed on a fresh Potato Dextrose Agar (PDA) plate and cultivated in the dark at a constant temperature of 25 ℃. After hyphae grow out, according to the color, texture and growth speed of the bacterial colony, a sterile inoculating needle is used for picking fresh hypha blocks of a single bacterial colony, and the bacterial colony is inoculated to a new PDA flat plate, and is cultured in a dark inversion mode in a constant temperature incubator at 25 ℃ for 3-5 days. Pure bacterial strains are obtained through multiple subcultures, and purified bacterial colonies are inoculated into slant culture and preserved at 4 ℃. To observe colony, mycelium and spore morphology of CNBG-PGPF-2 strain, corn meal medium (COM) was selected for cultivation.
PDA formula: 3g of potato soaked powder, 20g of glucose and 15g of agar, 1000mL of deionized water is added, and the mixture is sterilized by high-pressure steam at 121 ℃ for 20min.
COM medium formulation: corn steep powder 7g, 1000mL deionized water was added and autoclaved at 121℃for 20min. Agar 15g was added to the solid medium.
Experimental results: after 5 days of culture of CNBG-PGPF-2 on solid corn meal medium (COM), the colony was almost white, and the aerial hyphae were densely developed (FIG. 1). The strain has more branches of hyphae, multiple branches are distributed on the main handle, and the top of the hyphae grows singly (figure 2). After 3 days of culture on liquid corn meal medium (COM), the conidia of the strain were smooth, spherical or nearly spherical (FIG. 3), and had high spore yield, exceeding 10/μL.
Example 2 molecular biological characterization of endophytic fungi CNBG-PGPF-2
The fungi (3 mycelium blocks) cultured on the plate are obtained by a 5mm puncher, placed in a 2mL bacteria-preserving tube containing 500 mu L of 0.01M phosphate buffer (pH 7.2-7.4), and fully and uniformly vibrated. An appropriate amount of sterilized glass beads (dia. Glass beads, biospec) having diameters of 0.5mm and 0.1mm were added (mixing the two glass beads in equal proportions), and the mixture was ground with a shaker at a speed of 6.5m/s for 1min. Finally, the mixture was centrifuged at 12000rpm for 2min, and the supernatant was taken out in a 1.5mL sterilized centrifuge tube for later use.
PCR amplification was performed on the strain target gene (ITS) fragment using the above crude DNA solution as a template and ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3') as upstream and downstream primers. The PCR amplification reaction system was 50. Mu.L: 25. Mu.L of LEASyTaq Mix, 2. Mu.L of forward primer ITS1, 2. Mu.L of reverse primer ITS4, 3. Mu.L of DNA template, and 17. Mu.L of sterile water. Amplification conditions: pre-denaturation at 94℃for 5min; denaturation at 94℃for 30s, annealing at 55℃for 30s, extension at 72℃for 40s,36 cycles; extending at 72℃for 10min.
The PCR products were detected by 1% agarose gel electrophoresis and sent to the company for sequencing. The sequencing result is compared and analyzed with all ITS sequences in the database by using Blast program in Gen Bank database of NCBI, and 10-15 species with highest similarity are selected for analysis of the evolutionary tree.
Experimental results: and (3) splicing the sequencing results to obtain the ITS gene near-full-length sequence (shown as SEQ ID NO. 1) of the strain CNBG-PGPF-2, wherein the length is 566bp. The ITS gene sequence of the strain is subjected to homologous sequence search in GenBank database (NCBI), and the strain with highest similarity is found by BLAST analysis. As a result, it was found that the similarity of the strain to Paraphaeosphaeria sardoa CBS 501.71 was 97.81%. Meanwhile, the strain was found to have clustered with a plurality of Paraphaeosphaeria fungi using MEGA 7 software and maximum likelihood method (Maximum Likelihood methods), and was closest to Paraphaeosphaeria sardoa CBS 501.71 (FIG. 4), and it was determined that the strain was likely Paraphaeosphaeria sardoa. Then CNBG-PGPF-2 is stored in China general microbiological culture Collection center (CGMCC), address: no. 3 of North Chen Xili No.1, no. 22 of Beijing, chaoyang district, with 2023, CGMCC No.40636 and Paraphaeosphaeria sardoa.
EXAMPLE 3 Co-culture test of CNBG-PGPF-2 Strain 1/2MS Medium and Arabidopsis thaliana
(1) The arabidopsis seeds are sterilized firstly: 60-80 Arabidopsis seeds are put into a 2mL centrifuge tube for surface sterilization. Firstly, 1mL of sterile water is sucked for repeated blowing and shaking to clean seeds, and the supernatant is discarded after natural sedimentation. And then, sucking 1mL of 75% ethanol solution, repeatedly blowing and vibrating to disinfect seeds, lasting for 2-3 min, naturally settling, discarding the supernatant, sucking 1mL of sterile water, repeatedly blowing and vibrating to wash out ethanol, naturally settling, discarding the supernatant, and repeating for 2 times. Then 1mL of 10% sodium hypochlorite solution is sucked for repeated blowing and shaking to disinfect seeds, the duration is 6-8 min, and the supernatant is discarded after natural sedimentation. Then sucking 1mL of sterile water, repeatedly blowing, oscillating to wash out sodium hypochlorite, naturally settling, discarding the supernatant, repeating for 2 times, and finally sucking 1mL of sterile water to resuspend the seeds.
