CN111778171B - Bacillus belgii and application thereof - Google Patents
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Abstract
The invention provides a bacillus beilesensis zk1 with the preservation number: CGMCC NO. 17209. The invention takes the strain obtained by laboratory separation as an example, firstly provides the rot-causing effect of the Bacillus belgii on peach fruits, and detects that the strain produces toxic substances after infecting the chickpeas by utilizing metabonomics. The Bacillus belgii strain is used as a standard strain, and has an important effect on quality control of picked peach fruits.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a bacillus beilesiensis strain.
Background
Peach (amygdalusspersca Linn) belongs to Rosaceae, and has been cultivated in China for four thousand years. In 2017, the total cultivation area and the total yield of Chinese peaches account for 51.16% and 57.95% of the total amount of world peaches (FAOSTAST, 2017), respectively. The peach varieties bred in China are multiple, the distribution of the cultivation areas is wide, a plurality of provinces such as Shandong, Shaanxi, Yunnan, Jiangsu and Guangdong are covered, and the peach variety is an important component of local fruit tree planting industry. Fresh food is one of the main sales forms of peaches in the market of China.
Peaches for fresh eating need to be stored and transported for a while after being harvested, but during this process, the peaches are often rotted in batches. The preservation and freshness of the picked peaches are always difficult for the following reasons. Peach is a respiration jump type stone fruit, and the picked peach can have enzymatic reaction, so that the fruit is oxidized, aged, softened and browned. In addition to this, many pathogenic bacteria, such as, for example, Streptomyces fructicola (Monilinia spp.), Penicillium spp, Rhizopus spp and Botrytis Cinerea (Bortrytis Cinerea) etc., infect peach through wounds due to the perishable nature of peach fruit and susceptibility to decay pathogens, resulting in a large number of peach fruit decays. At present, the study on the pathogenic bacteria stored after peach picking at home and abroad mainly focuses on fungi. It is widely accepted in the industry that these decay caused by fungi are important factors limiting the preservation of peaches after harvest, and there is no report on the pathogenicity of bacteria to peach fruits for a while.
Disclosure of Invention
Aiming at the problem of bacterial decay of picked peaches, the inventor takes the chickpeas in the Guangdong Heiping area as a main research object, adopts a tissue culture method, uses a Nutrient Agar (NA) culture medium to separate a bacillus zk1 from decayed pulps, and proves that the bacillus zk1 has strong decay-causing effect on different varieties of peaches through a pathogenicity experiment that the strain is repeatedly infected with peaches.
The invention provides a Bacillus belgii zk1(Bacillus velezensis zk1) separated from rotten pulp of an olecranberry fruit, which is preserved in the common microorganism center of China Committee for culture Collection of microorganisms (address: No. 3 of West Lu No.1 of Suzuo No. 3 of Suzuo of Chaoyang district, Beijing City, China academy of sciences microbiology institute) within 1 month and 17 days in 2019, and the preservation number is CGMCC NO. 17209.
According to some embodiments of the invention, the bacillus belgii has the amino acid sequence of SEQ ID NO: 1.
According to some embodiments of the invention, the colony biology of bacillus belgii is: white round, irregular edge, fold in the middle of bacterial colony, easy picking up, thallus has stickness.
According to some embodiments of the invention, the method of activating bacillus belgii is: streaking culture is carried out on an NA solid culture medium, and then liquid culture is carried out in an NB liquid culture medium. Wherein the NA solid medium is an NA flat plate.
According to some embodiments of the invention, the NA solid medium is formulated as: 1L of distilled water, 8-12g of peptone, 4-6g of sodium chloride, 2.5-3.5g of beef extract powder and 12-18g of agar.
According to some embodiments of the invention, the NA solid medium is formulated as: 1L of distilled water, 10.0g of peptone, 5.0g of sodium chloride, 3.0g of beef extract powder and 15.0g of agar.
