CN111040974A - Banana endophytic Bacillus belgii and application thereof - Google Patents
Banana endophytic Bacillus belgii and application thereof Download PDFInfo
- Publication number
- CN111040974A CN111040974A CN201911413362.2A CN201911413362A CN111040974A CN 111040974 A CN111040974 A CN 111040974A CN 201911413362 A CN201911413362 A CN 201911413362A CN 111040974 A CN111040974 A CN 111040974A
- Authority
- CN
- China
- Prior art keywords
- banana
- strain
- bacillus
- bacillus belgii
- endophytic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses banana endophytic Bacillus belgii and application thereof. The strain is named as Bacillus velezensis (BS), and the preservation number is GDMCC No: 60930, the strain is preserved in Guangdong province microbial strain preservation center of No. 59 large institute of Mieli Zhonglu No. 100, Guangzhou city in 2019, 12 and 11 months. The bacterial strain belongs to banana endophytic bacteria, has no pathogenicity, and has a growth promoting effect on bananas; the bacterial strain and the biological preparation thereof have obvious inhibition effect on the banana vascular wilt FOC4, obviously alleviate the occurrence of banana vascular wilt symptoms, and the biological preparation has environment-friendly and nontoxic sources and small influence on ecological environment; the strain has different degrees of inhibition effects on plant pathogenic fungi of different genera; the culture condition is simple, the propagation speed is high, the thalli and the bacterial liquid can be used as main control bodies, the preservation and the transportation are easy, and the development and application prospect is good.
Description
Technical Field
The invention belongs to the technical field of biological control of plant diseases, and particularly relates to banana endophytic Bacillus belgii and application thereof.
Background
Bananas (Musa spp.) are plants of the genus Musa of the family musaceae, the plants being large herbaceous plants, which are grown in about 130 countries and regions worldwide with annual yields exceeding 1 million tons. China's banana producing area is mainly concentrated in Guangdong, Guangxi, Yunnan, Hainan, Fujian, Taiwan and other fields, and the planting area exceeds 36hm2Is an important economic crop in subtropical regions in China.
Diseases are main factors for restricting the development of the banana industry all over the world, and a plurality of diseases affecting the development of the banana industry are caused, wherein one of the most important diseases is banana vascular wilt. Banana vascular wilt, also known as banana panama disease, was first discovered in 1904 in hawaii of the united states and is a soil-borne fungal disease caused by vascular bundle necrosis due to Fusarium oxysporum cubeba specialized Form (FOC). Once the banana wilt occurs, the plant rate is 10% -40%, and the serious rate exceeds 90%, so that the yield of banana plantations is reduced, even the banana plantations are lost, the banana plantations are extremely destructive and difficult to radically cure, and serious economic loss is caused to global banana growers. In 12 months in 1995, banana vascular wilt caused by infection of No. 4 physiological race (FOC4) was first discovered in Guangdong province, and therefore, intensive research on prevention and treatment of banana vascular wilt is urgent. The control of banana wilt in production mainly takes chemical prevention and control as the main control. The use of a large amount of chemical pesticides brings potential risks to the problem of drug residue of bananas and accelerates the generation of drug resistance of pathogenic microorganisms. In biological control of plant diseases, Bacillus spp (Bacillus spp.) is widely used. The Bacillus velezensis is a novel broad-spectrum biocontrol Bacillus, widely exists in soil, air, water and other environments, exerts the biocontrol effect by producing substances such as antibacterial protein or antibacterial peptide and the like, has no influence on the normal growth of bananas, has very important effect on controlling banana wilt and is an ideal control method.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a strain of banana endophytic Bacillus belgii.
The invention also aims to provide application of the banana endophytic Bacillus belgii.
Still another object of the present invention is to provide a biological agent prepared based on the above described banana endophytic bacillus belgii.
The purpose of the invention is realized by the following technical scheme: the banana endophytic Bacillus belgii is named as Bacillus belgii (Bacillus velezensis) BS, and the deposit number is GDMCC No: 60930, the strain is preserved in Guangdong province microbial strain preservation center of No. 59 large institute of Mieli Zhonglu No. 100, Guangzhou city in 2019, 12 and 11 months.
