Use of a salt-tolerant branch of trichoderma in promoting plant growth
By synthesizing plant hormones using salt-tolerant Cladosporium 550W1, plant growth can be promoted, solving the problem of difficult plant growth in harsh environments such as saline-alkali land, and achieving efficient and environmentally friendly agricultural production increase and soil improvement.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- 江西省 中国科学院庐山植物园
- Filing Date
- 2025-09-25
- Publication Date
- 2026-05-15
AI Technical Summary
Existing technologies lack effective environmentally friendly microbial fertilizers, making it difficult to promote plant growth in harsh environments such as saline-alkali land, and traditional chemical fertilizers are harmful to the environment.
Using the salt-tolerant Cladosporium halotolerans 550W1 strain, this microbial preparation promotes plant growth by synthesizing plant hormones such as auxin (IAA), gibberellin (GA), and cytokinin (CTK), and can grow under a wide range of temperature and pH conditions, providing an environmentally friendly microbial preparation.
It significantly increases crop yield, promotes robust plant growth, reduces reliance on chemical fertilizers, improves soil health, and aligns with the development direction of green agriculture.
Smart Images
Figure CN121160476B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of microbial technology, specifically to the application of salt-tolerant Cladosporium in promoting plant growth. Background Technology
[0002] Plant growth-promoting microorganisms (PGPMs) are a group of microorganisms that interact with plants to promote plant growth and enhance stress resistance through various mechanisms. PGPMs improve soil physicochemical properties, increase soil fertility, enhance plant stress resistance, increase crop yield, and improve crop quality. In recent years, green, sustainable, and eco-friendly agriculture has gained increasing attention, and PGPMs, as microbial fertilizers, have broad application prospects. Therefore, developing environmentally friendly and sustainable agricultural microbial technologies is of great significance. Summary of the Invention
[0003] The purpose of this invention is to provide an application of salt-tolerant Cladosporium in promoting plant growth, so as to solve the problems mentioned in the background art.
[0004] To achieve the above objectives, the present invention provides the following technical solution: the application of salt-tolerant Cladosporium or its mycelial solution in promoting plant growth.
[0005] Furthermore, the salt-tolerant Cladosporium is a salt-tolerant Cladosporium ( Cladosporium halotolerans 550W1, deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 41943, deposited on May 20, 2025, at the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.
[0006] Furthermore, the culture conditions for the salt-tolerant Cladosporium are: temperature of 20-37℃ and pH of 4-11.
[0007] Furthermore, the plant is a plant of the Solanaceae family or the Brassicaceae family.
[0008] Furthermore, the Solanaceae plant in question is the potato.
[0009] Furthermore, the cruciferous plant in question is Arabidopsis thaliana.
[0010] Furthermore, the salt-tolerant Cladosporium promotes plant growth by synthesizing plant hormones.
[0011] Furthermore, the plant hormones include auxins, gibberellins, and cytokinins.
[0012] Another object of the present invention is to provide a microbial preparation for promoting plant growth, comprising an agriculturally acceptable carrier and the above-mentioned salt-tolerant Cladosporium or its inoculum.
[0013] The salt-tolerant Cladosporium 550W1 strain provided by this invention can promote plant growth by synthesizing plant hormones such as auxin, gibberellin and cytokinin. At the same time, the salt-tolerant Cladosporium 550W1 has outstanding advantages such as strong environmental adaptability, eco-friendliness and high safety, providing an excellent core strain resource for the development of new microbial fertilizers and the promotion of green and sustainable agriculture. Attached Figure Description
[0014] Figure 1 Phylogenetic tree of strain 550W1.
[0015] Figure 2 The figure shows the results of the potato growth promotion experiment. In the figure, A is the phenotype of the control group (CK) and the test-tube seedlings inoculated with salt-tolerant Cladosporium 550W1, and B is a comparison of potato yield between the control group (CK) and the potato seedlings inoculated with salt-tolerant Cladosporium 550W1.
[0016] Figure 3 Phenotypic characteristics of Arabidopsis plants inoculated with salt-tolerant Cladosporium 550W1, representing the control group (CK) and the control group (CK).