According to some embodiments of the invention, the NB liquid medium is formulated as: 1L of distilled water, 8-12g of peptone, 4-6g of sodium chloride and 2.5-3.5g of beef extract powder.
According to some embodiments of the invention, the NB liquid medium is formulated as: 1L of distilled water, 10.0g of peptone, 5.0g of sodium chloride and 3.0g of beef extract powder.
According to some embodiments of the invention, the streaking culture is performed at 35-40 ℃ for 20-30 h.
According to some embodiments of the invention, the liquid culture conditions are: culturing at constant temperature of 35-40 ℃ for 6-10h at 200-300 rpm.
In another aspect, the invention provides the use of bacillus beijerinckii according to the first aspect for quality control after fruit picking.
According to some embodiments of the invention, the quality control comprises detecting the presence, absence or amount of bacillus belgii in the fruit.
According to some embodiments of the invention, the fruit is a rosaceous fruit.
According to a preferred embodiment of the invention, the fruit is peach.
According to a further preferred embodiment of the present invention, the peach is selected from at least one of the group consisting of a eagle peach, a honey peach, a flat peach, a wild peach, a yellow peach and a nectarine.
The Bacillus belgii zk1(Bacillus velezensis zk1) provided by the invention has a strong rot-causing effect on peach fruits, and generates toxic metabolites when rotting peach flesh. The invention provides pathogenicity of the bacillus belief for fruits and vegetables for the first time and preserves the seeds of the fruits and vegetables. The strain is recommended to be used as a standard strain for monitoring and researching the quality control of the picked peach fruits.
Drawings
FIG. 1 shows the decay-causing effect of Bacillus belgii zk1 on eagle peach, the left panel is eagle peach inoculated with Bacillus belgii zk1, the right panel is blank control; the upper 3 holes (near the fruit tip) are infected without wound, and the lower 3 holes are infected with wound.
FIG. 2 shows the rot-causing effect of B.beijerinckii zk1 on other varieties of peaches, the left panel is peaches inoculated with B.beijerinckii zk1, and the right panel is blank control; 3 holes on the left side of the fruit are free from wound infection, and 3 holes on the right side of the fruit are wound infection; according to the sequence from left to right and from top to bottom, respectively, honey peach, flat peach, wild peach, yellow peach and nectarine are obtained.
FIG. 3 shows the colony morphology of Bacillus belgii zk 1.
FIG. 4 shows a phylogenetic tree of B.belgii zk 1.
Detailed Description
The technical solutions of the present invention are further described in detail with reference to the drawings and specific examples, which are provided for illustrative purposes only and do not limit the present invention in any way.
Performing isolation and identification of a strain
The method for screening the decay-causing bacteria of the picked chickpeas comprises the following steps:
1. olecranon collection
In the middle period of the olecranon fruit period, at least 30 rotten fruits of the olecranon fruits in different orchards are randomly collected in a peaceful area (one of main production areas of the olecranon fruits) of the river source city in Guangdong province. Sealed with sterile bags and transported to the laboratory over 24h at room temperature.
2. Cultivation and isolation of spoilage bacteria
Tissue culture method is adopted, small pieces (1cm × 1cm × 1cm) of tissue at the edge part of the putrefactive pulp are cut with a sterilized knife on a sterile operating platform, and are placed on Nutrient Agar (NA) culture medium, and cultured for 24h at 37 ℃. Numbering the cultured strains, picking strains with obviously different colony forms on the surface of a culture medium by using an inoculating loop on an aseptic operation table, carrying out streak separation in an aseptic culture dish, repeatedly carrying out two parallels on each strain, and culturing for 24 hours at 37 ℃.
