Biocontrol fungus Penicillium citreonigrum JSNL-B16, biocontrol agent, preparation method and application thereof

Abstract

The invention belongs to the technical field of strawberry biocontrol, and discloses a biocontrol fungus Penicillium chrysogenum ( Penicillium citronigrum )JSNL-B16, biocontrol agent and preparation method and application thereof. The biocontrol bacterium identified in the present invention is named Penicillium chrysogenum ( Penicillium lemon-black )JSNL-B16, deposited by China General Microbiological Culture Collection Committee, with a deposit number of CGMCC NO.40490. B.cinerea and C.siamense Both have good antagonistic activity against B.cinerea The inhibition rate of the strain reached 66.85%. C.siamense The inhibition rate of the strain reached 51.24%. In addition, the biocontrol fungus Penicillium flavoglossum JSNL-B16 provided by the present invention significantly reduced the severity of the diseases of strawberry gray mold and anthracnose, and the root length, plant height, fresh weight and other indicators of strawberry were increased.

Description

Technical Field

[0001] The invention belongs to the technical field of strawberry biocontrol, and relates to a biocontrol bacterium Penicillium citreonigrum JSNL-B16, a biocontrol agent, and a preparation method and application thereof. Background Art

[0002] Strawberry is a perennial herbaceous plant of the genus Fragaria in the family Rosaceae, with the characteristics of early fruiting, short growth cycle, strong adaptability and high yield. Strawberry is rich in calcium, sodium, magnesium, iron and other beneficial trace elements, and has high nutritional value. In addition, due to its good color, fragrance and taste, strawberry is deeply loved by consumers and also brings high economic benefits. However, during the growth process of strawberry, it is often infected by a variety of diseases, among which strawberry gray mold and anthracnose are the most common diseases in strawberry production in recent years.

[0003] Strawberry gray mold is caused by Botrytis cinerea. In addition to infecting strawberries, it can also harm grapes, cucumbers, tomatoes, leeks, peppers and other plants, with a wide host range. Gray mold is widely distributed, spreads rapidly, and causes serious damage, causing huge losses to the yield and quality of fruits and vegetables worldwide, ranking second among the top ten plant diseases in the world. Gray mold is the most serious fungal disease that causes damage to strawberries after flowering. The disease mainly infects fruits, and also infects leaves, fruit stalks, calyx, petals and petioles, causing flower and fruit rot. The diseased fruit rate of susceptible varieties is generally 30%-60%, and in severe cases, even a total crop failure occurs, causing huge losses to strawberry production.

[0004] Strawberry anthracnose is caused by the genus Colletotrichum (Colletotrichum spp.), including Colletotrichum gloeosporioides, C.acuta tum, C.fragariae and C.siamense. In addition to infecting strawberries, it can also infect grapes, apples, peppers, mangoes, citrus, peaches, pears and soybeans. When strawberry anthracnose occurs, strawberry leaves, petioles, stipules, runners, petals and fruits can all be affected. The disease is highly harmful and destructive, generally causing a 25%-30% reduction in strawberry production, up to 80% in severe cases, or even devastating losses. Strawberry anthracnose is the third largest disease restricting China's strawberry industry.

[0005] In the existing technology, the prevention and control of strawberry gray mold and anthracnose mainly rely on chemical agents such as pyrimidineamines, amides, pyridinamines, dicarboximides, triazoles and pyrroles. However, the large-scale use of chemical agents has led to the continuous enhancement of pathogen resistance, greatly reducing the prevention and control effect, and posing a great threat to the ecological environment and food safety, endangering human health, and is not conducive to the sustainable development of my country's green agriculture.

[0006] Therefore, there is an urgent need for an efficient and low-toxic control measure for gray mold and anthracnose. Plant endophytic fungi inhabit asymptomatic host plant tissues and are one of the key factors affecting plant growth, nutrition and health. They can help plants obtain nutrients, inhibit plant pathogens, and resist biotic and abiotic stresses. Since plant endophytic fungi are derived from plants, they are safe for plants, environmentally friendly, and not prone to drug resistance. They can overcome the drawbacks of chemical pesticide control. Therefore, plant endophytic fungi are a source of new compounds that promote the sustainable development of green agriculture.

