Salt-tolerant bacillus YH30 and application thereof

Abstract

The invention provides a salt-tolerant bacillus YH30 and application thereof, and relates to the technical field of microbial control. The salt-tolerant bacillus YH30 is classified and named as the salt-tolerant bacillus, the salt-tolerant bacillus YH30 is preserved in the China General Microbiological Culture Collection Center (CGMCC) on December 2, 2024, and the preservation number is CGMCC No: 32877; the salt-tolerant bacillus YH30 is applied to prevention and treatment of grapefruit penicilliosis, blueberry fruit gray mold and macadamia nut anthracnose. The method overcomes the defects in the prior art, can effectively control the harm of grapefruit fruit penicilliosis, blueberry fruit gray mold and macadimia nut leaf anthracnose, and promotes the improvement of the resistance level of grapefruit fruits, blueberry fruits and macadimia nut leaves; the method has a positive promotion effect on the sustainable and healthy development of the planting industry of grapefruits, blueberries and macadamia nuts.

Description

Technical Field

[0001] The present invention relates to the technical field of microbial control, and in particular to a halotolerant Bacillus YH30 and an application thereof. Background Art

[0002] Grapefruit is an evergreen tree in the Rutaceae family, a backcross between the sweet orange and pomelo. It is also known as grapefruit or red grapefruit. Its fruit is oblate to spherical, with a thin peel that is pale yellow or greenish-yellow when ripe, often with a reddish hue. The flesh is tender and juicy, mostly pale yellow or pink, with a sweet-sour flavor and a slightly bitter taste.

[0003] Currently, grapefruit fruit penicillium mold is mainly controlled through chemical means, with the preservatives used including carbendazim, thiophanate-methyl, imazalil, thiabendazole, and fufujing. These agents can effectively inhibit the growth and reproduction of penicillium mold and reduce the occurrence of the disease.

[0004] Blueberry is a perennial deciduous berry shrub of the genus Vaccinium in the Ericaceae family. The blueberry fruit is not only attractive in color but also has a unique flavor. It can be eaten fresh or processed into a variety of foods. However, the diseases encountered by blueberry fruits during transportation and storage mainly include storage rot. In addition, they may also be affected by diseases such as gray mold, which is caused by Botrytis cinerea ( Botrytis cinerea Gray mold is a widespread and devastating fungal disease of plants caused by botrytis cinerea that infects over 200 plant species, including vegetables, fruits, flowers, and ornamental plants, causing significant economic losses to global agricultural production. To prevent and control gray mold, resistant varieties should be selected. In autumn and winter, dead branches, fallen leaves, and diseased fruit should be removed. During the growing season, diseased fruit, vines, and leaves should be promptly removed and sprayed with pesticides. Watering should be strictly controlled, avoiding watering on rainy days. Ventilation and dehumidification should be increased to keep relative humidity below 65%. Chemical control measures include spraying the entire plant with a 1500-fold dilution of 50% boscalid water-dispersible granules before flowering, and focusing on spraying with a 1000-fold dilution of 400g / L pyrimethanil suspension concentrate during flowering.

[0005] Macadamia, also known as macadamia nut, Australian walnut, Queensland chestnut, etc., is a tall evergreen tree of the genus Macadamia in the family Proteaceae. Macadamia anthracnose is a common and highly destructive disease that primarily affects the leaves, young shoots, and fruits of macadamia. On leaves, it primarily infects the young or mature leaves of grafted or cutting seedlings; on young shoots, it causes new shoots to turn black; on fruits, black spots initially appear, and the surface of the fruit is covered with orange-yellow, needle-shaped fruiting bodies of the pathogen. As the disease worsens, the peel rots and spreads to the fruit stalk, causing a large number of fruits to drop before maturity. In the early stages of the disease, small black, water-soaked patches appear on the affected area. As the pathogen grows, the area of ​​the lesions expands, and black conidiophores appear in the shape of rings, surrounded by a yellow halo. In severe cases, the entire leaf turns brown and twists.

