Irregular mucor and use thereof

Abstract

The application discloses irregular mucor and application thereof, and the strain number of the irregular mucor (Mucor irregularis) Mucor irregularis ) is JSAFC 3408, which is preserved in the China General Microbiological Culture Collection Center on May 6, 2025, and the preservation number is CGMCC No.41982. The application obtains a kind of irregular mucor (Mucor irregularis) Mucor irregularis ) by separating and purifying soil around a pepper. The strain can inhibit the growth of C. globosum, promote the development of the pepper, regulate the growth of the plant, and has great significance for the development of a biocontrol agent for preventing and treating pepper root rot in the future.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology and relates to an irregular mold and its applications. Background Technology

[0002] Chili peppers, a widely cultivated economic crop and condiment globally, are extensively grown in both northern and southern my country. Their unique flavor and rich nutritional content make them an important category in agricultural production. In recent years, with increasing consumer demand, the scale of chili pepper cultivation has continued to expand, and intensive and large-scale planting models have gradually become popular. However, the accompanying disease problems have become increasingly prominent, among which root rot has become one of the main obstacles restricting the healthy development of the chili pepper industry. *Hylocereus huanghuaiense* (… Globisporangium huanghuaiense Different names for the same thing Pythium huanghuaiense This disease is one of the pathogens that causes root rot in chili peppers. In the early stages of the disease, the plant's roots turn brown and rot, significantly reducing their ability to absorb water and nutrients. This leads to symptoms such as yellowing, wilting, and drooping leaves. In severe cases, the entire plant withers and dies, resulting in reduced yield or even total crop failure. Therefore, developing a new biocontrol agent for the prevention and control of root rot in chili peppers is of paramount importance. Summary of the Invention

[0003] Purpose of the invention: The purpose of this invention is to provide an irregular mold ( Mucor irregularis ), and its application in inhibiting the growth of *Hylocereus huanghuaiense* and promoting the growth of chili peppers.

[0004] Technical solution: This invention provides an irregular mold ( Mucor irregularis The irregular mold ( Mucor irregularis The strain number is JSAFC 3408, which was deposited at the China General Microbiological Culture Collection Center on May 6, 2025, with accession number CGMCC No. 41982.

[0005] Furthermore, the nucleotide sequence of the ITS gene of the irregular mold is shown in SEQ ID NO.1.

[0006] The present invention also provides a biocontrol agent, wherein the active ingredient of the biocontrol agent is the aforementioned irregular mold (Mucor). Mucor irregularis ).

[0007] The present invention also provides the above-mentioned irregular mold ( Mucor irregularis Biocontrol agents inhibit the growth of *Cyclocarya paliurus* (Huanghuai) Globisporangium huanghuaiense Applications during growth.

[0008] The present invention also provides the above-mentioned irregular mold ( Mucor irregularis Application of biocontrol agents in promoting chili pepper growth.

[0009] Furthermore, the application specifically involves using irregularly shaped molds (Rhizopus) Mucor irregularis The spore suspension of JSAFC 3408 was used to drench the roots of peppers to promote plant height, leaf and root growth.

[0010] Further, the preparation method of the spore suspension is as follows: (1) Take the biocontrol fungus *Mucor irregularis* JSAFC 3408, and under aseptic conditions, pick up the hyphae with an inoculation needle and inoculate them into sterilized PDA medium for 5-7 days; (2) Under aseptic conditions, scrape off the hyphae, rinse the hyphae with sterile water and filter them with gauze; (3) Under aseptic conditions, adjust the spore concentration with sterile water.

[0011] Furthermore, the culture conditions described in step (1) are 25°C, 65% humidity, and dark culture.

[0012] Furthermore, the concentration of the spore suspension is 10. 5 / ml.

[0013] Furthermore, the spore suspension is applied at a rate of 5 mL / time / 2 days for 30 days.

[0014] Beneficial effects: Compared with the prior art, the present invention has the following outstanding advantages: This application obtained an irregular mold (Mucor) by isolating and purifying the soil around the chili pepper. Mucor irregularis This strain can inhibit the growth of *Hylocordia huanghuaiense* and promote pepper development, thus regulating plant growth. It is of great significance for the future development of biocontrol agents for the prevention and control of root rot. Attached Figure Description

[0015] Figure 1 : Mucor irregularis Phylogenetic tree of JSAFC 3408.

[0016] Figure 2 The biocontrol bacteria of the present invention Mucor irregularis JSAFC 3408 was used in confrontation culture with *Cytospora huanghuaiensis*.

[0017] Figure 3 The biocontrol bacteria of the present invention Mucor irregularis Effects of JSAFC 3408 on chili pepper root growth. Detailed Implementation

[0018] The technical solution of the present invention will be further described below with reference to the accompanying drawings.

[0019] In this invention, the term "biocontrol bacteria" refers to beneficial microorganisms that can prevent and control plant diseases, mainly including bacteria, fungi, and actinomycetes.

[0020] Example 1: Mucor irregularisIsolation and identification of JSAFC 3408

[0021] 1. Mucor irregularis Separation of JSAFC 3408

[0022] Soil from which the roots of chili peppers in Jurong were well grown was used to isolate JSAFC 3408. The colony characteristics are as follows: When cultured on PDA plates, the colonies are round, with abundant mycelial growth, and the mycelium is attached to the surface of the medium in a cotton-like manner.

[0023] 2. Mucor irregularis JSAFC 3408 Molecular Biological Identification

[0024] (1) Genomic DNA of strain JSAFC 3408 was extracted using a kit from TIANGEN.

[0025] (2) Amplify the ITS gene of genomic DNA.

[0026] DNA amplification was performed using a 30 μL reaction volume, containing 15 µL of 2×EasyTaq PCR SuperMix (+dye), 1 μL (10 μM) each of primer pairs ITS1 (SEQ ID NO.2: 5'-CTTGGTCATTTAGAGGAAGTAA-3') & ITS4 (SEQ ID NO.3: 5'-TCCTCCGCTTATTGATATGC-3'), 2 μL of template DNA, and 11 μL of ddH2O. The PCR amplification program conditions were as follows: 94℃ pre-denaturation for 5 min; 94℃ denaturation for 30 s, 54℃ annealing for 45 s, 72℃ extension for 60 s, for a total of 35 cycles; and a final extension at 72℃ for 10 min.

[0027] (3) After the PCR products obtained by amplification were subjected to 1% agarose gel electrophoresis, they were observed under ultraviolet light. The PCR products with target bands were sent to Nanjing Qingke Biotechnology Co., Ltd. for sequencing. The sequencing results were as follows: the ITS gene sequence of strain JSAFC 3408 was (SEQ ID NO.1:).

[0028] (4) Compare the SEQ ID NO.1 sequence with the NCBI database website and construct a phylogenetic tree, see [link to relevant documentation]. Figure 1 The comparison results are shown in Table 1. Based on the results in Table 1, it can be inferred that the biocontrol bacterium JSAFC 3408 of the present invention is... Mucor irregularis Named Mucor irregularis JSAFC 3408 was deposited on May 6, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 41982, and classified as *Mucor irregularis*. Mucor irregularis The depositary address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Beichen Courtyard 1, Chaoyang District, Beijing.

[0029] Table 1. Sequence alignment results of SEQ ID NO.1

[0030] gene strain base number similarity ITS IMCC2011 592 100%

[0031] Example 2: Mucor irregularis Inhibitory effect of JSAFC 3408 on Huanghuai coccidioidomyces

[0032] (1) On a clean bench, a 4 mm diameter mycelial disc of *Pythium huanghuaiense* sp. nov. isolated from soybean: morphology, molecular phylogeny and pathogenicity. *BiodiversityData Journal*, 9: e65227. was inoculated onto one side of a PDA medium plate. Mucor irregularis JSAFC 3408 mycelial discs with a diameter of 4 mm were inoculated onto the symmetrical side of PDA medium as the experimental group; and medium plates inoculated only with *Cyclocarya paliurus* were used as the control group.

[0033] (2) The experimental group and control group from step (1) were cultured upside down in a constant temperature incubator at 25°C. The culture conditions were 8 h of light followed by 16 h of dark culture.

[0034] (3) After the experimental group and control group culture media from step (2) were artificially cultured for 7 days under the above culture conditions, the colony radius of *Hylocereus huanghuaiense* was measured using the plate confrontation method, and the results were observed. Mucor irregularis The inhibitory effect of JSAFC 3408 on Huanghuai coccidioidomycetes.

[0035] The results are as follows Figure 2 As shown, the colony radius of *Cyclocarya paliurus* in the blank control group was 2.19 cm; in the experimental group, the average radius of the *Cyclocarya paliurus* pathogen, determined by the plate confrontation method, was only 0.58 cm, indicating that... Mucor irregularis JSAFC 3408 showed significant inhibitory effects against *Cyclocarya paliurus*.

[0036] Example 3: Mucor irregularis The effects of JSAFC 3408 on chili pepper growth

[0037] (1) Take a piece of the strain with a diameter of 5 mm using a punch. Mucor irregularisJSAFC 3408 mycelial blocks were inoculated into 9 cm diameter petri dishes (containing 15 ml of PDA medium) and cultured in the dark at 25°C and 65% humidity for 7 days.

[0038] (2) The cultured for 7 days Mucor irregularis JSAFC 3408 was processed under aseptic conditions. Mycelia were scraped, rinsed with sterile water, and filtered through gauze. The spore concentration was then adjusted to 10. 5 / ml.

[0039] (3) Select pots with a diameter of 12 cm, and transplant one healthy chili seedling of uniform growth into each pot. The experiment was conducted in two groups (control group and treatment group), with 10 replicates for each treatment. The experimental group consisted of watering each pot with 5 mL of [unspecified solution]. Mucor irregularis JSAFC 3408 spore suspension; control group: 5 mL of sterile water was poured into each pot.

[0040] (4) Water each group of chili seedlings every 2 days. After 30 days, count the height of the chili plants and observe the growth and development of the chili seedlings.

[0041] The results are as follows Figure 3 As shown in Table 2.

[0042] Table 2. Statistical results of chili plant height

[0043] control group experimental group plant height (cm) 13.82±0.15 16.03±0.08

[0044] The plant height in the table is expressed as: mean ± standard error.

[0045] As shown in Table 2, the plant height of the experimental group was significantly better than that of the control group. Mucor irregularis JSAFC3408 promotes the growth of chili peppers. Figure 3 It can be seen Mucor irregularis JSAFC 3408 can also promote the growth of chili leaves and roots.

Claims

1. An irregular type of Mucor ( Mucor irregularis ), characterized in that, The strain number of the irregular mucor (Mucor irregularis) Mucor irregularis is JSAFC 3408, which was preserved in the China General Microbiological Culture Collection Center on May 6, 2025, and the preservation number is CGMCC No. 41982.

2. A biocontrol agent, characterized in that, The active ingredient of the biocontrol agent is the irregular Mucor of claim 1 Mucor irregularis ).

3. The irregular mold as described in claim 1 ( Mucor irregularis The biocontrol agent according to claim 2 inhibits the growth of *Hylocereus huanghuaiense* (…). Globisporangium huanghuaiense Applications during growth.

4. The irregular Mucor (Mucor irregularis) of claim 1, Mucor irregularis application of the biocontrol bacteria agent of claim 2 in promoting growth of pepper.

5. The application according to claim 4, characterized in that, The application is specifically for promoting the growth of pepper plant height, leaf and root system by using spore suspension of irregular mucor (Mucor irregularis) JSAFC 3408 to root irrigation. Mucor irregularis JSAFC 3408 to root irrigation.

6. Use according to claim 5, characterized in that, The preparation method of the spore suspension is as follows: (1) Take the biocontrol fungus Rhizopus irregularis JSAFC 3408 strain, pick up the hyphae with an inoculation needle under sterile conditions, inoculate them into sterilized PDA medium, and culture for 5-7 days; (2) Under sterile conditions, scrape off the hyphae, rinse the hyphae with sterile water and filter with gauze; (3) Under sterile conditions, adjust the spore concentration with sterile water.

7. Use according to claim 6, characterized in that, The culture conditions described in step (1) are 25℃, 65% humidity, and dark culture.

8. The application according to claim 5, characterized in that, The concentration of the spore suspension is 10 5 / ml.

9. The application according to claim 5, characterized in that, The spore suspension was applied at a rate of 5 mL per application every 2 days for 30 days.

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