[0007] In summary, the prior art still lacks a green and highly effective biocontrol bacterium for preventing strawberry gray mold and anthracnose. Summary of the invention

[0008] In view of the above problems, the present invention provides a biocontrol fungus Penicillium citreonigrum JSNL-B16 for preventing and controlling strawberry gray mold and anthracnose, a biocontrol agent, and a preparation method and application thereof.

[0009] The present invention adopts the following technical solutions:

[0010] In a first aspect, the present invention provides a biocontrol bacterium for preventing and controlling gray mold and anthracnose of strawberry. The biocontrol bacterium is Penicillium citreonigrum JSNL-B16, and the depository unit is General Microbiology Center of China National Microbiological Culture Collection Committee, and the deposit number is CGMCC NO.40490.

[0011] Penicillium citreonigrum JSNL-B16, classified as Penicillium citreonigrum, was registered and deposited at the General Microbiology Center of the China Microorganism Culture Collection (CGMCC) on January 16, 2023. The deposit number is CGMCC NO.40490, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0012] After screening and identification, the ITS gene sequence of the genomic DNA of Penicillium citreonigrum JSNL-B16 was SEQ ID NO.1, and the BenA gene sequence was SEQ ID NO.2.

[0013] In a second aspect, the present invention provides a biocontrol agent for preventing and controlling strawberry gray mold and strawberry anthracnose, wherein the biocontrol agent comprises hyphae or conidia of Penicillium citreonigrum JSNL-B16, or a fermentation product of Penicillium citreonigrum JSNL-B16.

[0014] In a third aspect, the present invention further provides a method for preparing the biocontrol agent according to the second aspect, wherein the preparation method comprises any of the following steps:

[0015] 1) Penicillium citreonigrum JSNL-B16 with a preservation number of CGMCC NO.40490 is inoculated into PDA medium for cultivation; or

[0016] 2) The mycelium block of Penicillium citreonigrum JSNL-B16 with the preservation number of CGMCC NO.40490 was inoculated into PDB liquid culture medium for fermentation.

[0017] Furthermore, the fermentation conditions are: culturing in the dark at 22-26°C and 150-180 rpm for 4-6 days.

[0018] In a fourth aspect, the present invention further provides use of the biocontrol bacteria described in the first aspect or the biocontrol agent described in the second aspect in preventing and controlling strawberry gray mold and strawberry anthracnose.

[0019] Furthermore, the strawberry gray mold is caused by B. cinerea, and the strawberry anthracnose is caused by C. siamens.

[0020] In a fifth aspect, the present invention further provides a method for preventing and controlling gray mold and anthracnose of strawberry, the method comprising applying the biocontrol bacteria described in the first aspect or the biocontrol agent described in the second aspect to the plant growth environment.

[0021] Compared with the prior art, the present invention has the following technical effects:

[0022] 1) The biocontrol fungus Penicillium flavosum JSNL-B16 provided by the present invention has good antagonistic activity against B. cinerea and C. siamense, with an inhibition rate of 66.85% for B. cinerea strains and an inhibition rate of 51.24% for C. siamense strains. At the same time, the extracellular metabolites secreted by the strain JSNL-B16 have an inhibitory effect on both B. cinerea and C. siamense.

[0023] 2) The biocontrol fungus Penicillium flavosum JSNL-B16 provided by the present invention significantly reduces the disease severity of strawberry gray mold and anthracnose, increases its root length, plant height, fresh weight and other indicators, and has a growth-promoting effect on strawberry plants.

[0024] 3) The biocontrol fungus Penicillium flavosum JSNL-B16 provided by the present invention can induce strawberry to develop resistance to B. cinerea and C. siamense. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the specific implementation manner of the present invention or the technical solution in the prior art, the drawings required for describing the specific implementation manner are briefly introduced below.

[0026] Figure 1 This is a diagram showing the effect of Penicillium flavolumum JSNL-B16 in Example 1 on inhibiting the mycelial growth of B. cinerea (the 1st to 2nd plate from the left) and C. siamense (the 1st to 2nd plate from the right) on the plate;

[0027] Figure 2 The effect of the extracellular metabolites secreted by Penicillium flavolum JSNL-B16 on B. cinerea and C. siamense in Example 2;

[0028] Figure 3 The effect of Penicillium JSNL-B16 on strawberry plants in Example 3;

[0029] Figure 4 This is a graph showing the results of Penicillium flavosum JSNL-B16 inhibiting B. cinerea from infecting strawberry fruit in Example 4;

[0030] Figure 5 This is a graph showing the results of Penicillium flavosum JSNL-B16 inhibiting C. siamense from infecting strawberry leaves in Example 4. DETAILED DESCRIPTION

[0031] The embodiments of the technical solution of the present invention will be described in detail below in conjunction with the accompanying drawings. The following embodiments are only used to more clearly illustrate the technical solution of the present invention, and are therefore only used as examples, and the protection scope of the present invention cannot be limited by this. It should be noted that, unless otherwise specified, the technical terms or scientific terms used in this application should be the usual meanings understood by those skilled in the art to which the present invention belongs.

