CN111925973A - Litchi endophytic Burkholderia gladioli and application thereof in preventing and treating litchi anthracnose and litchi frost blight - Google Patents
Litchi endophytic Burkholderia gladioli and application thereof in preventing and treating litchi anthracnose and litchi frost blight Download PDFInfo
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Abstract
The invention belongs to the technical field of biological control of plant diseases, and discloses litchi endophytic Burkholderia gladioli and application thereof in control of litchi anthracnose and litchi frost blight. The strain is named as Burkholderia gladioli SZPT16, is preserved in Guangdong province microorganism strain preservation center, is abbreviated as GDMCC, and has the address: the microbial research institute of Guangdong province, No. 59 building, No. 5 building, of Mieli Zhonglu, Guangzhou City, has a preservation number of GDMCC No. 61168, and a preservation time of 2020, 8 and 25 days. The strain is derived from litchi endophytic bacteria, and the strain and the biological preparation thereof have strong inhibition effects on litchi colletotrichum and litchi peronophythora litchii and have excellent biocontrol effects; the source is environment-friendly and nontoxic, and the litchi chinensis juice is safer and more reliable to eat and processed products of litchi chinensis, and has good development and application prospects.
Description
Technical Field
The invention belongs to the technical field of biological control of plant diseases, and particularly relates to litchi endophytic Burkholderia gladioli and application thereof in control of litchi anthracnose and litchi frost blight.
Background
Litchi (lichi chinensis Sonn) is an important subtropical economic crop, has good fruit taste, beautiful color and rich nutrition, is a good name of Chinese precious fruit, has high economic value and is one of the most competitive fruits in the international market of China. The two major diseases of anthracnose and frost blight in litchi production seriously affect the yield and quality of litchi, the anthracnose of litchi is mainly caused by Siamese anthracnose bacterium Colletotrichum size, and the frost blight of litchi is caused by Peronophythora litchii. However, at present, the control of the two diseases in production mainly takes chemical agent control as the main control, and the use of a large amount of chemical pesticides not only destroys the ecological environment, reduces the fruit quality, improves the drug resistance of pathogenic bacteria, and the like. In order to search for a safe and effective method for preventing and treating litchi diseases, the method for preventing and treating biological control microorganisms can effectively make up for the deficiency of chemical prevention and treatment.
The plant endophytic bacteria and the plants can mutually and reciprocally symbiotically live in a long term and colonize in plant cell gaps or cells, so that the defect of poor colonization of rhizosphere bacteria can be effectively overcome. As a new microbial resource, because the endophytic bacteria system of the plant is distributed in the plant body, the living environment is good, the biocontrol effect is more easily exerted, and the microbial resource has good application prospects in the fields of biological control, biodegradation and plant growth promotion. The burkholderia species have significant control effects on plant diseases caused by pathogenic bacteria such as Botrytis cinerea (Botrytis cinerea), Fusarium oxysporum (Fusarium oxysporum), Rhizoctonia solani (Rhizoctonia solani) and the like. The patent (CN201510315978.1) discloses that a wild rice endogenous Burkholderia gladioli strain can effectively inhibit rhizoctonia solani, fusarium graminearum, sclerotinia sclerotiorum and the like. Bacteria of the genus burkholderia are widely distributed in plants and in the rhizosphere soil, and some species of burkholderia have been commensally symbiotic with their plant hosts. Therefore, the endophytic bacteria which are obtained by separating and screening from the litchi and have antagonistic action on litchi pathogenic bacteria are inoculated back to the host plant, so that the colonization effect is easier, and scientific basis and technical support are provided for green prevention and control of litchi diseases and benign development of litchi industry.
Disclosure of Invention
In order to overcome the defects and shortcomings of the prior art for preventing and treating litchi anthracnose and frost blight and develop the application of gladiolus Burkholderia, the invention mainly aims to provide the litchi endogenous gladiolus Burkholderia which is obtained by separating litchi endophytic microorganisms serving as screening objects and has obvious inhibition effect on litchi anthracnose and frost blight, is named as gladiolus Burkholderia SZPT16 and lays a foundation for biological prevention and treatment of litchi anthracnose and frost blight; in addition, the antagonistic endophyte screened from the litchi branches coexists with litchi for a long time, and is safe and reliable for litchi disease control.
