CN113881593A - Acer lawinii with biocontrol potential and application thereof - Google Patents

Abstract

The invention discloses a maple Rahnella anserine strain with biocontrol potential and application thereof. The Aceraceae has the strain number of ZF458, and the registration number of the Aceraceae ordinary microorganism center is CGMCC No. 22292. Experiments prove that the Aceraceae maple Rahnella ZF458 has wide antibacterial spectrum and can inhibit 16 pathogenic fungi and bacteria; the acer mono ZF458 can be applied to preventing and treating the Chinese rose root cancer and/or inhibiting the agrobacterium tumefaciens in actual production.

Description

Acer lawinii with biocontrol potential and application thereof

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a maple Rahnella with biocontrol potential and application thereof.

Background

China rose root cancer is a bacterial disease caused by Agrobacterium tumefaciens and occurs worldwide. According to the research, the disease reduces the stem thickness by 63 percent, reduces the yield by 36 percent and shortens the service life of plants (Li Xiaoli, Chenghiansen and Li Zheng, the prevention and treatment of the root cancer of cut flower China rose, Heilongjiang agricultural science 2009 (9): 77-78.). The Chinese rose root cancer pathogen can survive on tumor tissues and soil surfaces for months to more than one year. The nematode and grafting tool can be used for nematode, grafting tool and machine. If the humidity is too high, the wound is too large, the contact time with the soil is long, and the infection probability is increased. After disease attack, the tumor is nodular and woody, the diameter can reach several millimeters, and the plant after injury has poor growth, lack of vitality, small leaves, early yellow, early fallen leaves and thin flowers. The pathogenic bacteria have wide host range, and can simultaneously infect roses, peach trees and the like (occurrence and prevention technology of diseases of roses in the same city as solanum tuberosum. inner Mongolia forestry 2009, (06): 18-19.). Until now, no effective measures have been found to control this disease.

In view of the important economic and ornamental value of Chinese rose and the serious hazard of the root cancer, the exploration of effective prevention and treatment measures of the Chinese rose root cancer is of great significance. The research of Wanqilu shows that the agrobacterium tumefaciens separated from peach root cancer can be biologically controlled by using Bacillus as an antagonistic strain (Wanqilu. separation of peach root cancer pathogen and antagonistic bacteria screening research. Nanjing forestry university 2016, 37-41.); the actinomycetes separated from two poplar rhizosphere by the Jing has stronger bacteriostatic activity on poplar root cancer pathogenic bacteria, and the strain T-S-22 is selected as an antagonistic strain (the Jing, screening of poplar root cancer disease inoculation antibacterium and preliminary study of antagonistic activity thereof, Hebei agriculture university 2007, 33-39.); studies of Aspergillus niger xj (Aspergillus niger xj) and related spore powder products by nijiang showed that it is an antagonistic fungus with good control effect on agrobacterium tumefaciens (nijiang Aspergillus niger antibacterial active ingredient isolation and mechanism of action studies, 2017,57-59, university of guizhou); 2 Alcaligenes faecalis with obvious inhibition effect on root cancer are obtained by high bud and the like (high bud, plum blossom, GuoRongjun and the like. 2 peach rhizosphere bacteria Alcaligenes faecalis has inhibition effect on root cancer. fruit tree academy 2015,32(2):267 and 273.). At present, no report about the Acer maple Rahnella has the function of preventing and treating the Chinese rose root cancer.

Disclosure of Invention

The invention aims to solve the technical problem of how to prevent and treat Chinese rose root cancer or how to inhibit agrobacterium tumefaciens.

In order to solve the technical problems, the invention provides the Acerana maple. The Aceraceae is Aceraceae, the strain number is ZF458, and the registration number of the Aceraceae is CGMCC No. 22292. Hereinafter referred to as Acerana maple ZF 458.

The 16S rDNA nucleotide sequence of the Acer aizoon ZF458 can contain a DNA molecule shown as a sequence 1 in a sequence table. The nucleotide sequence of the gyrB gene of the Aceraceae ZF458 can be a sequence 2 in a sequence table. The nucleotide sequence of rpoB gene of Aceraria maple ZF458 can be sequence 3 in the sequence table. The nucleotide sequence of rpoD gene of Aceraria maple ZF458 can be sequence 4 in the sequence table.

In order to solve the technical problems, the invention also provides a culture of the Acer nigeri ZF458, wherein the culture can be obtained by culturing the Acer nigeri ZF458 in a microorganism culture medium (such as a fermentation liquid containing the Acer nigeri ZF458 and a substance secreted into a liquid culture medium, or a solid culture medium containing the Acer nigeri ZF 458).

In order to solve the technical problems, the invention also provides a microbial inoculum. The inoculant may comprise Aceraceae ZF458 or/and metabolites of Aceraceae ZF458 or/and culture of Aceraceae ZF 458.

The active ingredients of the fungicide can be Aceraceae maple Rahnella ZF458 or/and metabolites of Aceraceae maple Rahnella ZF458, the active ingredients of the fungicide can also contain other biological ingredients or non-biological ingredients, and other active ingredients of the fungicide can be determined by a person skilled in the art according to the effect of the fungicide.

The microbial inoculum may also include a carrier. The carrier may be a solid carrier or a liquid carrier. The solid carrier is a mineral material or a biological material; the mineral material may be at least one of grass peat, clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica, and diatomaceous earth; the biological material is at least one of straws, pine shells, rice straws, peanut shells, corn flour, bean flour, starch, grass peat and animal manure of various crops; the liquid carrier can be water; the bacterium agent, Aceraria maple ZF458 or/and Aceraria maple ZF458 metabolite can exist in the form of cultured living cells, fermentation liquor of living cells, filtrate of cell culture or mixture of cells and filtrate. The preparation formulation of the microbial inoculum can be various preparation formulations, such as liquid, emulsion, suspending agent, powder, granules, wettable powder or water dispersible granules.

According to the requirement, the microbial inoculum can also be added with a surfactant (such as Tween 20, Tween 80 and the like), a binder, a stabilizer (such as an antioxidant), a pH regulator and the like.

Above, the metabolite of the Aceraceae maple Rahnella ZF458 can be a fermentation broth of Aceraceae maple Rahnella ZF 458. The fermentation liquor of Aceraceae Aceranikowii ZF458 can be prepared by the following method: culturing Acer Rahneri ZF458 in liquid fermentation culture medium, and collecting fermentation liquid (containing Acer Rahneri ZF458 and substances secreted into the liquid culture medium), wherein the fermentation liquid is metabolite of Acer Rahneri ZF 458.

In the above, the microbial inoculum may be a microbial inoculum having at least one of the following characteristics:

A1) has phosphorus dissolving capacity;

A2) producing siderophore;

A3) inhibiting agrobacterium vitis;

A4) inhibiting agrobacterium chinensis;

A5) preventing and treating plant root cancer;

A6) inhibiting pathogenic bacteria of plant diseases.

The pathogenic pathogen of the plant root cancer described above may be agrobacterium roseum or agrobacterium vitis. The plant may be sunflower.

The phytopathogenic pathogen described above may be a bacterium or a fungus. The bacteria can be any one of root cancer bacteria, muskmelon fruit blotches, cucumber soft rot, tomato ulcers and tomato bacterial spot germs. The fungus can be any one of colletotrichum capsici, phytophthora capsici, corynespora cubensis and alternaria solani.

The agrobacterium tumefaciens described above may be agrobacterium vitis and/or agrobacterium rosa chinensis. The plant described above may be sunflower. The sunflower variety may be Sasa veitchii.

The invention also discloses at least one application of Aceraria maple ZF458, which belongs to the protection scope of the invention:

B1) inhibiting pathogenic bacteria of plant diseases;

B2) inhibiting agrobacterium vitis;

B3) inhibiting agrobacterium chinensis;

B4) preventing and treating plant root cancer.

