CN112760267A

CN112760267A - Lysobacter enzymogenes CX06 for antagonizing xanthomonas campestris and application thereof

Info

Publication number: CN112760267A
Application number: CN202110178171.3A
Authority: CN
Inventors: 李磊; 李宝聚; 张子玉; 谢学文; 石延霞; 柴阿丽
Original assignee: Institute of Vegetables and Flowers Chinese Academy of Agricultural Sciences
Current assignee: Institute of Vegetables and Flowers Chinese Academy of Agricultural Sciences
Priority date: 2021-02-09
Filing date: 2021-02-09
Publication date: 2021-05-07
Anticipated expiration: 2041-02-09
Also published as: CN112760267B

Abstract

The invention discloses an lysobacter enzymogenes CX06 for antagonizing xanthomonas campestris and application thereof, wherein the lysobacter enzymogenes CX06 is preserved in China general microbiological culture Collection center of China Committee for culture Collection of microorganisms at 07-08 days of 2020, and the preservation number is CGMCC No. 20321. The invention also discloses application of the lysobacter enzymogenes CX06 in inhibiting pathogenic bacteria. In the bacteriostatic spectrum determination test, the strain CX06 shows broad-spectrum resistance and has antagonistic effect on 7 pathogenic fungi and 5 pathogenic bacteria. In a greenhouse potting effect test, the prevention effect of the strain CX06 on cabbage black rot is as high as 90.7%, the prevention effect on cabbage stem basal rot is 62.8%, and the prevention effect on cabbage bacterial black rot is 79.98%, which shows that the strain CX06 has a good application prospect and provides a certain theoretical basis and technical support for further development and utilization.

Description

Lysobacter enzymogenes CX06 for antagonizing xanthomonas campestris and application thereof

Technical Field

The invention belongs to the technical field of biology, and particularly relates to an antagonistic xanthomonas campestris lysobacter enzymogenes CX06 and application thereof.

Background

At present, the prevention and treatment measures for the xanthomonas campestris in production mainly depend on disease resistance breeding and chemical prevention and treatment. Common pesticides for chemical control include some agricultural antibiotics (kasugamycin), copper preparations (oxine-copper), and the like. In general, the types of pesticides for preventing and treating the xanthomonas campestris diseases are few, the use of some pesticides can be inhibited, and the prevention and treatment effects of some pesticides are general, so that more novel, efficient and safe prevention and treatment measures are urgently needed to be developed in production. The biological control is a control method which utilizes beneficial microorganisms to inhibit pathogenic bacteria, reduce the number of the pathogenic bacteria and prevent the infection of the pathogenic bacteria so as to reduce the occurrence of diseases, is concerned by people due to the advantages of green, no pollution, low cost and the like, and has great development prospect in the aspect of controlling bacterial diseases.

Lysobacter enzymogenes (Lysobacter enzymogenes) belongs to the family Xanthomonas (Xanthomonadaceae), the order Xanthomonas (Xanthomonadales), the class γ -Proteobacteria (Gamma Proteobacteria), and the phylum Proteobacteria (Proteobacteria). The lysobacter enzymogenes has high antagonistic bacteria activity and biological control potential of plant diseases, and is a novel biological control bacterium for plant diseases. The lysobacter enzymogenes has strong activities of chitinase, protease and cellulase, and can dissolve gram-positive bacteria, gram-negative bacteria, filamentous fungi, yeast and algae. The disease prevention mechanism is as follows: (1) is rich in extracellular enzymes; (2) secreting a biosurfactant; (3) producing an antibiotic; (4) better colonization ability. Therefore, the lysobacter enzymogenes strain with antagonistic action on xanthomonas campestris is separated and identified, and antibacterial active substances are obtained from the lysobacter enzymogenes strain, so that the lysobacter enzymogenes strain with safe, efficient and broad-spectrum biocontrol potential and the construction of biocontrol resources, the ecological environment is protected, and the use of chemical pesticides is reduced.

Disclosure of Invention

The invention aims to provide lysobacter enzymogenes CX06 for antagonizing xanthomonas campestris and application thereof.

In order to achieve the above object, the present invention firstly provides Lysobacter enzymogenes (CX 06) with the preservation number of CGMCC No. 20321.

The Lysobacter enzymogenes CX06 provided by the invention has been preserved in China general microbiological culture Collection center (CGMCC for short; address: Beijing city, rising area, Beicheng West Lu No. 1, institute of microbiology, China academy of sciences; postal code: 100101) in 07 month 08 in 2020, and the preservation number is CGMCC No. 20321.

In order to achieve the above object, the present invention further provides a suspension or culture solution or fermentation broth or fermentation product of the above Lysobacter enzymogenes (CX 06) or a microbial preparation containing the same.

In order to achieve the purpose, the invention also provides application of the Lysobacter enzymogenes (CX 06) or bacterial suspension or culture solution or fermentation liquor or fermentation product thereof or microbial inoculum containing the same in inhibiting pathogenic bacteria.

The invention also provides application of the Lysobacter enzymogenes CX06 or bacterial suspension or culture solution or fermentation liquor or fermentation product thereof or a microbial inoculum containing the same in preparation of products for inhibiting pathogenic bacteria.

The invention also provides application of the Lysobacter enzymogenes CX06 or bacterial suspension or culture solution or fermentation liquor or fermentation product thereof or a microbial inoculum containing the same in preventing and treating plant diseases caused by pathogenic bacteria.

The invention also provides application of the Lysobacter enzymogenes CX06 or bacterial suspension or culture solution or fermentation liquor or fermentation product thereof or a microbial inoculum containing the same in preparing products for preventing and treating plant diseases caused by pathogenic bacteria.

The invention also provides application of the Lysobacter enzymogenes CX06 or bacterial suspension or culture solution or fermentation liquor or fermentation product thereof or a microbial inoculum containing the same in preventing and treating cabbage black rot and/or cabbage stem basal rot and/or cabbage bacterial black rot.

The invention also provides application of the Lysobacter enzymogenes CX06 or bacterial suspension or culture solution or fermentation liquor or fermentation product thereof or a microbial inoculum containing the same in preparing products for preventing and treating cabbage black rot and/or cabbage stem basal rot and/or cabbage bacterial black rot.

In order to achieve the above object, the present invention also provides a product, the active ingredient of which is the above Lysobacter enzymogenes (CX 06) or its suspension, culture solution, fermentation product, or microbial inoculum containing the same;

the product has any one of the following functions of a1) -a 3):

a1) inhibiting pathogenic bacteria;

a2) preventing and treating plant diseases caused by pathogenic bacteria;

a3) preventing and treating cabbage black rot and/or cabbage stalk base rot and/or cabbage bacterial black rot.