The invention also aims to provide a biological agent prepared on the basis of the litchi chinensis var gladioli burkholderia.
The invention also aims to provide application of the litchi endophytic Burkholderia gladioli in preventing and treating litchi anthracnose and frost blight.
The purpose of the invention is realized by the following scheme:
an endophytic Burkholderia gladioli strain of litchi is named as Burkholderia gladioli (SZPT 16) and is preserved in Guangdong province microorganism strain preservation center, GDMCC for short, and the address is as follows: the microbial research institute of Guangdong province, No. 59 building, No. 5 building, of Mieli Zhonglu, Guangzhou City, has a preservation number of GDMCC No. 61168, and a preservation time of 2020, 8 and 25 days.
The litchi endophytic Burkholderia gladioli is cultured on an LB culture medium at 28 ℃ for 24 hours, and the colony is creamy, moist and smooth.
A biological preparation is prepared based on the above endogenous Burkholderia gladioli of litchi.
The biological agent is prepared by performing liquid fermentation on the litchi chinensis var gladioli burkholderia, and preferably comprises the following steps: inoculating the litchi chinensis var gladioli Burkholderia to an LB liquid culture medium for culture to obtain the biological agent.
The pH value of the LB liquid culture medium is 6.8-7.0.
The culture refers to culturing for 24 hours at the temperature of 28-30 ℃ and the shaking speed of a shaking table of 180-200 rpm.
The invention also provides application of the biological agent in preventing and treating litchi anthracnose.
The invention also provides application of the biological agent in preventing and treating litchi frost blight.
The invention also provides application of the litchi endophytic Burkholderia gladioli in preventing and treating litchi anthracnose.
The invention also provides application of the litchi chinensis endogenous gladiolus burkholderia in preventing and treating litchi frost blight.
The litchi endogenous gladiolus burkholderia provided by the invention is derived from litchi endogenous bacteria, and is healthy and safe to human bodies and litchi. The litchi endogenous Burkholderia gladioli and the biological preparation thereof have strong inhibition effects on litchi anthracnose and phytophthora parasitica, have excellent biocontrol effects on litchi anthracnose and phytophthora parasitica, are environment-friendly and nontoxic in biological source, are sprayed on the surface of litchi to be used for preventing litchi anthracnose and phytophthora parasitica, have small influence on the ecological environment, and are safer and more reliable for eating of the litchi and processing products of the litchi; the culture condition requirement is low, and the method has good development and application prospects.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. the Burkholderia gladioli obtained by the invention is derived from endophytic bacteria of litchi branches, and is healthy and safe to human bodies and litchi; the application of the composition in preventing and treating litchi anthracnose and frost blight is safer and more reliable for litchi eating and processed products;
2. the gladiolus burkholderia and the biological preparation thereof have strong inhibition effects on litchi anthracnose and phytophthora parasitica, have excellent biological control effects on litchi anthracnose and phytophthora parasitica, are environment-friendly and nontoxic in biological source, and have small influence on ecological environment;
3. the Burkholderia gladioli obtained by the invention has low requirement on culture conditions and has good development and application prospects.
Drawings
FIG. 1 is a single colony of Burkholderia gladioli SZPT16 on LB medium.
FIG. 2 shows the plate-faced culture of Burkholderia gladioli SZPT16 and Colletotrichum litchi, wherein T is Colletotrichum litchi (Colletotrichum siamense), a is blank control, and b is experimental group.
FIG. 3 shows the plate-faced culture of Burkholderia gladioli SZPT16 and Phytophthora litchi, wherein F is Phytophthora litchi (Peronophythora litchii), a is blank control, and b is experimental group.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the embodiments of the present invention are not limited thereto. The reagents mentioned in the examples are commercially available.