3. Pathogenicity verification of putrefying bacteria
Taking healthy chickpea fruits, wiping the surface of the peel with 75% alcohol for 3 times, punching 3 round holes (diameter is 2mm and depth is 1cm) on the equator of each chickpea by using a sterile iron nail after the alcohol is volatilized, digging and sticking the single bacterial colony cake obtained by separation on the wound of the chickpea, sticking the bacterial cake close to the tip of the chickpea to perform non-invasive infection, and performing 3 holes each time. Meanwhile, a blank control group is made of sterile fungus cakes. Subpackaging each fruit with sterile bag, spraying sterile water for moisturizing, culturing at 37 deg.C for 48 hr, observing and recording putrefactive effect. And (3) streaking and separating the bacterial strains with the putrefactive effect from the wounds of the hawk-bill peaches again, comparing the bacterial colony morphology before inoculation, and streaking the bacterial strains with basically consistent morphology for seed preservation.
The separated bacteria were infected with other varieties of peaches as described above.
4. Identification of Bacillus belgii zk1
Through the steps, the inventor separates a strain of bacteria from the decayed chickpea flesh, and the bacteria can cause decay on wounds and non-wounds of the chickpea, the juicy peach, the flat peach, the wild peach, the yellow peach and the nectarine (fig. 1 and 2), and performs morphological observation, physiological and biochemical experiments and strain identification on the bacillus zk 1.
(1) Morphological characteristics of culture
On the nutrient agar medium, the colonies of the bacillus zk1 are white circles, the edges are irregular, wrinkles are formed in the middle of the colonies, the colonies are easy to pick up, and the thalli have viscosity, as shown in figure 3.
(2) Physiological and biochemical experiment
Note: "+" indicates positive reaction, and "-" indicates negative reaction.
(3)16S rDNA sequencing and sequence analysis
And (3) performing PCR amplification on a 16SrDNA sequence of the bacteria by taking the extracted bacterial genome DNA as a template, wherein primers are 27 f: 5'-AGAGTTTGATCCTGGCTCAG-3', respectively; 1492 r: 5'-ACGGCTACCTTGTTACGACTT-3' are provided. And (3) PCR reaction system: 2xT5 super PCR mix uL, upstream and downstream primers (10. mu. mol/L) each 1 uL, DNA template 1 uL, adding ultrapure water to 20 uL. PCR amplification procedure: pre-denaturation at 95 ℃ for 65 min; denaturation at 95 ℃ for 30s, annealing at 55 ℃ for 3s, extension at 72 ℃ for 1min, and 35 cycles; extension at 72 ℃ for 7 min. The PCR amplification product was detected by 1.0% agarose gel electrophoresis. Sequencing was performed by south core medical limited.
The sequencing results for 16S rDNA are as follows:
the sequencing results are compared in GenBank and compared with the homology of 16S rDNA genes of other strains, and the strain zk1 has the highest sequence similarity with Bacillus velezensis in the result of the highest matching degree, so the strain is named as Bacillus velezensis zk 1. And a phylogenetic tree (figure 4) is established, MEGA4.1 software is adopted to construct by using an adjacency method, Bootstrap evolution analysis is carried out, and a self-development test confidence value is set to be 1000 times. The strain is preserved in the China general microbiological culture Collection center (address: No. 3 of West Lu No.1 of Beijing Korean district, microbial research institute of Chinese academy of sciences) at 17.1.2019, with the preservation number of CGMCC NO. 17209.
Carrying out metabonomic research on infection of chickpeas by Bacillus diberelis zk1
1. Strain activation
After the strain zk1 was cultured on NA solid medium at 37 ℃ for 24 hours, a single colony was taken and cultured in NB liquid medium at 37 ℃ and 200rpm until logarithmic phase (8 hours). The bacterial cells were collected by centrifugation at 3000rpm for 15min and adjusted to a bacterial suspension having a bacterial concentration of 106CFU/mL using sterile physiological saline for further use.