To achieve the above object, the present invention finally provides any one of the following methods b1) -b 3):

b1) a method of suppressing a pathogenic bacteria comprising the steps of: treating pathogenic bacteria with the Lysobacter enzymogenes (CX 06) or its suspension, culture solution, fermentation liquid, fermentation product or bacteria preparation containing the same;

b2) a method of controlling plant diseases caused by pathogenic bacteria, comprising the steps of: treating plants with the above Lysobacter enzymogenes (CX 06) or its suspension, culture solution, fermentation broth, fermentation product, or microbial preparation containing the same;

b3) a method for preventing cabbage black rot and/or cabbage stalk base rot and/or cabbage bacterial black rot comprises the following steps: treating cabbage or Chinese cabbage with the above Lysobacter enzymogenes (CX 06) or its suspension, culture solution, fermentation broth, fermentation product, or microbial inoculum containing the same.

Any of the above-mentioned pathogenic bacteria are plant pathogenic bacteria. The plant pathogenic bacteria can be pathogenic fungi or pathogenic bacteria.

The pathogenic fungi may specifically include Corynespora cassiicola (Corynespora cassiicola), Fusarium oxysporum (Fusarium oxysporum), Phytophthora capsici (Phytophthora capsicii), Botrytis cinerea (Botrytis cinerea), Rhizoctonia solani (Rhizoctonia solani), Phytophthora solani (Stemphylium solani), and Chaetomium citrulli (Ascochyta citrullina).

The pathogenic bacteria may specifically include Xanthomonas campestris pathovar campestris (Xanthomonas campestris pv. campestis), Corynebacterium michiganensis (Corynebacterium microprensis subsp. sepedonicus), Pseudomonas syringae lachrynensis (Pseudomonas syringae pv. lachrynans), Agrobacterium vitis (Agrobacterium vitis), Corynebacterium michiganensis (Corynebacterium microprensis subsp. microprensis).

Any of the plant diseases described above includes black rot and basal rot. The black rot can be cabbage black rot or Chinese cabbage bacterial black rot. The basal rot can be a cabbage stem basal rot.

Any one of the above plants may be Brassica oleracea or Brassica oleracea.

The bacterial concentration of the Lysobacter enzymogenes CX06 in any one of the above bacterial suspension, culture solution, fermentation broth, fermentation product or microbial inoculum can be 1 × 10⁷cfu/mL-1×10⁹cfu/mL, specifically 1X 10⁸cfu/mL。

In a specific embodiment, the specific preparation method of the bacterial suspension or culture solution or fermentation product or microbial inoculum can comprise the following steps: inoculating Lysobacter enzymogenes CX06 in LB liquid culture medium, and culturing at 28 deg.C and 180r/min under shaking to concentration of 1 × 10⁸cfu/mL。

In another embodiment, the specific preparation method of the bacterial suspension or culture solution or fermentation product or microbial inoculum can comprise the following steps: inoculating Lysobacter enzymogenes CX06 in LB liquid culture medium, and culturing at 28 deg.C and 180r/min under shaking to concentration of 1 × 10⁸cfu/mL, then diluted to 100 volumes with distilled water.

The invention separates a strain CX06 with obvious antagonism to wild rape yellow single cell pathogenic variety from the rhizosphere soil of crops. The strain CX06 is preliminarily identified as Lysobacter enzymogenes (Lysobacter enzymogenes) by analyzing the morphological characteristics, Biolog detection, electron microscope scanning, physiological and biochemical indexes and multigene phylogenetic tree of the strain. On different culture media, the strain CX06 grows very fast, and on NA culture media, LB culture media and TSA culture media, bacterial colonies are all in faint yellow, have protruding surfaces, are moist and glossy, and are not easy to pick up; the strain CX06 has certain influence on hyphae of pathogenic fungi, and can cause the enlargement, distortion and deformity of the hyphae and influence the normal growth and reproduction of the hyphae. The enzyme activity test is carried out on the strain CX06, and the metabolite of the strain CX06 is determined to contain cellulase, chitinase and protease. In the bacteriostatic spectrum determination test, the strain CX06 shows broad-spectrum resistance and has antagonistic effect on 7 pathogenic fungi and 5 pathogenic bacteria. In a greenhouse potting effect test, the prevention effect of the strain CX06 on cabbage black rot is as high as 90.7%, the prevention effect on cabbage stem basal rot is 62.8%, and the prevention effect on cabbage bacterial black rot is 79.98%, which shows that the strain CX06 has a good application prospect and provides a certain theoretical basis and technical support for further development and utilization.

Drawings

FIG. 1 shows 8 strains isolated and screened from soil samples. A: CX 01; b: CX 02; c: CX 03; d: CX 04; e: CX 05; f: CX 06; g: CX 07; h: CX 08.

FIG. 2 shows the morphological characteristics of strain CX06 on different media. A: LB culture medium; b: TSA medium; c: NA culture medium; d: 10% TSA medium.

FIG. 3 is an electron micrograph of strain CX 06.

FIG. 4 is a phylogenetic tree of strain CX06 based on multiple gene sequences.

FIG. 5 shows the analysis of bacterial inhibition spectrum of strain CX06 and the effect on the hypha of pathogenic fungus. Fig. 5a is the analysis of the bacterial inhibition spectrum of strain CX06 fungus, wherein, a: fusarium oxysporum (Fusarium oxysporum); b: botrytis cinerea (Botrytis cinerea); c: rhizoctonia solani (Rhizoctonia solani); d: corynebacterium polystachyum (Corynespora cassiicola); e: ascochyta citrulli (Diaporthe batatas); f: stemphylium solani (stemphyllium solani); g: phytophthora capsici (Phytophthora capsicii). FIG. 5b shows the effect of strain CX06 on the hyphae of pathogenic fungi.

FIG. 6 shows the bacterial inhibition spectrum analysis of strain CX 06. A: corynebacterium michiganensis subsp. sepedonicus; b: corynebacterium michiganensis canker pathogenic variant of corynebacterium michiganensis (Clavibacter michiganensis subsp.); c: agrobacterium vitis (Agrobacterium vitas); d: wild rape Xanthomonas campestris pathovar campestris (Xanthomonas campestris pv. campestris); e: pseudomonas syringae lachryrrans (Pseudomonas syringae pv. lachryrrans).

FIG. 7 shows the enzyme activity assay of strain CX 06. A: cellulase Celluase; b: chitinase; c: protease.

FIG. 8 is a growth curve of strain CX 06.

FIG. 9 is a pH change curve of strain CX 06.

