New strain melanospora sp. And application thereof

Abstract

The invention discloses a new strain of melanospora sp. And application thereof, the new strain of melanospora sp. Is preserved in China General Microbiological Culture Collection Center on September 26, 2024, the preservation number is CGMCC No.41569, the melanospora sp. Is a microbial inoculum of a W18 strain, the preservation number is CGMCC No.41569, and the preservation number is CGMCC No.41569. The melanospora sp. Is applied to preparation of a plant growth promoter. The application of the melanospora sp. To plants comprises the following steps: 1, preparing a plant growth promoter; the method comprises the following steps: S1, preparing a spore suspension: inoculating a PDA culture medium with a strain, culturing until spores are completely produced, scraping the spores, and preparing the spore suspension by using a conditioning solution; s2, seedling raising: soaking plant seeds in sterile deionized water for one day in advance, and then soaking the plant seeds in sterile water at 50-70 DEG C for 1.5-3 hours; s3, seedling transplanting. According to the application, the relative water content, the chlorophyll content and the auxin content of plant seedling leaves can be remarkably improved, the plant growth promoting effect is achieved, and the plant is excellent in character and has wide application value in agriculture.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and in particular relates to a new strain of black spore mold and application thereof. Background Art

[0002] Hazelnut, as one of the four most famous nuts in the world, contains extremely rich nutrients, such as high content of protein, vitamin E, folic acid, eight essential amino acids for the human body and resveratrol. Hazelnut belongs to the genus Corylus in the Betulaceae family in plant classification, and there are about 20 species worldwide. Wild hazelnuts are mainly distributed in temperate regions such as Asia, Europe and North America, among which Turkey, Italy, Spain, the United States and Portugal are the main producing countries. Although hazelnuts have strong environmental adaptability and can grow under different climatic conditions, they prefer cool climates and require specific temperature and humidity conditions. However, with the trend of global warming, the growth environment of some hazelnuts has been affected, which has a significant impact on their economic yield and fruit quality.

[0003] Nigrospora, a member of the phylum Ascomycotina, class Sordariomycetes, order Xylariales, and family Apiosporaceae, is a fungus with a wide host range and plays a key role in a variety of ecosystems. In particular, when Nigrospora is used as an endophytic fungus in plants, it exhibits significant effects in promoting host growth, enhancing development, and improving resistance to biotic and abiotic stresses. Therefore, exogenous addition of Nigrospora fungicides has become an effective strategy to enhance the adaptability of hazel trees, promote their growth and development, and improve the quality of hazelnuts.

[0004] Therefore, the screening and research and development of strains have continuous research value. The discovery of beneficial strains has a positive effect on enhancing the adaptability and resistance of host plants, optimizing soil quality, improving soil fertility, and promoting yield and quality improvement. This application focuses on screening multifunctional microorganisms that have a significant promoting effect on hazel growth. The screening of this strain is of great significance to ensuring the normal growth of hazel, increasing yield and optimizing quality. Summary of the invention

[0005] The purpose of the present invention is to provide a new strain of black spore mold and its application, which has a good growth-promoting effect and can improve the photosynthesis efficiency of plants. At the same time, it can also stimulate plants to produce more auxin, thereby promoting the growth and development of plants.

[0006] In order to achieve the above object, a technical solution provided by a specific embodiment of the present invention is as follows:

[0007] A new strain of Black Sporophycetes, with the deposit number CGMCC No.41569, was deposited in the General Microbiology Center of China National Microbiological Culture Collection Administration on September 26, 2024.

[0008] In one or more embodiments of the present invention, the black spore mold is a fungal agent of the W18 strain.

