CN117683644B - 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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Abstract
The invention discloses a plant endophytic fungus (Paraphaeosphaeria sardoa) and application thereof in plant growth promotion, and 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 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 improved by 36.70 percent compared with that of a control group, the dry weight of the underground part is improved by 76.74 percent, the content of soluble protein is improved by 56.65 percent, and the content of soluble sugar is improved 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.
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
The endophytic fungi (endophytic fungi) are fungi which are parasitic in the plant body, participate in the whole life history of the host plant and have no obvious harm to the host plant. 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.
Fungi Paraphaosphaeria are a new genus found in recent years and have no exact chinese name. Studies have shown that Paraphaosphaeria genus fungi can improve plant disease resistance, and studies on whether they can promote plant growth have not been reported yet.
Disclosure of Invention
The invention aims to: in order to solve the defects in the prior art, the technical problem to be solved by the invention is to separate a plant endophyte (Paraphaeosphaeria sardoa) capable of promoting plant growth from the root system of the apocarya, develop the research on the plant growth promoting effect, and provide a foundation for the subsequent deep development of microbial fertilizers and the reduction of the use 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 month 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 the aspect of promoting plant growth.
Wherein the concentration of the endophytic fungi (Paraphaeosphaeria sardoa) CNBG-PGPF-2 spores in the fermentation broth filtrate is 5 multiplied by 10 4 to 2 multiplied by 10 5 spores per 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-cultivated on a 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 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 improved by 36.70 percent compared with that of a control group, the dry weight of the underground part is improved by 76.74 percent, the content of soluble protein is improved by 56.65 percent, and the content of soluble sugar is improved 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 the colony morphology of CNBG-PGPF-2 strain on maize meal medium (COM), with the left side of the figure being front side and the right side being back side.
FIG. 2 shows the mycelium morphology of CNBG-PGPF-2 strain on maize meal medium (COM).
FIG. 3 shows spore morphology of CNBG-PGPF-2 strain in maize meal medium (COM).
FIG. 4 is a phylogenetic tree of CNBG-PGPF-2 strains.
FIG. 5 shows the pro-active effect of CNBG-PGPF-2 strain 1/2MS medium co-cultured with Arabidopsis thaliana, with the top panel on the front side and the bottom panel on the back side (Control group, CNBG-PGPF-2 treatment group).
FIG. 6 shows the pro-active effect of CNBG-PGPF-2 strain 1/2MS medium co-cultured with tomato (Control as Control group, CNBG-PGPF-2 as treatment group).
FIG. 7 shows the effect of CNBG-PGPF-2 strain 1/2MS medium co-culture with tomato on tomato seedling growth index, wherein the left side of the figure is tomato seedling fresh weight data, and the right side is tomato seedling root length data (Control is Control group, CNBG-PGPF-2 is treatment group).
FIG. 8 shows the colonization of CNBG-PGPF-2 strain on tomato root system (Control, treatment CNBG-PGPF-2).
FIG. 9 shows the effect of CNBG-PGPF-2 strain pot growth promoting test on tomato seedling growth index, wherein the upper left part of the graph shows tomato seedling height growth rate data, the upper right part shows tomato seedling aerial part dry weight data, the lower left part shows tomato seedling root length data, and the lower right part shows tomato seedling underground part dry weight data (Control is Control group, CNBG-PGPF-2.10%, CNBG-PGPF-2.20%, CNBG-PGPF-2.50% is treatment group).
FIG. 10 shows the effect of CNBG-PGPF-2 strain pot growth-promoting test on tomato seedling physiological index, wherein the left side of the figure shows tomato seedling soluble protein content data, and the right side shows tomato seedling soluble sugar content data (Control is Control group, CNBG-PGPF-2%, CNBG-PGPF-2% 20%, 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 endophytic fungi CNBG-PGPF-2
Taking root system of strong plant at planting base (Jurong, jiangsu) of apocarya, 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: CNBG-PGPF-2 after 5 days of culture on solid corn meal medium (COM), the colonies were 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 identification 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 μ LEasyTaq Mix, 2 μL forward primer ITS1, 2 μL reverse primer ITS4, 3 μL DNA template, 17 μL 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 result 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, by constructing a phylogenetic tree using MEGA 7 software and using the maximum likelihood method (Maximum Likelihood methods), it was found that the strain was clustered with a plurality of Paraphaeosphaeria fungi, which were 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: north Star Xilu No.1, 3 of the Chaoyang district of Beijing, the preservation date is 2023, 5 and 22 days, the preservation number is CGMCC No.40636, and the classification is Paraphaeosphaeria sardoa.
EXAMPLE 3CNBG-PGPF-2 Strain 1/2MS Medium Co-culture experiments with 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 mycelium block (right side) with the thickness of CNBG-PGPF-2, 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 the arabidopsis seedlings of the control group, especially the number of lateral roots of the arabidopsis seedlings was significantly higher than that of the control group (fig. 5).
