Mucor circinelloides em-27, microbial inoculum and application
By using the Mucor EM-27 strain and its microbial inoculant, the problem of soil quality decline caused by chemical fertilizers was solved, wheat growth was significantly promoted, and plant growth was significantly improved.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- CHINA AGRI UNIV
- Filing Date
- 2025-06-11
- Publication Date
- 2026-06-19
Smart Images
Figure CN120555205B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of microbial application technology, and in particular to an elegant strain of Mucor EM-27, a microbial agent, and its application. Background Technology
[0002] The application of chemical fertilizers has effectively ensured grain yield, but excessive or unbalanced application of fertilizers has led to unfavorable developments in the physical, chemical, and biological indicators of the soil. This is mainly due to the reduction of soil organic matter content, the deterioration of physical and chemical properties, and changes in soil microbial activity and community structure, which in turn affect the soil's ability to supply and retain fertilizer and its mineralization and decomposition of soil nutrients.
[0003] Microbial inoculants can not only transform insoluble phosphorus and potassium compounds in the soil, but also enhance crop resistance to prevent and control pests and diseases, significantly promoting crop growth. Increased microbial reproduction, metabolism, and enzyme activity promote the mineralization and decomposition of soil organic matter and the release of nutrients, providing a favorable root zone environment for crop growth and improving crop quality.
[0004] *Actinomucor elegans* is a filamentous fungus belonging to the phylum Mucoromycota, class Mucoromycetes, order Mucorales, family Mucoraceae, and genus *Actinomucor*. Its main characteristics include a colony morphology similar to *Mucor*, cottony in appearance, not exceeding 1 cm in height, initially white or light orange-yellow, later turning dark bluish-brown-yellow. It possesses creeping hyphae and rhizoids. The sporangiophores are erect, with branches concentrated at the apex. The main branch has a large sporangium at its tip, below which are 3-8 whorled branches, each branch apex bearing a sporangium. The main sporangiophore is approximately 30 micrometers in diameter, and each branch typically has a transverse septum. The sporangia are spherical, generally no more than 80 micrometers in diameter on the main branch, but sometimes reaching 120 micrometers, while those on branches are 20-50 micrometers in diameter. After maturation, the sporangium walls dissolve or crack, leaving behind a collar. The sporangium axis is inconsistent, appearing as an oval to pear shape within larger sporangia, measuring 50-60 × 30-40 μm. *Mucor davidii* is widely distributed in nature, found in soil and on plant roots. Currently, *Mucor davidii* is mainly used in cheese fermentation and is one of the main *Mucor* species used in fermented bean curd production in my country. There are currently no reports of *Mucor davidii* being used to promote plant growth. Summary of the Invention
[0005] The purpose of this invention is to provide an elegant *Mucor radiata* strain EM-27, a microbial agent, and its application, in order to solve the problems existing in the prior art.
[0006] To achieve the above objectives, the present invention provides the following solution:
[0007] One of the technical solutions of this invention is a strain of Actinomucor elegans EM-27, which was deposited on April 24, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC NO.41924.
[0008] The second technical solution of the present invention is a microbial inoculant, including the aforementioned *Mucor radiata* EM-27.
[0009] The third technical solution of the present invention is the application of the *Rhizopus radiata* EM-27 or the microbial agent in the preparation of products that promote plant growth.
[0010] The fourth technical solution of the present invention is a product that promotes plant growth, comprising the aforementioned *Mucor radiata* EM-27 or the aforementioned microbial agent; the plant comprises wheat.
[0011] The fifth technical solution of the present invention is the application of the *Rhizopus radiata* EM-27 or the microbial agent in the preparation of products that antagonize bacteria.