2. Sample preparation
In a triangular lake orchard in Neng-Ping county, Heyuan, Guangdong province, 30 olecranberry trees with the age of 10 years are selected, mature olecranberry fruits with no damage to the surfaces and uniform size and color are picked, and the ripe olecranberry fruits are transported to a laboratory within 12 hours at the temperature of 4 ℃ in a cold chain. Washing the surface of semen Ciceris Arietini with sterile distilled water for 5min, and scrubbing with 75% ethanol by using sterile alcohol cotton ball (scrubbing for 3 times after the ethanol is volatilized). All the sterilized chickpeas are placed on a sterile operating platform to be dried, round holes (each fruit has only one wound, the diameter is 3mm, the depth is 10mm) are punched at the equator position of the fruit by using sterilized iron nails, and 0.1mL of bacterial suspension is immediately injected (the solution is prepared and used within 10 min). All fruits are individually sealed and packed in plastic bags in a sterile operating table, and sterile distilled water is sprayed into the bags to keep the humidity above 80% and a certain amount of air is left. The treated chickpeas were randomly divided into 5 groups of 10 parallel groups and stored at 37 ℃. The method comprises the following steps of taking the chickpeas stored for 15min as a blank control group (day 0), and taking the chickpeas putrefactive pulps stored for 1 day, 2 days, 3 days and 4 days as experimental groups respectively. 1g of peach flesh is dug into a freezing tube within the radius of 40mm by taking the wound as the center of a circle for each fruit, and the peach flesh is immediately frozen and stored by liquid nitrogen. All samples were stored in a-80 ℃ freezer prior to testing.
3. Metabonomics experimental procedure
Thawing at room temperature, cutting semen Ciceris Arietini tissue with small scissors, homogenizing with homogenizer, and performing ultrasonic treatment for 10min (ice water bath), wherein the process is repeated 4 times. Refrigerating at-20 deg.C for 120 min. Centrifuging at 13000rpm for 15min at 4 deg.C, collecting 200 μ L supernatant, and vacuum drying at room temperature. Dissolve with 200. mu.L acetonitrile: water (1:1, v/v), vortex for 30s, sonicate for 10min, centrifuge again at 13000rpm, 4 ℃ for 15 min. 150 μ L of the supernatant was loaded onto a machine, and 10 μ L of each sample was mixed to prepare QC.
The assay used ultra-high pressure liquid phase (Agilent 1290, Agilent Technologies) and high resolution mass spectrometry (Agilent 6545 QTOF, Agilent Technologies) in combination with chromatography columns (ACQUITY UPLC HSST 31.8 μm, 2.1X 100 mm; Waters) for metabolite detection. The column temperature was 35 ℃ and the loading was 2. mu.L. The positive ion mode is a ═ water: formic acid (1000:1), B ═ acetonitrile: formic acid (1000: 1). The anion mode is A ═ water +2mM CH3COONH4, B ═ acetonitrile. The sample flow rate was 500. mu.L/min, 95% A was maintained for 0.5min, the drop at 2min was 90%, and at 8min was 40%. Part A remained at 4% at 9 to 11min, rose to 95% at 12min and was maintained for 3min until the end. The Agilent 6545A QTOF mass spectrometer carries out primary and secondary mass spectrum Data Acquisition based on an Auto MS/MS mode under the control of control software (LC/MS Data Acquisition, Version B.08.00), and the mass scanning range m/z (50-1100). Respectively adopting positive and negative ion modes for collection; ESI ion source dry Gas temperature (Gas Temp) was 320 ℃, nitrogen Flow rate (Gas Flow) was 8l/min, sheath Gas Flow rate (sheathGasflow) was 12l/min, sheath Gas temperature (sheathGasTemp) was 350 ℃, capillary voltage (VCap) was 3500V (negative ion mode) and 4000V (positive ion mode).
And (4) performing principal component analysis on all peaks detected by the QC sample and the test sample. And performing data processing such as peak searching, peak alignment and the like through software MS-DIAL, and matching the primary and secondary maps with the compound map of the database to obtain an identification result. For the mass spectrometric results, coefficient of variation (CV, RSD) calculations were performed using QC samples, excluding peaks with CV greater than 30% and excluding peaks that were not detected in QC samples, and then the sample peaks were normalized using the R kit MetaboAnalystR. The normalized data were subjected to statistical tests (T-test; more than two Kruskal-Wallis H-test) and median fold difference calculations (two-set comparisons) on a group-by-group basis. To determine differentially expressed metabolites, analysis of variance and Variable Impact Projection (VIP) values were used to determine statistical significance. The difference was statistically significant when P <0.05, VIP > 1. Differential analysis and enrichment analysis were performed using the metaboanalyst4.0 software. Hierarchical Cluster Analysis (HCA) and partial least squares discriminant analysis (PLS-DA) were performed on the final differential compounds. And searching a database (version V2.0) which independently integrates Metlin, MassBank, MoNA and HMDB based on the primary map and the secondary map to obtain an identification result.
4. Toxic metabolite identification results
In the rotten peach flesh on day 4, a significant up-regulation of N, N-dimethyl-1, 4-phenylenediamine (P < 0.001) was detected. N, N-dimethyl-1, 4-phenylenediamine is a toxic compound, generally from the chemical industry, but has not been reported in food. The experiment may detect the structural analogue of N, N-dimethyl-1, 4-phenylenediamine. It is not clear which substances in peach pulp are utilized by bacillus belgii zk1 and produce N, N-dimethyl-1, 4-phenylenediamine or its structural analogs, and it is noted that fruits infected with bacillus belgii zk1 are not recommended for consumption.
The Bacillus belgii zk1 provided by the invention is separated from rotten chickpea fruit pulp, and has strong pathogenicity on picked peach fruits through pathogenicity verification, so that the pulp can be quickly rotten in the shelf life of the fruits. Bacillus belgii is generally studied as a biocontrol bacterium. Different from the previous research, the invention takes the strain obtained by laboratory separation as an example, firstly proposes the rot-causing effect of the Bacillus belgii on peach fruits, and detects that the strain produces toxic substances after infecting the chickpeas by utilizing metabonomics. The strain of Bacillus velezensis zk1 is used as a standard strain, and has an important effect on the quality control of peach fruits after picking.
It should be noted that the above-mentioned embodiments are only for explaining the present invention, and do not constitute any limitation to the present invention. The present invention has been described with reference to exemplary embodiments, but the words which have been used herein are words of description and illustration, rather than words of limitation. The invention can be modified, as prescribed, within the scope of the claims and without departing from the scope and spirit of the invention. Although the invention has been described herein with reference to particular means, materials and embodiments, the invention is not intended to be limited to the particulars disclosed herein, but rather extends to all other methods and applications having the same functionality.
Claims (4)
1. The Bacillus belgii has the preservation number of CGMCC NO. 17209.
2. Use of bacillus beijerinckii according to claim 1 for quality control after fruit picking; the fruit is peach.
3. Use according to claim 2, wherein said quality control comprises detecting the presence, absence or amount of said Bacillus belgii in said peaches.
4. Use according to claim 2 or 3, wherein the peach is selected from at least one of the group consisting of an eagle peach, a honey peach, a flat peach, a wild peach, a yellow peach and a nectarine.
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| CN105199996A (en) * | 2015-11-03 | 2015-12-30 | 河北省农林科学院植物保护研究所 | Bacillus amyloliquefaciens for controlling tomato gray mould and application of bacillus amyloliquefaciens |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105199996A (en) * | 2015-11-03 | 2015-12-30 | 河北省农林科学院植物保护研究所 | Bacillus amyloliquefaciens for controlling tomato gray mould and application of bacillus amyloliquefaciens |
| CN105985922A (en) * | 2016-01-20 | 2016-10-05 | 华南农业大学 | Bacillus aryabhattai J5 and application thereof |
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| Title |
|---|
| 微生物产生的酶抑制剂研究1.蛋白酶抑制剂的筛选方法探讨;刘华珍等;《抗生素》;19831231;第8卷(第5期);全文 * |
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