[0032] As described in the present invention, the term "biocontrol bacteria" refers to beneficial microorganisms that can prevent and control plant diseases, mainly bacteria, fungi and actinomycetes.

[0033] As described in the present invention, the term "B. cinerea" is classified as Fungi, Ascomycota, Leotiomycete, Helotiales, Sclerotiniaceae, Botryotini a. It is mainly distributed in the United States, Sweden, Germany, France, China, Brazil, Japan, South Korea, Argentina, India, Chile and other countries.

[0034] As described in the present invention, the term "C. siamense" is classified as: Eumycota, Deuteromyeotina, Coelomycetes, Melanconiales, Melanconiaceae, Colletotrichum. It is mainly distributed in the United States, Canada, Australia, Japan, the United Kingdom, China, Belgium, Germany, India, Vietnam, South Korea, Brazil, Panama, Mexico and other countries.

[0035] Example 1 Screening of Penicillium flavolum JSNL-B16

[0036] 1. Isolation of potential biocontrol bacteria

[0037] 1.1. Obtaining experimental materials: Select healthy strawberry plants with good growth from the field (Biancheng Town, Jurong City) and take their root tissues.

[0038] 1.2. Isolation and culture of endophytic fungi in strawberry roots: Cut several pieces of strawberry root tissue, add 100 mL of sterile water and 2 drops of Tween 20, shake at 220 rpm for 20 min at 25 ° C, treat with sterile water for 20 s, soak in 75% (v / v) ethanol for 30 s, soak in 2.0% (v / v) sodium hypochlorite solution for 2 min, and finally rinse with sterile water 3-4 times, and dry the surface moisture with sterile absorbent paper. Under sterile conditions, the root segments were further cut into shorter segments (0.25 cm), and 5 pieces were randomly selected from each sample and placed on PDA plate culture medium containing ampicillin (50 mg / L) and rifampicin (50 mg / L), with three replicates. The culture dish was sealed and cultured at 25 ° C in the dark for 1-2 days. When the mycelium appeared from the root tissue, the small piece of culture medium growing on the edge of the culture medium was carefully transferred to a new PDA plate together with the mycelium.

[0039] 2. Screening of antagonistic bacteria by plate confrontation method

[0040] The PDA plate confrontation method was used. B. cinerea and C. siamense strain cakes (5 mm in diameter) and the test endophytic fungi cakes that had been cultured for 3-5 days were symmetrically inoculated 2 cm from the edge of a 9 cm diameter plate containing 15 mL of PDA culture medium. The plate inoculated with only B. cinerea and C. siamense strain cakes was used as a control. The plates were cultured at 25°C in the dark. After 5 days, the colony radius of B. cinerea and C. siamense was measured and the inhibition rate was calculated.

[0041] Finally, a strain with good antagonistic activity against B. cinerea and C. siamense on the plate was screened out, which was specifically manifested by strong inhibition of the growth of pathogenic fungi hyphae. It was numbered JSNL-B16.

[0042] like Figure 1 As shown, the 1st and 2nd plates from the left are JSNL-B16 and B. cinerea strains confronting each other. Compared with the control, the colony of the B. cinerea strain in the experimental group is significantly smaller, and the inhibition rate reaches 66.85%. The 1st and 2nd plates from the right are JSNL-B16 and C. siamense strains confronting each other. Compared with the control, the colony of the C. siamense strain in the experimental group is significantly smaller, and the inhibition rate reaches 51.24%, indicating that the hyphae growth of both B. cinerea and C. siamense strains is inhibited.

[0043] 3. Molecular biological identification of the strains obtained

[0044] 3.1. The genomic DNA of strain JSNL-B16 was extracted using the kit (DP320-03) from TIANGEN.