[0017] Figure 4 Figure 1 shows the experimental results of verifying the mechanism of action of salt-tolerant Cladosporium 550W1 in promoting plant growth; in the figure, A is the experimental results of phosphorus solubilization, potassium solubilization, and calcium solubilization, and B is the experimental results of hormone synthesis. Detailed Implementation
[0018] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.
[0019] In one embodiment of the present invention, a strain 550W1 is provided, which was isolated from Lushan hemp seed potato. No more than 50 mg of 550W1 mycelium was scraped under aseptic conditions, placed in a 2 mL sterile centrifuge tube, and rapidly ground into powder in a liquid nitrogen bath. Fungal DNA was extracted using a BioFlux fungal genomic DNA extraction kit according to the kit's instructions. The extracted total DNA from the strain was used as a template to amplify the ITS sequence. PCR amplification was performed using universal fungal primers ITS1 and ITS4.
[0020] The nucleotide sequence of primer ITS1 is 5'-TCCGYTAGGTGAACCTGCGG-3' (as shown in SEQ ID NO.1 in the sequence listing); the nucleotide sequence of primer ITS4 is 5'-TCCTCCGCTTATTGATATGC-3' (as shown in SEQ ID NO.2 in the sequence listing).
[0021] The PCR reaction system consisted of: 2 µL template DNA, 1 µL primer ITS1, 1 µL primer ITS4, 15 µL 2×SuperTaq PCRStarMix (Dye), and ddH2O to a final volume of 30 µL.
[0022] The PCR reaction conditions were: 95℃, 5 min; 95℃, 30 s, 55℃, 30 s, 72℃, 1 min, 30 cycles; 72℃, 5 min.
[0023] The PCR products obtained from PCR amplification were detected by electrophoresis on a 1.0% agarose gel, and the amplified products were sequenced. The sequencing results are shown in SEQ ID NO.3 of the sequence listing. The sequencing results were compared with the NCBI database, and phylogenetic analysis was performed with ITS sequences that showed high similarity, such as... Figure 1 As shown, strain 550W1 is related to salt-tolerant cladosporium ( Cladosporium halotolerans It is most similar to , so it is named Salt-tolerant Cladosporium ( Cladosporium halotolerans )550W1, and deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC No.41943, deposited on May 20, 2025, at No.3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences.
[0024] In another embodiment of the present invention, the application of the above-mentioned salt-tolerant Cladosporium 550W1 or its bacterial solution in promoting plant growth is also provided; the salt-tolerant Cladosporium 550W1 has plant growth-promoting effects and can synthesize plant hormones such as auxin (IAA), gibberellin (GA) and cytokinin (CTK).
[0025] In practical applications, the culture conditions for the aforementioned salt-tolerant Cladosporium 550W1 are as follows: normal growth is achieved at temperatures of 20-37℃ and pH values of 4-11 on MS or PDA medium. The aforementioned plants are either Solanaceae or Brassicaceae plants; preferably, the Solanaceae plant is potato; and the Brassicaceae plant is Arabidopsis thaliana.
[0026] In another embodiment of the present invention, a microbial preparation for promoting plant growth is also provided, comprising an agriculturally acceptable carrier and the aforementioned salt-tolerant Cladosporium 550W1 or its bacterial suspension. It should be noted that an agriculturally acceptable carrier includes, but is not limited to, water, buffer solutions, or culture media.
[0027] In this embodiment of the invention, the salt-tolerant Cladosporium 550W1 can promote plant growth by synthesizing plant hormones such as auxin, gibberellin and cytokinin; at the same time, the salt-tolerant Cladosporium 550W1 has outstanding advantages such as strong environmental adaptability, eco-friendliness and high safety, providing excellent core strain resources for the development of new microbial fertilizers and the promotion of green and sustainable agriculture.
[0028] The following embodiments are implementation examples of the technical solution of the present invention in practical applications, but are not limited thereto. Unless otherwise specified, all materials and reagents involved are commercially available products; unless otherwise specified, all experimental methods used are conventional methods.