FIG. 10 shows the potting test of strain CX06 against cabbage black rot. A: strain CX 06; b: kasugamycin 1000 times liquid; c: blank control (distilled water); d: healthy control.

FIG. 11 shows a potting test of strain CX06 for stem rot of cabbage. A: strain CX 06; b: 4% jinggangmycin 500 times liquid; c: blank control (distilled water).

FIG. 12 is a comparative test of the control effect of strain CX06 and other reagents. A: CX 06; b: polyoxin; c: paenibacillus polymyxa; d: bacillus cereus; e: pseudomonas fluorescens; f: bacillus firmus; g: bacillus licheniformis; h: a zhongshengmycin; i: copper hydroxide; j: b, bacillus subtilis; k: kasugamycin; l: zinc thiazole; m: ethylicin; n: copper (succinate + glutarate + adipate); o: copper quinolinate; p: saildew copper; q: tetramycin; r: blank control (distilled water).

Deposit description

The name of Chinese: lysobacter enzymogenes

Latin name: lysobacter enzymogenes

The strain number is as follows: CX06

The preservation organization: china general microbiological culture Collection center

The preservation organization is abbreviated as: CGMCC (China general microbiological culture Collection center)

Address: xilu No. 1 Hospital No. 3 of Beijing market facing Yang district

The preservation date is as follows: year 2020, month 07 and 08

Registration number of the preservation center: CGMCC No.20321

Detailed Description

The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.

The experimental procedures in the following examples, unless otherwise indicated, are conventional and are carried out according to the techniques or conditions described in the literature in the field or according to the instructions of the products. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified. The medium in the following examples is naturally pH unless otherwise specified. Unless otherwise stated, the quantitative tests in the following examples were performed in triplicate, and the results were averaged.

The test media and test solutions referred to in the following examples are as follows:

the PDA culture medium is used for antagonism tests, and the specific formula of the PDA culture medium is as follows: 200g of potato, 20g of glucose, 15g of agar and 1000mL of distilled water.

The CMC culture medium is used for the determination of the cellulase, and the specific formula of the CMC culture medium is as follows: magnesium sulfate 0.1g, ammonium sulfate 1g, 10 XPhosphate buffer (dipotassium hydrogen phosphate 70g, potassium dihydrogen phosphate 20g, distilled water 1000mL)100mL, yeast extract 5g, glycerol 2mL, sodium carboxymethylcellulose 1g, agar 8g, distilled water 900 mL.

The LB culture medium, the NB culture medium and the WA culture medium are used for measuring bacterial inhibition spectra, and the specific formula of the LB liquid culture medium is as follows: 10g of tryptone, 5g of yeast extract, 10g of NaCl and 1000mL of distilled water. The specific formula of the NB culture medium is as follows: 10g of peptone, 3g of beef powder, 5g of NaCl, 1000mL of distilled water and pH 7.0. The specific formula of the WA culture medium is as follows: 5g of agar and 1000mL of distilled water, and subpackaging the obtained mixture into test tubes with each volume of 5 mL.

The protease culture medium is used for measuring protease, and the specific formula of the protease culture medium is as follows: NA culture medium (10 g of peptone, 3g of beef powder, 5g of NaCl, 15g of agar, 980mL of distilled water, pH7.0), 20mL of skimmed milk, boiling at high temperature, and adding into the melted NA culture medium.

The chitinase culture medium is used for chitinase determination, and the specific formula of the chitinase culture medium is as follows: casein peptone (vitamin-free) 5g, chitin salt 3g, 5% dipotassium hydrogen phosphate aqueous solution 7mL, MgSO₄·7H₂5mL of O (1: 10000), 15g of agar, water and water are added to make the volume reach 1000mL, the pH value is 7.0, the temperature is 116 ℃, and the autoclave sterilization is carried out for 15 min.

Congo red solution: 0.2% (w/v). Congo red powder 2g, distilled water 1000 mL. A small amount of alcohol can be added to help the dissolution to be uniform.

NaCl solution: 1M. NaCl 58.5g, distilled water 1000 mL.

Example 1 isolation, identification and preservation of Strain CX06

Isolation of Strain CX06

1. Strain isolation

Taking rhizosphere soil samples of different crops, weighing 10g of each soil sample, adding the soil samples into 90mL of sterilized water, and placing the mixture in a constant temperature shaking table at 28 ℃ for shaking culture for 30 min. Mixing the soil sample according to the proportion of 10^-1To 10^-8Performing gradient dilution to gradient 10^-6、10^-7、10^-8The soil sample was plated, 100. mu.L of each gradient was pipetted, plated on LB plates, 3 plates each, and incubated at 28 ℃ for 2 days. Different types of colonies were picked and purified on LB plates, and after 3 times of purification, they were stored in a-80 ℃ cryopreservation tube. Finally, 8 strains of bacteria were isolated from rhizosphere soil of different crops and named CX01-CX08 (FIG. 1).

2. Strain screening

Using 8 strains obtained in the step 1 as test strains, using Xanthomonas campestris pathovar campestris (Xanthomonas campestris pv. Campesris) as an indicator strain, and screening 1 antagonistic biocontrol strain CX06 by a double-layer culture method.

The specific method comprises the following steps: selecting single colony of strain, inoculating in LB liquid culture medium, performing shaking culture at 28 deg.C and 180r/min to obtain 1 × 10⁸cfu/mL, inoculating 5 mu L of bacterial suspension of the strain to be detected in the center of the PDA plate, culturing at 28 ℃ for 24h, inverting the culture dish, adding 3mL of chloroform into each culture dish in a fume hood, and standing for 12h to inactivate antagonistic bacteria. Inoculating pathogenic bacteria into NB culture medium, performing shake culture at 28 deg.C and 180r/min for 36h, adding 100 μ L bacterial suspension into 4mL 5% (m/v) WA culture medium, mixing, and pouring into PDA plate as upper layer. Culturing for 24h in an incubator, and observing and measuring the size of the inhibition zone.

II, identification of Strain CX06

1. Physiological and biochemical identification

Referring to the method of the handbook of identifying common bacteria systems, the strain CX06 is subjected to the following physiological and biochemical tests respectively: gram staining test, growth temperature test, salt tolerance test, motility test, catalase test, V-P test, starch hydrolysis test, citrate utilization test and gelatin liquefaction test.

The results of physiological and biochemical measurements are shown in Table 1, and the results show that the strain CX06 is a gram-negative bacterium, the optimal growth temperature is 28-37 ℃, the strain can normally grow in 1% NaCl, and the strain 4% NaCl inhibits the growth and has mobility. Can hydrolyze chitin, gelatin and Tween 20, can not hydrolyze starch and cellulose, and can utilize citrate as carbon source.