[0045] 3.2. Amplify the ITS and BenA genes of genomic DNA respectively. DNA amplification uses a 30 μL reaction volume, including 15 μL 2×EasyTaqPCR SuperMix (+dye), 1 μL of primer F / R (10 μM), 2 μL template DNA and 11 μL ddH2O. The PCR amplification program conditions are as follows: 94°C pre-denaturation for 5 min; 94°C denaturation for 30 s, 54°C annealing for 45 s, 72°C extension for 60 s, a total of 35 cycles; and finally 72°C extension for 10 min.

[0046] 3.3. The amplified PCR products were subjected to 1% agarose gel electrophoresis and observed under ultraviolet light. The PCR products with target bands were sent to Beijing Qingke Biotechnology Co., Ltd. for sequencing. The sequencing results were as follows: the ITS gene sequence of the strain JSNL-B16 screened in step 2 was SEQ ID NO.1, and the BenA gene sequence was SEQ ID NO.2.

[0047] 3.4. Compare the two sequences of SEQ ID NO.1 and SEQ ID NO.2 on the NCBI database website. The comparison results are shown in Table 1. According to the results in Table 1, it can be inferred that the biocontrol bacteria JSNL-B16 of the present invention is Penicillium citreonigrum, and is named Penicillium citreonigrum JSNL-B16.

[0048] Table 1 Alignment results of SEQ ID NO.1 and SEQ ID NO.2

[0049]

[0050] Example 2 Effects of extracellular metabolites of Penicillium flavolum JSNL-B16 on B. cinerea and C. siamense

[0051] 1. Inoculate B. cinerea and Penicillium flavosum JSNL-B16 into 9 cm diameter culture dishes containing 15 mL PDA medium, place in a 25°C incubator, and culture for 5 days in the dark. When B. cinerea and C. siamense strains have grown all over the culture dishes, use a hole puncher (5 mm in diameter) to punch holes at the edge of the colonies to prepare bacterial plates.

[0052] 2. Acquisition of extracellular metabolites secreted by strain JSNL-B16

[0053] A water filter membrane (pore size 0.22 μm, diameter 5 cm) was covered on a 6 cm diameter PDA medium plate, and the filter membrane was closely attached to the surface of the PDA plate. Then, a JSNL-B16 bacterial plate (diameter 5 mm) of Penicillium chrysanthemi was inoculated on the filter membrane, and a blank PDA medium was used instead of the JSNL-B16 bacterial plate (diameter 5 mm) for control, and cultured at 25°C in the dark.

[0054] 3. After 1 day of cultivation, carefully remove the filter membrane with the culture of Penicillium JSNL-B16 on it with tweezers, and tear off the filter membrane and blank PDA block as a control. A plate containing extracellular metabolites of Penicillium JSNL-B16 and a blank PDA plate were prepared.

[0055] 4. In a clean bench, use an inoculation needle to pick up B. cinerea and C. siamense bacterial discs (5 mm in diameter) and inoculate them into the above-prepared plates, with the mycelium facing downwards. Set up 3 replicates for each.

[0056] 5. Place the inoculated plate in a 25°C incubator, incubate in dark for 3-5 days, and measure the colony size using the cross method. The inhibition rate is calculated according to the formula: inhibition rate = (colony diameter of the control group - colony diameter of the treatment group) / colony diameter of the control group × 100%.

[0057] The results are shown in Table 2.

[0058] Table 2 Inhibitory effect of extracellular metabolites secreted by JSNL-B16 on B. cinerea and C. siamense

[0059]

[0060] The numerical expression of colony diameter in the table is: mean ± standard error.

[0061] from Figure 2 As shown in the data of Table 2, the extracellular metabolites secreted by strain JSNL-B16 have inhibitory effects on both B. cinerea and C. siamense.

[0062] Example 3 Effect of Penicillium flavosum JSNL-B16 on the growth of strawberries

[0063] 1. Inoculate the strain Penicillium flavolumum JSNL-B16 into a 9-cm diameter culture dish and culture it at 25°C in the dark for 5 days.

[0064] 2. Cut 3 mm × 3 mm mycelium blocks at the edge of the Penicillium JSNL-B16 colony, transfer 30 mycelium blocks into 200 mL PDB liquid culture medium under sterile conditions, and culture them at 25 ° C, dark, and 160 rpm for 5 days. After the strain is fully fermented, the strain and the fermentation liquid are crushed to make a bacterial suspension.