[0012] The sixth technical solution of the present invention is a product for antagonizing bacteria, comprising the aforementioned *Rhizopus radiata* EM-27 or the aforementioned microbial agent; the bacteria include *Ferrovibrio*, *Ectopseudomonas*, *Pseudomonas*, *Kribbella*, *Ochrobactrum*, *Bosea*, *Bradyrhizobium*, *Niabella*, *Agromyces*, *Rheinheimera*, *Variovorax*, *Microbacterium*, *Protaetiibacter*, *Arthrobacter*, *Agrobacterium*, *Cellulosimicrobium*, *Ramlibacter*, *Streptomyces*, *Brevibacillus*, *Polaromonas*, *Actinotalea*, and *Nocardioides*.
[0013] Based on the above technical solution, the present invention has the following technical effects:
[0014] This invention isolated a strain of *Rhizopus yazhi* from the rhizosphere of wheat. Antagonistic experiments revealed that 8.7% of rhizosphere bacteria exhibited antagonistic effects against it, and 91.3% of rhizosphere bacteria could coexist with *Rhizopus yazhi*. Furthermore, application of *Rhizopus yazhi* significantly promoted wheat growth and development. Therefore, *Rhizopus yazhi* shows promising application prospects in improving plant growth and development. Attached Figure Description
[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0016] Figure 1 This is a colony morphology diagram of *Mucor radiata* EM-27.
[0017] Figure 2 To construct a phylogenetic tree based on ITS gene sequences.
[0018] Figure 3 This is a diagram showing the antagonistic relationship between strain EM-27 and rhizosphere bacteria.
[0019] Figure 4 The EM-27 strain promotes wheat plant height.
[0020] Figure 5 To promote wheat dry weight using strain EM-27. Detailed Implementation
[0021] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.
[0022] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.
[0023] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.
[0024] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be obvious to those skilled in the art. This application specification and embodiments are merely exemplary.
[0025] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.
[0026] Unless otherwise specified, the technical solutions described in this invention are all conventional solutions in the field, and the reagents or raw materials used are all purchased from commercial channels or are publicly available unless otherwise specified.
[0027] This invention provides a strain of Actinomucor elegans EM-27, which was deposited on April 24, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC NO.41924.
[0028] Embodiments of the present invention also provide microbial agents, including the aforementioned *Mucor radiata* EM-27.
[0029] This invention also provides the application of the *Mucor radiata* EM-27 or the microbial agent in the preparation of products that promote plant growth.
[0030] In some specific implementations, the plant includes wheat.
[0031] This invention also provides a product for promoting plant growth, comprising the aforementioned *Mucor radiata* EM-27 or the aforementioned microbial inoculant; the plant comprising wheat.
[0032] This invention also provides the application of the *Mucor radiata* EM-27 or the microbial agent in the preparation of products antagonistic to bacteria.
[0033] In some specific implementations, the bacteria include Ferrovibrio, Ectopseudomonas, Pseudomonas, Kribbella, Ochrobactrum, Bosea, Bradyrhizobium, Niabella, Agromyces, Rheinheimera, Variovorax, Microbacterium, Protaetiibacter, Arthrobacter, Agrobacterium, Cellulosimicrobium, Ramlibacter, Streptomyces, Brevibacillus, Polaromonas, Actinotalea, and Nocardioides.
[0034] This invention also provides a product for antagonizing bacteria, comprising the aforementioned *Rhizopus radiata* EM-27 or the aforementioned microbial agent; the bacteria include *Ferrovibrio*, *Ectopseudomonas*, *Pseudomonas*, *Kribbella*, *Ochrobactrum*, *Bosea*, *Bradyrhizobium*, *Niabella*, *Agromyces*, *Rheinheimera*, *Variovorax*, *Microbacterium*, *Protaetiibacter*, *Arthrobacter*, *Agrobacterium*, *Cellulosimicrobium*, *Ramlibacter*, *Streptomyces*, *Brevibacillus*, *Polaromonas*, *Actinotalea*, and *Nocardioides*.