TABLE 1 analysis results of physiological and biochemical characteristics of Strain CX06

Note: + indicates that the test result is positive; -indicating that the test result is negative.

2. Biolog assay

A single colony of the strain CX06 is selected and inoculated on a slant of a test tube of an NA culture medium, and the slant is cultured for 24 hours at 28 ℃. The strain CX06 was assayed for the utilization of a sole carbon source by the China center for agricultural microbial cultures Collection using the BIOLOG GEN III kit (according to the instructions of the kit).

The results of the Biolog assay are shown in Table 2 and indicate that strain CX06 can utilize D-maltose, alpha-D-lactose, D-melibiose, D-mannose, D-trehalose and L-rhamnose, but cannot utilize glucan, stachyose, D-mannitol and minocycline. The strain CX06 was preliminarily identified as a lysobacter.

TABLE 2 determination of the sole carbon Source utilization of Strain CX06 Using BIOLOG GENIII reagent strips

Note: positive; -, negative; w, weak positive

3. Morphological identification

1) Morphological observation

Taking 3 μ L CX06 bacterial liquid, respectively spotting in the center of 10% TSA, NA, LB culture medium, culturing at 28 deg.C for 2-4d, and observing morphology.

The results are shown in FIG. 2 and show that: the colony morphology of the strain CX06 on the NA culture medium, the LB culture medium and the TSA culture medium is light yellow, the colony surface is prominent, and the thallus is moist, glossy and sticky and is not easy to pick up; on 10% TSA culture medium, the colony morphology appears milky white, the surface is flat, and the colony is not easy to pick up.

2) Observation by electron microscope

The strain CX06 is streaked on the NA culture medium, and in the fresh growing place of the bacterial colony, the bacterial colony is washed by sterilized water, and the culture medium is slightly shaken and repeatedly washed to prepare bacterial suspension. Dripping 1-2 drops of fresh PTA compound dye solution on the paraffin plate, taking the new copper mesh sheet to adsorb the bacterial suspension, and placing on clean filter paper for drying. And when the bacterial suspension on the copper mesh is dried, contacting the surface of the copper mesh, which adsorbs the bacterial suspension, with the negative dye solution, and carrying out floating dyeing for 15 s. The copper mesh was removed from the negative dye solution, the negative dye solution was blotted dry with filter paper, and dried under a fluorescent lamp for 45 seconds. The sample was placed under a transmission electron microscope at 80 kv to observe the cell morphology. The results are shown in FIG. 3.

4. Molecular identification

After the genomic DNA of the strain CX06 was extracted in a small amount using a common bacterial genome extraction kit, the DNA was purified by using the 16S rDNA universal primer 27F: 5'-AGAGTTTGATCCTGGCTCAG-3', 1492R: 5'-TACGGCTACCTTGTTACGACTT-3', the 16S rDNA sequence of strain CX06 was amplified by PCR.

And (3) PCR running program: pre-denaturation at 95 ℃ for 5 min; denaturation at 95 ℃ for 30s, annealing at 55 ℃ for 30s, and extension at 72 ℃ for 45s for 34 cycles; extension at 72 ℃ for 10 min. The obtained product is subjected to sequence determination by Bomaide biology company, and the sequencing result shows that: the 16S rDNA gene sequence of the strain CX06 is shown as a sequence 1 in a sequence table.

And (3) downloading the rest sequences by the NCBI website, constructing a phylogenetic tree by using MEGA7.0 through a maximum likelihood method, and analyzing the genetic relationship of the phylogenetic tree.

As a result, as shown in FIG. 4, this strain was clustered with Lysobacter enzymogenes YC36, and it was confirmed that the strain CX06 was Lysobacter enzymogenes.

The strain CX06 is determined to belong to Lysobacter enzymogenes (Lysobacter enzymogenes) by combining the results of morphological identification, physiological and biochemical identification, Biolog determination and molecular identification.

Third, preservation of Strain CX06

Lysobacter enzymogenes (CX 06) has been deposited in China general microbiological culture Collection center (CGMCC; address: Beijing, the West Lu No. 1 of the sunward Suzhong, China academy of sciences, Microbiol research institute; postal code: 100101) at year 2020, month 07 and day 08, with the collection number of CGMCC No. 20321.

Example 2 determination of the bacterial inhibition Spectrum of Strain CX06

Firstly, determination of fungus bacteriostasis spectrum and influence of bacterial strain CX06 on hypha of pathogenic fungus

1. Determination of fungal inhibition spectra

And (3) determining the antibacterial spectrum of the fungi by adopting a plate confronting method.

Pathogenic fungi: corynebacterium polytrichum (Corynespora cassiicola), Fusarium oxysporum (Fusarium oxysporum), Phytophthora capsici (Phytophtora capsicii), Botrytis cinerea (Botrytis cinerea), Rhizoctonia solani (Rhizoctonia solani), Rhizopus solani (Stemphylium solani), and Chaetomium citrulli (Ascochyta citrullina).

Corynebacterium polystachyum (Corynespora cassicola) has been described in the literature "homosala, lipamopsis, eucrypti, schwerwinia". pathogenic differentiation of corynebacterium polystachyum in cucumber, tomato and eggplant hosts. 465- & 470 ", the public can obtain from the vegetable flower research institute of Chinese academy of agricultural sciences to repeat the experiments of the present application and cannot be used for other purposes.

Fusarium oxysporum (Fusarium oxysporum) has been described in the literature "Charantia, Zhang Xiaohui, Xueyaro, Xiechuan, Chailali, Li Bao Poly. pyrazolopyrimidine derivative BDO-1 induces resistance of cucumber to blight. 877- & 890 ", publicly available from the vegetable and flower institute of Chinese academy of agricultural sciences to repeat the experiments of the present application, and not usable for other purposes.

Phytophthora capsici (Phytophthora capsicii) has been described in the literature "Chengyongchao, Kanghuajun, Cabernet, Zhanghongjie, Xianan, Li Bao poly. establishment and application of Phytophthora capsici RT-PCR detection technology. horticultural proceedings, 2018, 45 (05): 997-.

Staphylium solani (stemphyllium solani) has been described in the literature "li xinyu, li yu liei, chenlida, shiyanxia, chairali, schoenzhi, li bao j. screening and identification of staphylium solani sigatoka antagonistic bacteria. horticulture proceedings, 2020, 47 (04): 741 and 748, publicly available from the vegetable and flower institute of Chinese academy of agricultural sciences to repeat the experiments of this application, and not for other uses.