In order to solve the technical problems, the invention also provides at least one application of the microbial inoculum, which is described above, as follows:

B1) inhibiting pathogenic bacteria of plant diseases;

B2) inhibiting agrobacterium vitis;

B3) inhibiting agrobacterium chinensis;

B4) preventing and treating plant root cancer.

The phytopathogenic pathogen described above may be a bacterium or a fungus. The bacteria can be any one of root cancer bacteria, muskmelon fruit blotches, cucumber soft rot, tomato ulcers and tomato bacterial spot germs. The fungus can be any one of colletotrichum capsici, phytophthora capsici, corynespora cubensis and alternaria solani.

The agrobacterium tumefaciens described above may be agrobacterium vitis and/or agrobacterium rosa chinensis.

The pathogenic pathogen of the plant root cancer described above may be agrobacterium roseum or agrobacterium vitis. The plant may be sunflower.

In order to solve the technical problems, the invention also provides a method for preparing the microbial inoculum. The method may include the step of culturing Aceraceae maple Rahnella ZF458 in a microbial culture medium.

The Acer maple Rahnella bacteria ZF458(Rahnella aceris) is obtained by screening soil samples collected from a marsh land, and experiments prove that the Acer maple Rahnella ZF458 has a wide antibacterial spectrum and can inhibit 16 pathogenic fungi and bacteria; the bacillus subtilis has better prevention effect on the Chinese rose agrobacterium tumefaciens, shows good prevention effect on the Chinese rose agrobacterium tumefaciens in a living test, and does not attack all sunflowers inoculated with antagonistic bacterium maple Rahnella ZF458 compared with the control of two pathogenic bacteria inoculated with the Chinese rose agrobacterium and the agrobacterium vitis. In actual production, the Acer maple Rahnella ZF458 can be applied to preventing and treating Chinese rose root cancer and/or inhibiting Agrobacterium tumefaciens, and has wide application prospect.

Deposit description

The strain name is as follows: aceraria acearum

Latin name: rahnella acids

The strain number is as follows: ZF458

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: 2021 year 05, 08 months

Registration number of the preservation center: CGMCC No.22292

Drawings

FIG. 1 is a strain morphology observation. (A) Strain ZF458 PDA plate back; (B) strain ZF458 single colony; (C) observing morphological characteristics of the thalli by a transmission electron microscope; (D) and observing the morphological characteristics of the thallus by a scanning electron microscope.

FIG. 2 shows construction of ZF458 multigene phylogenetic tree based on 16S rDNA, gyrB, rpoB and atpD.

FIG. 3 shows the determination of the bacterial inhibition spectrum of the strain ZF458 on pathogenic fungi. The first row of graphs is, from left to right, a tomato gray mold pathogen (Botrytis cinerea), a tomato gray leaf spot pathogen (Stemphylium lycopersici (Enjoji) Yamamoto), a cucumber Rhizoctonia solani (Rhizoctonia solani), a cucumber gummy stem blight pathogen (Ascochyta citrilinia), a pepper anthracnose pathogen (Colletotrichum capsaci (Syd.) E.J.Butler & Bisby.), a cucumber root rot pathogen (Fusarinm solani), a Phytophthora capsici pathogen (Phytophthora capsici), a cucumber target spot pathogen (Corynespora cassiopola), and the second row of graphs is, from left to right, a corresponding control strain without addition of biocontrol agents.

Fig. 4 shows the determination of the bacterial inhibition spectrum of the strain ZF458 on pathogenic bacteria. The first row of graphs are sequentially from left to right of tomato bacterial wilt pathogenic bacteria (Ralstonia solanacearum), melon fruit spot pathogenic bacteria (Acidoorax avenae subsp. citrus), Chinese rose root cancer pathogenic bacteria (Agrobacterium tumefaciens), tomato ulcer pathogenic bacteria (Clavibacterium micondum subsp. Microbacterium), tomato bacterial spot pathogenic bacteria (Pseudomonas syringae pv. tomato), potato ring rot pathogenic bacteria (Clavibacterium micobacterium subsp. sepedonic), cucumber angular leaf spot pathogenic bacteria (Pseudomonas syringae pv. lachryman), cucumber soft rot pathogenic bacteria (Pectibacter carotovorum), and the second row of graphs are sequentially from left to right of corresponding controls without adding biocontrol strains.

Fig. 5 is the analysis of the biocontrol related factor of the strain ZF 458. (A) ZF458 phosphorus solubilizing ability; (B) ZF458 siderophore capacities.

FIG. 6 is a graph of the growth curve of the strain ZF458, the abscissa is the shake culture time of the strain, and the ordinate is the absorbance OD of the bacterial suspension_600nmThe value is obtained.

FIG. 7 is a curve of pH value change of the strain ZF458, the abscissa is strain shake culture time, and the ordinate is pH value of bacterial suspension.

Fig. 8 shows the in vivo control effect of strain ZF458 on rose root cancer. (A) Inoculation strain ACCC 19185; (B) inoculating a mixed solution (1:1) of 1500 times of 46% copper hydroxide water dispersible granules and the strain ACCC 19185; (C) inoculating a mixed solution (1:1) of the strain ZF458 and the strain ACCC 19185; (D) inoculating sterilized water; (E) inoculating a strain K308; (F) inoculating a 1500-time liquid mixture (1:1) of 46% copper hydroxide water dispersible granules and a strain K308; (G) inoculating mixed liquor (1:1) of a strain ZF458 and a strain K308; (H) and inoculating sterilized water.

FIG. 9 shows the control effect of bacterial suspension of strain ZF458 and supernatant on Chinese rose root cancer. (A, B) cell suspension results; (C, D) centrifugation of the supernatant of the cell suspension.

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 tomato variety to be tested in the following examples is "Zhongza 201" from the institute for vegetables and flowers, academy of agricultural sciences of China.

The media in the following examples are as follows:

the NB liquid medium (for shake culture propagation of bacteria) had the composition: 10g of peptone, 3g of beef powder, 5g of NaCl5g, 20g of agar and 1000mL of distilled water.

The composition of the NA liquid medium (for activated preservation of bacteria) is: 10g of peptone, 3g of beef powder, 5g of NaCl and 1000mL of distilled water.

The composition of the WA medium (for determining bacterial inhibition spectrum) is as follows: agar 5g and distilled water 1000mL, and the mixture was dispensed into 5mL test tubes.

The PDA medium (for fungal antagonism experiments) had the composition: 200.0g of potato, 20.0g of glucose, 20.0g of agar and 1000mL of distilled water.

The composition of the PD medium (for the fungal propagation shake culture test) was: 200.0g of potato, 20.0g of glucose and 1000mL of distilled water

The composition of NBRIP inorganic phosphorus medium (for phosphorus solubilizing ability assay) was: ca₃(PO₄)₂5g, glucose 10g, (NH)₄)₂SO₄ 0.15g，KCl₂·6H₂O 0.5g，MgSO₄·7H₂O0.25 g, distilled water 1000 mL.

The composition of the CAS medium (for siderophore assay) was: 0.5mg CAS dissolved in 50mL water; l mM FeCI₃·6H ₂0 in 10mM HCI, pH 2.0; 72.9mg HDTMA was dissolved in 40mL water, pH was adjusted to 7.0, volume was adjusted to 100mL, sterilized and mixed with 900mL (agar 20g, glucose 9g, peptone 4.5g, beef extract 2.7g, sodium chloride 4.5g)

The composition of the NaCl solution was: 58.5g of 1M NaCl, 1000mL of distilled water.

Example 1 isolation, screening, identification and preservation of strains

Firstly, separation and purification of strains

A soil sample is collected from Sichuan marsh lands, and the specific sampling method comprises the following steps: sampling points are randomly selected from the plot by adopting a five-point sampling method, 0-10cm of soil is collected by a soil collector, and each treatment is repeated for 3 times. The collected soil samples are uniformly mixed and sealed by a polyethylene sterile bag, and the mixture is placed in an ice box and taken back to a laboratory and stored at the temperature of minus 20 ℃ for later use.