[0009] In one or more embodiments of the present invention, the black spore mold is used in the preparation of a plant growth promoter, and the application of the black spore mold on plants includes the following steps: preparing spores of strain W18 into a suspension and then using the suspension to irrigate the roots of cultivated plants, and other management methods can be conventional management methods;

[0010] S1. Preparation of spore suspension: inoculate the strain on PDA medium, culture until complete spore production, scrape spores, and use a conditioning solution to prepare a spore suspension;

[0011] S2. Seedling cultivation: Soak the plant seeds in sterile deionized water for one day in advance, then soak them in 50-70℃ sterile water for 1.5-3h, soak them in 75% alcohol for 20-40min, rinse them with sterile water for 3-5 times, soak them in 2% sodium hypochlorite solution for 15-30min, rinse them with sterile water for 3-8 times, and evenly spread the sterilized seeds on sterilized sand for germination for 20-30 days;

[0012] S3. Transplanting seedlings: Sterilize peat soil, vermiculite, perlite and sand, mix them evenly in proportion and place them in plastic pots. Transplant the seedlings into the pots containing the mixed soil. After 6-12 days of acclimatization, irrigate the roots with a spore suspension of Black Sporoderm. Inoculate the roots of each seedling with a spore suspension of Black Sporoderm W18.

[0013] In one or more embodiments of the present invention, the plant is hazelnut.

[0014] In one or more embodiments of the present invention, the plant is a pear tree.

[0015] In one or more embodiments of the present invention, the spore concentration of the suspension is 8*106 cfu / ml

[0016] In one or more embodiments of the present invention, the temperature for culturing spores in step S1 is 20-36° C., and the culturing time is 5-10 days.

[0017] In one or more embodiments of the present invention, the conditioning solution is sterile water.

[0018] In one or more embodiments of the present invention, the mixing ratio of peat soil, vermiculite, perlite and sand in step S3 is 6:3:3:1.

[0019] In one or more embodiments of the present invention, in step S3, 10 ml of a spore suspension of Nigrosporium W18 is inoculated at the root of each seedling.

[0020] Compared with the prior art, the new strain of black spore mold and its application of the present invention have the following benefits:

[0021] 1) It has the effect of increasing the relative water content, chlorophyll content and auxin content of hazel seedling leaves;

[0022] 2) The W18 strain has good adaptability, is convenient for laboratory research and large-scale cultivation, and has a good growth-promoting effect on plants, and has potential application value as a biofertilizer. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0024] Figure 1 The morphological picture of strain W18 growing on PDA medium and the microscopic morphological pictures of spores and conidia;

[0025] Figure 2 is the phylogenetic tree of strain W18;

[0026] Figure 3 This is the relative water content of leaves under strain W18 treatment;

[0027] Figure 4 This is the chlorophyll content diagram under the treatment of strain W18;

[0028] Figure 5 This is a graph of auxin content under strain W18 treatment. DETAILED DESCRIPTION

[0029] In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0030] Example 1

[0031] like Figure 1As shown, a new strain of Black Sporophyceae and its application in one embodiment of the present invention, wherein a new strain of Black Sporophyceae, with a preservation number of CGMCC No.41569, was deposited in the General Microbiological Center of China Microorganism Culture Collection Administration on September 26, 2024, with an address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the name of the deposited organism is confirmed to be Black Sporophyceae.

[0032] The black spore mold is a fungicide of the W18 strain.

[0033] Black spore mold is used in the preparation of plant growth promoters, and the application of black spore mold on plants includes the following steps:

[0034] S1. Preparation of spore suspension: inoculate the TB2 strain on PDA medium, culture until complete spore production, scrape the spores, and use a conditioning solution to prepare a spore suspension;

[0035] The spore concentration of the suspension is 8*106 cfu / ml, the spore production temperature in step S1 is 20-36°C, and the culturing time is 5-10 days. The conditioning liquid is sterile water.

[0036] Preferably, the spore production temperature is 28°C, and the culture is carried out for 7 days until the spores are fully produced.

[0037] S2. Seedling cultivation: Soak the plant seeds in sterile deionized water for one day in advance, then soak them in 50-70℃ sterile water for 1.5-3h, soak them in 75% alcohol for 20-40min, rinse them with sterile water for 3-5 times, soak them in 2% sodium hypochlorite solution for 15-30min, rinse them with sterile water for 3-8 times, and evenly spread the sterilized seeds on sterilized sand for germination for 20-30 days;

[0038] Preferably, the seeds are soaked in 60°C sterile water for 2 hours, soaked in 75% alcohol for 30 minutes, rinsed with sterile water for 3 to 5 times, soaked in 2% sodium hypochlorite solution for 20 minutes, and rinsed with sterile water for 5 times. The sterilized hazel seeds are evenly spread on sterilized sand and germinated for 20 to 30 days.