(2) Uniformly spreading the sterilized Arabidopsis seeds on a 1/2MS solid culture medium, sucking off excessive sterile water on the culture medium, properly drying in an ultra-clean bench, sealing by a sealing film, transferring to an illumination culture room, and vertically culturing at 22 ℃ in 16h illumination/8 h darkness until the seeds germinate.
(3) The left side and the right side of the 1/2MS solid culture medium are respectively provided with a sterile PDA block (left side) with the diameter of 5mm and the thickness of 3mm and a CNBG-PGPF-2 mycelium block (right side) at a position 0.9cm away from the edge of the culture dish, and the volumes of the PDA block and the mycelium block are close to 60mm 3 . After the seeds germinate, selecting seedlings with consistent growth vigor, uniformly and transversely placing the seedlings on a culture dish, sealing the seedlings by sealing films on the left side and the right side, transferring the seedlings to an illumination culture room, vertically culturing the seedlings for 7-10 days at 22 ℃ in 16h illumination/8 h darkness, and collecting and photographing the Arabidopsis seedlings when obvious differences exist.
Experimental results: plant-endophytic fungi interaction experiments were performed on 1/2MS solid medium using Arabidopsis thaliana and endophytic fungi CNBG-PGPF-2. After a period of time, the Arabidopsis plants inoculated with CNBG-PGPF-2 were larger, the leaves were greener, and the root system was developed, compared to Arabidopsis seedlings of the control group, and especially the number of lateral roots of Arabidopsis seedlings was significantly higher than that of the control group (FIG. 5).
EXAMPLE 4 Co-culture test of CNBG-PGPF-2 Strain 1/2MS Medium and tomato
(1) Firstly, sterilizing tomato seeds: 60-80 tomato seeds are put into a 15mL centrifuge tube for surface sterilization. Firstly, 10mL of sterile water is sucked, the seeds are repeatedly blown and cleaned by shaking, and the supernatant is removed after natural sedimentation. And then, absorbing 10mL of 75% ethanol solution, repeatedly blowing and vibrating to disinfect seeds for 2-3 min, naturally settling, removing supernatant, absorbing 10mL of sterile water, repeatedly blowing and vibrating to wash out ethanol, naturally settling, removing supernatant, and repeating for 2 times. Then 10mL of 10% sodium hypochlorite solution is sucked for repeated blowing and shaking to disinfect seeds, the duration is 6-8 min, and the supernatant is removed after natural sedimentation. Then 10mL of sterile water is sucked for repeated blowing and shaking to wash out sodium hypochlorite, the supernatant is removed after natural sedimentation, and after repeating for 2 times, 10mL of sterile water is sucked for resuspension of the seeds.
(2) Uniformly spreading the sterilized tomato seeds on a 1/2MS solid culture medium, absorbing and discarding excessive sterile water on the culture medium, properly drying in an ultra clean bench, sealing by a sealing film, marking by a marker pen, transferring to an illumination culture chamber, and vertically culturing at 25 ℃ for 12h in an illumination/12 h dark until the tomato seeds are exposed to white.
(3) The left side and the right side of the 1/2MS solid culture medium are respectively provided with a sterile PDA block (left side) with the diameter of 5mm and the thickness of 3mm and a CNBG-PGPF-2 mycelium block (right side) at a position 0.9cm away from the edge of the culture dish, and the volumes of the PDA block and the mycelium block are close to 60mm 3 . After the seeds are exposed and white, selecting tomato seeds with consistent growth vigor, uniformly and transversely placing the tomato seeds on a culture dish, sealing the tomato seeds by using sealing films, transferring the tomato seeds to an illumination culture room, vertically culturing the tomato seeds for 7-12 days in the dark at 25 ℃ for 12h, collecting tomato seedlings when obvious differences exist, measuring, photographing and counting fresh weight and root length.
Experimental results: plant-endophytic fungi interaction experiments were performed on 1/2MS solid medium using tomato and endophytic fungi CNBG-PGPF-2. After 12 days, as shown in fig. 6 and 7, after the CNBG-PGPF-2 strain is inoculated, the root system of the tomato seedlings is obviously developed, the fresh weight and the root length are obviously higher than those of a control group, the fresh weight and the root length are respectively improved by 175.97 percent and 44.01 percent, and the effect of promoting the growth is obvious.
The hypha of CNBG-PGPF-2 was observed by an inverted microscope to closely contact with the root of tomato seedlings, and the hypha was wound around the surface of tomato root and colonized inside the root (FIG. 8), indicating that CNBG-PGPF-2 can successfully establish an interaction relationship with tomato.
EXAMPLE 5 potted plant growth-promoting test of CNBG-PGPF-2 Strain
(1) Tomato (variety: red tomato) grows seedlings in advance, seedlings with the same size are selected after one week and transplanted into flowerpots (10 cm multiplied by 10 cm) containing nutrient soil, one seedling is transplanted into one flowerpot, 5 flowerpots are placed into each tray to serve as 5 repetitions, and two trays are arranged for 10 repetitions in total for each treatment of tomato plants.