Weighing 2g of uniformly mixed soil sample, adding the uniformly mixed soil sample into a centrifugal tube filled with 18mL of sterile water, oscillating and uniformly mixing the uniformly mixed soil sample by using a vortex oscillator to prepare a soil suspension, and sequentially diluting the soil suspension by a 10-time gradient until the dilution multiple is 10^-7. Respectively diluting by 10 times by using a micropipette^-4、10^-5、10^-6、10^-7The soil suspension was pipetted 100. mu.L and spread on LB plates, 28. mu.L, respectivelyCulturing at inverted temperature for 24 h. Single colonies of different morphologies were picked and placed in 50 wt% sterile glycerol and stored at-80 ℃ until use.

Secondly, identifying strains

2.1 morphological identification

Morphological feature observation was performed on the strain ZF 458.

The results are shown in FIG. 1 and show that: the strain ZF458 is milky white and translucent on a PDA solid culture medium, and the periphery of a colony is wet. The strain observed under transmission electron microscope is rod-shaped, with a long diameter of about 4 μm and a short diameter of about 1 μm, and has round ends (as shown in FIG. 1).

2.2 physiological and biochemical identification

The following physiological and biochemical tests were carried out on the strain ZF458 with reference to the methods in "textbook for identification of common bacteria systems" (Dongxu bead, Chuiamiao English. 2001. Manual for identification of common bacteria systems. Beijing: scientific Press.) and "Bojie's Manual for identification of bacteria" (Bukannan R E.1984. Bojie's Manual for identification of bacteria. Beijing: scientific Press.): gram test, growth temperature test, salt tolerance test, gelatin liquefaction test, acid production test and the like.

The results are shown in table 1 and show that: the strain ZF458 is gram-negative bacteria, can normally grow at 4-41 deg.C and NaCl concentration of 1%, and can utilize glycerol, arabinose, mannitol, inositol, etc. The ZF458 was preliminarily determined to be of the genus Rahnella in combination with the above.

TABLE 1 identification of physiological and biochemical characteristics

Note: + is positive; -is indicated as negative.

2.3 Biolog detection

ZF458 single colonies are picked and inoculated on a KB culture medium test tube inclined plane, the culture is carried out for 24h at the constant temperature of 28 ℃, and the determination of the utilization of the unique carbon source is carried out by using a BIOLOG GEN III kit (operated according to the kit specification) by the China agricultural microbial strain collection center.

The use of the unique carbon source by strain ZF458 on the BIOLOG GENIII reagent strip is shown in Table 2.

TABLE 2 determination of sole carbon Source utilization of Strain ZF458 using BIOLOG GENIII reagent strips

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

2.4 molecular characterization

After extracting the strain ZF458 genomic DNA in small amounts using the TIANGEN bacterial genomic DNA extraction kit, 16S rDNA, gyrB, rpoB, and rpoD genes were selected for amplification and sequencing (table 3). The PCR amplification system is 25 mu L, and comprises 0.5 mu L of each of the upstream primer and the downstream primer and 1 mu L of template DNA; 12.5 mu L of T-Taq Mix; ddH₂O10.5. mu.L. PCR reaction procedure: pre-denaturation at 95 ℃ for 3 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.

TABLE 3 ZF458 common conserved gene primer sequence information table

Name of Gene	Upstream primer sequence (5 '-3')	Downstream primer sequence (5 '-3')	Fragment size
				16S rDNA	AGAGTTTGATCCTGGCTCAG	AAGGAGGTGATCCAGCCGCA	1543bp
gyrB	GACAAGCTGGTGTCTTCCGA	CATCTCGCCCAGACCTTTGT	2416bp
				atpD	CACGGTTGAGCACATCCTCT	GGAGAGTACTTCGCGAGTCG	1383bp
rpoB	GTCCGACTTCACTGTGAGCA	GTGAAAAAGCCAGCGACGTT	4029bp

The resulting PCR product was sequenced by Bomader Biometrics. The sequencing result shows that: the 16SrDNA, gyrB, rpoB and atpD gene sequences of the strain ZF458 are respectively shown as sequences 1 to 4 in a sequence table. And (3) downloading the rest sequences by the NCBI website, connecting and comparing the 16S rDNA, gyrB, rpoB and atpD gene sequences of different strains by using MEGA 7.0, Sequence Matrix and Seaview4 in Sequence, constructing a phylogenetic tree by adopting a maximum likelihood method, and analyzing the classification status.

Construction of ZF458 multigene phylogenetic tree based on the rDNA, gyrB, rpoB and atpD gene sequences of the strains ZF 45816S, the results are shown in FIG. 2, and the results show that: the strain ZF458 is gathered in one branch with Aceraceae SAP-19.

And determining that the strain ZF458 belongs to Aceraria maple (Rahnella aceri) according to the results of morphology, physiological and biochemical characteristics, Biolog detection and molecular identification.

Third, preservation of Strain ZF458

Aceraceae (Rahnella aceriana) ZF458 has been deposited in China general microbiological culture Collection center (CGMCC; address: Beijing Kogyo No. 1, West Lu 1, Kyoho, China academy of sciences, Microbiol research institute; postal code: 100101) at 08.05.2021, and the preservation number is CGMCC No.22292 (Aceraceae ZF 458).

Example 2 determination of the bacterial inhibition Spectroscopy of Strain ZF458

8 common vegetable pathogenic fungi and 8 pathogenic bacteria are selected to determine the bacteriostasis spectrum of the Aceraceae maple Rahnella ZF 458.

The specific appearance of the 8 vegetable pathogenic fungi and 8 pathogenic bacteria are shown in table 3, and are as follows:

colletotrichum capsicii (Syd.) E.J. Butler & Bisby has been disclosed in the document Dugongfu, Shigella, Li Bao, Guo Ying lan, Popul, Hainan winter vegetable anamorphic fungi research 2016,14(03): 142-.

Botrytis cinerea (Botrytis cinerea) has been disclosed in the document "Shiyanxia, Tangming, jin Ji Wen, Xiechu, Chailali, Li Bao poly. vegetable crop Botrytis cinerea" for evaluation of resistance to different types of fungicides Chinese vegetable 2016,3:60-65 ", publicly available from the vegetable and flower institute of the applicant's Chinese academy of agricultural sciences for repeating the present invention and not for other uses.

The corynebacterium polystachyum (corynebacterium cassicola) has been disclosed in the literature "bulrush, ligupo, cabbagel, schwerewingii. corynebacterium polystachyum is pathogenic in cucumber, tomato and eggplant, horticulture proceedings 2011,38(3): 465-.

Phytophthora capsici (Phytophthora capsicii) is disclosed in the literature "Zhuhui, Wang Happy, Li Bao Ju, Cabernet Sauvignon, Chili root rot type epidemic disease pathogen identification and control agent screening, plant protection academy 2007, (4): 373) 378.

Rhizoctonia solani (Rhizoctonia solani) has been disclosed in the document "antagonism of Rhizoctonia solani and Fusarium oxysporum of Cucumis sativus and Fusarium oxysporum" Chinese vegetable 2008, (6):9-12 ", and is available to the public from vegetable and flower research institute of Chinese academy of agricultural sciences for repeating the present invention and is not useful for other applications.

Fusarium solani (Fusarium solani) has been disclosed in "cardia-aster, Shiyanxia, Xiechuan, Chailali, Li Bao-poly. calcium cyanamide soil disinfection control effect on cucumber root rot and soil pathogenic bacteria. Garden proceedings 2016,43(11): 2173-.