The nucleotide sequence of 16S rRNA of the banana endophytic Bacillus belgii is shown in SEQ ID No.1, and the sequence length is 1426 bp; the nucleotide sequence of gyrB is shown in SEQ ID No.2, and the sequence length is 1169 bp.
The banana endophytic Bacillus beleisi is applied to prevention and control of banana fusarium wilt, Peronospora litchi, litchi colletotrichum, longan leaf blight, guava branch blight and guava dry rot; preferably the application in preventing and treating banana wilt.
The Fusarium oxysporum F.sp.cubense (FOC) is FOC4 which is a No. 4 physiological race of Fusarium oxysporum f.sp.cubense.
A biological agent is prepared based on the banana endophytic Bacillus belgii.
The biological agent is obtained by inoculating Bacillus belgii in LB liquid culture medium for culture.
The culture is carried out for 12h under the conditions of 28 ℃ and 180 rpm.
The biological agent is applied to preventing and treating banana wilt pathogen, litchi downy mildew, litchi anthracnose pathogen, longan leaf blight pathogen, guava branch blight pathogen and guava dry rot pathogen; preferably the application in preventing and treating banana wilt.
The Fusarium oxysporum F.sp.cubense (FOC) is FOC4 which is a No. 4 physiological race of Fusarium oxysporum f.sp.cubense.
Compared with the prior art, the invention has the following advantages and effects:
1. the Bacillus belgii obtained by the invention belongs to banana endophytic bacteria, has no pathogenicity, and has a certain growth promoting effect on bananas.
2. The Bacillus belgii strain BS has an excellent inhibition effect on the banana vascular wilt FOC4, remarkably alleviates the occurrence of banana vascular wilt symptoms, and has important practical significance for preventing and treating banana vascular wilt.
3. The biological preparation of the Bacillus belgii strain BS has an excellent inhibiting effect on the banana fusarium wilt FOC4, is environment-friendly and nontoxic in source, and has little influence on the ecological environment.
4. The Bacillus belgii strain BS has simple culture condition and high propagation speed. The thallus and the bacterial liquid can be used as main bodies for prevention and control, are easy to store and transport, and have good development and application prospects.
5. The Bacillus beilai strain BS strain of the invention also has antagonistic action with different degrees on other plant pathogenic fungi of different genera, including phytophthora litchi (Peronohthora litchii), Colletotrichum litchi (Colletotrichum siamense), longan leaf blight (Phomopsis longana), Psidium guajava (Neofusicoccum parvum) and Psidium guajava (Nalanthamla psidii). Therefore, the microbial composite material is a more ideal biocontrol microbial resource and has application prospect in the field of biological control of plant diseases.
Drawings
FIG. 1 is a graph showing the inhibitory effect of Bacillus belgii strain BS on FOC4 of Fusarium oxysporum f.sp.
FIG. 2 is a graph showing the inhibitory effect of B.beijerinckii strain BS on several plant pathogenic fungi; wherein, the graph A is a graph of the inhibition effect on Peronopthhora litchii; FIG. B is a graph showing the inhibitory effect on Colletotrichumsiamense of litchi colletotrichum; FIG. C is a graph showing the inhibitory effect on Phomopsis longana of longan leaf blight fungus; FIG. D is a graph showing the inhibitory effect on Neofusicoccum parvum of Psidium guajava; FIG. E is a graph showing the inhibitory effect on Psidium guajava dry rot fungus Nalanthamala psidii.
FIG. 3 is a graph showing the inhibitory effect of a biological agent of Bacillus belgii strain BS on FOC4 of Fusarium oxysporum; wherein, the picture A is a colony morphology picture of FOC4 normal growth of banana vascular wilt bacteria; FIG. B is a graph showing the inhibitory effect of a biological agent of Bacillus belgii strain BS on FOC4 of Fusarium oxysporum f.sp.
FIG. 4 is a graph showing the inhibitory effect of a biological agent of Bacillus belgii strain BS on Colletotrichumsiamense litchi; wherein, the picture A is a picture of colony morphology of the normal growth of the litchi anthracnose pathogen; FIG. B is a graph showing the inhibitory effect of a biological agent of Bacillus belgii strain BS on Colletotrichum litchi.
FIG. 5 is a phylogenetic tree of Bacillus beleisi strain BS based on the 16S rRNA and gyrB gene sequences.