The watermelon Ascochyta citrullina has been reported in the literature "Liushulin, Guxingfang, Shinya, Li Bao Ju, Miao break, Wangmei, Wanghei, Zhangsheng Ping, melon vegetable tendril blight research overview. Chinese vegetables, 2013 (18): 1-10 ", publicly available from the vegetable and flower institute of Chinese academy of agricultural sciences to repeat the experiments of the present application, and not for other uses.

Botrytis cinerea (Botrytis cinerea) has been described in the literature "chardonnay, tang ming, jin zhi, xie, chairali, li bao. 60-65, publicly available from the vegetable and flower institute of Chinese academy of agricultural sciences to repeat the experiments of the present application, and not for other uses.

Rhizoctonia solani (Rhizoctonia solani) has been described in the literature "antagonism of trichoderma viride against Rhizoctonia solani and fusarium oxysporum f.cucumeri, sorrel, grandsond, sauvigna delavayi,. trichoderma viride, chinese vegetables, 2008, (6): 9-12 ", publicly available from the vegetable and flower institute of Chinese academy of agricultural sciences to repeat the experiments of the present application, and not for other uses.

The specific method comprises the following steps: inoculating 5mm selected fungus target bacteria slice in the center of 90mm PDA plate, and culturing at 28 deg.C for 2 d. Inoculating the strain in liquid LB culture medium, performing shake culture at 28 deg.C and 180r/min for 16h, and adjusting the bacterial suspension concentration to 10⁸cfu/mL. 5 mu L of bacterial suspension of the strain is inoculated at 4 opposite points 10mm away from the edge of the culture dish, the inoculated liquid LB culture medium is used as a blank control, and the control growth amount (colony radius) and the treatment growth amount (growth radius after inoculation of bacteria) of the target bacteria are measured after the culture is carried out for 5 days at 28 ℃, and are expressed by the bacteriostasis rate. The bacteriostatic rate (%) (control growth amount-treated growth amount)/control growth amount × 100.

The results are shown in table 3 and fig. 5a, and show that: the strain CX06 can effectively inhibit the hypha growth of the polyspora, phytophthora capsici, fusarium oxysporum, botrytis cinerea, stemphylium solani, ascochyta citrullus and rhizoctonia solani. The inhibition rate of rhizoctonia solani is more than 50%, and the inhibition rate of rhizoctonia solani on other pathogenic fungi is more than 20%.

TABLE 3 fungal inhibition

Note: data are mean ± sem, with different small letters indicating significant differences at the 0.05 level.

2. Effect of Strain CX06 on hyphae of pathogenic fungi

And picking up hyphae at the boundary of the opposite plate bacterial colony in the determination of the fungus bacteriostasis spectrum, manufacturing a lactophenol oil slide, observing under an optical microscope, and taking a picture.

The results are shown in FIG. 5b and show that: compared with normal hyphae, the hyphae treated by the strain CX06 and pathogenic fungi in opposition are abnormal in shape, twisted, deformed, thickened on the cell wall at the top end, expanded in local cells, uneven in distribution of cell contents in the hyphae, disordered in growth, thickened on the cell wall, increased in abnormal branches and the like.

Second, determination of bacterial inhibition spectrum

And (3) performing bacterial inhibition spectrum determination by adopting a double-layer culture method.

Pathogenic bacteria: yellow rape wild rape pathogenic variant (Xanthomonas campestris pv. campestis), Corynebacterium melaleucum potato pathogenic variant (Clavibacter microorganismis subsp. sepedonicus), Pseudomonas syringae lachrynia pathogenic variant (Pseudomonas syringae pv. lachrynans), Agrobacterium vitis (Agrobacterium vitis), Corynebacterium glutamicum tomato canker pathogenic variant (Clavibacter microorganismis subsp. microorganischenensis).

Wild rape Xanthomonas campestris pathovar campestris (Xanthomonas campestris pv. campestris) has been described in the literature "Zhang Yang, Li jin Nu, Zhou Hui Min, Li Bao Ju. 23-25 ", publicly available from the vegetable and flower institute of Chinese academy of agricultural sciences to repeat the experiments of the present application, and not for other uses.

Corynebacterium michiganensis potato ring rot pathogenic variety (Clavibacter michiganensis subsp. sepedonicus) has been disclosed in "Leyizhao, Yi Yibanu, Zheng Fei, Shiyanxia, Chaihai, Xiuwen, Li Bao Ju." screening and prevention effect of disease-preventing bacteria of potato black nevus. report of plant pathology, 2020, https:// doi.org/10.13926/j.cnki.apps.000357 ", publicly available from vegetable and flower institute of Chinese academy of agricultural sciences to repeat the experiments of this application, and is not applicable for other uses.

Pseudomonas syringae lachryrna lachryrrans has been described in the literature "li huai ling, li bao poly doctor's hand-note (fifty-three) for the symptomatic diversity and comprehensive control of cucumber bacterial angular leaf spot disease, chinese vegetables, 2012, (21): 23-25 ", publicly available from the vegetable and flower institute of Chinese academy of agricultural sciences to repeat the experiments of the present application, and not for other uses.

Agrobacterium vitis (Agrobacterium vitis) has been described in "zhao yi banyan, li yu yi, xie weng, xianya, bairali, sun guangye, li bao pi.beleisi bacillus ZF2 has control effect on polyspora corynomycosis. 217 + 225 ", publicly available from the vegetable and flower institute of Chinese academy of agricultural sciences to repeat the experiments of the present application and not for other uses.

Corynebacterium michiganensis tomato canker pathogenic variants (Clavibacter michiganensis subsp. michiganensis) have been described in "li yizhao, yi, zheng fei, shi xian, chai li, xiweng, li bao ju. screening and control effects of bacillus apium soft rot antagonist, No. 2020, 36 (03): 388- & 395 ", the public is available from the vegetable and flower institute of Chinese academy of agricultural sciences to repeat the experiments of the present application and not be used for other purposes.

The specific method comprises the following steps: inoculating the strain in the liquidIn an LB culture medium, shaking and culturing at 28 ℃ and 180r/min for 16h, and adjusting the concentration of the bacterial suspension to 10⁸cfu/mL. mu.L of bacterial suspension of the strain was inoculated into the center of a 90mm PDA plate, and cultured at 28 ℃ for 24 hours in a hood using an inoculated liquid LB medium as a blank, 3mL of chloroform was added to the inverted dish, and the mixture was allowed to stand for 12 hours. Inoculating pathogenic bacteria in NB culture medium, performing shake culture at 28 deg.C for 36 hr, and adjusting bacterial suspension concentration to 10⁸cfu/mL. Add 100. mu.L of the pathogenic bacteria suspension to 4mL of 5% (m/v) WA medium, mix well and pour into PDA plate as the upper layer. Culturing for 48h in an incubator, and observing and measuring the size of the inhibition zone. The bacteriostatic rate (%) (control growth amount-treated growth amount)/control growth amount × 100.