The diagnosis and control of the leaf spot of the tomato Stemphylium roseosporus (Enjoji) Yamamoto is disclosed in the document "plum blossoms, perigory aromas, li jinju, xi scholaria". chinese vegetable 2010, (23):24-26 ", publicly available from vegetable and flower research institute of chinese academy of agricultural sciences to repeat the present invention and not usable for other purposes.

The species Ascochyta citrullina is disclosed in the literature "SHI Yanxia, MENG Shanshan shan, XIE Xuewen, CHAI Ali, LI Baoju. Dry heat treatment processes of the occurrence of Cladosporumcumerinum, Ascochyta citrullina, and Colpolytrichum orbiculare on the surface and interior of the cell section. Horticultural Plant Journal 2016,2(01): 35-40", publicly available from vegetable and flower research institute of the national academy of agricultural sciences, to repeat the present invention, and not available for other uses.

The specific origins of 8 pathogenic bacteria are as follows:

corynebacterium michiganensis subsp. michiganensis has been disclosed in the document "li huan, huixia, xietui, li bao, occurrence rule and prevention and treatment technique of tomato canker, chinese vegetables 2011,23: 24-27", publicly available from the vegetable and flower institute of the applicant's chinese academy of agricultural sciences, to repeat the present invention, and not usable for other applications.

The Agrobacterium tumefaciens (Agrobacterium tumefaciens) is derived from China agricultural microbial culture collection management center, and the strain number is ACCC 19185.

The lilac Pseudomonas tomato pathogenic variety (Pseudomonas syringae pv. tomato) is disclosed in the literature "Chailali, Paiguoli, Guowawa, Cabernet, Xianauxia, Schimmer plum, Li Bao poly. establishment and application of a real-time fluorescence quantitative PCR detection method for tomato bacterial spot pathogen.

The Clavibacter michiganensis subsp (sepedonicus) is disclosed in the literature "Maria Yan, Zhansen, Wang Li, Gao Zhong Qiang, Yu hong Jun, Jiang Wei Jie, Jiang Wei doctor, focused production of early spring arched shed potato high-efficiency cultivation technique in the first line of fourteen Teng City, China vegetable 2015, (08):70-73.

Solanaceous Ralstonia solanacearum has been disclosed in the literature "quadruple PCR detection method for china army, caylaria, cabbages, the xue article, the Yuan army sea, the Li Bao poly. tomato bacterial spot pathogen, ulcer pathogen, Ralstonia solanacearum and scab pathogen. horticulture bulletin 2018,45(11): 2254-.

The generation rule and prevention and treatment of bacterial fruit blotch of the watermelon subspecies of the Acidovorax avenae (Acidovorax avenae subsp. citrulli) 'Fanxgxi, Jinwei, Zhou Hui Ming, Li Bao Ju doctor's hand-note (forty one) melon are disclosed in Chinese vegetable 2011, (21) 26-29.

Pseudomonas syringae cucumber angular leaf spot pathogenic variant (Pseudomonas syringae pv. lachrymans) "Penyue, Fangxingfang, Zhang Shengping, Li Bao Jue, Shehiwen, Bokeliang, Dongshuo, Miao break, research progress on cucumber bacterial angular leaf spot, Chinese vegetables 2021(03): 28-35", publicly available from vegetable and flower research institute of Chinese academy of agricultural sciences, to repeat the present invention, and is not available for other uses.

Carrot soft rot Pectobacterium (Pectobacterium carotovorum) "Li Lei, Zhao Yi Banyan, Zhengfei, Shiyanxia, Chaarali, Thanksgang, Li Bao Jue. screening and preventing effects of antagonistic bacillus for soft rot of celery are disclosed in China biological prevention and control institute 2020,36(03):388 one 395", the public can obtain from vegetable and flower research institute of Chinese academy of agricultural sciences to repeat the invention and can not be used as other purposes.

The inhibition effect of the Merremia maplensis ZF458 on pathogenic bacteria is measured by adopting a plate confronting method (fungi) and a double-layer culture method (bacteria) respectively. The plate confrontation method (fungus) for measuring the inhibition effect of the Aceraria maplensis ZF458 on pathogenic bacteria comprises the following specific steps: inoculating 5mm diameter pathogenic fungus cake in the middle of PDA plate, and dripping 5 μ L of the OD at a distance of 2.5cm from the center by cross method_600nmThe Acer maple Rahnella ZF458 bacterial suspension (experimental treatment) of 0.8 is compared with the Acer maple Rahnella ZF458 bacterial suspension, and cultured at 28 ℃, and the plate is investigated after the control plate grows to the edge of the plate. Each treatment was repeated 3 times. The control colony diameter and the experimental treatment colony diameter of the target bacteria are measured and expressed by the bacteriostasis rate. Bacteriostatic ratio (%) - (control colony diameter-diameter of experimental treated colony)/diameter of control colony]X 100. Double-layer culture method (bacteria) for measuring maple treeA method for measuring the inhibitory effect of Endomyces gracilis ZF458 on pathogenic bacteria, which comprises the following steps: taking Aceraria maple ZF458 glycerol tube from-80 deg.C refrigerator, sucking 1mL, adding into 250mL conical flask containing 100mL NB culture medium, shaking at 28 deg.C and 180r/min for 24 hr, and measuring OD_600nmValue and adjustment of OD Using blank NB liquid Medium_600nmWhen the strain is equal to 0.8 (NB liquid medium is used as a blank control), a suspension of Merremia maple ZF458 is obtained, 5 mu L of the suspension of Merremia maple ZF458 is inoculated in the center of a PDA plate, and the plate is incubated at 28 ℃ for 24h and then inactivated by fumigation with chloroform to serve as a lower auxin layer (experimental treatment). The control was made of a suspension of Acer amurensis ZF 458. 5mL of the suspension containing 100. mu.L of OD was used_600nmWA medium of pathogenic bacteria of 0.8 as the upper layer. After culturing at 28 ℃ for 48h, the diameter of the inhibition zone is measured, the inhibition rate is calculated, and each treatment is repeated for 3 times. Bacteriostatic ratio (%) - (control colony diameter-diameter of experimental treated colony)/diameter of control colony]×100。

The results are shown in table 4, fig. 3 and fig. 4, and indicate that: the Aceraceae ZF458 can effectively inhibit the growth of hyphae of pepper anthracnose (pathogenic bacteria are pepper anthracnose bacteria in a table 4), phytophthora capsici, cucumber tendril-leaved blight (pathogenic bacteria are watermelon ascochyta in a table 4), cucumber target spots (pathogenic bacteria are multi-main corynebacterium sp in a table 4) and tomato gray leaf spots (pathogenic bacteria are tomato stolonifera in a table 4), does not inhibit pathogenic fungi such as tomato gray mold (pathogenic bacteria are botrytis cinerea in a table 4), cucumber rhizoctonia solani in a table 4) and melon fusarium wilt (pathogenic bacteria are fusarium solani in a table 4), and has an inhibition rate of the tomato gray leaf spots of about 36.68%. The Aceranibacter mapleianensis ZF458 has a good inhibition effect on pathogenic bacteria, the inhibition diameter on Agrobacterium tumefaciens is more than 7.0cm, the inhibition diameter on muskmelon fruit spots (the pathogenic bacteria are Acidophila avenae in Table 4), cucumber soft rot (the pathogenic bacteria are Peucedanum dauricum in Table 4), tomato ulcer (the pathogenic bacteria are Corynebacterium cremorium michiganense in Table 4), tomato bacterial spots (the pathogenic bacteria are Pseudomonas syringae and tomato pathogenic variants in Table 4) is more than 5.0cm, and the growth of potato ring rot (the pathogenic bacteria are Corynebacterium cremorium ring rot subspecies in Table 4), cucumber angular spot (the pathogenic bacteria are Pseudomonas syringae and cucumber angular spot pathogenic variants in Table 4) and tomato bacterial wilt (the pathogenic bacteria are Ralstonia solanacearum in Table 4) can be inhibited.