FIG. 6 is a photograph showing the effect of Bacillus belgii strain BS on banana vascular wilt caused by banana vascular wilt FOC 4; wherein panel a is the CK control group; fig. B is a BS processing group; panel C is the FOC4 treatment group; panel D is the BS and FOC4 processing set.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to embodiments, but it will be understood by those skilled in the art that the following embodiments and examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. Those who do not specify the conditions are performed according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Banana wilt (Fusarium oxysporum f.sp. cubense) (chikun. Guangdong fruit tree fungus disease journal [ M ]. Beijing: Chinese agriculture Press, 2000: 24-37.); peronophora litchii (Peronophora litchii) (chikura, Panxuanping, Liu. litchi Fr. blight research I. identification of pathogenic bacteria and infection process [ J ] Phytopathology report, 1984(02): 113-; lichee anthracnose pathogen (Colletotrichum siamense) (chiffon, Guangdong fruit tree fungus disease journal [ M ]. Beijing: Chinese agriculture Press, 2000: 38-42.); longan leaf blight (Phomopsis longana) (chiffon, Guangdong fruit tree fungus disease journal [ M ]. Beijing: Chinese agriculture Press, 2000: 132-; guava branch blight (Neofusicoccum parvum) (chiffon, Guangdong fruit tree fungal disease J [ M ]. Beijing: Chinese agriculture Press, 2000: 65-74.); psidium guajava dry rot (Nalanthamala psidi) (Zhonhaoping, Chen Yi Jun. early, soil actinomycetes and plant avoidance application in preventing and controlling guava diseases [ J ]. academic newspaper of agricultural improvement field in high male areas, 2017, 20 (02): 1-10.).
Example 1 isolation and purification of Bacillus belgii Strain BS
(1) Preparing a PDA culture medium: firstly, washing and peeling potatoes, weighing 200g of potatoes, cutting into small pieces, adding water, boiling for 20-30 min, and filtering with eight layers of gauze. Adding glucose 20g, adding water to 1000mL, stirring, adding agar 15g, heating to dissolve completely, packaging into 250mL triangular flask, sterilizing at 121 deg.C for 20min, and storing.
(2) Preparing an LB culture medium: weighing 10g of tryptone, 5g of yeast extract and 10g of sodium chloride, adding water to 1000mL, uniformly stirring, adding 15g of agar, fully heating and dissolving, subpackaging into 250mL triangular bottles, sterilizing for 20min at 121 ℃ by moist heat, and storing for later use.
(3) The banana tissue culture seedling samples are collected in Guangzhou city, Guangdong province, brought back to a laboratory, cleaned and surface sterilized, and separated and cultured by a tissue isolation method to obtain the strains. The method comprises the following specific steps: after the collected banana tissue culture seedling samples are washed by sterile water, the base parts of the pseudostems are cut into tissue blocks, sterilized in 75% (v/v) ethanol for 30s respectively, then sterilized in 3% (v/v) sodium hypochlorite for 1min, and washed 3 times by the sterile water. Air-drying surface water, inoculating to PDA culture medium plate, and culturing at 28 deg.C for 3-4 days. And (3) selecting bacteria to carry out single colony culture, and purifying the strain on an LB solid medium plate by using a plate marking method to obtain a single colony for later use.
Example 2 screening of Bacillus belgii Strain BS
(1) Determination of the Activity of Bacillus belgii BS by plate confrontation
Taking banana vascular wilt No. 4 physiological race FOC4 as a target, placing banana vascular wilt No. 4 physiological race FOC4 fungus cakes with the diameter of 7mm on a PDA plate culture medium, dipping a little bacterial liquid of the strain obtained in the step (3) of the example 1 by using a sterilization toothpick, marking at a position about 5cm away from the fungus cakes, and inversely culturing in an incubator at 28 ℃ for 4-5 days. A strain which has obvious bacteriostasis to the target bacteria is screened out, and the strain is named as BS. The results show that the Bacillus belgii BS has obvious inhibiting effect on the fusarium oxysporum FOC4 (figure 1). According to the above method, phytophthora litchi (Peronopthora litchii), Colletotrichum litchi (Colletotrichum siamense), Phytophthora longans (Phomopsis longana), Psidium guajava (Neofusicoccum parvum) and Psidium guajava (Nalanthamala psidii) are cultured by plate-confrontation. The results show that the strain BS has obvious inhibiting effect on the pathogenic bacteria (figure 2).