[0006] Anthracnose can be controlled through agricultural, physical, chemical, and biological methods. Agricultural control includes pruning drooping branches and maintaining ventilation and light to reduce the number of pathogens. In early spring, weeds and dead branches and leaves in the fields should be removed and burned or buried to eliminate overwintering adults and nymphs. Resistant varieties should be selected and covered with composted fruit husks. Physical control involves exploiting the tendency of thrips to move towards blue. Blue sticky traps should be set up in the fields at the same height as the crop to attract and kill adult insects. Chemical control includes spraying with a 700-800 times dilution of 80% anthracnose thiophanate or an 800-1000 times dilution of 70% thiophanate-methyl. Annually, during the infestation period, spray leaves, young shoots, and young fruit with an 800-900 times dilution of 50% carbendazim, a 700-750 times dilution of 80% anthracnose thiophanate, or a 250-500 times dilution of thiophanate-methyl.

[0007] Based on this, existing technologies for the prevention and control of grapefruit blue mold, blueberry gray mold and macadamia anthracnose mostly use chemical control methods. However, the use of chemical agents may pollute the soil, water sources and other environments, and destroy the ecological balance. Compared with chemical control, biological control is less likely to cause pests and diseases to develop drug resistance, can maintain the stability of control effects, and reduce the increased difficulty and cost of control caused by enhanced drug resistance. Therefore, research on microbial control methods is crucial for the growth and storage of fruits and nuts. Summary of the Invention

[0008] In response to the shortcomings of the existing technology, the present invention provides a salt-tolerant Bacillus YH30 and its application. The YH30 strain can effectively control the damage caused by blue mold of grapefruit fruit, gray mold of blueberry fruit, and anthracnose of macadamia nut leaves, while promoting the improvement of the resistance level of grapefruit fruit, blueberry fruit, and macadamia nut leaves, and has a positive promoting effect on the sustainable and healthy development of the grapefruit, blueberry, and macadamia nut planting industries.

[0009] To achieve the above objectives, the technical solution of the present invention is implemented through the following technical solutions: A halodurable Bacillus spp. Bacillus halotolerans ) YH30, the classification of the halodurable Bacillus YH30 is named as halodurable Bacillus Bacillus halotolerans , and was deposited in the General Microbiology Center of China Culture Collection Administration on December 2, 2024, with the deposit number CGMCC No: 32877.

[0010] Halophilous Bacillus YH30 is used to control grapefruit blue mold, blueberry gray mold and macadamia anthracnose.

[0011] Preferably, the grapefruit Penicillium pathogenic fungus is Penicillium italicum ( Penicillium italicum ), Penicillium digitatum ( Penicillium digitatum )、Penicillium expansum( Penicillium expansu ) any one of the following; the blueberry gray mold pathogen is Botrytis cinerea ( Botrytis cinerea ); The macadamia nut anthrax pathogenic fungus is anthrax ( Colletotrichum aeschynomenes ).

[0012] Preferably, the application method is to prepare a spore concentration of 1×10 6 -1×10 8 The product with a concentration of 100mg / mL can prevent and control grapefruit blue mold, blueberry gray mold and macadamia anthracnose.