TABLE 4 bacteriostatic effect of Aceraceae Rahnella ZF458 on pathogenic bacteria

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

Example 3 measurement of Merremia maple ZF458 biocontrol factor

1. Determination of phosphorus solubilizing ability

Respectively dotting purified Aceraria maple ZF458 to NBRIP inorganic phosphorus culture medium by adopting a transparent ring method, repeating each group for 3 times, culturing for 14d at 28 ℃, measuring the diameter of a phosphorus-dissolving transparent ring and the diameter of a bacterial colony, and calculating the ratio HD/CD of the diameter (HD) of the transparent ring to the diameter (CD) of the bacterial colony.

2. Assay for siderophore

After Aceraria maple ZF458 activation, the Aceraria maple is spotted on a CAS solid plate and placed at 28 ℃ for inverted culture. If Aceraria mapleianensis ZF458 produces an obvious orange-yellow transparent ring on a CAS plate, the strain is proved to have the capability of secreting siderophilin; the size and the color of the transparent circle can indicate that the siderophilic capacity of the strain ZF458 is strong and weak, and the larger the transparent circle is, the darker the color is, which indicates that the siderophilic capacity of the Aceranibacter mapleianus ZF458 is strong.

TABLE 5 determination of antagonistic bacteria ZF458 biocontrol related factors

The results are shown in table 5 and fig. 5, and show that: the Aceraria maple Ralstonia ZF458 generates a phosphorus dissolving ring on an NBRIP plate, the total diameter of the phosphorus dissolving ring is 17mm, and the colony diameter of the Aceraria maple ZF458 is 7 mm; the Aceranibacter mapleianus ZF458 produces an obvious orange-yellow transparent ring on a CAS plate, the total diameter of the orange-yellow transparent ring is 10mm, the diameter of a colony of the Aceranibacter mapleianus ZF458 is 8mm, and the siderophin secretion capacity (the total diameter of the orange-yellow transparent ring/the diameter of the colony) is 1.25.

The result of the biocontrol factor determination shows that the Aceraria maple ZF458 can utilize inorganic phosphorus and siderophin and has the growth promoting potential.

Example 4 Aceraceae Rahnella ZF458 growth and pH Change Curve determination

Selecting single colony of Aceraria acearum ZF458 in PD liquid culture medium, performing shake culture at 28 deg.C and 180r/min for 16h, and regulating bacterial suspension concentration to OD with blank PD liquid culture medium_600nm0.6, in a PD broth (100mL/250 mL). Shaking and culturing at 28 deg.C and 180r/min for 48h, and measuring OD every 4h_600nmValue and pH.

OD for time-dependent growth of Aceraria maple ZF458_600nmThe values and the pH values are measured to know the growth rule of the strain ZF458 and the pH change value of the bacterial liquid of the strain ZF 458. The results are shown in fig. 6 and fig. 7, and show that the acer maple Raylen bacterium ZF458 continuously grows along with the increase of time, and the growth lag phase of the acer maple Raylen bacterium ZF458 is 0-4h after inoculation; the strain enters a logarithmic phase within 4-20 hours, and the number of the strains is logarithmically increased; the growth of the thallus is kept stable after 20h, and the OD is kept at 40h_600nmThe value finally reaches 1.305. The pH of the Aceraria maple ZF458 bacterial liquid is rapidly reduced after 0-12h after inoculation, the pH is changed from alkalinity to acidity, the pH of the bacterial liquid is kept stable after 12h, and the pH value is finally 3.28 after 40 h.

Example 5 in vivo efficacy assay of Aceraria maple ZF458 (as antagonistic bacterium) against root cancer of Rosa chinensis

Inoculating Aceraria acearum ZF458 into a 250mL conical flask containing 100mL NB culture medium, performing shake culture at 28 deg.C and 180r/min for 28h, collecting the fermentation broth, and determining OD of the fermentation broth_600nmValue andadjustment to OD Using blank NB liquid Medium_600nm0.8 (blank NB broth) to give a suspension of maple rahnella ZF 458. The method comprises inoculating tested root cancer bacteria (Agrobacterium tumefaciens ACCC19185 (obtained from China agricultural microbial culture Collection center) and Agrobacterium vitis (Agrobacterium vitis) (obtained from China agricultural microbial culture Collection center, with the strain number of ACCC19185, and the strain number of ACCC19185 for short) and Agrobacterium tumefaciens K308 (obtained from China agricultural microbial culture Collection center, and the related documents including Ficus benghaiensis, Lei, Xian Wen, Shi Yanxia, Chaarali, Sunyu, Li Bao poly, Bacillus belezii ZF2 for preventing and treating multiple main Coryneba bacteria, China biological prevention and treatment Commission 2019,35(02): 217) 225 ″, which can be obtained from applicant, used only for repeating the present invention, and can not be used for other purposes) as pathogenic bacteria control strains into NB culture medium, shaking and collecting fermentation broth, and determining OD of the fermentation broth_600nmValue and adjust to OD using blank NB broth_600nmControl suspensions of two pathogens were obtained at 0.8.

Respectively mixing the bacterial suspensions of two pathogenic bacteria controls and the Acer nigeri ZF458 bacterial suspensions in the volume of 1:1 to obtain a mixed solution (1:1) of a strain ZF458 and a strain ACCC19185 and a mixed solution (1:1) of the strain ZF458 and a strain K308, and treating the mixed solution as the Acer nigeri ZF458 bacterial suspension; simultaneously, mixing 1500 times of solution of 46 percent copper hydroxide water dispersible granules (DuPont, USA, registration number: PD20110053) with two bacterial suspensions of pathogenic bacteria control by volume of 1:1 to obtain 1500 times of solution of 46 percent copper hydroxide water dispersible granules and mixed solution (1:1) of a strain ACCC19185 and mixed solution (1:1) of 1500 times of solution of 46 percent copper hydroxide water dispersible granules and strain K308 for control treatment of two medicaments; respectively and independently inoculating the two pathogenic bacteria control bacterial suspensions as pathogenic bacteria control; sterilized water was inoculated as a clear water control. Pricking micro-wound on the stem of sunflower (from Shandong shogao, Shanghai, China) with sterilized cotton wool, wrapping the wound with sterilized absorbent cotton dipped with inoculum (Acerana maple ZF458 suspension treatment, drug control treatment, pathogenic bacteria control and clear water control), repeating the treatments for 8 times, and culturing in greenhouse for 10-15 days to observe symptoms. And (4) investigating a result: cutting 2cm stem with tumor and adjacent stem with equal length (control stem), weighing, calculating tumor weight and preventing diseases. Tumor weight (g) ═ tumor-bearing stem segment-control stem segment; control effect (%) ═ control tumor weight-treated tumor weight)/control tumor weight × 100.

The results are shown in table 6 and fig. 8, and show that: the sunflowers inoculated with the two pathogenic bacteria control (A and E in figure 8) of the Chinese rose agrobacterium (ACCC19185) and the grape agrobacterium (K308) are all attacked, and the inoculation positions are expanded and are in a tumor shape; sunflower seedlings inoculated with the mixed solution of the antagonistic bacteria Acer nikoense ZF458 and two pathogenic bacteria (C and G in figure 8), and inoculated with the mixed solution of the chemical agent (46% of copper hydroxide water dispersible granules of 1500 times) and the two pathogenic bacteria (B and F in figure 8) have no disease, and the control effect of the antagonistic bacteria Acer nikoense ZF458 (88.5 +/-4.25 and 90.12 +/-3.24) is higher than that of the control chemical agent 46% of copper hydroxide water dispersible granules (52.0 +/-1.28 and 60.2 +/-3.57) (Table 6).

TABLE 6 determination of in vivo prevention of root cancer of Rosa chinensis by antagonistic bacterium ZF458

Example 6 measurement of Effect of Aceraceae Rahnella ZF458 (as antagonistic bacterium) cells and supernatant on prevention of bacterial leaf blight of Rosa chinensis

Respectively inoculating Aceraceae Aceranivorus ZF458 and Chinese rose root cancer pathogenic bacteria, namely Chinese rose Agrobacterium (ACCC19185), into NB culture medium, shake culturing at 28 deg.C and 180r/min for 24 hr, collecting fermentation broth, and measuring OD of the fermentation broth_600nmValue and adjustment of OD using blank NB broth_600nm0.8 (blank NB broth) to give a suspension of maple rahnella ZF 458. Sucking 100 mu L of Chinese rose agrobacterium suspension, coating a PDA (personal digital Assistant) plate, and putting the Chinese rose agrobacterium suspension into a constant-temperature incubator at 28 ℃ for 24h to obtain a Chinese rose agrobacterium inoculation plate. Centrifuging Aceraria maple ZF458 bacterial suspension, sucking supernatant, filtering with a filter element, placing an Oxford cup in the center of an Agrobacterium chinensis inoculation flat plate, sucking 20 mu L of filtered supernatant, putting into the Oxford cup, culturing at 28 ℃ for 24h, and observing whether a bacteriostatic zone is generated. Sucking 1mL of sterilized water and injecting into the container after centrifugationIn Acer mono-maple Ralstonia ZF458 thalli, the thalli are fully suspended to obtain Acer mono-maple ZF458 thalli suspension; sucking 20 mu L of thallus suspension, putting into an oxford cup in the center of the Chinese rose agrobacterium inoculation flat plate, culturing for 24h at 28 ℃, and observing whether a bacteriostatic zone is generated.

The results are shown in table 7 and fig. 9, and show that: the suspension supernatant of the antagonistic strain ZF458 has no prevention and treatment effect on the root cancer pathogenic bacteria, and the suspension of the antagonistic strain Merremia maple ZF458 has prevention and treatment effect on the root cancer pathogenic bacteria; the bacteriostatic diameter of the thallus suspension is more than 2.42cm, and the bacteriostatic rate is more than 26%.

TABLE 7 bacteriostasis rate of antagonistic bacteria Acera maple Rahnella ZF458 thallus suspension and supernatant to Chinese rose root cancer germ

Note: "-" has no inhibitory effect.

In conclusion, the maple Rahnella anserine ZF458 obtained by screening has a wide antibacterial spectrum, and can inhibit 16 pathogenic fungi and bacteria; the compound has good control effect on the Chinese rose agrobacterium tumefaciens and shows good control effect on the Chinese rose agrobacterium tumefaciens in a living body test.

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> Acer lawinii strain with biocontrol potential and application thereof

<130> GNCSQ212078

<160> 4

<170> PatentIn version 3.5

<210> 1

<211> 1543

<212> DNA

<213> Aceraceae)

<400> 1

taaggaggtg atccaaccgc aggttcccct acggttacct tgttacgact tcaccccagt 60

catgaatcac aaagtggtaa gcgccctccc gaaggttaag ctacctactt cttttgcaac 120

ccactcccat ggtgtgacgg gcggtgtgta caaggcccgg gaacgtattc accgtagcat 180

tctgatctac gattactagc gattccgact tcatggagtc gagttgcaga ctccaatccg 240

gactacgaca tactttatga ggtccgcttg ctctcgcgag ttcgcttctc tttgtatatg 300

ccattgtagc acgtgtgtag ccctactcgt aagggccatg atgacttgac gtcatcccca 360

ccttcctccg gtttatcacc ggcagtctcc tttgagttcc caccattacg tgctggcaac 420

aaaggataag ggttgcgctc gttgcgggac ttaacccaac atttcacaac acgagctgac 480

gacagccatg cagcacctgt ctcagagttc ccgaaggcac taagccatct ctggcgaatt 540

ctctggatgt caagagtagg taaggttctt cgcgttgcat cgaattaaac cacatgctcc 600

accgcttgtg cgggcccccg tcaattcatt tgagttttaa ccttgcggcc gtactcccca 660

ggcggtcgac ttaacgcgtt agctccggaa gccacgcctc aagggcacaa cctccaagtc 720

gacatcgttt acagcgtgga ctaccagggt atctaatcct gtttgctccc cacgctttcg 780

cacctgagcg tcagtctttg tccagggggc cgccttcgcc accggtattc ctccagatct 840

ctacgcattt caccgctaca cctggaattc tacccccctc tacaagactc tagcttgcca 900

gtttcaaatg cagttcccac gttaagcgcg gggatttcac atctgactta acaaaccgcc 960

tgcgtgcgct ttacgcccag taattccgat taacgcttgc accctccgta ttaccgcggc 1020

tgctggcacg gagttagccg gtgcttcttc tgcgagtaac gtcaatcacc acacgtatta 1080

agtatgatgc cttcctcctc gctgaaagtg ctttacaacc ctaaggcctt cttcacacac 1140

gcggcatggc tgcatcaggc ttgcgcccat tgtgcaatat tccccactgc tgcctcccgt 1200

aggagtctgg accgtgtctc agttccagtg tggctggtca tcctctcaga ccagctaggg 1260

atcgtcgcct aggtgagcca ttacctcacc tactagctaa tcccatctgg gcacatccga 1320

tggcgtgagg tccgaagatc ccccactttg ctctttcgag gtcatgcggt attagctacc 1380

gtttccagta gttatccccc tccatcaggc agtttcccag acattactca cccgtccgcc 1440

gctcgccggc aaagtagcaa gctacttccc gctgccgctc gacttgcatg tgttaggcct 1500

gccgccagcg ttcaatctga gccatgatca aactcttcaa tta 1543

<210> 2

<211> 2415

<212> DNA

<213> Aceraceae)