(2) Growth rate method for determining activity of Bacillus belgii BS biological agent
Dipping a small amount of bacterial liquid of the bacterial strain BS obtained by screening in the step (1) into a 250mL conical flask containing 150mL of liquid LB culture medium, and placing the conical flask in a shaking table at 28 ℃ and shaking the bacterial liquid at 180rpm for 12 hours. And (4) centrifuging the bacterial liquid at 3000rpm for 10min, and filtering and removing thalli by using a bacterial filter to obtain the biological preparation of the strain BS. Biological preparation of the strain BS and PDA culture medium are mixed according to the proportion of 1: 9, and pouring the mixture. Respectively connecting banana vascular wilt FOC4 bacterial cake and litchi anthracnose bacterial cake with diameter of 7mm at the center of the plate, culturing at 28 deg.C until the control group grows over the culture dish, replacing with sterile water, and repeating for three times. And measuring the diameter of the test group after the control group bacterial colony grows over the culture dish, and calculating the bacteriostasis rate. The result shows that the biological preparation of the Bacillus belgii BS has obvious inhibition effect on the banana fusarium wilt FOC4, and the bacteriostasis rate is 26.49% (figure 3); the biological agent of the Bacillus belgii BS has an obvious inhibition effect on litchi anthracnose, and the bacteriostasis rate is 13.32% (figure 4).
Inhibition (%) - (control colony diameter-treatment colony diameter)/(control colony diameter-cake diameter) × 100%
Example 3 identification of Bacillus belgii Strain BS
A single colony was picked up in 3mL LB liquid medium, cultured overnight on a shaker at 28 ℃ and 180rpm, and the bacterial DNA was extracted by the method of OMEGAD3350-1 Bacteria DNA Kit and stored at-20 ℃ for further use. PCR amplification was performed using 16S rRNA universal primers 27F/1492R, and sequencing was performed by Competition Biotechnology engineering (Shanghai) Ltd after the detection was passed. The nucleotide sequence of 16S rRNA is shown in SEQ ID No. 1.
27F:5’-AGAGTTTGATCCTGGCTCAG-3’；
1492R:5’-GGTTACCTTGTTACGACTT-3’。
The 16S rRNA sequencing result is submitted to NCBI for sequence comparison, and the result shows that the strain BS has higher homology with strains such as Bacillus velezensis K2, B.velezensis BH21, B.velezensis DH82, Bacillus amyloliquefaciens TUL308, B.amyloliquefaciens WH1G and the like, and the homology is up to 99 percent. Therefore, it is difficult to determine the classification status of the strain BS simply by the 16S rRNA sequence.
In order to clarify the classification status of the specific species of the strain BS, the gyrB gene sequence was further analyzed. Conserved gene gyrB primer UP1/UP2S is used for PCR amplification, and sequencing is carried out by Comptopetition Biotechnology engineering (Shanghai) corporation. The nucleotide sequence of gyrB is shown in SEQ ID No. 2.
UP1:5’-GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA-3’；
UP2S:5’-AGCAGGGTACGGATGTGCGAGCCRTCNACRTCNGCRTCNGTCAT-3’。
The 16S rRNA sequences and the conserved gene gyrB sequences of the strain BS and other Bacillus strains are used for constructing a phylogenetic tree (figure 5) by adopting MEGA-X software, and the result shows that the strain BS is determined to be the Bacillus velezensis DH-82 when the genetic relationship between the strain BS and the Bacillus velezensis DH-82 is nearest.