[0065] 3. Select pots with a diameter of 12 cm. The experiment was set up in 2 groups. The experimental group was as follows: 50 mL of the bacterial suspension of Penicillium JSNL-B16 and sterilized vermiculite were added to each pot and mixed evenly. The mixture was added to the pot to 1 / 3. Three healthy strawberry seedlings (the variety is Hongyan) with the same growth were transplanted into each pot, and then covered with a thin layer of the mixture of bacterial suspension and vermiculite. The control group was as follows: 50 mL of PDB liquid culture medium and sterilized vermiculite were added to each pot and mixed evenly. The mixture was added to the pot to 1 / 3. Three healthy strawberry seedlings (the variety is Hongyan) with the same growth were transplanted into each pot, and then covered with a thin layer of the mixture of PDB liquid culture medium and vermiculite. Three replicates were set for each treatment.

[0066] 4. Water each group of strawberries with sterile water, and count the root length, plant height and fresh weight of the strawberries after 10 days.

[0067] The results are as follows Figure 3 And as shown in Table 3.

[0068] Table 3 Statistical results of strawberry root length, plant height and fresh weight

[0069]

[0070] Among them, the numerical expressions of root length, plant height and fresh weight in the table are: mean ± standard error.

[0071] From the data in Table 3, we can see that the plant height and fresh weight of the experimental group are better than those of the control group, and the root length is significantly increased. The above results show that Penicillium JSNL-B16 has a growth-promoting effect on strawberry plants.

[0072] Example 4 Penicillium JSNL-B16 induces strawberry resistance

[0073] 1. Effect of Penicillium flavosum JSNL-B16 on strawberry gray mold caused by B. cinerea

[0074] 1.1. Inoculate B. cinerea and Penicillium flavosum JSNL-B16 in 9 cm diameter culture dishes containing 15 mL PDA medium, place in a 25°C incubator, and culture in the dark for 5 days. Use a cork puncher (5 mm in diameter) to punch holes at the edges of the B. cinerea and JSNL-B16 colonies to prepare bacterial plates.

[0075] 1.2. Select healthy strawberry fruits (the variety is Hongyan), wash them with tap water, disinfect the surface with 70% (v / v) ethanol twice, and finally rinse the surface of the fruit with sterile water three times, and prick the fruit surface lightly 3-5 times with a sterilized inoculation needle. Place the above-mentioned JSNL-B16 bacterial disc on the wound on the fruit surface, with the mycelium side facing down. Use PDA blank culture medium to make a bacterial disc instead of JSNL-B16 for control, and moisturize with sterile wet absorbent cotton. Place the inoculated fruit in a sterilized tray, lay two layers of sterile gauze in the tray and wet it with sterile water, and cover the tray with plastic wrap to keep it moist. Place it at 25℃ and dark conditions for 24 hours.

[0076] 1.3 Remove the JSNL-B16 and PDA blank culture medium plates on the fruit, replace them with B. cinerea plates, with the mycelium side facing down, replace with new sterile wet absorbent cotton, cover with plastic wrap and seal the tray to keep moisture. Place at 25℃ in the dark for 3 days.

[0077] 1.4 Check the disease condition of strawberry fruits, record the diameter of lesions using the cross method, and calculate the average value.

[0078] The results are as follows Figure 4 And as shown in Table 4.

[0079] Table 4 Statistical results of lesion diameters on fruits inoculated with B. cinerea

[0080]

[0081] The numerical expression of colony diameter in the table is: mean ± standard error.

[0082] like Figure 4As shown, the right side shows the inoculation of strain JSNL-B16 24 hours later and then inoculated with B. cinerea strain, and the left side shows the inoculation of blank PDA 24 hours later and then inoculated with B. cinerea strain. It can be seen that the lesions in the experimental group are significantly smaller than those in the control group, indicating that the strawberry fruits in the experimental group have developed resistance to B. cinerea.

[0083] 2. Effect of Penicillium flavosum JSNL-B16 on strawberry anthracnose caused by C. siamens

[0084] 2.1. C.siamense and Penicillium flavosum JSNL-B16 were inoculated into 9 cm diameter culture dishes containing 15 mL of PDA medium, placed in a 25°C incubator, and cultured in the dark for 5 days. Use a cork puncher (5 mm in diameter) to punch holes at the edges of the C.siamense and JSNL-B16 colonies to prepare bacterial plates.

[0085] 2.2. Select healthy strawberry leaves (the variety is Hongyan), wash them with tap water, disinfect them with 70% (v / v) ethanol twice, and finally rinse the leaf surface with sterile water for 3 times. Use a sterilized inoculation needle to lightly prick the left and right sides of the leaf back 3-5 times. Place the above-mentioned JSNL-B16 bacterial disc on the wound on the right side of the leaf, with the mycelium facing down. Use a PDA blank medium bacterial disc (diameter 5mm) on the left side as a control. At the same time, place PDA blank medium on both sides of the leaf as a blank control, and moisturize with sterile wet absorbent cotton. The inoculated leaves are placed in a sterilized tray, two layers of sterile gauze are laid in the tray and wetted with sterile water, and the tray is covered with plastic wrap to keep it moist. Place it at 25℃ and dark conditions for 24 hours.

[0086] 2.3 Remove the JSNL-B16 and PDA blank culture medium plates on both sides of the leaves and replace them with C.siamense culture plates with the mycelium side facing down. Replace with new sterile wet absorbent cotton and cover the sealed tray with plastic wrap to keep moisture. Place at 25℃ in the dark for 6 days.

[0087] 2.4 Check the disease condition of strawberry leaves, record the diameter of lesions using the cross method, and calculate the average value.

[0088] The results are as follows Figure 5 And as shown in Table 5.

[0089] Table 5 Statistical results of lesion diameter on leaves inoculated with C.siamense

[0090]

[0091] The numerical expression of colony diameter in the table is: mean ± standard error.

[0092] like Figure 5As shown, the left side of the front of the leaf was inoculated with strain JSNL-B16 24 hours later and then inoculated with C.siamense strain, and the right side was inoculated with blank PDA 24 hours later and then inoculated with C.siamense strain. It can be seen that the lesions in the experimental group are significantly smaller than those in the control group, indicating that the strawberry leaves in the experimental group have developed resistance to C.siamense.

[0093] Unless otherwise specifically stated, the numerical value set forth in these embodiments does not limit the scope of the present invention. In all examples shown and described here, unless otherwise specified, any specific value should be interpreted as merely exemplary, rather than as restriction, and therefore, other examples of exemplary embodiments may have different values.

Claims

1. A biocontrol bacterium for preventing and controlling gray mold and anthracnose of strawberry, characterized in that: The biocontrol bacteria is Penicillium chrysogenum ( Penicillium citreonigrum )JSNL-B16, deposited by: China General Microbiology Culture Collection Center, deposit number: CGMCC NO. 40490.

2. A biocontrol agent for preventing and controlling strawberry gray mold and strawberry anthracnose, characterized in that: The biocontrol agent includes the yellow dark Penicillium ( Penicillium citreonigrum ) Mycelium or conidia of JSNL-B16, the strawberry gray mold is caused by Botrytis cinerea ( Botrytis cinerea ) is caused by Colletotrichum siamensis ( Colletotrichum siamens )cause.

3. A method for preparing the biocontrol agent according to claim 2, characterized in that: The preparation method adopts any of the following: 1) Penicillium chrysogenum with the accession number of CGMCC NO. 40490 ( Penicillium citreonigrum ) JSNL-B16 was inoculated and cultured in PDA medium; or 2) Penicillium chrysogenum with the accession number of CGMCC NO. 40490 ( Penicillium citreonigrum ) JSNL-B16 mycelium blocks were inoculated into PDB liquid culture medium for fermentation.

4. The preparation method according to claim 3, characterized in that: The culture conditions are: 22-26°C, 150-180 rpm, dark culture for 4-6 days.

5. Use of the biocontrol bacteria according to claim 1 or the biocontrol agent according to claim 2 in preventing and controlling strawberry gray mold and strawberry anthracnose, characterized in that: The strawberry gray mold is caused by Botrytis cinerea ( Botrytis cinerea ) is caused by Colletotrichum siamensis ( Colletotrichum siamens )cause.

6. A method for preventing and controlling gray mold and anthracnose of strawberry, characterized in that: The control method comprises applying the biocontrol bacteria of claim 1 or the biocontrol agent of claim 2 to the plant growth environment.

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