<400> 2

atgtcgaatt cttatgactc ctcaagtatc aaggtattaa aagggctgga cgcggtgcgt 60

aagcgccccg gcatgtatat cggcgatacc gatgacggca ctggtctgca ccacatggta 120

ttcgaggttg tggacaacgc tatcgacgaa gccctcgcgg gccactgtaa agagattcag 180

gtcacgatcc atgcggataa ctctgtttcc gtacaggatg atggccgtgg tattccgacc 240

ggcattcatg aagaagaggg cgtttctgct gctcaggtca ttatgaccgt tcttcatgct 300

ggtggtaaat tcgacgataa ctcgtataaa gtctccggcg gtctgcatgg cgtaggtgtt 360

tccgtcgtta atgccttgtc ggaaaaactg gaactggtta tccgccgtga aggcaaagtg 420

cacactcaga cttacgtgca cggcgagccg caggatccgc tgaaagtgat cggcgatact 480

gacgtaaccg gtaccacggt acgtttctgg ccgagcttca acaccttcac caatcacaca 540

gaattcgagt atgagattct ggcgaaacgc ctgcgtgaat tgtcattcct gaactccggc 600

gtggcaattc gtctgctgga taaacgtgat ggtaagaatg atcacttcca ttatgaaggc 660

ggtatcaaag ccttcgtgga atacctgaac aaaaacaaaa ccccaatcca tccgaccgta 720

ttctatttct ctacgatgaa ggacgatatt ggcgttgagg ttgcgttgca gtggaatgat 780

ggtttccagg aaaacattta ctgcttcacc aacaatattc cacagcgcga tggcgggact 840

cacttagccg gtttccgttc ggcaatgacc cgtaccctga acgcttacat ggataaagaa 900

ggttacagca agaaatccaa aatcagcgcc accggtgacg atgcccgtga aggtttgatt 960

gctgtggtgt ctgtgaaggt accggatcct aagttctctt cacaaaccaa agacaaactg 1020

gtttcttctg aagtgaaaac ggccgttgaa acgctgatga acgagaagct ggtggattac 1080

ctgatggaaa atccgtcaga cgccaaaatc gttgtgggga aaatcatcga cgctgcgcgc 1140

gcccgtgaag cagcccgtaa agcccgtgaa atgacccgcc gtaaaggtgc gctggacctg 1200

gctggcttgc caggcaaact ggcggactgt caggaacgtg acccggctca ttccgaactg 1260

tacttagtgg aaggggactc agcgggcggc tcagcaaaac aaggccgtaa ccgtaagaac 1320

caggcgattc tgccgttgaa aggtaaaatc ctcaacgtag aaaaagcgcg cttcgacaaa 1380

atgctctctt ctcaggaagt ggcaacactg attactgcgc tcggttgcgg cattggccgt 1440

gacgagtaca acccggacaa actgcgctat cacagcatca tcatcatgac cgatgccgac 1500

gtcgatggtt cacacatccg taccctgttg ctgacgttct tctaccgtca gatgcctgaa 1560

attgtcgagc gtggccacgt gtttatcgca cagcctccat tgtacaaagt gaaaaaaggt 1620

aagcaggaac agtacattaa agatgatgaa gcgatggatc agtatcagat ttccattgcg 1680

atggacgggg caacgttaca cgccaacgcc catgcaccag ccctggcggg tgaaccgctg 1740

gagaaactgg tggctgaaca tcacagcgtg cagaaaatga ttggccgcat ggaacgtcgt 1800

tatccgcgtg cgctgctgaa caacctgatc tatcagccga ccctgccggg gtcagatctg 1860

accgatcagg cgaaagttca gacctggata gaatcgctgg tcgcgcgtct caatgagaaa 1920

gagcagcacg gcagttctta cagcgcgatc gtgcgtgaaa accgtgaaca tcagcttttc 1980

gaaccggttc tgcgtatccg tacacacggt gttgatacgg attacgatct cgatgcagac 2040

ttcattcagg gtggcgaata ccgcaaaatc tgtgcgctgg gcgaacagct gcgcggcctg 2100

atcgaagaag atgccttcat cgaacgtggc gaacgccgtc agccagtgac cagcttcgag 2160

caggcgctgg aatggctggt gaaagagtcc cgtcgtggtc tgtcgattca gcgatacaaa 2220

ggtctgggcg aaatgaaccc tgatcagctg tgggaaacca ccatggatcc tgaacaacgt 2280

cgcatgttac gtgtgaccgt gaaagatgcc atcgctgctg accagttgtt cactacgctg 2340

atgggcgatg cggttgaacc gcgtcgcgcc ttcatcgaag agaatgccct gaaagcggcc 2400

aatatcgata tctga 2415

<210> 3

<211> 4029

<212> DNA

<213> Aceraceae)