[0029] Example 1: This example provides an experiment on the growth-promoting effect of halophilic cephalosporin 550W1 on potatoes, as detailed below:
[0030] S1. Preparation of salt-tolerant Cladosporium 550W1 bacterial suspension: The salt-tolerant Cladosporium 550W1 bacterial suspension, which has been cultured in PDB medium at 28℃ and 200rpm in the dark for three days, is centrifuged at 20℃ and 5000rpm for 10min. The supernatant is removed, and the suspension is washed and resuspended with PBS buffer to obtain the salt-tolerant Cladosporium 550W1 bacterial suspension for inoculation. The PBS buffer has a concentration of 0.01M and a pH of 7.4. It is prepared as follows: Dissolve 8g of NaCl, 0.2g of KCl, 1.44g of Na2HPO4 and 0.24g of KH2PO4 in deionized water, and bring the volume up to 1L with deionized water to obtain the PBS buffer.
[0031] S2. Potato plantlets were inoculated with the above-mentioned salt-tolerant Cladosporium 550W1 bacterial suspension, and a control group (CK) with only PBS buffer was added. The results are as follows: Figure 2 As shown in the figure; it can be seen from the figure that the leaves of the test-tube seedlings inoculated with salt-tolerant Cladosporium 550W1 are significantly greener and the growth is more vigorous than that of the control group (e.g. Figure 2(As shown in A). Additionally, in an outdoor experiment, inoculating the roots of 10 potato plants with a salt-tolerant Cladosporium 550W1 bacterial solution resulted in a 20.6% increase in yield compared to uninoculated plants (e.g., ...). Figure 2 (As shown in B).
[0032] Example 2: This example provides a growth-promoting experiment of halophilic cladosporium 550W1 on the model plant Arabidopsis thaliana, as detailed below:
[0033] S1. Preparation of salt-tolerant Cladosporium 550W1 bacterial suspension: The salt-tolerant Cladosporium 550W1 bacterial suspension, which has been cultured in PDB medium at 28℃ and 200rpm in the dark for three days, is centrifuged at 20℃ and 5000rpm for 10min. The supernatant is removed, and the suspension is washed and resuspended with PBS buffer to obtain the salt-tolerant Cladosporium 550W1 bacterial suspension for inoculation. The PBS buffer has a concentration of 0.01M and a pH of 7.4. It is prepared as follows: Dissolve 8g of NaCl, 0.2g of KCl, 1.44g of Na2HPO4 and 0.24g of KH2PO4 in deionized water, and bring the volume up to 1L with deionized water to obtain the PBS buffer.
[0034] S2. After surface disinfection, Arabidopsis thaliana seeds were sown on MS plates under sterile conditions. After 4 leaves emerged (about 10 days), they were transplanted into the substrate and covered with plastic wrap to keep them moist for 3 days. After 14 days, they were inoculated. 3 mL of the above-mentioned salt-tolerant Cladosporium 550W1 bacterial solution was added to each pot in the experimental group, and 3 mL of PBS solution was added to each pot in the control group (CK).
[0035] Experimental results are as follows Figure 3 As shown in the figure, the plants inoculated with salt-tolerant Cladosporium 550W1 grew significantly better than the uninoculated control group, indicating that salt-tolerant Cladosporium 550W1 can promote the growth of the model plant Arabidopsis thaliana.
[0036] Example 3: This example provides an experimental verification of the mechanism of action of salt-tolerant Cladosporium 550W1 in promoting plant growth, as detailed below:
[0037] Phosphorus, potassium, and calcium solubilization experiments: *Cladosporium halophilum* 550W1 was cultured in existing potassium-solubilizing liquid medium + agar, existing modified inorganic phosphorus liquid medium + agar, and existing PDA medium + 20% CaCO3, respectively, to analyze its phosphorus, potassium, and calcium solubilization abilities; the results are as follows: Figure 4 As shown in Figure A, it can be seen from the figure that *Cladosporium halophilum* 550W1 does not have the ability to solubilize phosphorus, potassium, or calcium.