[0039] S3. Transplanting: Sterilize peat soil, vermiculite, perlite and sand in a ratio of 6:3:3:1 and evenly mix them in a plastic pot. Select hazel seedlings with consistent growth and transplant them into pots containing mixed soil. After 6-12 days of seedling acclimatization, irrigate the roots with black spore suspension. Inoculate the roots of each pear seedling with 10 ml of black spore suspension W18 (inoculation concentration is 8*10 6 cfu / ml), and the blank control CK was set as the non-inoculated treatment. The harvest was carried out after 2 months of cultivation.

[0040] S4. Harvesting seedlings: After 2 months of cultivation, the physiological indices of the hazel seedlings, such as leaf water content, leaf green color and auxin content, were measured.

[0041] Among them, the best treatment is to irrigate the roots with a suspension of black spores after the seedlings have been grown for 10 days.

[0042] Preferably, the plant is one of the hazelnut or pear trees.

[0043] After the experiment, the results are as follows: Figure 3 As shown, it can be seen that the inoculated strain W18 significantly increased the water content of the leaves of hazel seedlings, thereby improving the drought resistance of hazel seedlings;

[0044] like Figure 4 As shown, it can be seen that the inoculated strain W18 significantly increased the chlorophyll a and chlorophyll b contents in the leaves of hazel seedlings, enhanced the light energy utilization rate and organic matter synthesis rate of hazel seedlings, and provided strong support for the growth and development of hazel seedlings.

[0045] like Figure 5 As shown, it can be seen that the inoculated strain W18 significantly increased the auxin content in the roots, stems and leaves of hazel seedlings, which can promote the growth and development of hazel seedlings in many aspects, and provide strong technical support for the high-yield and high-quality cultivation of hazelnuts.

[0046] Example 2

[0047] Isolation, Screening and Identification of Nigrospora sp. W18

[0048] S11. Isolation of strains: Healthy and vigorous hazel (Corylus heterophylla) stem samples were collected from the mycorrhizal seedling cultivation center (Guizhou Institute of Biology). The collected fragments were sterilized.

[0049] The specific disinfection steps are: 75% ethanol treatment for 30 seconds, sterile water rinse, 5% sodium hypochlorite treatment for 30 seconds, rinse again, and 75% ethanol treatment for 30 seconds again, followed by three rinses. Then, the fragments were wiped dry with sterile paper and inoculated on potato dextrose agar (PDA).

[0050] S12. Morphological identification: After 3 days of growth, collect one mycelial tip and transfer it to PDA and culture it in the dark at 25°C for 7 days. Observe the growth and development with a stereoscope and microscope. The results are as follows: Figure 1 As shown. The colony spreads flat on the PDA medium, cotton-like, loose, white at first, and then turns gray-brown due to a large number of spores. The mycelium is transparent to light brown, smooth, septate, and branched. The conidiophores are similar to the vegetative mycelium, short and sparse. The conidial cells are spherical or subspherical, light brown, smooth, single-bud type, and determinative. The spores are spherical, subspherical or elliptical, without buds, smooth, shiny, without septa, and black.

[0051] S13. Molecular identification: Based on the comparison of ITS sequences and phylogenetic tree analysis of TUB2 and TEF1-α gene fragments, the results obtained are as follows Figure 2 As shown in the figure, it can be seen that the W18 strain formed a unique lineage in the joint gene phylogenetic tree, which is closely related to N. chinensis and has good support. Based on comprehensive morphological identification, the strain is considered to be a new strain of Black Sporophytum.

[0052] The strain W18 screened by the present application was isolated from plant seedlings. After morphological and molecular biological identification, the strain was identified as a new strain Nigrospora sp. W18 (Nigrospora. sp. W18). By treating plant seedlings with the microbial agent of the new strain Nigrospora sp. W18 (Nigrospora. sp. W18), the relative water content of hazelnut seedling leaves increased by 18.03%, the chlorophyll a and chlorophyll b contents increased by 5.65% and 10.95% respectively, and the auxin contents of roots, stems and leaves increased by 140.19%, 20.37% and 145.55% respectively, indicating that the new strain Nigrospora sp. W18 (Nigrospora. sp. W18) provided by the present invention can significantly increase the relative water content, chlorophyll content and auxin content of plant seedling leaves, has a growth-promoting effect on plants, is a strain with excellent traits, and has broad application value in agriculture.

[0053] In summary, the strain W18 screened by the present invention has good growth performance on PDA medium, has good adaptability, is convenient for laboratory research and large-scale cultivation, and is found by inoculating hazelnut seedlings: the strain also has a good growth-promoting effect on plants, can improve the photosynthesis efficiency of plants, and increase the chlorophyll content and relative water content of leaves, so that plants can better absorb and utilize light energy; at the same time, it can also stimulate plants to produce more auxin, thereby promoting plant growth and development. This shows that the W18 strain of the present application is a strain with excellent traits, has broad application prospects and potential utilization value, and is expected to make greater contributions to agricultural production and ecological environmental protection.

[0054] It will be apparent to those skilled in the art that the invention is not limited to the details of the exemplary embodiments described above and that the invention can be implemented in other specific forms without departing from the spirit or essential features of the invention. Therefore, the embodiments should be considered exemplary and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations falling within the meaning and scope of the equivalent elements of the claims be included in the invention. Any reference numeral in a claim should not be considered as limiting the claim to which it relates.

[0055] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A new strain of black spore mold, characterized in that Including the deposit number CGMCC No.41569.

2. A new strain of black spore mold according to claim 1, characterized in that The black spore mold is a bacterial agent of the W18 strain.

3. An application of a new strain of black spore mold, characterized in that: The black spore mold is used in the preparation of a plant growth promoter, and the application of the black spore mold on plants comprises the following steps: S1. Preparation of spore suspension: inoculate the strain on PDA medium, culture until complete spore production, scrape spores, and use a conditioning solution to prepare a spore suspension; S2. Seedling cultivation: Soak the plant seeds in sterile deionized water for one day in advance, then soak them in 50-70℃ sterile water for 1.5-3h, soak them in 75% alcohol for 20-40min, rinse them with sterile water for 3-5 times, soak them in 2% sodium hypochlorite solution for 15-30min, rinse them with sterile water for 3-8 times, and evenly spread the sterilized seeds on sterilized sand for germination for 20-30 days; S3. Transplanting seedlings: Sterilize peat soil, vermiculite, perlite and sand, mix them evenly in proportion and place them in plastic pots. Transplant the seedlings into the pots containing the mixed soil. After 6-12 days of acclimatization, irrigate the roots with a spore suspension of Black Sporoderm. Inoculate the roots of each seedling with a spore suspension of Black Sporoderm W18.

4. The use of a new strain of Black Sporophycetes according to claim 3, characterized in that: The plant is hazelnut.

5. The use of a new strain of Black Sporophycetes according to claim 3, characterized in that: The plant is a pear tree.

6. The use of a new strain of Black Sporophycetes according to claim 4 or 5, characterized in that: The spore concentration of the suspension is 8*106 cfu / ml.

7. A new strain of Black Sporophycetes and its application according to claim 4 or 5, characterized in that: The spore production temperature in step S1 is 20-36° C. and the spore production time is 5-10 days.

8. The use of a new strain of Black Sporophycetes according to claim 4 or 5, characterized in that: The conditioning solution is sterile water.

9. The use of a new strain of Black Sporophycetes according to claim 4 or 5, characterized in that: The mixing ratio of peat soil, vermiculite, perlite and sand in step S3 is 6:3:3:

1.

10. The use of a new strain of Black Sporophycetes according to claim 9, characterized in that: In the step S3, 10 ml of the spore suspension of Nigrosporium W18 was inoculated at the root of each seedling.

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