<400> 3

ttactcgtct tccagctcga tgttgatacc cagagaacgg atttctttca acagtacgtt 60

gaaggattcc ggcatgcctg gttccatacg atgatcgcca tctacgatgt ttttatacat 120

cttagtacgg ccattcacgt catcggactt aacagtcagc atttcctgca gggtatatgc 180

ggcaccatat gcttccagtg cccacacttc catctcaccg aagcgctgac caccgaactg 240

agctttacca cccagcggct gctgagtaac aaggctgtaa gaaccggtag aacgcgcgtg 300

catcttatcg tcaacaaggt ggttcagttt cagcatgtac atgtagccga cggtaacctg 360

acgctcgaat tgctcgccgg tacggccgtc aaacagtgtg atctgaccag aggttgggat 420

tccgcccatt tcaagcagtt gcttgatctc tgtctctttc gcaccgtcga agactggtgt 480

cgcgattggc ataccttttt tcaggttctc agccagacgc aatacttcgt catcggtaaa 540

ggtggtcaga tccactttct gacgtacgtc gtcgcccaga tcataggcac gctggatgaa 600

ctcgcgcagc ttagaaactt cttcctgttt cttcagcatg gcattgattt tttcaccaat 660

acctttcgcg gccataccca ggtgggtttc caaaatctga ccaatgttca tacgtgatgg 720

aacgcccagc gggttcagta cgatatcaac aggagtcccg ttttcatcgt aaggcatatc 780

ttcgatcggg ttgatcttgg agataacacc tttgttaccg tggcggcctg ccattttgtc 840

acccggctgg atttgacgtt taacggccaa atacacttta acgattttca gcacacctgg 900

tgccagatca tcgccctgag tgattttacg acgcttagct tcgagttttt tctcaaactc 960

ggatttcagt tcgtcgtact gttcagcaag ctgttccaac tgattctgtt tgtcttcgtc 1020

agtcagtgac agttctaacc aacgatcgcg aggcaatttg cccagcttgt cagcttcaac 1080

accgccagca accagcgcag actggatacg tgcaaacaga ccggcttcca ggatctgcag 1140

ttcttcagtc aggtctttct tcgcctgctt cagctgcatc tcttcgattt ccaacgcacg 1200

cttatctttt tccacgccat cgcgggtgaa gacttgcacg tcgataaccg taccggacac 1260

gccgtttggt acacgcagag aagagtcttt aacgtcagac gctttctcac cgaagatcgc 1320

acgcagcagt ttctcttccg gcgtcagttg ggtttcacct ttaggcgtca ccttacccac 1380

cagaatgtca ccaccggtca cttcagcacc gatataaacg ataccggatt catccagttt 1440

ggagagcgca gcttcaccca cgttagggat gtcagcggtg atctcttcag gccctaactt 1500

ggtgtcacgg gacacacatg ccagttcctg gatatggatg gtcgtgaagc ggtcttcctg 1560

cacaacacgt tcggagacca agatggagtc ttcgaagttg taaccgttcc aaggcatgaa 1620

cgcgacacgc atgttctgac ccagagccag ttcgccaaga tctgtagacg gaccatctgc 1680

cagcacgtcg ccgcgctcga ttggctcgcc cagattcaca cacggcatct ggttgatgca 1740

ggtgttctgg ttagaacggg tgtatttggt caggttataa atgtcgatac ctgcttcgcc 1800

cgggtacatc tcttcttcgt taacacgaat aacgatacgg gatgcatcca cgtactgaac 1860

aataccgcca cgtttggcta cggcagtaac accggagtca actgctacag cacgttccat 1920

accagtacct accagcggct tatcagcgcg cagagtcgga actgcctgac gttgcatgtt 1980

cgcacccatc aatgcacggt tggcgtcatc gtgttccagg aatggaatca atgaagcacc 2040

aacggataca acctgttggg tggaaacgtc catgtagtca acctgatcgc ggctgaagag 2100

gcttgattcg cctttgctac gacaagtgac caggtcttca atgaagcgcc cttcttcgtc 2160

caggttggag ttcgcctgag cgattacgaa gttgccttct tcaatggcag acagatagtt 2220

gatttcatca gtcaccagac catcacgcac gcgacggtaa ggggtttcca ggaaaccgta 2280

ctcattggtc tgtgcataga cagacaagga gttgatcaga ccgatgtttg gaccttctgg 2340

cgtttcgatt ggacatacgc ggccgtagtg agtcgggtgt acgtctcgaa cttcaaagcc 2400

agcacgttca cgggtcaaac cgcccgggcc caatgcagag atacgacgct tgtgcgtgat 2460

ctcagacaat gggttgttct ggtccataaa ttgtgacaac tggcttgagc cgaagaattc 2520

tttcaccgca gccgaaatcg gcttagcgtt gatcatatcc tgaggcatca gtgtgtcgag 2580

atcgcccaga gacagacgct ctttaacagc acgctcaaca cgaaccagac caacacggaa 2640

ttggttttca gccatttcgc cgacggaacg aatacgacgg ttgcccaagt ggtcgatatc 2700

atccacttcg ccctggccgt tacgaatgcc gatcagcttt ttcattacct gaatgatgtc 2760

gtctttgctc aggatacctg aaccttcgat ttcatcacgc aacagagaac ggttgaactt 2820

catacgacct acagcagaca ggtcgtaacg atcttcagag aagaacaggt tctcaaacag 2880

gctttccgcc gcttcacgcg ttggtggctc accaggacgc atcattcggt agatctcaac 2940

cagtgcgctc aggcgatcgc tggttgggtc gacgcgaatg gtctcggaca tgtacgcgcc 3000

atgatcgagg tcgttggtga acagcgtttc aatggttttg tgaccggact ggctcagttt 3060

agcaagcagg tccagagaca attccatgtt agccgggacg atcagctcac cggtattggt 3120

gtcgatgtag tctttagaga ccactttccc tgcgatgtat tcaacaggaa cttcgatgcg 3180

ctgaatttcg tctttttcaa gctgacgaat atgacgggct gtgatgcggc ggcctttttc 3240

tacatagact ttgccgtcag cttcgatatc aaaggaagct gtttcaccac gcaggcgctc 3300

aggtaccagt tccatctgca acttgttgtc gcggatctgg taagtcacct tgtcaaagaa 3360

caggtcaagg atctgttcag tcgtgtaatt taatgcacgc agaatgatgg tcgcaggcaa 3420

tttacggcga cggtcaatac gtacaaacag gttgtctttc gggtcaaact cgaaatctaa 3480

ccatgaaccg cggtaaggga tgatacgcgc gttataaagc actttacccg aagagtgggt 3540

tttaccctta tcgctgtcga agaatacgcc cggactacgg tgcagctgag atacgataac 3600

cctctcagta ccgttgatca caaaggtacc attttcggtc atgagcggaa tttcgcccat 3660

atagacttct tgttccttga tgtctttgac cgtaccttct ggtgcttcac gttcgtagat 3720

taccaggcgc agttttacgc gcagtggcgc ggagaacgtc acaccacgga tctggcattc 3780

tttaacgtcg aatacaggct caccaagacg gtagctaacg tattgcagct cagagttacc 3840

gctgtagctc ttgataggga atacagaacg gaatgcagct tccagaccgt actggccttc 3900

cggatcttgc tcgataaact tctggaacga gtcaagctgg atagaaagga gatatggaat 3960

gtccaaaact tgtggacgtt taccaaaatc cttacgaatg cgttttttct cggtatagga 4020

gtaaaccat 4029

<210> 4

<211> 1383

<212> DNA

<213> Aceraceae)

<400> 4

atggctactg gaaagattat ccaggtaatc ggcgccgtgg tggacgtcga gttccctcag 60

gatgcagtac cgaacgtgta caatgctctt gaggtagaaa acggtacctc caaactggtg 120

ctggaagtac agcaacagtt aggcggcggc gttgttcgtt gtatcgcaat gggtacctca 180

gacggcctgc gtcgcggtct gaaagtgaac aacctggaac acccaattga agtaccggtg 240

ggtaaagcga ctctgggtcg tatcatgaac gtattgggtg aaccaatcga catgaaaggt 300

gcgatcggcg aagaagaacg tcgtgcgatc caccgcgctg cgccttctta tgaagagctg 360

gcaaactccc aggaactgct ggaaaccggt atcaaagtta tggacctgat gtgtccgttc 420

gctaagggcg gtaaagtcgg tctgttcggt ggtgcgggtg ttggtaagac tgtaaacatg 480

atggaactga tccgtaacat cgcgatcgag cactccggtt attctgtgtt tgcaggtgtg 540

ggtgaacgta ctcgtgaggg taacgacttc tatcacgaaa tgaccgactc caacgttatc 600

gacaaagttt ccctggtgta tggtcagatg aatgagccac caggtaaccg tctgcgcgtt 660

gcactgaccg gcctgaccat ggcggagaaa ttccgtgacg aaggtcgtga cgtactgctg 720

ttcgttgaca acatttaccg ttacaccctg gccggtaccg aagtgtccgc acttctgggc 780

cgtatgccat cagcggtagg ttatcagcca acgctggcgg aagagatggg cgctctgcaa 840

gaacgtatca cctcgaccaa aagtggttca atcacctccg tacaggccgt ttacgttcct 900

gcggatgact tgactgaccc atctccagct accaccttcg ctcacttaga tgcgaccgtg 960

gttctgagcc gtcagatcgc gtcactgggt atttacccgg cggttgaccc actggattcc 1020

accagccgtc agctggatcc actggttgtt ggtcaggagc actacgatgt ggctcgtggc 1080

gtgcagtcta ttctgcaacg ttaccaggaa ctgaaagaca tcatcgcgat cctgggtatg 1140

gacgagttgt cagaagatga caaactggtt gtatcccgtg cgcgtaaaat tcagcgcttc 1200

ctgtcccagc cattcttcgt ggccgaagtc ttcaccggtt ctccgggcaa gttcgtatcg 1260

ctgaaagata ccattcgtgg tttcaaaggc attatggacg gcgactacga tcacctgccg 1320

gaacaagcgt tctacatggt tggcaccatt gatgaagcag tggaaaaagc caagaaactg 1380

taa 1383

Claims (10)

1. Aceransertia maple, characterized in that: the Aceraceae is Aceraceae, the strain number is ZF458, and the registration number of the Aceraceae is CGMCC No. 22292.

2. The maple Rahnella of claim 1, wherein: the 16SrDNA nucleotide sequence of the Acerana maple contains a DNA molecule shown as a sequence 1 in a sequence table; the nucleotide sequence of the gyrB gene of the Aceransella maplensis is a sequence 2 in a sequence table; the nucleotide sequence of rpoB gene of the Aceransella maple is sequence 3 in the sequence table; the nucleotide sequence of the rpoD gene of the Aceranei is a sequence 4 in a sequence table.

3. The culture of Acerana maple of claim 1 or 2, which is obtained by culturing the Acerana maple of claim 1 or 2 in a microbial culture medium.

4. A microbial inoculum, which is characterized in that: the microbial inoculum contains the Acerana maple of claim 1 or 2 or/and the metabolite of the Acerana maple of claim 1 or 2 or/and the culture of claim 3.

5. The microbial inoculum according to claim 4, characterized in that: the microbial inoculum has at least one of the following characteristics:

A1) has phosphorus dissolving capacity;

A2) producing siderophore;

A3) inhibiting agrobacterium vitis;

A4) inhibiting agrobacterium chinensis;

A5) preventing and treating plant root cancer;

A6) inhibiting pathogenic bacteria of plant diseases.

6. The microbial inoculum of claim 5, wherein: the plant disease pathogenic bacteria are bacteria or fungi; the bacteria are any one of root cancer bacteria, muskmelon fruit blotches, cucumber soft rot, tomato ulcers and tomato bacterial spot germs; the fungus is any one of colletotrichum capsici, phytophthora capsici, corynespora cubensis and alternaria solani.

7. The maple Rahnella of claim 1 or 2 for use in at least one of:

B1) inhibiting pathogenic bacteria of plant diseases;

B2) inhibiting agrobacterium vitis;

B3) inhibiting agrobacterium chinensis;

B4) preventing and treating plant root cancer.

8. Use of the inoculant of claim 4 or 5 for at least one of:

B1) inhibiting pathogenic bacteria of plant diseases;

B2) inhibiting agrobacterium vitis;

B3) inhibiting agrobacterium chinensis;

B4) preventing and treating plant root cancer.

9. Use according to claim 7 or 8, characterized in that: the plant disease pathogenic bacteria are bacteria or fungi; the bacteria are any one of root cancer bacteria, muskmelon fruit blotches, cucumber soft rot, tomato ulcers and tomato bacterial spot germs; the fungus is any one of colletotrichum capsici, phytophthora capsici, corynespora cubensis and alternaria solani.

10. A method for producing the microbial agent of claim 4 or 5 or 6, comprising the step of culturing the Aceraceae maple naenier as claimed in claim 1 or 2 in a microbial culture medium.

Images