EXAMPLE 4CNBG-PGPF-2 Strain 1/2MS Medium Co-culture experiments with 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 mycelium block (right side) with the thickness of CNBG-PGPF-2, 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 CNBG-PGPF-2 strains are 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 mycelia of CNBG-PGPF-2 were found to be in close contact with the roots of tomato seedlings by observation with an inverted microscope, and the mycelia were entangled on the surface of the tomato roots and colonized inside the roots (FIG. 8), indicating that CNBG-PGPF-2 can successfully establish an interaction relationship with tomatoes.
Example 5CNBG-PGPF-2 potted plant growth promoting test
(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 perforator, the mycelium blocks had a volume of about 60mm 3, and the mycelium blocks were placed in 200mL PDB medium and cultured in the dark at 25℃at 160rpm for 7 days, after which the fermentation broth was collected. The fermentation broth was first filtered with gauze and then diluted one time to 400mL with sterile distilled water as the fermentation broth filtrate, with a spore concentration of 1 x 10 5/mL. Four treatments were set up for tomato, namely sterile distilled water, 10% filtrate (spore concentration 1×10 4/mL), 20% filtrate (spore concentration 2×10 4/mL), 50% filtrate spores (concentration 5×10 4/mL), 40mL solution was irrigated along the plant root for each pot during treatment, 200mL solution was required for each tray, and 0mL, 400ml×10% = 40mL, 400ml×20% = 80mL, 400ml×50% = 200mL broth filtrate were required for each treatment.
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, 10% CNBG-PGPF-2 of the filtrate treatment of the fermentation broth increases the plant height growth rate of tomato plants by 23.52% compared with the control group; 10% CNBG-PGPF-2 of the filtrate treatment of the fermentation broth significantly increased the dry weight of the aerial parts of tomato plants, 36.70% compared with the control group. 20% CNBG-PGPF-2 of the filtrate treatment of the fermentation liquor obviously improves the root length of tomato plants, and is improved by 19.73% compared with a control group; 10%, 20% and 50% CNBG-PGPF-2 of the broth filtrate treatments 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 treatment of 20% CNBG-PGPF-2 fermentation liquor filtrate improves the soluble protein content of tomato plants, and is improved by 56.65% compared with a control group; treatment with 10%, 20% and 50% CNBG-PGPF-2 broth filtrate significantly increased the soluble sugar content of tomato plants, as compared to control, 144.26%, 156.96% and 83.48%, respectively (fig. 10).
Claims (9)
1. A plant endophyte Paraphaeosphaeria sardoa CNBG-PGPF-2, which is characterized in that the plant endophyte Paraphaeosphaeria sardoa CNBG-PGPF-2 is deposited in the China general microbiological culture Collection center of the culture Collection center of microorganisms, with a deposit number of 2023, 5 months and 22 days: CGMCC No. 40636.
2. A fermentation broth filtrate, characterized in that it contains endophyte Paraphaeosphaeria sardoa CNBG-PGPF-2 according to claim 1.
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: each 1000 mL deionized water contains 3: 3 g potato extract powder, 20 g glucose, and 20: 20 min by high pressure steam sterilization at 121deg.C.
5. A microbial preparation comprising endophyte Paraphaeosphaeria sardoa CNBG-PGPF-2 according to claim 1 or comprising the fermentation broth filtrate according to claim 2.
6. Use of endophyte Paraphaeosphaeria sardoa CNBG-PGPF-2 of claim 1, a fermentation broth filtrate of claim 2 or a microbial preparation of claim 5 for promoting plant growth, said plant being arabidopsis thaliana or tomato.
7. The use according to claim 6, wherein the concentration of endophytic fungi Paraphaeosphaeria sardoa CNBG-PGPF-2 spores in the broth filtrate is 5 x 10 4/mL to 2 x 10 5/mL.
8. The application according to claim 6, characterized in that it comprises in particular: plant and plant endophytic fungi Paraphaeosphaeria sardoa CNBG-PGPF-2 were co-cultivated on a solid medium.
9. The use of claim 8, wherein the solid medium comprises MS medium.
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| CN114292769A (en) * | 2021-12-01 | 2022-04-08 | 陕西省生物农业研究所 | Saline-alkali-resistant tomato leaf endophyte, fermentation liquor, preparation method and application |
| CN116601289A (en) * | 2020-12-16 | 2023-08-15 | 科莱恩产品(德国)公司 | Method for producing filamentous fungal whole broth enzyme compositions with low biomass formation and high protein yields |
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| CN116601289A (en) * | 2020-12-16 | 2023-08-15 | 科莱恩产品(德国)公司 | Method for producing filamentous fungal whole broth enzyme compositions with low biomass formation and high protein yields |
| CN114292769A (en) * | 2021-12-01 | 2022-04-08 | 陕西省生物农业研究所 | Saline-alkali-resistant tomato leaf endophyte, fermentation liquor, preparation method and application |
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