Example 4 determination of controlling Effect of Bacillus belgii BS on banana vascular wilt
2-3 cakes of banana fusarium oxysporum FOC4 were picked, placed in a 250mL sterilized Erlenmeyer flask containing 150mL YPD medium, and shaken in an incubator at 28 ℃ for 2 d. Filtering the hyphae and collecting spores. Diluting the spore with sterile water to a concentration of 107Spore suspension of one/mL, and storing for later use. Dipping a little single colony of BS by using an inoculating loop, placing the single colony in a 250mL sterilized conical flask containing 150mL LB liquid culture medium, shaking the single colony in an incubator at 28 ℃ for 12h, and diluting the BS bacterial liquid to OD by using sterile water600About 0.1. Taking 12 banana seedlings with basically the same growth vigor, properly damaging roots of the banana seedlings by using sterilizing scissors, soaking the banana seedlings in a spore suspension for 30min, and planting the banana seedlings in a sterilizing flowerpot with a certain soil content. 6 pots of banana seedlings inoculated with FOC4 and soaked in the spore suspension were watered with BS culture liquid until the soil was wet (BS and FOC4 treated group), and 6 other banana seedlings inoculated with FOC4 were watered with sterile water (FOC4 treated group). Another 12 banana seedlings not inoculated with FOC4 were planted in flowerpots, 6 of which were watered with BS bacterial liquid until the soil was wet (BS treated group), and the other 6 were watered with sterile water as control (CK control group). The growth of banana seedlings was observed every day.
After the banana seedlings grow for 35 days, the BS treatment group has no pathogenicity effect on the bananas and has an obvious growth promotion effect compared with a CK control group. Compared with the FOC 4-treated group, the BS and FOC 4-treated group had significantly reduced incidence of banana vascular wilt (fig. 6). The result shows that the Bacillus belgii BS has obvious inhibition effect on banana wilt, reduces the incidence of banana diseases and has obvious growth promotion effect on bananas.
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> southern China university of agriculture
<120> banana endophytic Bacillus belgii and application thereof
<160>6
<170>SIPOSequenceListing 1.0
<210>1
<211>1426
<212>DNA
<213> Bacillus belgii (Bacillus velezensis)
<220>
<223>16S rRNA nucleotide sequence
<400>1
cttcggcggc tggctccata aaggttacct caccgacttc gggtgttaca aactctcgtg 60
gtgtgacggg cggtgtgtac aaggcccggg aacgtattca ccgcggcatg ctgatccgcg 120
attactagcg attccagctt cacgcagtcg agttgcagac tgcgatccga actgagaaca 180
gatttgtggg attggcttaa cctcgcggtt tcgctgccct ttgttctgtc cattgtagca 240
cgtgtgtagc ccaggtcata aggggcatga tgatttgacg tcatccccac cttcctccgg 300
tttgtcaccg gcagtcacct tagagtgccc aactgaatgc tggcaactaa gatcaagggt 360
tgcgctcgtt gcgggactta acccaacatc tcacgacacg agctgacgac aaccatgcac 420
cacctgtcac tctgcccccg aaggggacgt cctatctcta ggattgtcag aggatgtcaa 480
gacctggtaa ggttcttcgc gttgcttcga attaaaccac atgctccacc gcttgtgcgg 540
gcccccgtca attcctttga gtttcagtct tgcgaccgta ctccccaggc ggagtgctta 600
atgcgttagc tgcagcacta aggggcggaa accccctaac acttagcact catcgtttac 660
ggcgtggact accagggtat ctaatcctgt tcgctcccca cgctttcgct cctcagcgtc 720
agttacagac cagagagtcg ccttcgccac tggtgttcct ccacatctct acgcatttca 780
ccgctacacg tggaattcca ctctcctctt ctgcactcaa gttccccagt ttccaatgac 840
cctccccggt tgagccgggg gctttcacat cagacttaag aaaccgcctg cgagcccttt 900
acgcccaata attccggaca acgcttgcca cctacgtatt accgcggctg ctggcacgta 960
gttagccgtg gctttctggt taggtaccgt caaggtgccg ccctatttga acggcacttg 1020