[0013] The present invention provides a halotolerant Bacillus subtilis YH30 and its application, which have the following advantages over the prior art: The present invention provides a salt-tolerant Bacillus YH30 strain, which can effectively inhibit the growth of grapefruit pathogenic fungi Penicillium expansum, blueberry pathogenic fungi Botrytis cinerea, and macadamia nut anthracnose, and plays a good role in preventing and controlling grapefruit fruit penicillium mold, blueberry fruit gray mold, and macadamia nut leaf anthracnose. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Figure A shows the colony morphology of strain YH30 on LB medium in Example 1 of the present invention; Figure B shows the Gram staining and spore staining results of strain YH30; Figure 2 The growth results of the plate confrontation test in Example 2 of the present invention after 3 days of inoculation are shown, where A is the YH30 group and B is the control group; Figure 3 This is a histogram showing the growth of the diameter of Penicillium colonies expanded at different times in the YH30 group and the control group in Example 2 of the present invention; Figure 4 The growth results of the plate confrontation test in Example 3 of the present invention after 2 days of inoculation are shown, where A is the YH30 group and B is the control group; Figure 5 This is a histogram of the growth of Botrytis cinerea colony diameters at different times in the YH30 group and the control group in Example 3 of the present invention; Figure 6 The growth results of the plate confrontation test in Example 4 of the present invention after 3 days of inoculation are shown, where A is the YH30 group and B is the control group; Figure 7 The figure is a bar graph showing the growth of anthrax colony diameters at different times in the YH30 group and the control group in Example 4 of the present invention; Figure 8 This is a schematic diagram comparing the decay conditions of the control group (A) and the YH30 group (B) in the grapefruit Penicillium expansum control experiment 10 days after Example 5 of the present invention; Figure 9This is a bar graph comparing the lesion diameters of the control group and the YH30 group at different times in the experiment on controlling Penicillium expansum in grapefruit in Example 5 of the present invention; Figure 10 This is a bar graph showing the incidence of blueberries at different times in the control group and the YH30 group during the blueberry Botrytis cinerea control experiment in Example 6 of the present invention; Figure 11 This is a bar graph comparing the diameters of blueberry lesions in the control group and the YH30 group at different times in the blueberry Botrytis cinerea control experiment in Example 6 of the present invention; Figure 12 This is a schematic diagram of the disease conditions of blueberries in the control group (A) and the YH30 group (B) at 8 days of the blueberry Botrytis cinerea control experiment in Example 6 of the present invention; Figure 13 This is a bar graph showing the incidence of macadamia nut leaves at different times in the control group and the YH30 group during the experiment on the prevention and control of anthracnose on macadamia nut leaves in Example 7 of the present invention; Figure 14 This is a bar graph showing the comparison of the diameters of macadamia nut leaf lesions at different times in the control group and the YH30 group in the experiment on the control of anthracnose on macadamia nut leaves in Example 7 of the present invention; Figure 15 This is a schematic diagram of the lesions of the control group (A) and the YH30 group (B) at 10 days after the experiment on the control of anthracnose on macadamia leaves in Example 7 of the present invention. DETAILED DESCRIPTION

[0015] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention are clearly and completely described below in conjunction with 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. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0016] The following halotolerant Bacillus strain YH30 was isolated from the leaves of a healthy grapefruit plant in Xishuangbanna, Yunnan Province, and deposited in the General Microbiology Center of the China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. It is classified as halotolerant Bacillus Bacillus halotolerans , strain number is YH30, preservation time is: December 2, 2024, preservation number is: CGMCC No: 32877. Example 1:

[0017] Identification of the species of halotolerant Bacillus YH30: 1. Morphological identification LB medium formula: peptone 10 g, yeast powder 5 g, sodium chloride 10 g, agar powder 15 g, dilute to 1000 mL with distilled water, sterilize at 121 °C for 20 min, and set aside.

[0018] Inoculate the halodurable Bacillus strain YH30 onto an LB plate and invert at 28°C for 24 hours. Record and describe the colony morphology, color, and other characteristics. Select a small number of cells for Gram staining and observe the staining and bacterial characteristics using an optical microscope.

[0019] like Figure 1 As shown, the strain forms yellowish opaque colonies with a rough, raised surface, irregular edges, and a certain thickness in LB solid medium ( Figure 1 A), Gram staining showed purple, indicating that the strain was a Gram-positive bacterium, and spore staining results showed that the strain could produce spores, indicating that it had the ability to produce spores ( Figure 1 B, the bacteria are blue, the spores are red, and the yellow arrows point to the spores. Bar = 20 μm).

[0020] 2. Molecular Biology Identification DNA of Bacillus halodurans YH30 was extracted using a bacterial DNA extraction kit (Cat: YZ-D3350, Beijing Solebeau Technology Co., Ltd.). 16S rDNA universal primers were used for PCR amplification. The amplified products were detected by gel electrophoresis and sent to Kunming Shuoqing Biotechnology Co., Ltd. for bidirectional sequencing. The sequencing results were compared with sequences of similar model strains using the NCBI website (https: / / www.ncbi.nlm.nih.gov). A phylogenetic tree of the 16S rDNA gene sequences was constructed using the Newton-Jones method using MEGA11 software.

[0021] The 16SrDNA sequence of the halodurable Bacillus subtilis YH30 is shown in SEQ ID NO.1: SEQ ID NO.1: Example 2:

[0022] Plate confrontation culture test of the effect of halotolerant Bacillus sp. YH30 on the pathogenic fungus Penicillium expansum of grapefruit penicillium disease ( Penicillium expansu ) inhibition results: 10 μL of the purified Penicillium expansum spore suspension was inoculated on the filter paper in the center of the PDA culture dish. Four sterile filter paper discs were placed 3 cm away from the center. 10 μL of the fermentation suspension of Bacillus halophilus YH30 (concentration of 1×10 7 The treatment with sterile water was used as the control group. Three independent replicates were set for each treatment. The colony diameters were counted after culturing at 28°C in the dark for 3 days, and the inhibition rate was calculated.