Example 1: separation, purification and preservation of litchi endogenous gladiolus burkholderia
(1) Preparation of LB medium: weighing 10g of Tryptone (Tryptone, Oxoid LTD LP0042, England), 5g of Yeast extract (Yeast extract, Oxoid LTD LP0021, England) and 10g of sodium chloride (NaCl, national chemical group chemical reagent Co., Ltd., 10019318), adding 1000mL of water, stirring uniformly, adding 15g of agar, heating and dissolving completely, subpackaging, sterilizing at 121 ℃ for 20min, and storing for later use. The LB liquid culture medium is prepared by the same formula as the LB solid culture medium except that agar is not added, the weighed medicament is fully dissolved and then is subpackaged into conical flasks (each flask is 100mL), and the conical flasks are plugged and bound, sterilized at 121 ℃ for 20min, cooled and stored for later use.
(2) Separation and purification of litchi endogenous gladiolus burkholderia:
and collecting samples in 6 months in 2019 in a Shenzhen Xili fruit field litchi base, quickly filling the collected samples into a plastic bag, and carrying the plastic bag back to a laboratory for separation at 4 ℃.
Cutting branch tissue block, soaking in 70% alcohol for 1min, soaking in 3% sodium hypochlorite for 3-5min, and cleaning with sterile water for 3 times. (0.1 mL of the last washing solution is taken and coated on an LB culture medium, or 0.1mL of the last washing solution is taken and added into an LB liquid culture medium, shaking culture is carried out at the temperature of 30 ℃, and if no microorganism grows after 48 hours, the surface is considered to be disinfected). Culturing the tissue blocks with sterilized surfaces on an LB flat plate, observing the growth condition of bacterial colonies regularly, purifying and separating the obtained bacterial strains by adopting a flat plate scribing method, and storing the bacterial strains at the temperature of minus 80 ℃ and 30% of glycerol for later use.
The purified strains were subjected to 16S rDNA sequence analysis, wherein the sequence of a strain isolated from litchi shoots (shown below) showed 99.72% homology with Burkholderia gladioli with accession number AY268163 in GenBank, confirming that the strain was classified and named as Burkholderia gladioli (Burkholderia gladioli) and SZPT 16. The sequence fragments are as follows:
CTGACGGCATGCTTAACATGCAGTCGAACGGCAGCACGGGTGCTTGCACCTGGTGGCGAGTGGCGAACGGGTGAGTAATACATCGGAACATGTCCTGTAGTGGGGGATAGCCCGGCGAAAGCCGGATTAATACCGCATACGATCTACGGATGAAAGCGGGGGACCTTCGGGCCTCGCGCTATAGGGTTGGCCGATGGCTGATTAGCTAGTTGGTGGGGTAAAGGCCCACCAAGGCGACGATCAGTAGCTGGTCTGAGAGGACGACCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATTTTGGACAATGGGCGAAAGCCTGATCCAGCAATGCCGCGTGTGtGAAGAaGgCcTtCGGGtTGTAAAGCACTTTtGTCCGGAAaGAAaTCcTGAgGGCTAATATCCTTCGGGGATGACGGTACCGGAAGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTTAATCGGAATTACTGGGCGTAAAGCGTGCGCAGGCGGTTTGTTAAGACCGATGTGAAATCCCCGGGCTCAACCTGGGAACTGCATTGGTGACTGGCAAGCTAGAGTATGGCAGAGGGGGGTAGAATTCCACGTGTAGCAGTGAAATGCGTAGAGATGTGGAGGAATACCGATGGCGAAGGCAGCCCCCTGGGCCAATACTGACGCTCATGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGTCAACTAGTTGTTGGGGATTCATTTCCTTAGTAACGTAGCTAACGCGTGAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGATGATGTGGATTAATTCGATGCAACGCGAAAAACCTTACCTACCCTTGACATGGTCGGAATCCTGGAGAGATCCGGGAGTGCTCGAAAGAGAACCGATACACAGgTGCTGCATgGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCCTTAGTTGCTACGCAAGAGCACTCTAGGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGGGTAGGGCTTCACACGTCATACAATGGTCGGAACAGAGGGTCGCCAACCCGCGAGGGGGAGCTAATCCCAGAAAACCGATCGTAGTCCGGATTGCACTCTGCAACTCGAGTGCATGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTTTACCAGAAGTGGCTAGTCTAACCGCAAGGAGGACGGTCACCACGGTAGATAG
the Burkholderia gladioli SZPT16 is cultured on LB culture medium at 28 deg.C for 24 hr, and the colony is creamy, moist and smooth. (see FIG. 1).