(2) Fresh cultured endophytic fungi (3-5 mycelium blocks) were obtained with a 5mm punch, the mycelium blocks having a volume of about 60mm 3 The fermentation broth was collected after being placed in 200mL of PDB medium and cultured in the dark at 160rpm at 25℃for 7 days. Filtering the fermentation broth with gauze, diluting with sterile distilled water to 400mL to obtain fermentation broth filtrate with spore concentration of 1×10 5 And each mL. The tomato is treated with four treatments, respectively sterile distilled water, 10% filtrate (spore concentration 1×10) 4 individual/mL), 20% filtrate (spore concentration 2X 10) 4 individual/mL), 50% filtrate spores (concentration 5X 10) 4 Each per mL), 40mL of solution was irrigated along the main root of the plant for each pot, 200mL of solution was required for each tray, and 0mL, 400mL x 10% = 40mL, 400mL x 20% = 80mL, 400mL x 50% = 200mL broth filtrate was required for each of the four treatments.
PDB culture medium formula: 3g of potato soaked powder, 20g of glucose, 1000mL of deionized water and high-pressure steam sterilization at 121 ℃ for 20min.
(3) Root irrigation treatment is carried out after tomato transplanting is carried out for one week, then treatment is carried out for 10 days, and the steps are repeated for three times, and plant height and other data are measured in the period; plant materials are collected when growth potential difference is visible between treatments, and data such as root length, fresh weight and the like are measured. After the plants are cleaned, the plants are deactivated for 30min at 105 ℃, and then are baked to constant weight in an oven at 80 ℃, and the data such as dry weight, nutrient element content and the like are measured.
Experimental results: as shown in fig. 9, the treatment of 10% CNBG-PGPF-2 fermentation broth filtrate increased the plant height growth rate of tomato plants by 23.52% compared to the control group; the treatment of 10% CNBG-PGPF-2 fermentation broth filtrate significantly increased the dry weight of the aerial parts of tomato plants, 36.70% compared to the control group. The 20% CNBG-PGPF-2 fermentation liquor filtrate treatment obviously improves the root length of tomato plants, and is improved by 19.73% compared with a control group; treatment with 10%, 20% and 50% CNBG-PGPF-2 broth filtrate significantly increased the dry weight of the underground part of the tomato plant by 69.77%, 67.44% and 76.74% respectively compared to the control group.
According to measurement, the soluble protein content of tomato plants is improved by 56.65% compared with a control group by treating 20% CNBG-PGPF-2 fermentation broth filtrate; treatment with 10%, 20% and 50% CNBG-PGPF-2 broth filtrate significantly increased the soluble sugar content of tomato plants, 144.26%, 156.96% and 83.48% respectively, compared to the control group (fig. 10).
Claims (10)
1. Endophyte strainParaphaeosphaeria sardoa) CNBG-PGPF-2, characterized in that the endophyte is [ ] of the plantParaphaeosphaeria sardoa) CNBG-PGPF-2 was deposited at the China general microbiological culture Collection center, with accession number: CGMCC No.40636.
2. A fermentation broth filtrate comprising the endophyte of claim 1Paraphaeosphaeria sardoa）CNBG-PGPF-2。
3. A process for the preparation of a fermentation broth filtrate as claimed in claim 2, comprising the steps of: obtaining fresh cultured endophytic fungus mycelium blocks by using a puncher, putting the mycelium blocks into a PDB (PDB) culture medium, culturing in the dark for 7-10 days, collecting fermentation liquor, filtering the fermentation liquor by using gauze, and diluting the fermentation liquor by using sterile distilled water to obtain fermentation liquor filtrate.
4. A method of preparing a fermentation broth filtrate according to claim 3, wherein the PDB medium comprises the following composition: every 1000 parts of mL deionized water contains 3-g parts of potato soaked powder, 20g of glucose and is sterilized by high-pressure steam at 121 ℃ for 20min.
5. A microbial preparation comprising the endophyte of claim 1Paraphaeosphaeria sardoa) CNBG-PGPF-2 or a broth filtrate comprising the composition of claim 2.
6. The endophyte of claim 1Paraphaeosphaeria sardoa) Use of CNBG-PGPF-2, the broth filtrate of claim 2, or the microbial preparation of claim 5 for promoting plant growth.
7. The use according to claim 6, wherein the endophytic fungi in the filtrate of the fermentation brothParaphaeosphaeria sardoa) The concentration of CNBG-PGPF-2 spores was 5×10 4 Per mL to 2X 10 5 And each mL.
8. The use according to claim 6, wherein the plant comprises one or both of arabidopsis thaliana and tomato.
9. The application according to claim 6, characterized in that it comprises in particular: plant and plant endophytic fungi are treatedParaphaeosphaeria sardoa) CNBG-PGPF-2 was co-cultured with plant-endophytic fungi on solid medium.
10. The use according to claim 9, wherein the solid medium comprises MS medium.
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