Fusarium lateritium and its use

Abstract

The application discloses a fusarium lateritium and application thereof, belongs to the technical field of plant disease prevention and the field of plant growth promotion, and separates a new endophytic fungus from a cyclobalanopsis glauca seed, and the new endophytic fungus is identified as the fusarium lateritium through morphology and molecular biology, and is shown to have antagonistic and bacteriostatic activity on botrytis cinerea, can secrete an iron carrier and promote lateral root growth of arabidopsis thaliana. It is indicated that the strain can be used for a biocontrol agent for plant botrytis disease and a plant growth regulator, and is applied to the fields of plant disease and growth promotion.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of plant disease prevention and control and the field of plant growth promotion, and particularly relates to a seed endophytic biocontrol fungus Fusarium lateritium of Quercus glauca and application thereof. BACKGROUND

[0002] Plant endophyte refers to a kind of microorganism that lives in the tissues or cells of growing and healthy plants at a certain stage or the whole stage of its life history, and does not cause obvious disease symptoms to the host plants. In the long-term evolution, endophyte and host plant interact to form a mutualistic symbiotic relationship. The plant provides a living environment for the endophyte, and the endophyte can promote the growth and development of the host plant by producing chemical substances affecting plant growth or enhancing the resistance of the host plant to pathogenic bacteria and the tolerance to surrounding environmental factors.

[0003] In the recent research boom of endophyte, the research on endophyte of woody plants is relatively less, especially the seed, as the most important reproductive organ of plants, which should contain many endophytic microorganisms that colonize in plants for a long time. It is a transmission medium for endophyte from the mother plant to the offspring, and beneficial endophyte is passed down from generation to generation, which has a direct or indirect influence on the growth traits of plants and plays an important role in the growth process of plants.

[0004] Plant diseases have always been a serious threat to agricultural and forestry production. Botrytis cinerea, also known as gray mold, is a plant pathogen with a wide host range, strong destructive and spreading ability. At present, the prevention and control of the pathogen mainly focuses on chemical methods. However, long-term and large-scale use of chemical pesticides not only causes serious environmental pollution, but also makes the gray mold resistant and weakens the control ability. Therefore, finding microorganisms with antagonistic ability to gray mold is conducive to the popularization of biological control methods for gray mold and the reduction of negative effects of chemical pesticides.

[0005] Quercus glauca is a evergreen broad-leaved tree of Fagaceae Quercus, which has the characteristics of cold, drought and poor tolerance, strong ecological adaptability, and wide distribution. It is an important landscaping tree species. Its wood is tough and can be used for pile, boat, tool handle and other timber. The seed contains 60-70% of starch, which can be used as feed, wine making and ethanol fuel. It is also an important economic and timber tree species. The main disease of Quercus glauca plantation is Quercus glauca mildew, which is caused by Botrytis cinerea. The disease occurs on leaves, and at the early stage, scattered round black spots appear on the leaves. In severe cases, stone sulfur mixture is used for prevention and control. The use of biological pesticides (microorganisms) instead of chemical pesticides to control Quercus glauca mildew is of great significance to the green and healthy sustainable development of Quercus glauca plantation.

[0006] At present, there is little research on the endophyte of Quercus glauca, and there is no report on the isolation and identification of the endophyte in Quercus glauca seed and the functional research thereof. SUMMARY

[0007] The technical problem solved by the present application is to provide a Quercus glauca endophytic biocontrol fungus and application thereof.

[0008] The technical solution of the present application is a Fusarium lateritium QGZJ08, which is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.40429.

[0009] A microbial preparation contains the above-mentioned Fusarium lateritium QGZJ08.

[0010] The above-mentioned Fusarium lateritium QGZJ08 or microbial preparation is applied to the prevention and treatment of gray mold caused by Botrytis cinerea.

[0011] The above-mentioned Fusarium lateritium QGZJ08 or microbial preparation is applied to the promotion of plant growth.

[0012] Further, the promotion of plant growth refers to the promotion of iron absorption of plants.

[0013] Further, the promotion of plant growth refers to the promotion of lateral root growth of plants.

[0014] Further, the plant is Arabidopsis thaliana.

[0015] Compared with the prior art, the present application has the following beneficial effects:

[0016] 1. The Quercus glauca seed endophytic fungus Fusarium lateritium QGZJ08 of the present application has good antagonistic activity to Botrytis cinerea, and can be used for the prevention and treatment of gray mold.