(3) Preservation of burkholderia gladioli SZPT 16: the strain is preserved in Guangdong province microorganism culture collection center, GDMCC for short, at 8 months and 25 days in 2020, address: the microbial research institute of Guangdong province, No. 59 building, No. 5 building, of Mingzhou Mieli Dazhou No. 100, Guangzhou city, the number of which is GDMCC 61168.
Example 2: determination of Burkholderia gladioli activity by confrontation culture method
(1) LB medium, LB liquid medium were prepared as in example 1;
(2) preparation of PDA culture medium: weighing 200g of potato, cutting into strips, boiling, filtering with 16 layers of gauze, adding water into the filtrate to 1000mL, adding 15g of agar, fully heating to dissolve, subpackaging in conical bottles (200 mL per bottle), sterilizing at 121 ℃ for 20min, cooling and storing for later use.
(3) Preparation of PDA + carrot culture medium: weighing 200g of carrot, fully juicing by using a juicer, filtering by using 16 layers of gauze, supplementing water to 1000mL of filtrate, adding 15g of agar, fully heating and dissolving, mixing in the prepared PDA culture medium in equal volume, subpackaging in conical flasks (200 mL of culture solution in each flask), sterilizing at 121 ℃ for 20min, cooling and storing for later use.
(4) Preparation of a single colony of burkholderia gladioli: and (3) streaking and inoculating the Burkholderia gladioli to an LB culture medium plate for activation culture for 24h, and then picking out a single colony for streaking and storing.
(5) Activation culture of litchi anthrax pathogenic strain: inoculating litchi anthrax strain stored at 4 deg.C onto PDA culture medium plate for activation, and uniformly beating with sterilized puncher into round cake with diameter of 5mm from outer edge of colony after fungus grows over the plate.
(6) Activation culture of peronophythora litchi pathogenic strains: inoculating the peronophythora litchi strain stored at 20 ℃ to a PDA + carrot culture medium plate for activation, and uniformly beating the strain into a circular fungus cake with the diameter of 5mm from the outer edge of a colony by using a sterilized puncher after the fungus grows over the plate.
(7) The inhibition effect of Burkholderia gladioli on the growth of colletotrichum litchii is determined: inoculating the fungus cake obtained in the step (5) to a position 25mm away from the edge of a PDA culture medium plate, inoculating the activated Burkholderia gladioli strain to be detected in the step (4) at a position about 50mm away from the opposite direction, taking the non-inoculated Burkholderia gladioli strain as a control, repeating the steps for 4 times, and culturing at a constant temperature of 25 ℃ for 7 days. In the treatment group, the growth radius of the litchi colletotrichum between the edge of the anthracnose bacterium cake and the center of the litchi endophytic bacterium line is measured as the treatment growth radius; in the control group, the growth radius of the litchi colletotrichum was the control growth radius. Finally, the bacteriostatic rate was calculated according to the following formula.
Inhibition (%) - (control growth radius-treatment growth radius)/control growth radius x 100
(8) The inhibition effect of burkholderia gladioli on the growth of peronophythora litchi is determined: inoculating the fungus cake obtained in the step (6) to a position 25mm away from the edge of a PDA + carrot culture medium plate, inoculating the activated Burkholderia gladioli strain to be detected in the step (4) at a position about 50mm away from the opposite direction, taking the non-inoculated Burkholderia gladioli strain as a control, repeating the steps for 4 times, and culturing at a constant temperature of 25 ℃ for 7 days. In the treatment group, the growth radius of the peronophythora litchi between the edge of the peronophythora litchi strain cake and the center of the endophytic bacteria line of the litchi is measured as the treatment growth radius; in the control group, the growth radius of phytophthora litchi was the control growth radius. Finally, the bacteriostatic rate was calculated according to the following formula.
Inhibition (%) - (control growth radius-treatment growth radius)/control growth radius x 100
(9) And (4) analyzing results: as can be seen from the figure 2, the litchi endophytic Burkholderia gladioli SZPT16 has an obvious inhibition effect on the growth of litchi anthracnose pathogen, and the inhibition rate of the produced Burkholderia gladioli SZPT16 on the litchi anthracnose pathogen is 52.9% by the confrontation culture method.
(10) And (4) analyzing results: as can be seen from the figure 3, the endogenous Burkholderia gladioli SZPT16 of litchi has an obvious inhibition effect on the growth of peronophythora litchi, and the inhibition rate of the Burkholderia gladioli SZPT16 on the peronophythora litchi is 61.1% through the confrontation culture method.
Example 3: indoor litchi endophytic Burkholderia gladioli SZPT16 for preventing and treating litchi anthracnose and frost blight
(1) LB medium and LB liquid medium were prepared as in example 1. PDA medium, PDA + carrot medium were prepared as in example 2.
(2) Culturing seed bacterial liquid: inoculating litchi endogenous Burkholderia gladioli SZPT16 into LB liquid culture medium, performing shake culture at 28 ℃ for 24h at 200rpm, sampling and measuring OD values at 600nm in an ultraclean workbench every 3h after 16h, zeroing with culture solution without inoculation in the measuring process until the OD values of seed bacteria liquid are all between 0.5-0.8, and obtaining seed bacteria liquid;
(3) preparation of Burkholderia gladioli suspension (biological preparation): mixing the seed bacterial liquid prepared in the step (2) with a mixture of 1: fermenting and culturing in LB liquid culture medium at 500 volume ratio, and performing shaking culture at 28 deg.C and 200rpm for 36h to obtain biocontrol microbial inoculum with viable bacteria concentration of 2-3 × 107-108CFU/mL。
(4) Preparation of litchi colletotrichum spore suspension: the procedure of the same procedure as in (5) of example 2 was repeated for 9 days, and then sterile water was added to the plate to prepare a spore suspension with a concentration of 105one/mL.
(5) Preparing a peronophythora litchi sporangium suspension: the same procedure as in (6) and (7) of example 2 was followed for the cultivation of Phytophthora litchid, adding sterile water into the plate, washing the sporangia to prepare sporangia suspension with the concentration of 104one/mL.
(6) Indoor control evaluation of gladiolus burkholderia SZPT16 on litchi anthracnose: picking healthy litchi fruits (the variety 'Feizixiao', the healthy 'Feizixiao' fruits from Guangdong Shenzhen Xili fruit field, the maturity of the fruits is about 85 percent) with consistent maturity and uniform size, washing the fruits in running water, and airing the fruits. The healthy litchi fruits were treated as follows: spraying fruit with the suspension (biological preparation) of the burkholderia gladioli SZPT16 prepared in the step (3); group B (control) fruits were sprayed with LB liquid medium at equal dilution. Each treatment is repeated for 3 times, 30 fruits are treated repeatedly, and 300mL of the bacterial liquid and the control group are sprayed for each treatment (3 times). And (3) after 24 hours of spray treatment, spraying and inoculating the litchi anthracnose pathogen spore suspension prepared in the step (4), culturing for 5 days under the conditions of average temperature of 25 ℃ and relative humidity of 80%, and then beginning to investigate the disease index and calculate the biocontrol effect, wherein the results are shown in table 1.