ttcttcccta acaacagagc tttacgatcc gaaaaccttc atcactcacg cggcgttgct 1080
ccgtcagact ttcgtccatt gcggaagatt ccctactgct gcctcccgta ggagtctggg 1140
ccgtgtctca gtcccagtgt ggccgatcac cctctcaggt cggctacgca tcgtcgcctt 1200
ggtgagccgt tacctcacca actagctaat gcgccgcggg tccatctgta agtggtagcc 1260
gaagccacct tttatgtctg aaccatgcgg ttcagacaac catccggtat tagccccggt 1320
ttcccggagt tatcccagtc ttacaggcag gttacccacg tgttactcac ccgtccgccg 1380
ctaacatcag ggagcaagct cccatctgtc cgctcgaact gcatgt 1426
<210>2
<211>1169
<212>DNA
<213> Bacillus belgii (Bacillus velezensis)
<220>
<223> nucleotide sequence of gyrB gene
<400>2
gggcagcgga tataagtatc cggcggtctt cacggtgtag gggcatctgt cgtaaacgcc 60
ttgtcgacca ctcttgacgt tacggttcat cgtgacggaa aaatccacta tcaggcgtac 120
gagcgcggtg tacctgtggc cgatcttgaa gtgatcggtg atactgataa gaccggaacg 180
attacgcact tcgttccgga tccggaaatc ttcaaagaaa caatcgtata cgactatgat 240
ctgctttcaa accgtgtccg ggaattggcc ttcctgacaa aaggcgtaaa catcacgatt 300
gaagacaaac gtgaaggaca agaacggaaa aacgagtacc actacgaagg cggaatcaaa 360
agctatgttg agtacttaaa ccgttccaaa gaagtcgttc atgaagagcc gatttatatc 420
gaaggcgaga aagacggcat aacggttgaa gttgcattgc aatacaacga cagctataca 480
agcaatattt attctttcac gaataatatc aacacatacg aaggcgggac gcacgaagcc540
ggatttaaaa ccggtctgac ccgtgtcata aacgactatg caagaagaaa agggattttc 600
aaagaaaatg atccgaattt aagcggggat gatgtgagag aagggctgac ggccattatt 660
tcaattaagc accctgatcc gcaattcgaa gggcagacga aaaccaagct cggcaactcc 720
gaagcgagaa cgatcactga tacgctgttt tcttctgcgc tggaaacatt ccttcttgaa 780
aatccggact cagcccgcaa aatcgttgaa aaaggtttaa tggccgcaag agcgcggatg 840
gcagcgaaaa aagcgcggga attgacccgc cgcaaaagtg cgcttgagat ttccaatctg 900
ccgggcaaac tggcggactg ttcttctaaa gatccgagca tttccgagct gtatatcgta 960
gagggtgact ctgcgggcgg atcagcgaaa cagggacggg accgtcattt ccaagccatt 1020
ctgccgctgc gcggtaagat tctgaacgtt gagaaagcca gacttgataa gattctctca 1080
aacaatgagg tcagatcaat gatcacggcc ctcggaacag gaatcggaga agattttaat 1140
ctgaaaaagc gcgttatcat agacagtgt 1169
<210>3
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<223>16s rRNA rRNA Universal primer 27F
<400>3
agagtttgat cctggctcag 20
<210>4
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<223>16s rRNA Universal primer 1492R
<400>4
ggttaccttg ttacgactt 19
<210>5
<211>41
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<223> Gene gyrB primer UP1
<220>
<221>misc_feature
<222>(27)..(27)
<223>n is a, c, g, t or u
<220>
<221>misc_feature
<222>(30)..(30)
<223>n is a, c, g, t or u
<220>
<221>misc_feature
<222>(33)..(33)
<223>n is a, c, g, t or u
<400>5
gaagtcatca tgaccgttct gcaygcnggn ggnaarttyg a 41
<210>6
<211>44
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<223> Gene gyrB primer UP2S
<220>
<221>misc_feature
<222>(27)..(27)
<223>n is a, c, g, t or u
<220>
<221>misc_feature
<222>(33)..(33)
<223>n is a, c, g, t or u
<220>
<221>misc_feature
<222>(39)..(39)
<223>n is a, c, g, t or u
<400>6
agcagggtac ggatgtgcga gccrtcnacr tcngcrtcng tcat 44
Claims (8)
1. An endophytic Bacillus belgii of banana, which is characterized in that: the Bacillus belgii (Bacillus velezensis) BS has the preservation number GDMCC No: 60930, the strain is preserved in Guangdong province microbial strain preservation center of No. 59 large institute of Mieli Zhonglu No. 100, Guangzhou city in 2019, 12 and 11 months.
2. The application of the banana endophytic Belleville bacillus of claim 1 in preventing and treating banana wilt, litchi downy mildew, litchi colletotrichum, longan leaf blight, guava branch blight and guava dry rot.
3. Use according to claim 2, characterized in that: the fusarium oxysporum f.sp.cubense, FOC is No. 4 physiological race FOC4 of fusarium oxysporum f.sp.cubense.