[0038] Synthetic hormone experiment: Salt-tolerant Cladosporium 550W1 was placed in PDB medium and cultured by shaking at 28℃ and 200 rpm in the dark. Plant hormones were then measured. The method for determining plant hormones was as follows: a single colony of Salt-tolerant Cladosporium 550W1 cultured in PDB medium at 28℃ and 200 rpm in the dark was used as a sample. The concentration of plant hormones was determined using a commercially available enzyme-linked immunosorbent assay (ELISA) kit, following the kit's instructions. The results are as follows: Figure 4 As shown in Figure B, it can be seen from the figure that *Cladosporium halophilum* 550W1 can simultaneously synthesize plant hormones such as IAA, GA, and CTK; the kit numbers used for the determination of each type of plant hormone are shown in Table 1 below.
[0039] Table 1
[0040]
[0041] In summary, the salt-tolerant Cladosporium 550W1 provided in the embodiments of the present invention has the following beneficial technical effects:
[0042] 1. Significantly increases crop yield: Inoculation with this salt-tolerant Cladosporium 550W1 can increase the yield of crops such as potatoes by 20.6%, which is of great significance for ensuring food security and improving agricultural production efficiency.
[0043] 2. Promote plant vegetative growth: Potato test-tube seedlings and Arabidopsis plants inoculated with salt-tolerant Cladosporium 550W1 showed significant advantages in morphological indicators such as plant height, stem diameter, and number of leaves. The plants were more robust and lush, and the leaves were greener.
[0044] 3. Salt-tolerant Cladosporium 550W1 can simultaneously synthesize three key plant hormones:
[0045] IAA: Directly promotes cell elongation and root development, and enhances the plant's ability to absorb water and nutrients;
[0046] CTK: Promotes cell division, delays leaf senescence, and maintains vigorous physiological activity of the plant;
[0047] GA: Promotes stem elongation, breaks dormancy, and accelerates the overall growth process;
[0048] This multi-hormone synergistic mechanism can regulate plant growth and development in all aspects, with a more stable and comprehensive effect than a single hormone mechanism.
[0049] 4. Wide environmental adaptability: Salt-tolerant Cladosporium 550W1 can grow normally in a wide range of acidity and alkalinity conditions, ranging from 20-37℃ to pH 4-11. This means that it has good colonization and survival potential in different seasons and soils of different properties in most regions, and its application range is wide.
[0050] 5. Reduce dependence on chemical fertilizers: This salt-tolerant Cladosporium 550W1 directly stimulates plant growth through hormones, rather than through traditional microbial fertilizer mechanisms such as phosphorus, potassium, and calcium solubilization. This provides a new, alternative, or complementary path for increasing agricultural production, which helps reduce the amount of chemical fertilizers used and is in line with the development direction of green agriculture.
[0051] 6. Environmentally friendly and highly safe: As a naturally occurring fungus, halophilic 550W1 is safer than chemically synthesized agents and genetically modified crops, and has higher public acceptance. At the same time, the use of microbial agents helps to improve the soil microbial community, maintain soil health, and reduce non-point source pollution caused by fertilizer runoff.
[0052] Based on the above-described preferred embodiments of the present invention, and through the foregoing description, those skilled in the art can make various changes and modifications without departing from the inventive concept. The technical scope of this invention is not limited to the contents of the specification.
Claims
1. The application of a salt-tolerant Cladosporium or its mycelial solution in promoting plant growth, characterized in that, The salt-tolerant Cladosporium is a salt-tolerant Cladosporium ( Cladosporium halotolerans 550W1 is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC No. 41943.
2. The application according to claim 1, characterized in that, The culture conditions for the salt-tolerant Cladosporium are: temperature 20-37℃, pH 4-11.
3. The application according to any one of claims 1-2, characterized in that, The plant in question is either a member of the Solanaceae family or a member of the Brassicaceae family.
4. The application according to claim 3, characterized in that, The plant in question belongs to the Solanaceae family and is the potato.
5. The application according to claim 3, characterized in that, The cruciferous plant in question is Arabidopsis thaliana.
6. The application according to any one of claims 1-2, characterized in that, The salt-tolerant Cladosporium promotes plant growth by synthesizing plant hormones, including auxins, gibberellins, and cytokinins.
7. A microbial preparation for promoting plant growth, comprising an agriculturally acceptable carrier, characterized in that, It also includes salt-tolerant Cladosporium or its culture; the salt-tolerant Cladosporium is a salt-tolerant Cladosporium ( Cladosporium halotolerans 550W1 is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC No. 41943.