The results are shown in table 4 and fig. 6, and show that: the strain CX06 has good inhibitory effect on pathogenic bacteria, and has the bacteriostatic diameters of more than 1.70cm on Xanthomonas campestris pathogenic variant, Corynebacterium michiganensis canker pathogenic variant, Agrobacterium vitis, Pseudomonas syringae lacrimation pathogenic variant and Corynebacterium michiganensis potato ring rot pathogenic variant, wherein the bacteriostatic rate on Xanthomonas campestris pathogenic variant is more than 30%.

TABLE 4 bacteria inhibition Rate

Example 3 detection of enzymatic Activity of Strain CX06

Detection of proteases

Selecting strain CX06 single colony, inoculating in LB liquid culture medium, shaking culturing at 28 deg.C and 180r/min to concentration of 1 × 10⁸cfu/mL. The strain CX06 bacterial suspension was inoculated at the central position of the protease culture medium, each treatment was repeated 3 times, and the digestion circle was observed after incubation at 28 ℃ for 24 hours.

Secondly, detection of cellulase

Selecting strain CX06 single colony, inoculating in LB liquid culture medium, shaking culturing at 28 deg.c and 180r/minTo a concentration of 1X 10⁸cfu/mL. Inoculating strain CX06 bacterial suspension at the center of the CMC culture medium, culturing at 28 ℃ for 24h, dyeing with 3mL Congo red dye for 30min, decolorizing with 5mL of 1mol/L NaCl solution for 15min, repeating each treatment for 3 times, and observing digestion circles.

Tri, chitinase assay

Selecting strain CX06 single colony, inoculating in LB liquid culture medium, shaking culturing at 28 deg.C and 180r/min to concentration of 1 × 10⁸cfu/mL. The bacterial strain CX06 bacterial suspension is inoculated at the central position of the chitinase culture medium, each treatment is repeated for 3 times, and the digestion circle is observed after the culture is carried out for 24 hours at the temperature of 28 ℃.

The results are shown in FIG. 7 and show that: the strain CX06 has obvious transparent circles in a protease culture medium, forms yellow transparent circles after being dyed and decolored in a CMC culture medium, and has very obvious transparent circles in a chitinase culture medium. Indicating that strain CX06 has the ability to produce proteases, cellulases and chitinases.

Example 4 growth Curve and pH determination of Strain CX06

Selecting single bacterial colony of the strain, inoculating in LB liquid culture medium, performing shake culture at 28 deg.C and 180r/min for 16h, and adjusting bacterial suspension concentration to 1 × 10⁸cfu/mL. The bacterial suspension is prepared according to the following steps of 1: 1000 parts of the mixture was added to LB liquid medium and cultured with shaking at 28 ℃. OD of 48h strain was continuously measured every 4h₆₀₀The values are repeated 3 times, together with the number of viable bacteria and the pH value.

The results are shown in fig. 8 and 9, and show that: under the condition of 28 ℃, the bacterial number of the strain CX06 rapidly increases at the initial stage of culture, reaches the maximum value after 30h, and OD is₆₀₀The value was 2.12, the number of plate colonies was 2.7X 10¹²Per mL; then the number of the thalli is gradually stabilized. In addition, the pH of strain CX06 gradually increased with the increase of the culture time, and the pH was 8.14 after 48 hours.

Example 5 detection of Effect of Strain CX06 on prevention and treatment of blue-and-white rot

And (4) carrying out cabbage black rot prevention effect determination by adopting a spray inoculation method.

The specific method comprises the following steps: selecting pathogenic variants of Brassica napus (Xanthomonas campestris pv. campuses)tris) single colony was inoculated in NB medium, and shaking-cultured at 28 ℃ and 180r/min to a concentration of 1X 10⁸cfu/mL, seedlings of Brassica oleracea (Zhonggan No. 21) were pretreated by spray inoculation. Selecting strain CX06 single colony, inoculating in LB liquid culture medium, shaking culturing at 28 deg.C and 180r/min to concentration of 1 × 10⁸cfu/mL, diluting by 100 times, spraying and inoculating to cabbage seedlings 24h after inoculation of pathogenic bacteria. Each treatment was repeated 3 times for 10 strains. Thereafter, cabbage development was observed every day. And calculating the morbidity, disease index and prevention and treatment effect. The experiment was set up with 3 treatments: (1) 100 times of bacterial liquid (1X 10) of strain CX06⁸cfu/mL); (2) 4% kasugamycin 1000 times liquid; (3) blank control (distilled water); (4) healthy control.

4% kasugamycin wettable powder (registered No. PD20100329, Mawei Biotech Co., Ltd., Mount Rushan).

Incidence (%) was 100 × total number of diseased leaves/total number of examined leaves.

Disease index is 100 × ∑ (number of diseased leaves at each stage × relative stage value)/(total number of investigated leaves × 9). Classification criteria of disease grade: level 0: no disease spots; level 1: the lesion area accounts for less than 5% of the whole leaf area; and 2, stage: the lesion area accounts for 6 to 25 percent of the whole leaf area; and 3, level: the lesion area accounts for 26-50% of the whole leaf area; 4, level: the lesion area accounts for 51 to 75 percent of the whole leaf area; and 5, stage: the lesion area accounts for more than 75% of the whole leaf area.

The control effect (%) is 100 x (control disease index-treatment disease index)/control disease index.

The results are shown in table 5 and fig. 10, and show that: the strain CX06 has good control effect on cabbage black rot caused by yellow-cell pathogenic variety of wild rape under greenhouse condition. The disease index of the cabbage plants inoculated with strain CX06 was 8.4, which was 82.2 lower than the disease index of the control test group inoculated with the Xanthomonas campestris pathovar. From the control effect, the control effect of the strain CX06 on cabbage black rot is 90.7%, which is higher than the control effect of kasugamycin.

TABLE 5 potted plant control of cabbage Black rot by Strain CX06

Treatment of	Concentration of	Index of disease condition	Control effect (%)
				CX06	1×10⁸cfu/mL	8.4	90.7
Kasugamycin	1000 times liquid	14.4	84.1
				Control CK	-	90.6	-

Example 6 detection of Effect of Strain CX06 on controlling cabbage Stem base rot

And (4) carrying out the determination of the cabbage stalk basal rot prevention effect by adopting a wheat grain inoculation method.

The specific method comprises the following steps: pricking caulis et folium Brassicae Capitatae (Zhonggan No. 21) stem base with needle to make wound, inoculating with wheat grain with Rhizoctonia solani mycelium, treating for 1d, irrigating root, selecting strain CX06, inoculating into LB liquid culture medium, performing shake culture at 28 deg.C and 180r/min to obtain 1 × 10 strain⁸cfu/mL, 10mL for each plant, 10 plants each, after which cabbage development was observed daily. And calculating the morbidity, disease index and prevention and treatment effect. The experiment set up 3 treatments: (1) strain CX06 (1X 10)⁸cfu/mL); (2) 4% validamycin 500 times liquid; (3) blank control (distilled water).

4% validamycin aqua (Tongda chemical plant, Jining, Shandong province, PD 20142295).

The results are shown in table 6 and fig. 11, and show that: the strain CX06 has good control effect on the cabbage stalk basal rot caused by rhizoctonia solani under the greenhouse condition. The disease index of the cabbage plant inoculated with the strain CX06 is 31.25, and is reduced by 53.45 compared with a control test group inoculated with rhizoctonia solani. From the control effect, the control effect of the strain CX06 on the cabbage stalk base rot is 62.8 percent, and is higher than the control effect of 4 percent of jinggangmycin.

TABLE 6 potted plant control of cabbage stalk rot by Strain CX06

Treatment of	Concentration of	Index of disease condition	Control effect (%)
				CX06	1×10⁸cfu/mL	31.25	62.8
4% validamycin aqua	500 times liquid	44.4	47.2
				Control CK	-	84.1	-

Example 7 control of lysobacter enzymogenes CX06 with other drugs

The in vivo control effect of the strain CX06 and other medicaments on the bacterial black rot of the white vegetables is determined by a spraying method.

The specific method comprises the following steps: selecting a single colony of Xanthomonas campestris pathovar campestris (Xanthomonas campestris pv. campestris) and inoculating to NB medium, and culturing at 28 deg.C and 180r/min under shaking to a concentration of 1 × 10⁸cfu/mL, and spraying and inoculating on the Chinese cabbage seedlings. After the pathogenic bacteria are inoculated for 24 hours, respectively diluting the control medicament and CX06 bacterial suspension by 100 times, spraying and inoculating the diluted control medicament and CX06 bacterial suspension on Chinese cabbage seedlings, normally culturing, and investigating the disease condition index and the prevention effect of other treatment groups after the plants of the blank control treatment group are completely diseased. Distilled water was used as a blank (CK) for the test article. Repeat 3 times for each 15 plants treated. The preparation method of the CX06 bacterial suspension comprises the following steps: selecting strain CX06 single colony, inoculating in LB liquid culture medium, shaking culturing at 28 deg.C and 180r/min to concentration of 1 × 10⁸cfu/mL。

The control agents were: 10 hundred million CFU/g Paenibacillus polymyxa wettable powder (Guangdong Shangfeng biological science Co., Ltd., registration number: PD20181264) 350-fold liquid, 1000 hundred million spores/g Bacillus subtilis wettable powder (Wuhan Keno biological science Co., Ltd., registration number: PD20140209) 1400-fold liquid, 8 hundred million spores/g waxy Bacillus wettable powder (Shandong Tainuo medicine Co., registration number: PD20094534) 100-fold liquid, 100 hundred million spores/g Bacillus firmus wettable powder (Jiangxi Shuquan biological science Co., registration number: PD20184023) 100-fold liquid, 80 hundred million live spores/ml Bacillus licheniformis water solution (Guangxi Swingzi pesticide Co., registration number: PD20140122) 90-fold liquid, 5 hundred million spores/g fluorescent pseudomonas wettable powder (Shandong Tainuo medicine Co., registration number: PD 0874) 50-fold liquid, a 3% zhongshengmycin wettable powder (Fujian Kaili biological products Co., Ltd., registration number: PD20110113)650 times liquid, a 4% kasugamycin wettable powder (Shandong province Rushan Korea biological science Co., registration number: PD20100329)1000 times liquid, an 80% ethylicin emulsifiable solution (Kaifeng Dacron Biotech Co., registration number: PD20101285)2200 times liquid, a 0.3% tetramycin aqueous solution (Liaoning Microengineering Co., registration number: PD 345)1100 times liquid, a 10% polyoxin wettable powder (Shandong province Derong province Biotechnology Co., registration number: PD20100857)1000 times liquid, a 30% copper (Heilongqiziqi Kaihahu chemical engineering Co., registration number: PD20097367)275 times liquid, a 46% copper hydroxide water dispersible granule (U.S. Du Pongzhou PD 0051800 registration number: PD 0053) 2011 Jiang river Queen Quyou chemical engineering Co., registration number: PD20097367)275 times liquid, a 46% quinoline suspension (Olympha Co., U.S. Biotechnology Co., registration number: Otsuma technology Co., Ltd., registration certificate number: PD20200743)1500 times liquid, 1.6% benziothiazolinone liniment (shanxi da huate science and technology industries ltd., registration number: PD20120375), 700 times of solution, 20% thiazole zinc suspending agent (zhejiang new agro-chemical industry ltd, registration no: PD20096932)1000 times of the solution.

Classification criteria of disease grade: level 0: no disease spots; level 1: the lesion area accounts for less than 5% of the whole leaf area; and 2, stage: the lesion area accounts for 6 to 25 percent of the whole leaf area; and 3, level: the lesion area accounts for 26-50% of the whole leaf area; 4, level: the lesion area accounts for 51 to 75 percent of the whole leaf area; and 5, stage: the lesion area accounts for more than 75% of the whole leaf area. Incidence (%) was 100 × total number of diseased leaves/total number of examined leaves. Disease index is 100 × ∑ (number of diseased leaves at each stage × relative stage value)/(total number of investigated leaves × 9). The control effect (%) is 100 x (control disease index-treatment disease index)/control disease index.

The results are shown in table 7 and fig. 12, and show that: the prevention effect of the strain CX06 on the bacterial black rot of the white vegetables reaches 79.98 percent, which is obviously higher than that of other medicaments.

Table 7, results of comparison of control effects of lysobacter enzymogenes CX06 and other agents

Example 7 level of resistance of Strain CX06 to antibiotics

10 antibiotics were selected: ampicillin, carbenicillin, kanamycin, chloramphenicol, streptomycin, tetracycline, gentamicin, rifampicin, vancomycin and spectinomycin, 3 antibiotic gradient plates of 50. mu.g/mL, 60. mu.g/mL and 70. mu.g/mL were prepared, and CX06 bacterial suspension was inoculated to observe growth conditions for 3 replicates. The preparation method of the CX06 bacterial suspension comprises the following steps: selecting strain CX06 single colony, inoculating in LB liquid culture medium, shaking culturing at 28 deg.C and 180r/min to concentration of 1 × 10⁸cfu/mL。

The results are shown in table 8 and show that: the strain CX06 has higher tolerance capability to various antibiotics, the tolerance levels to ampicillin, carboxin, chloramphenicol, kanamycin, streptomycin, gentamicin and spectinomycin all reach more than 50 mu g/mL, the tolerance level to tetracycline reaches 30 mu g/mL, and the tolerance levels to rifampicin and vancomycin are below 50 mu g/mL, which indicates that the strain CX06 is only sensitive to rifampicin and vancomycin and is not sensitive to the rest 8 antibiotics.

TABLE 8 antibiotic resistance level of strain CX06(μ g/mL)

The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific embodiments, it will be appreciated that the invention can be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is possible within the scope of the claims attached below.

Sequence listing

<110> vegetable and flower institute of Chinese academy of agricultural sciences

<120> lysobacter enzymogenes CX06 for antagonizing xanthomonas campestris and application thereof

<160> 1

<170> PatentIn version 3.5

<210> 1

<211> 1326

<212> DNA

<213> Artificial Sequence

<400> 1

tgcttctggt gcaacaaact cccatggtgt gacgggcggt gtgtacaagg cccgggaacg 60

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tgcagactcc aatccggact gagatagggt ttctgggatt ggcttgccct cgcgggtttg 180

cagccctctg tccctaccat tgtagtacgt gtgtagccct ggccgtaagg gccatgatga 240

cttgacgtca tccccacctt cctccggttt gtcaccggcg gtctccttag agttcccacc 300

attacgtgct ggcaactaag gacaagggtt gcgctcgttg cgggacttaa cccaacatct 360

cacgacacga gcacgacagc catgcagcac ctgtgttcga gttcccgaag gcaccaatcc 420

atctctggaa agttctcgac atgtcaaggc caggtaaggt tcttcgcgtt gcatcgaatt 480

aaaccacata ctccaccgct tgtgcgggcc cccgtcaatt cctttgagtt tcagtgcgac 540

cgtacttccc aggcggcgaa cttaacgcgt tagcttcgat actgagggcc aagttgcccc 600

caacatccag ttcgcatcgt ttagggcgtg gactaccagg gtatctaatc ctgtttgctc 660

cacgctttcg tgcctcagtg tcagtgctgg tccaggtagc cgccttcgcc acagatgttc 720

ctcccgatat ctacgcattt cactgctaca ccgggaattc cgctaccctc taccgcactc 780

tagtaagcca gtttccaatg ccattcccag gttgagccca gggctttcac atcagactta 840

acaaaccacc tacgcacgct ttacgcccag taattccgag taacgcttgc acccttcgta 900

ttaccgcggc tgctggcacg aagttagccg gtgcttattc ttccggtacc gtcatgactc 960

aaggttatta accctaagct tttctttccg gacaaaagtg ctttacaacc cgaaggcctt 1020

cttcacacac gcggcatggc tggatcaggc ttgcgcccat tgtccaatat tccccactgc 1080

tgcctcccgt aggagtctgg accgtgtctc agttccagtg tggctgatca tcctctcaga 1140

ccagctacgg atcgtcgcct tggtgggcct ttaccccgcc aactagctaa tccgacgtcg 1200

gctcatctat ctgcgtgagg ccttgcggtc ccccactttc acccgtaggt cgtatgcggt 1260

attagcgtaa gtttccctac gttatccccc acaaataggc agattccgac gtattcctca 1320

cccgtc 1326

Claims

1. Lysobacter enzymogenes (CX 06) with preservation number of CGMCC No. 20321.

2. The bacterial suspension or culture solution or fermentation broth or fermentation product of Lysobacter enzymogenes (CX 06) according to claim 1, or a microbial preparation containing the same.

3. Use of Lysobacter enzymogenes (CX 06) according to claim 1, or a bacterial suspension or culture solution or fermentation broth or fermentation product thereof, or a microbial preparation containing the same, for inhibiting pathogenic bacteria.

4. Use of Lysobacter enzymogenes (CX 06) according to claim 1, or a bacterial suspension or culture solution or fermentation broth or fermentation product thereof, or a microbial preparation containing the same, for producing a product for inhibiting pathogenic bacteria.

5. Use of Lysobacter enzymogenes (CX 06) according to claim 1, or a bacterial suspension or a culture solution or a fermentation broth or a fermentation product thereof, or a microbial agent containing the same, for controlling a plant disease caused by a pathogenic bacterium.

6. Use of Lysobacter enzymogenes (CX 06) according to claim 1, or a bacterial suspension or a culture solution or a fermentation broth or a fermentation product thereof, or a microbial inoculum containing the same, for the preparation of a product for controlling plant diseases caused by pathogenic bacteria.

7. Use of Lysobacter enzymogenes (CX 06) according to claim 1, or a bacterial suspension, a culture solution, a fermentation broth, a fermentation product, or a microbial inoculum containing the same, for preventing and treating cabbage black rot and/or cabbage stem basal rot and/or cabbage bacterial black rot.

8. Use of Lysobacter enzymogenes (CX 06) according to claim 1, or a bacterial suspension, a culture solution, a fermentation broth, a fermentation product or a microbial inoculum containing the same, for preparing a product for preventing and treating cabbage black rot and/or cabbage stem basal rot and/or cabbage bacterial black rot.

9. A product, wherein the active ingredient is Lysobacter enzymogenes (CX 06) of claim 1, or a bacterial suspension or culture solution or fermentation broth or fermentation product thereof, or a microbial preparation containing the same;

the product has any one of the following functions of a1) -a 3):

a1) inhibiting pathogenic bacteria;

a2) preventing and treating plant diseases caused by pathogenic bacteria;

10. Any one of the following b1) -b 3):

b1) a method of suppressing a pathogenic bacteria comprising the steps of: treating pathogenic bacteria with Lysobacter enzymogenes (CX 06) according to claim 1, or a bacterial suspension or culture solution or fermentation broth or fermentation product thereof, or a microbial preparation containing the same;

b2) a method of controlling plant diseases caused by pathogenic bacteria, comprising the steps of: treating a plant with Lysobacter enzymogenes (CX 06) according to claim 1, or a suspension or a culture solution or a fermentation broth or a fermentation product thereof, or a microbial preparation containing the same;

b3) a method for preventing cabbage black rot and/or cabbage stalk base rot and/or cabbage bacterial black rot comprises the following steps: treating cabbage or Chinese cabbage with Lysobacter enzymogenes (CX 06) according to claim 1, or a suspension, a culture solution, a fermentation broth, a fermentation product thereof, or a microbial preparation containing the same.