4. A biological agent characterized by: prepared based on the banana endophytic Bacillus belgii of claim 1.
5. The biological agent according to claim 4, characterized in that: is obtained by inoculating Bacillus belgii in LB liquid culture medium and culturing.
6. The biological agent according to claim 5, characterized in that: the culture is carried out for 12h under the conditions of 28 ℃ and 180 rpm.
7. The use of the biological agent of any one of claims 4 to 6 for the control of banana vascular wilt, Peronospora litchi, litchi anthracnose, longan leaf blight, Psidium guajava branch blight and Psidium guajava dry rot.
8. Use according to claim 7, characterized in that: the fusarium oxysporum f.sp.cubense, FOC is No. 4 physiological race FOC4 of fusarium oxysporum f.sp.cubense.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911413362.2A CN111040974B (en) | 2019-12-31 | 2019-12-31 | Banana endophytic Bacillus belgii and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911413362.2A CN111040974B (en) | 2019-12-31 | 2019-12-31 | Banana endophytic Bacillus belgii and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111040974A true CN111040974A (en) | 2020-04-21 |
| CN111040974B CN111040974B (en) | 2022-03-25 |
Family
ID=70242448
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911413362.2A Active CN111040974B (en) | 2019-12-31 | 2019-12-31 | Banana endophytic Bacillus belgii and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111040974B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112522144A (en) * | 2020-12-09 | 2021-03-19 | 华南农业大学 | Bumper-fragrant endogenous Bacillus belgii and application thereof |
| CN112899201A (en) * | 2021-03-16 | 2021-06-04 | 云南省农业科学院农业环境资源研究所 | Bacillus belgii, application thereof and method for preventing and treating banana wilt |
| CN113061554A (en) * | 2021-04-16 | 2021-07-02 | 中国热带农业科学院热带生物技术研究所 | Bacillus belgii for preventing and treating banana wilt and application thereof |
| CN113502246A (en) * | 2021-07-08 | 2021-10-15 | 广西科学院 | Compound microbial agent and preparation method and application thereof |
| CN113801826A (en) * | 2021-10-29 | 2021-12-17 | 广西科学院 | Bacillus belgii strain HYL-1 and application thereof |
| CN114164120A (en) * | 2021-11-18 | 2022-03-11 | 海南大学 | Microbial preparation for preventing and treating plant diseases and preparation method and application thereof |
| CN115369064A (en) * | 2022-09-30 | 2022-11-22 | 海南师范大学 | Development and application of multifunctional composite microbial inoculum for enhancing rhizosphere colonization and control effect of biocontrol bacillus belgii |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101199931B1 (en) * | 2010-11-12 | 2012-11-09 | 한국화학연구원 | Promotion of nitrogen mineralization of organic fertilizers and control of plant diseases using bacillus velezensis KRICT934 |
| CN109022304A (en) * | 2018-07-04 | 2018-12-18 | 华南农业大学 | One plant of Bei Laisi bacillus and its broad-spectrum antibacterial effect and the application in prevention and treatment banana disease |
| CN110283742A (en) * | 2019-06-17 | 2019-09-27 | 北京农业生物技术研究中心 | The Bei Laisi bacillus of one plant of broad-spectrum disease resistance and its application |
-
2019
- 2019-12-31 CN CN201911413362.2A patent/CN111040974B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101199931B1 (en) * | 2010-11-12 | 2012-11-09 | 한국화학연구원 | Promotion of nitrogen mineralization of organic fertilizers and control of plant diseases using bacillus velezensis KRICT934 |
| CN109022304A (en) * | 2018-07-04 | 2018-12-18 | 华南农业大学 | One plant of Bei Laisi bacillus and its broad-spectrum antibacterial effect and the application in prevention and treatment banana disease |
| CN110283742A (en) * | 2019-06-17 | 2019-09-27 | 北京农业生物技术研究中心 | The Bei Laisi bacillus of one plant of broad-spectrum disease resistance and its application |
Non-Patent Citations (2)
| Title |
|---|
| JIANFENG HUANG等: "Suppression of Fusarium wilt of banana by combining acid soil ameliorant with biofertilizer made from Bacillus velezensis H-6", 《EUROPEAN JOURNAL OF PLANT PATHOLOGY》 * |