(7) The disease investigation method comprises the following steps: level 0: no disease symptoms; level 1: the inoculation spot has slight lesion spots, and the necrotic area is less than 5%; and 3, level: the necrotic area is 5% -10%; and 5, stage: the necrotic area is 10% -25%; and 7, stage: the necrotic area is 25% -50%; and 9, stage: the necrotic area is over 50%. The statistical calculation method of the disease index and the biocontrol effect comprises the following steps: disease index (%) ═ Σ (number of disease stages × number of fruits of the disease stage)/(highest number of fruits of the disease stage × total number of fruits) × 100; biocontrol effect (%) (control disease index-treatment group disease index)/control disease index × 100.
(8) And (4) analyzing results: as can be seen from Table 1, the index of disease of the litchi fruits treated by the Burkholderia gladioli SZPT16 biological agent is lower than that of the litchi fruits treated by only inoculating phytophthora litchi, the litchi fruits are well prevented, and the average prevention effect reaches 56.54 percent, which shows that the Burkholderia gladioli biological agent has obvious prevention and treatment effect on anthracnose of the picked litchi fruits. Meanwhile, the control observation is carried out by only connecting with the biocontrol Burkholderia gladioli biological agent without spraying the litchi colletotrichum gloeosporioides spore suspension, the litchi morbidity is not caused, and the strain SZPT16 of the Burkholderia gladioli is not the litchi pathogenic bacteria and can be used as a post-harvest application strain for preventing and treating the litchi anthracnose.
TABLE 1 indoor control effect of Burkholderia gladioli SZPT16 on litchi fruit anthrax
| SZPT16 | Control | |
| Disease index (%) | 31.52±1.35 | 72.54±1.34 |
| Control effect (%) | 56.54 | -- |
(9) Indoor prevention evaluation of gladiolus burkholderia SZPT16 on frost blight of litchi: the source and the biocontrol bacteria treatment of the litchi fruits are the same as the step (6). And (3) after 24h of spray treatment, spraying and inoculating the phytophthora litchi sporangium suspension prepared in the step (5), culturing for 48h under the conditions that the average temperature is 25 ℃ and the relative humidity is 80%, and then beginning to investigate the disease index, and calculating the biocontrol effect, wherein the results are shown in table 2.
(10) The disease investigation method comprises the following steps: level 0: no disease symptoms; level 1: the inoculation spot has slight lesion spots, and the necrotic area is less than 5%; and 3, level: the necrotic area is 5% -10%; and 5, stage: the necrotic area is 10% -25%; and 7, stage: the necrotic area is 25% -50%; and 9, stage: the necrotic area is over 50%. The statistical calculation method of the disease index and the biocontrol effect comprises the following steps: disease index (%) ═ Σ (number of disease stages × number of fruits of the disease stage)/(highest number of fruits of the disease stage × total number of fruits) × 100; biocontrol effect (%) (control disease index-treatment group disease index)/control disease index × 100.
(11) And (4) analyzing results: as can be seen from Table 2, the index of disease of the litchi fruits treated by the Burkholderia gladioli SZPT16 biological preparation is lower than that of the litchi fruits treated by only inoculating peronophythora litchi, the litchi fruits are well prevented, the average prevention effect reaches 71.21%, and the Burkholderia gladioli biological preparation has an obvious prevention and treatment effect on the post-harvest peronophythora litchi.
TABLE 2 indoor control effect of Burkholderia gladioli SZPT16 on litchi fruit frost blight
| SZPT16 | Control | |
| Disease index (%) | 24.62±2.43 | 85.52±2.89 |
| Control effect (%) | 71.21 | -- |
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Sequence listing
<110> Shenzhen profession and technology college
<120> litchi endophytic Burkholderia gladioli strain and application thereof in preventing and treating litchi anthracnose and litchi frost blight
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1431
<212> DNA
<213> Burkholderia gladioli glakholderia gladioli)
<220>
<223> 16S rDNA sequence of Burkholderia gladioli (Burkholderia gladioli) SZPT16
<400> 1
ctgacggcat gcttaacatg cagtcgaacg gcagcacggg tgcttgcacc tggtggcgag 60
tggcgaacgg gtgagtaata catcggaaca tgtcctgtag tgggggatag cccggcgaaa 120
gccggattaa taccgcatac gatctacgga tgaaagcggg ggaccttcgg gcctcgcgct 180
atagggttgg ccgatggctg attagctagt tggtggggta aaggcccacc aaggcgacga 240
tcagtagctg gtctgagagg acgaccagcc acactgggac tgagacacgg cccagactcc 300
tacgggaggc agcagtgggg aattttggac aatgggcgaa agcctgatcc agcaatgccg 360
cgtgtgtgaa gaaggccttc gggttgtaaa gcacttttgt ccggaaagaa atcctgaggg 420
ctaatatcct tcggggatga cggtaccgga agaataagca ccggctaact acgtgccagc 480
agccgcggta atacgtaggg tgcgagcgtt aatcggaatt actgggcgta aagcgtgcgc 540
aggcggtttg ttaagaccga tgtgaaatcc ccgggctcaa cctgggaact gcattggtga 600
ctggcaagct agagtatggc agaggggggt agaattccac gtgtagcagt gaaatgcgta 660
gagatgtgga ggaataccga tggcgaaggc agccccctgg gccaatactg acgctcatgc 720
acgaaagcgt ggggagcaaa caggattaga taccctggta gtccacgccc taaacgatgt 780
caactagttg ttggggattc atttccttag taacgtagct aacgcgtgaa gttgaccgcc 840
tggggagtac ggtcgcaaga ttaaaactca aaggaattga cggggacccg cacaagcggt 900
ggatgatgtg gattaattcg atgcaacgcg aaaaacctta cctacccttg acatggtcgg 960
aatcctggag agatccggga gtgctcgaaa gagaaccgat acacaggtgc tgcatggctg 1020
tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa cccttgtcct 1080
tagttgctac gcaagagcac tctagggaga ctgccggtga caaaccggag gaaggtgggg 1140
atgacgtcaa gtcctcatgg cccttatggg tagggcttca cacgtcatac aatggtcgga 1200
acagagggtc gccaacccgc gagggggagc taatcccaga aaaccgatcg tagtccggat 1260
tgcactctgc aactcgagtg catgaagctg gaatcgctag taatcgcgga tcagcatgcc 1320
gcggtgaata cgttcccggg tcttgtacac accgcccgtc acaccatggg agtgggtttt 1380
accagaagtg gctagtctaa ccgcaaggag gacggtcacc acggtagata g 1431
Claims (9)
1. An endophytic Burkholderia gladioli strain of litchi is named as Burkholderia gladioli SZPT16 and is preserved in Guangdong province microorganism culture collection center, GDMCC for short, and the address is as follows: the microbial research institute of Guangdong province, No. 59 building, No. 5 building, of Mieli Zhonglu, Guangzhou City, has a preservation number of GDMCC No. 61168, and a preservation time of 2020, 8 and 25 days.
2. A biological agent, which is prepared based on the litchi chinensis var gladioli burkholderia as claimed in claim 1.
3. The biological agent according to claim 2, characterized by being prepared by a process comprising the steps of: inoculating the litchi chinensis var gladioli burkholderia as claimed in claim 1 into an LB liquid culture medium for culture to obtain a biological agent.
4. The biological agent according to claim 3, characterized in that: the pH value of the LB liquid culture medium is 6.8-7.0.
5. The biological agent according to claim 3, characterized in that: the culture refers to culturing for 24 hours at the temperature of 28-30 ℃ and the shaking speed of a shaking table of 180-200 rpm.
6. The application of the litchi chinensis var gladioli burkholderia as claimed in claim 1 in preventing and treating litchi anthracnose.
7. The application of the litchi chinensis var gladioli burkholderia as claimed in claim 1 in preventing and treating litchi frost blight.
8. Use of a biological agent according to any one of claims 2 to 5 for the control of litchi anthracnose.
9. The use of the biological agent of any one of claims 2-5 for controlling litchi peronophytosis.
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