[0017] 2、The endophytic fungus Fusarium lateritium QGZJ08 in Cyclobalanopsis glauca seeds can secrete siderophores and promote the growth of plant lateral roots, and has good growth-promoting effect. Compared with using chemical agents to promote plant growth, the biological method can greatly reduce the pollution to the ecological environment, and protect the health of human beings and the green and sustainable development of agriculture and forestry.

[0018] Strain preservation: On November 21, 2022, Fusarium lateritium QGZJ08 was preserved in the China General Microbiological Culture Collection Center, No. 3, Beichen West Road, Chaoyang District, Beijing, China, with the preservation number CGMCC No. 40429. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 is a flat culture effect diagram of the endophytic strain QGZJ08; the left picture is the front, and the right picture is the back;

[0020] Figure 2 is the antibacterial effect of the endophytic fungus Fusarium lateritium QGZJ08 on the mycelial growth of Botrytis cinerea for 5 days; the left picture is the control group without the endophytic fungus Fusarium lateritium QGZJ08, and the right picture is the treatment group with the endophytic fungus Fusarium lateritium QGZJ08;

[0021] Figure 3 is the detection result of the siderophore secreted by the endophytic fungus Fusarium lateritium QGZJ08;

[0022] Figure 4 is the promotion effect of the endophytic fungus Fusarium lateritium QGZJ08 on the lateral root growth of Arabidopsis thaliana for 16 days; the left picture is the control group with PDA medium, and the right picture is the treatment group with the endophytic fungus Fusarium lateritium QGZJ08. DETAILED DESCRIPTION

[0023] The experimental methods in the following examples are all conventional methods unless otherwise specified. The test materials used in the following examples are all purchased from commercial channels unless otherwise specified.

[0024] Example 1 Isolation and identification of endophytic fungus strain

[0025] Healthy Cyclobalanopsis glauca seeds with shells were placed in 50 mL sterile centrifuge tubes, first surface sterilized with 75% ethanol for 2 min, rinsed with sterile water 3 times, then soaked with 20% sodium hypochlorite for 20 min, rinsed with sterile water 7 times, then spread and placed on the ultraclean workbench to turn on the ultraviolet lamp for irradiation for 40 min, turn over after 20 min of irradiation, ensure that the seed surface is disinfected clean, finally rinse with sterile water again 3 times, retain the last time of washing sterile water to be coated on the PDA flat plate culture medium as a control to determine whether the seed surface is disinfected completely.

[0026] Tissue separation method: under sterile conditions, the seeds with completed surface sterilization were peeled off the shell and inner seed coat, cut into 1 cm*1 cm, 2 mm thick pieces, inoculated on PDA flat plate culture medium, sealed with sealing film, then cultured at room temperature. When the seed edge began to form colonies, the well-grown mycelium at the edge of different morphological fungal colonies was picked and inoculated at the center of a new PDA flat plate culture medium, sealed with sealing film, then the flat plate was cultured at room temperature, when the strain grew to 2 / 3 of the flat plate, the well-grown mycelium at the edge was picked again and inoculated at the center of a new PDA flat plate culture medium, sealed with sealing film, then the flat plate was cultured at room temperature, until a single strain was obtained. Figure 1 The plate culture effect diagram of the obtained endophytic fungal strain QGZJ08 is shown in FIG. 1, and the colony is white and velvety, with irregular edges.

[0027] The ITS gene sequencing (SEQ ID No. 1) of the isolated endophytic fungal strain QGZJ08 was performed, and the sequencing results were subjected to BLAST comparison on NCBI, and the similar strain name was Fusarium lateritium (Table 1), so the strain was Fusarium lateritium QGZJ08.

[0028] Table 1: ITS sequencing identification results

[0029] Accession QGZJ08 Domain Eukaryota Phylum Ascomycota Class Sordariomycetes Order Hypocreales Family Nectriaceae Genus Fusarium Reference species Fusarium lateritium

[0030] Strain preservation: on November 21, 2022, Fusarium lateritium QGZJ08 was preserved in the China General Microbiological Culture Collection Center, No. 3, Beichen West Road, Chaoyang District, Beijing, with the preservation number CGMCC No. 40429.

[0031] Example 2: Antagonistic experiment of endophytic fungi

[0032] In this example, Fusarium lateritium QGZJ08 was subjected to plate confrontation experiment against plant pathogenic fungus Botrytis cinerea, and the inhibitory activity of endophytic fungi was observed.

[0033] 1. Flat plate confrontation method:

[0034] A 7mm sterile puncher was used to punch a Botrytis cinerea block (block size Φ0.7cm) on a Botrytis cinerea plate in a vigorous growth stage, which was then inoculated upside down to the center of a new PDA plate, and a sealing film was used to seal the plate, which was then cultured at 25°C for 2 days. Then, a Fusarium lateritium QGZJ08 block (block size Φ0.7cm) was inoculated on the Botrytis cinerea plate at a distance of 3cm from the center, and two blocks of the same strain were inoculated symmetrically on each plate. A sealing film was used to seal the plate, which was then cultured at 25°C for 5 days to observe the growth of Botrytis cinerea.

[0035] 2. Confrontation experiment results:

[0036] (1) Measurement method:

[0037] A ruler was used to measure the diameter of the Botrytis cinerea colony, and the measurement path needed to pass through the colony center point, and the measurement direction was the straight line connecting the centers of the two endophytic fungal colonies.

[0038] Table 2: Botrytis cinerea colony diameter (CM)

[0039]

[0040] (2) Inhibition rate calculation

[0041] Inhibition rate (%) = (control colony diameter - treated colony diameter) / (control colony diameter) x 100%

[0042] The measurement data in Table 2 was substituted into the inhibition rate formula, and it was calculated that the inhibition rate of the Fusarium lateritium QGZJ08 of the present application on the plant pathogen Botrytis cinerea was 54.2%.

[0043] Studies have shown that at the fifth day, the mycelial growth of Botrytis cinerea on both sides of Fusarium lateritium QGZJ08 was significantly inhibited, as shown in Figure 2 , where the left is the control group and the right is the treatment group.

[0044] Therefore, the endophytic fungal biocontrol agent of the present application, Fusarium lateritium QGZJ08 of the Qinggang seed, can be applied to plant disease control, and has good effect.

[0045] Example 3: Detection of endophytic fungal secreted siderophores

[0046] In this example, the growing endophytic fungus Fusarium lateritium QGZJ08 cake was inverted and connected in the middle of the CAS plate medium, and cultured for 3 days for observation. A yellow halo appeared around the cake, indicating that the endophytic fungus had the ability to secrete iron carriers. See Figure 3 .

[0047] Example 4: Endophytic fungus promotes growth of Arabidopsis thaliana

[0048] 1. Experimental method:

[0049] Arabidopsis thaliana seeds were inoculated on 1 / 2MS plates after sterilization, and after 5 days of growth, endophytic fungus Fusarium lateritium QGZJ08 cake was inverted and inoculated under the plate. The control group was inoculated with clean PDA medium, and the growth state of Arabidopsis thaliana in the treatment group and the control group was observed after 11 days of culture.

[0050] 2. Experimental results

[0051] Compared with the control group, the main root length of Arabidopsis thaliana in the treatment group inoculated with endophytic fungus Fusarium lateritium QGZJ08 was inhibited, but the number and length of lateral roots were significantly increased. See Figure 4 , where the left is the control group and the right is the treatment group.

[0052] In summary, lateral roots are important organs for plants to absorb water and nutrients, and the endophytic fungus Fusarium lateritium QGZJ08 provided by the present application can promote the growth of plant lateral roots, thereby having a certain effect in promoting plant growth.

Claims

1. A strain of Fusarium lateritium QGZJ08, deposited in the General Microbiology Center of China Culture Collection Administration, with the deposit number CGMCC No. 40429.

2. A microbial preparation, characterized in that: Contains the Fusarium lateritium QGZJ08 according to claim 1.

3. Use of the Fusarium lateritium QGZJ08 according to claim 1 or the microbial preparation according to claim 2 in preventing and treating gray mold caused by Botrytis cinerea.

4. Use of the Fusarium lateritium QGZJ08 according to claim 1 or the microbial preparation according to claim 2 in promoting plant growth.

5. The use according to claim 4, characterized in that The promoting plant growth refers to promoting the absorption of iron by plants.

6. The use according to claim 4, characterized in that The promoting plant growth refers to promoting the growth of plant lateral roots.

7. The use according to claim 6, characterized in that The plant is Arabidopsis thaliana.

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