[0035] Example 1
[0036] Isolation, identification and preservation of strain EM-27
[0037] The *Actinomucor elegans* EM-27 strain described in this invention was isolated from the rhizosphere of potted wheat. The soil was collected from Quzhou County, Handan City, Hebei Province, and the wheat variety was Zhongmai 578. The wheat roots were completely removed, and the soil adhering to the roots was shaken off. The rhizosphere soil adhering to the root surface was washed with PBS buffer. A 10-fold serial dilution method was used: 1g of soil was weighed and poured into an Erlenmeyer flask containing 90mL of sterile water, thoroughly shaken to mix, and the supernatant was diluted to a concentration of 10. -3 10 -4 10 -5 10 -6Soil suspension. Fungal isolation was performed using the dilution plating method. 100 μL of the dilution was plated onto PDA medium (containing 1% Bengal red and 1% streptomycin). After incubation at 28°C for 3-5 days, colonies with different morphological characteristics were selected from the plates and purified repeatedly on PDA plates.
[0038] Fungal genomes were extracted using a fungal genome extraction kit. Using genomic DNA as a template, PCR amplification was performed using universal primers for fungal ITS sequences: upstream primer ITS1 (SEQ ID NO.1: 5'-TCC GTAGGT GAA CCT GCG G-3') and downstream primer ITS4 (SEQ ID NO.2: 5'-TCC TCC GCT TAT TGATAT GC-3').
[0039] PCR was performed in a 50 μL reaction system, specifically including: 2 μL template DNA, 2 μL upstream primer (0.25 μM), 2 μL downstream primer (0.25 μM), 25 μL PCR Master-Mix (2×), and 19 μL sterile water. The PCR reaction conditions were as follows: 95℃ for 5 min; 95℃ for 30 sec, 53℃ for 30 sec, 72℃ for 30 sec, for a total of 30 cycles; 72℃ for 10 min.
[0040] The amplification results were sequenced, and the nucleotide sequence of the ITS of strain EM-27 is shown in SEQ ID NO.3.
[0041] SEQ ID NO. 3: GATTATATAACTTGAGGGGAACTGGGCTTACGGGCTTGGTTTT.
[0042] Sequencing results were compared with BLAST in the GenBank nucleic acid database. The results showed that strain EM-27 had 98.81% homology with Actinomucor elegans. Figure 2 ).
[0043] Biological characteristics of the isolated strain EM-27 were observed. After culturing on PDA medium at 28℃ for 5 days, the colony characteristics were as follows: The colony morphology in PDA medium resembled *Mucor*, appearing cottony, initially white or light orange-yellow, later turning dark bluish-brown-yellow, with creeping hyphae and rhizoids. Figure 1 ).
[0044] Based on the above identification results, strain EM-27 was confirmed to be Actinomucor elegans. This strain was deposited on April 24, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC NO.41924.
[0045] Example 2
[0046] EM-27 bacterial antagonism test
[0047] The activated *Mucor radiata* EM-27 mycelial pellet was inoculated into the center of a PDA solid plate and incubated at 28°C for 5 days until the colonies covered the plate. Spores on the medium were washed with sterile water, and the spore suspension was collected. Spores were counted under a microscope using a hemocytometer, and the total concentration of the spore suspension was calculated. The suspension was then diluted to a concentration of 1×10⁻⁶. 7 Spore suspension. Inoculate 1% (v:v) of the spore suspension into melted PDA medium at approximately 50°C, mix well, and pour into plates. After the plates solidify, use sterile toothpicks to pick up colonies of 448 bacterial strains isolated from wheat rhizosphere and inoculate them onto plates containing *Rhizopus dauricum*. Each treatment is replicated three times. Incubate at 28°C for 3 days to observe the antagonistic effect of the bacteria on EM-27.
[0048] The results showed that all 448 bacterial strains isolated from the wheat rhizosphere could grow on plates containing strain EM-27, with significant inhibition zones observed in 39 of these strains. Figure 3 Of the 39 strains, 8.7% were antagonistic to EM-27.
[0049] Table 1. Antagonistic effects of rhizosphere bacteria and strain EM-27
[0050]
[0051] Example 3
[0052] Experiment on the growth-promoting effect of Elegant Mucor EM-27 in potted plants
[0053] Preparation method of Mucor EM-27 spore suspension: EM-27 strain was inoculated onto PDA solid medium and incubated in a 28℃ light incubator for 5 days. After incubation, approximately 5 ml of sterile water was added to the medium, and spores were scraped off using a spreader to obtain a spore suspension. The number of spores was counted using a hemocytometer, and the suspension was diluted to a concentration of 1×10⁻⁶. 7 CFU / mL spore suspension.
[0054] Soil was collected from Quzhou, Hebei Province, with 2 kg of sterilized soil per pot. Zhongmai 578 wheat was used in the experiment. Wheat seeds were disinfected by soaking in a 2% (m / v, g / mL) sodium hypochlorite solution for 10 minutes, then rinsed several times with sterile water, and spread evenly on sterile filter paper for germination. The wheat hypocotyl was exposed, and two seedlings of similar growth were sown in each pot. Two treatments were set up (three replicates per treatment): CK (no inoculation) and EM-27 (inoculated with *Mucor radiata* EM-27). When the wheat reached the seedling stage, 20 mL of spore suspension was applied to the roots. Plant growth indicators were measured two weeks later.
[0055] Table 2 Effects of strain EM-27 on wheat plant height and dry weight
[0056]
[0057] The results show that, from Figure 4 , Figure 5 It can be seen that inoculation with *Mucor radiata* EM-27 significantly increased wheat plant height and dry weight. Wheat plant height increased by 18.2%, and plant dry weight increased by 14.6%.
[0058] In summary, the *Mucor radiata* strain EM-27 provided by this invention can significantly promote the growth and development of wheat and has good application prospects.
[0059] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. For those skilled in the art, other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively describe all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.
Claims
1. An elegant strain of *Mucor* ( Actinomucor elegans EM-27, characterized in that, This strain was deposited on April 24, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC NO. 41924.
2. Microbial inoculant characterized in that, Includes the elegant radioactive mucor EM-27 as described in claim 1.
3. The application of the *Mucor radiata* EM-27 as described in claim 1 or the microbial inoculant as described in claim 2 in the preparation of products that promote plant growth, characterized in that... The plant in question is wheat.
4. A product that promotes plant growth, characterized in that, It includes the *Mucor radiata* EM-27 of claim 1 or the microbial inoculant of claim 2; the plant is wheat.
5. The application of the *Mucor radiata* EM-27 as described in claim 1 or the microbial agent as described in claim 2 in the preparation of products antagonistic to bacteria, characterized in that... The bacteria are selected from Ferrovibrio, Ectopseudomonas Pseudomonas, Kribbella, Ochrobactrum, Bosea, Bradyrhizobium, Niabella, Agromyces, Rheinheimera, Variovorax, Microbacterium, Protaetiibacter, Arthrobacter, Agrobacterium, Cellulosimicrobium, Ramlibacter, Streptomyces, Brevibacillus, Polaromonas, Actinotalea and Nocardioides At least one of them.
6. A product that antagonizes bacteria, characterized in that, Includes the *Mucor radiata* EM-27 of claim 1 or the microbial inoculant of claim 2; the bacteria are selected from... Ferrovibrio, Ectopseudomonas, Pseudomonas, Kribbella, Ochrobactrum, Bosea, Bradyrhizobium, Niabella, Agromyces, Rheinheimera, Variovorax, Microbacterium, Protaetiibacter, Arthrobacter, Agrobacterium, Cellulosimicrobium, Ramlibacter, Streptomyces, Brevibacillus, Polaromonas, Actinotalea and Nocardioides At least one of them.