[0023] The antagonistic ability to Penicillium expansu was observed and determined, and the colony diameter was determined by the cross method.

[0024] Specific results can be found in Figure 2 and Figure 3 As shown, Figure 2 Three days after inoculation in A, a clear inhibition zone appeared in the halotolerant Bacillus YH30. Figure 2 B is the control group, with no inhibition zone, that is, Bacillus halogenide YH30 can significantly inhibit the growth of Penicillium expansum, and according to Figure 3 It can be seen that on the 10th day, the diameter of the Penicillium expansum lesion was only 1.58 cm. Example 3:

[0025] Plate confrontation culture test of the effect of salt-tolerant Bacillus YH30 on the blueberry gray mold pathogen Botrytis cinerea ( Botrytis cinerea ) inhibition results: 10 μL of the purified Botrytis cinerea spore suspension was inoculated onto the filter paper in the center of the PDA culture dish. Four sterile filter paper discs were placed 3 cm away from the center. 10 μL of the fermentation suspension of Bacillus halophilus YH30 (concentration of 1×10 7 Each treatment was repeated three times independently, and the colony diameters were counted after culturing at 28°C in the dark for 2 days.

[0026] The antagonistic ability to Botrytis cinerea was observed and determined, and the colony diameter was determined by the cross method.

[0027] Specific results such as Figure 4 and Figure 5 As shown, Figure 4 After 2 days of inoculation in A, the salt-tolerant Bacillus YH30 showed an obvious inhibition zone and could significantly inhibit the growth of Botrytis cinerea. Figure 5It can be seen that the diameter of the lesion was only 1.7 cm on the 8th day. Example 4:

[0028] Plate confrontation culture test of the effect of halotolerant Bacillus sp. YH30 on the pathogenic fungus Colletotrichum anthracnose of macadamia nuts ( Colletotrichum aeschynomenes ) inhibition results: The purified anthrax bacteria were inoculated on the filter paper in the center of the PDA culture dish. Four sterile filter papers were placed 3 cm away from the center. 10 μL of the fermentation suspension of Bacillus halophilus YH30 (concentration of 1×10 7 Each treatment was repeated three times independently, and the colony diameters were counted after culturing at 28°C in the dark for 3 days.

[0029] The antagonistic ability to anthrax was observed and determined, and the colony diameter was determined by the cross-cross method.

[0030] Specific results such as Figure 6 and Figure 7 As shown, Figure 6 After 3 days of inoculation in A, the salt-tolerant Bacillus YH30 showed an obvious inhibition zone, which could significantly inhibit the growth of anthrax pathogenic fungi. Figure 7 The figure shows that the diameter of the lesion was only 3.61 cm on the 10th day. Example 5:

[0031] Effect of spore suspension of Bacillus halodurans YH30 on the growth of Penicillium expansum in grapefruit ( Penicillium expansu )’s control effect: 1. Fruit Selection: Grapefruit fruits of the Rio Red variety were purchased from the Donghua Farmers' Market in Kunming, Yunnan Province. Undamaged fruits of uniform size, firmness, and maturity, uniform color and gloss, and luster were selected for testing. The surfaces of the test samples were pre-washed with tap water, disinfected with 75% alcohol, and then dried at room temperature.

[0032] 2. Preparation of test culture: Penicillium expansum strains were previously isolated from diseased postharvest grapefruit and stored in the laboratory. The pathogenic strain was activated by incubation on potato dextrose agar (PDA) at 28°C for 7 days. Spores were first rinsed from the plate with sterile distilled water and transferred to a sterile centrifuge tube (filled with 2 mL of sterile water). After thorough shaking, the spore concentration was adjusted to 1×10 using a hemocytometer. 5 The halodurable Bacillus subtilis YH30 used in the test should be activated at least twice in LB liquid medium before use, and a spore suspension should be prepared. The concentration should be adjusted to 1×10 using a hemocytometer before use. 7 / mL, set aside.

[0033] 3. The experiment consisted of treatment and control groups. In the treatment group, uniformly spaced holes (approximately 5 mm deep and 4 mm wide) were made at the equator of the fruit using a sterile pipette tip. Each hole was inoculated with 20 μL of a spore suspension of Bacillus halodurans YH30. In the control group, each hole was inoculated with an equal volume of sterile water. Three hours later, 20 μL of a spore suspension of Penicillium expansum was injected into each wound. After inoculation, the fruit was placed in its packaging at room temperature. After five days, the fruit wounds were observed for infection, and the diameter of the lesions was recorded. Lesion diameters greater than 15 mm were determined using the cross-hatch method. Nine fruits were included in each treatment, for a total of 18 fruits in two groups, with two replicates.

[0034] like Figure 8 As shown in the figure, 10 days after inoculation with the spore suspension of Bacillus halophilus YH30 and pathogens, it was found that the spore suspension of Bacillus halophilus YH30 significantly inhibited the expansion of the rot diameter of grapefruit fruit ( Figure 8 B), effectively controlling post-harvest rot caused by Penicillium expansum in fruit wounds. Figure 9 As shown, pre-inoculation with Bacillus halodurans YH30 significantly reduced the lesion diameter of grapefruit fruit, with the lesion diameter reaching 2.63 cm on the 10th day. Example 6:

[0035] Effect of spore suspension of salt-tolerant Bacillus sp. YH30 on blueberry gray mold (Botrytis cinerea Botrytis cinerea )’s control effect: 1. Fruit Selection: Blueberries of the "Jewel" variety were purchased from the Donghua Farmers' Market in Kunming, Yunnan Province. Undamaged blueberries of uniform size, firmness, and maturity, uniform color and gloss, and undamaged were selected for testing. The surface of the test samples was pre-washed with tap water, disinfected with 75% alcohol, and then dried at room temperature.

[0036] 2. Preparation of test culture: A strain of Botrytis cinerea, previously isolated from diseased postharvest blueberry fruit, was stored in the laboratory. The pathogenic strain was activated by incubation on potato dextrose agar (PDA) at 28°C for 7 days. Spores were washed from the plate with sterile distilled water and transferred to a sterile centrifuge tube (filled with 2 mL of sterile water). After thorough shaking, the spore concentration was adjusted to 1×10 using a hemocytometer. 5 The halodurable Bacillus subtilis YH30 used in the test should be activated at least twice in LB liquid medium before use, and a spore suspension should be prepared. The concentration should be adjusted to 1×10 using a hemocytometer before use. 7 / mL, set aside.

[0037] 3. Soak the blueberry fruits in 2% sodium hypochlorite for 1 minute for disinfection. After air drying, divide the fruits into 2 groups, 9 fruits in each group, for a total of 18 fruits. Use a sterilized punch to punch a hole with a diameter and depth of 4 mm at the equator of the blueberry fruit. After the juice in the hole has drained, quantitatively inject 5 μL of the halodurable Bacillus YH30 treatment solution into each hole of the fruit. For the CK group, sterile water was used. After 2 hours, 5 μL of the gray mold spore suspension (concentration of 1×10 5 CFU / mL). After the treatment solution was completely absorbed, individual fruits were packaged in ziplock bags and stored in plastic baskets at room temperature. Each treatment consisted of nine fruits. After inoculation, the fruits were placed in packaging boxes at room temperature. After five days, the fruit wounds were observed for infection and the lesion diameter was recorded. When the diameter was greater than 5 mm, the lesion diameter was determined using the cross-hatch method. Each treatment consisted of nine fruits, with two groups totaling 18 fruits, repeated twice. Calculate the inoculation incidence rate (%) = total number of diseased holes in the fruit / total number of inoculated holes in the fruit × 100%.

[0038] The incidence of blueberry fruit was investigated and counted. Figure 10-12 As shown, Figure 10 and Figure 11 It can be shown that inoculation with the spore suspension of Halophilus Bacillus YH30 significantly inhibited the expansion of the rot diameter of blueberry fruits and the increase in the incidence of fruit, and effectively controlled the post-harvest rot caused by Botrytis cinerea in the fruit wounds. Among them, pre-inoculation with Halophilus Bacillus YH30 significantly reduced the diameter of the lesions of blueberry fruits. The diameter of the lesions was 1.23 cm on the 10th day. When all the CK group were diseased (the 8th day of storage, the incidence rate was 100%), the incidence rate was 83% after inoculation with Halophilus Bacillus YH30, and the storage conditions of the blueberries on the 8th day were as follows. Figure 12 As shown ( Figure 12 A is CK group, Figure 12 B is the YH30-treated group). Example 7:

[0039] The control effect of spore suspension of halotolerant Bacillus subtilis YH30 on anthracnose of macadamia leaves: 1. Test Fruit Selection: Macadamia leaves were collected from the hillside behind Southwest Forestry University in Kunming, Yunnan Province. Macadamia seedlings of the "Guireyin 3" variety were selected. Uniformly grown, disease-free, and mechanically damaged macadamia leaves were selected for testing. The test samples were pre-soaked in 75% alcohol for 30 seconds and then dried at room temperature.

[0040] 2. Preparation of test bacteria: The anthracnose fungus isolated from the diseased leaves of macadamia nuts was prepared. Colletotrichum aeschynomenes) strains, stored in the laboratory. The pathogenic strain was activated by culturing on potato dextrose agar (PDA) at 28°C for 7 days before use. The halodurable Bacillus subtilis YH30 used in the experiment was activated at least twice in LB liquid medium and prepared as a spore suspension. The spores were counted using a hemocytometer and the concentration was adjusted to 1×10 before use. 7 / mL, set aside.

[0041] 3. Use a sterile inoculating needle to prick each macadamia leaf twice along the veins on both sides of the leaf, ensuring that each wound is of uniform size. The leaves are then divided into two groups of nine leaves each, for a total of 18 leaves. The experiment consists of a control group and a treatment group.

[0042] The control group was treated with pathogenic mycelial disks with a diameter of 5 mm, and the treatment group was treated with 10 ml of 1×10 7 cells mL -1 10mL of 1X10 7 cells mL - 1 After treatment with a suspension of 100 μg / cm2, the leaves were inoculated with 5 mm diameter pathogenic mycelial disks. Lesion size was measured starting on the fifth day. Each treatment was replicated three times, and the experiment was repeated twice. Inoculation incidence (%) = total number of diseased wells on the leaf / total number of inoculated wells on the leaf × 100%.

[0043] Specific circumstances such as Figure 13-14 As shown, 2 days after inoculation with the spore suspension of Bacillus halophilus YH30 and pathogens, the incidence of macadamia nuts was investigated and statistically analyzed. It was found that the inoculation with the spore suspension of Bacillus halophilus YH30 significantly inhibited the expansion of the rot diameter of macadamia nut leaves and the increase in the incidence of leaves, and effectively controlled the rot caused by anthrax in leaf wounds. Among them, the pre-inoculation with Bacillus halophilus YH30 significantly reduced the lesion diameter of macadamia nut leaves. On the 10th day, the lesion diameter was 0.39 cm. When all the CK group were diseased (incidence rate 100%), the incidence rate was 83% after inoculation with Bacillus halophilus YH30. And as Figure 15 As shown, on the 10th day of storage, there was a significant difference in the growth of lesions between the control group (CK) and the treatment group ( Figure 15 A is the lesion growth of CK group, Figure 15 B shows the lesion growth of the YH30 treatment group).

[0044] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A strain of halotolerant Bacillus ( Bacillus halotolerans ) YH30, characterized by: The classification of the halogen-tolerant Bacillus subtilis YH30 is named as halogen-tolerant Bacillus subtilis Bacillus halotolerans , and was deposited in the General Microbiology Center of China Culture Collection Administration on December 2, 2024, with the deposit number CGMCC No: 32877.

2. A use of the halotolerant Bacillus YH30 according to claim 1 in preventing and controlling grapefruit penicillium mold, blueberry gray mold and macadamia anthracnose.

3. The use according to claim 2, characterized in that: The grapefruit Penicillium pathogenic fungus is Penicillium italicum ( Penicillium italicum ), Penicillium digitatum ( Penicillium digitatum )、Penicillium expansum( Penicillium expansu ) any one of the following; the blueberry gray mold pathogen is Botrytis cinerea ( Botrytis cinerea ); The macadamia nut anthrax pathogenic fungus is anthrax ( Colletotrichum aeschynomenes ).

4. The use according to claim 2, characterized in that: The application method is to prepare a spore concentration of 1×10 6 -1×10 8 The product with a concentration of 100mg / mL can prevent and control grapefruit blue mold, blueberry gray mold and macadamia anthracnose.

Images