| 刘雪娇等: "贝莱斯芽孢杆菌3A3-15生防和促生机制", 《河北大学学报(自然科学版)》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112522144A (en) * | 2020-12-09 | 2021-03-19 | 华南农业大学 | Bumper-fragrant endogenous Bacillus belgii and application thereof |
| CN112899201A (en) * | 2021-03-16 | 2021-06-04 | 云南省农业科学院农业环境资源研究所 | Bacillus belgii, application thereof and method for preventing and treating banana wilt |
| CN113061554A (en) * | 2021-04-16 | 2021-07-02 | 中国热带农业科学院热带生物技术研究所 | Bacillus belgii for preventing and treating banana wilt and application thereof |
| CN113502246A (en) * | 2021-07-08 | 2021-10-15 | 广西科学院 | Compound microbial agent and preparation method and application thereof |
| CN113801826A (en) * | 2021-10-29 | 2021-12-17 | 广西科学院 | Bacillus belgii strain HYL-1 and application thereof |
| CN113801826B (en) * | 2021-10-29 | 2023-02-03 | 广西科学院 | Bacillus belgii strain HYL-1 and application thereof |
| CN114164120A (en) * | 2021-11-18 | 2022-03-11 | 海南大学 | Microbial preparation for preventing and treating plant diseases and preparation method and application thereof |
| CN115369064A (en) * | 2022-09-30 | 2022-11-22 | 海南师范大学 | Development and application of multifunctional composite microbial inoculum for enhancing rhizosphere colonization and control effect of biocontrol bacillus belgii |
| CN115369064B (en) * | 2022-09-30 | 2023-07-07 | 海南师范大学 | Development and application of multifunctional composite microbial inoculum for enhancing rhizosphere colonization and control effect of biocontrol bacillus bailii |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111040974B (en) | 2022-03-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111040974B (en) | Banana endophytic Bacillus belgii and application thereof | |
| CN112899201B (en) | Bacillus belgii, application thereof and method for preventing and treating banana wilt | |
| CN112746046B (en) | Bacillus belgii and application thereof in prevention and treatment of cucumber bacterial angular leaf spot | |
| CN113249242B (en) | Paenibacillus polymyxa and application thereof in prevention and treatment of various soil-borne diseases | |
| CN113005056B (en) | Bacillus belgii HY19 and application thereof | |
| CN109456921B (en) | Paenibacillus polymyxa, application thereof, microbial agent, powder and granules | |
| CN113717901A (en) | Bacillus belgii and application thereof in prevention and treatment of various vegetable diseases | |
| CN111763643A (en) | Compound flora for preventing and treating peanut root rot and application thereof | |
| CN108865900B (en) | Isaria javanicus strain and application thereof | |
| CN107699526B (en) | Actinomycete strain for preventing and treating gray mold and application thereof | |
| CN103109870B (en) | Application of fermentation supernatant liquid of pseudomonas | |
| CN111073827B (en) | Bacillus beilesensis JTB8-2 and application thereof in biological control of orobanche meloidogyne | |
| CN110317747B (en) | Bacillus amyloliquefaciens JT68 and application thereof in prevention and treatment of tea anthracnose | |
| CN115261283A (en) | Bacillus cereus and application thereof in prevention and control of dry farming potato diseases | |
| CN108570432B (en) | Bacillus amyloliquefaciens and application thereof | |
| CN113444651B (en) | Saffron endophytic fungus and application thereof in preventing and treating bulb rot | |
| CN110484478A (en) | A kind of bacillus subtilis JZ2-1-12 and its application | |
| CN114196585A (en) | Burkholderia for preventing and treating tomato bacterial wilt and application thereof | |
| CN113862156A (en) | Fusarium oxysporum (Fusarium oxysporum) K2018-1418 and application thereof | |
| CN113801804A (en) | Banana wilt biocontrol antagonistic strain and application thereof | |
| CN113403205A (en) | Penicillium beijerinckii MP6 with bacteriostatic action and application thereof | |
| CN115960777B (en) | Bacillus pseudomycoides and application thereof in prevention and treatment of vegetable epidemic disease | |
| CN108441443B (en) | Strain for preventing and treating plant nematodes and application thereof | |
| CN108998395B (en) | Bacillus amyloliquefaciens and application thereof | |
| CN113832038B (en) | Fusarium equisetum (Fusarium equiseti) K2017-696 and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |