CN115678811A - Streptomyces fusiformis LS159 and application thereof - Google Patents
Streptomyces fusiformis LS159 and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of microorganisms, and particularly relates to streptomyces fusiformis and application thereof in the field of biological control of plant diseases and food preservation and preservation. The LS159 strain obtained by screening of the invention has a preservation number of CGMCC No.19503. Has broad-spectrum antibacterial property, can be used for biocontrol bacteria and preventing food from putrefaction, and has wide application value.
Description
Technical Field
The invention belongs to the technical field of microorganisms, particularly relates to a microbial strain and application thereof in the biological control field of plant diseases and food preservation and preservation field, and more particularly relates to a streptomyces fusiformis strain and application thereof.
Background
Microorganisms generally and widely exist in the nature, most of the microorganisms are harmless to human or beneficial, not only can live bacteria cells be applied to food fermentation and vaccine development, but also can produce organic acids, enzyme preparations, antibiotic metabolites and the like by utilizing the physiological metabolism of the microorganisms, and play an important role in the recycling and resource utilization of natural substances. However, a small part of microorganisms can cause plant diseases, cause food spoilage and cause harm and loss to human beings. For harmful microorganisms causing plant diseases and food spoilage, the application of chemical pesticides and chemical preservatives has been the main measure. With the improvement of living standard of people, the consciousness of food safety and ecological environment protection is gradually improved, and the application demand of prevention and treatment measures of green and low toxicity is continuously increased.
Actinomycetes, as a class of gram-positive prokaryotic microorganisms, widely exist in soil, water, plant surfaces, interior and other environments, and many species can produce antibacterial performance, and are a species commonly used in plant disease control and food preservation. Research shows that some actinomycetes inhibit the growth of harmful microorganisms through producing antibiotics and chitinase or limit and prevent the growth of pathogenic bacteria through ecological niches and nutrition competition, so that disease infection and harm are reduced. Streptomyces (Streptomyces)Streptomycesspp.), e.g. Streptomyces microflavus (S. Microflavus: (S. Flavus)S. microflavus) Streptomyces kanamycini: (S. kanamyceticus) Streptomyces rimosus (A)S. rimosus) Streptomyces globisporus (A), (B)S. globisporus) Etc., but with respect to Streptomyces fusiformis (S. netropsis) There have been few reports on the control of harmful microorganisms and related properties. Chinese patent (publication No. CN 107058131A) discloses a Streptomyces fusiformis strain for preventing and treating wheat scab and reducing gibberellin content in wheat, which is obtained by separating and screening soil collected from a rice field. A large number of researches show that the biological characteristics of different strains of the same microorganism and the antagonistic effect on different diseases are greatly different, so that more strains capable of preventing and treating other diseases need to be continuously screened, and more strain resources are provided for the application of the strains.
The fermentation of the microbial inoculum needs to add proper nutrient substances such as carbon, nitrogen source and the like, in order to reduce the cost, the wastes after industrial and agricultural production are usually considered to be used when selecting a culture medium, the wastes contain a certain amount of substances such as carbon source, nitrogen source and the like, and the sources of the excellent culture medium which can be used for the fermentation of the microbial inoculum are properly adjusted. Wherein, the beer production process generates a large amount of waste water through the working procedures of saccharification, fermentation, filling and the like, and the beer waste water contains rich organic matters such as sugar, protein and the like. China is a large country for beer production and consumption, 3-10 tons of wastewater can be generated when 1 ton of beer is produced, the discharge amount of the beer wastewater is large, and a large amount of capital is spent in breweries every year due to the fact that the beer wastewater is treated, and a large amount of energy is consumed. From another point of view, organic matters (including sugars, proteins and the like) in the beer wastewater are good nutrient substrates for the growth of microorganisms, and can be made into a microorganism culture medium for producing microorganism products through fermentation.
Disclosure of Invention
Based on the method, the streptomyces fusiformis capable of inhibiting the plant pathogenic microorganisms and the food spoilage microorganisms is screened, simultaneously, the physiological characteristics and the molecular identification are carried out, and the streptomyces fusiformis is subjected to fermentation culture by utilizing the beer wastewater, so that the method has important significance for reducing the introduction of chemical agents, reducing the environmental pollution, improving the food safety and realizing the sustainable development of agriculture. The inventor finally screens and identifies to obtain a strain of streptomyces fusiformisStreptomycesnetropsis) LS159, and the function thereof has been intensively studied, thereby completing the present invention.
The invention firstly provides a streptomyces fusiformisStreptomycesnetropsis) LS159 strain, the preservation number is CGMCC No.19503.
The research result shows that the LS159 strain can treat the pathogenic bacteria of the Chinese chestnut epidemic disease: (Cryphonectria parasitica) Rhizoctonia solani of poplar: (Rhizoctonia solani) Carrot soft rot pathogen (A)Pectobacterium carotovorum) And tomato bacterial wilt bacterium: (Ralstonia solanacearum) Has strong inhibiting effect; in addition, against the root rot pathogenic bacteria of Chinese cabbage (Fusarium oxysporum) And bacterial Populus tremula ulcer pathogenic bacteria: (Botryosphaeria dothidea) Also has certain inhibiting effect. And compared with the same streptomyces fusiformis strain (strain number LS 286)The antibacterial spectrum and the antibacterial effect of the 8 pathogenic bacteria are better. Compared with other streptomycetes separated from the same region, the bacteriostatic spectrum and the bacteriostatic effect are also different. Therefore, the invention further provides the application of the LS159 strain as a biocontrol bacterium. More particularly, it is used for preventing and treating Chinese chestnut epidemic disease pathogenic bacteria (A)Cryphonectria parasitica) Poplar and poplar rhizoctonia solani bacteria (A), (B)Rhizoctonia solani) Carrot soft rot pathogen (A)Pectobacterium carotovorum) Or tomato bacterial wilt bacterium (Ralstonia solanacearum) Resulting fungal diseases; or against the root rot pathogenic bacteria of Chinese cabbage (Fusarium oxysporum) Bacterial pathogenic bacteria of poplar ulcer: (A), (B)Botryosphaeria dothidea) Resulting in bacterial diseases. More particularly, it is used for disease control of chestnut, poplar, carrot, tomato or Chinese cabbage.
Further studies showed that the LS159 strain antagonizes 8 common food spoilage causing fungi and bacteria, among which Penicillium expansum (A), (B)Penicillium expansum) Staphylococcus aureus (1)Staphylococcus aureus) And Escherichia coli (Escherichia coli) Has the best antagonistic effect on Aspergillus flavus (Aspergillus flavus) ((R))Aspergillusflavus) Alternaria alternata (Alternaria alternata) And Bacillus cereus (B.cereus) (B.cereus)Bacillus cereus) Also has good inhibition effect. Therefore, the invention further provides the application of the LS159 strain in preventing and treating food spoilage. More particularly, it is useful for inhibiting Aspergillus nigerAspergillus niger) Aspergillus flavus (A) andAspergillusflavus) Penicillium expansum (Penicillium expansum) Alternaria alternata (B)Alternaria alternata) Or Staphylococcus aureus (Staphylococcus aureus) Escherichia coli (E.coli)Escherichia coli) Bacillus cereus (B.cereus)Bacillus cereus) Bacillus coagulans bacterium (A), (B) and (C)Bacillus coagulans）。
Further research of the invention shows that the LS159 strain can better utilize a culture medium taking beer wastewater as a main component, compared with an ISP2 culture medium, the growth speed of the streptomyces fusiforme fermented by the beer wastewater is slower than that of the fermentation treatment of the ISP2 culture medium in 1-3 days of the initial culture period, but the growth amounts of the streptomyces fusiforme and the ISP culture medium are gradually close to the same in the middle and later fermentation periods. Therefore, the invention also provides a method for fermenting the LS159 strain by using beer wastewater. Specifically, the LS159 strain is fermented by a fermentation culture solution taking beer waste water as a basic component. More specifically, the fermentation culture solution is prepared by mixing beer wastewater and tap water according to a volume ratio of 1:3-8, then adding glucose and yeast extract powder, adjusting the pH value to 7.0-7.4, sterilizing and cooling. The fermentation conditions are 30-34 ℃ and 100-200 rpm for 3-7d. Preferably, the fermentation culture solution is prepared by mixing beer wastewater and tap water according to a volume ratio of 1:5, mixing, adding 0.5% of glucose and 0.5% of yeast extract powder, adjusting the pH value to 7.2, sterilizing and cooling; the culture conditions were 32 ℃ and 150 rpm for 5 days.
Moreover, researches show that the antibacterial function of the Streptomyces fusiformis LS159 bacterial liquid fermented by the beer wastewater is consistent with the effect of the bacterial liquid cultured on an ISP2 culture medium, which indicates that the Streptomyces fusiformis can be antagonised by the beer wastewater fermentation. Therefore, the invention also provides a bacterial liquid obtained by the fermentation method. Furthermore, the invention provides application of the bacterial liquid as a biocontrol bacterium or in preventing and treating food spoilage.
The LS159 bacterial strain obtained by screening has broad-spectrum antibacterial property, can be used for biocontrol bacteria and preventing food from putrefaction, and has wide application value.
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FIG. 1 antagonizes the bacteriostatic effect of different strains of actinomycetes on Fusarium solani FD 1.
FIG. 2 LS159 Strain 16S rDNA phylogenetic Tree.
FIG. 3 shows the effect of fermentation of Streptomyces fusiformis LS159 by beer wastewater.
Biological material preservation information: the LS159 strain of the present invention is named as Streptomyces fusiformis (Streptomycesnetropsis) And was deposited in the general microbiological culture collection center of the China Committee for culture Collection of microorganisms (Unit Address: west road No.1, north west of the township, beijing, ministry of sciences, china, institute of microbiology, zip code: 100101 CGMCC for short, and the preservation number is CGMCC No.19503.
Detailed Description
The invention is illustrated below by means of specific examples for the best understanding of the invention, without however being restricted thereto.
Example 1 isolation and screening of Streptomyces having antagonistic function
(1) Source of soil sample
The soil is taken from a peony planting base in Xiangyuan county of Changzhi, shanxi, selecting healthy peony plants for 5 years of oil, pulling up the plants with roots, brushing the soil adhered to the roots (namely rhizosphere soil) by using a sterile brush after shaking gently, mixing samples of 5 plants into a soil sample, filling the soil into a sterile plastic bag, and taking the sterile plastic bag back to a laboratory to be stored in a refrigerator at 4 ℃.
(2) Isolation of Streptomyces strains
Taking a 10 g rhizosphere soil sample, putting the sample in 90 mL sterile distilled water, uniformly oscillating for 30 min, and respectively diluting the soil suspension to 10 -3 ，10 -4 ，10 -5 And (4) doubling. mu.L of the diluted solution was applied to a solid medium of Goodpasture I (synthetic medium of Goodpasture I37.5 g,3% potassium dichromate (aq) 3mL, distilled water 1000 mL, pH 7.4-7.7). Inversely culturing in a constant temperature incubator at 30 deg.C, observing the plate after 3-7 days, selecting colony with different shape, color and size, streaking on the Gao's I culture medium plate, and storing the purified strain in a refrigerator at 4 deg.C for further screening of antagonistic bacteria.
(3) Screening of antagonistic function of strain
Screening the antagonistic effect of the separated strain by using a plate confronting culture method. Fusarium solani (F. Solani) with pathogenic bacteria preserved in laboratory and strong pathogenicity to peony for oilFusariumsolani) FD1, in modified PDA medium (i.e. PDA medium mixed with ISP2 medium 1:1 in proportion: yeast extract powder 2 g, malt extract powder 5 g, glucose 2 g, naCl 5 g, yeast extract powder 2.5 g, peptone 5 g, distilled water 1000 mL) in the center of a flat plate, placing activated Fusarium solani FD1 in the center of the flat plate by using a sterilization puncher, taking a circular pathogenic bacterium block with the diameter of 0.5 cm, inoculating actinomycetes to be detected in a position 2.5 cm away from the pathogenic bacterium block, and inoculating each flat plate with the circular pathogenic bacterium block3 kinds of actinomycetes are cultured in an incubator at 28 ℃ for 5-7 d, the inhibition of the actinomycetes on pathogenic bacteria is observed and recorded every day, the diameters and antagonistic radiuses of the pathogenic bacteria are measured by a vernier caliper, and the inhibition rate is calculated. Bacteriostatic rate = (control pathogen diameter-treated pathogen diameter)/control pathogen diameter × 100%.
The results show that 650 pure-breed microorganisms are separated from the root soil of the oil peony collected in the field of the oil peony in the Changzhi area of Shanxi province, and 42 actinomycetes are determined according to the colony morphology. The actinomycetes have 20 strains which have antagonism on Fusarium solani FD1, the strains with the bacteriostasis rate of more than 50 percent have 5 strains which are LS2, LS62, LS159, LS164 and LS286 respectively, wherein the strains with the highest bacteriostasis rate are LS159 and LS164, and the specific figure is 1.
Example 2 physicochemical characterization of Streptomyces LS159 and identification of 16S rRNA molecules
According to the manual of 'common bacteria system identification', morphological observation and physiological and biochemical identification are carried out on the strain LS159 with the best effect of antagonizing fusarium solani, and meanwhile, the strain is subjected to 16S rRNA gene amplification and sequencing analysis.
The results of the physicochemical characteristics of the strain LS59 are as follows: on ISP2 solid culture medium (yeast extract powder 4 g, malt extract powder 10 g, glucose 4 g, agar 20 g, distilled water 1L, pH 7.4-7.6), the strain has light yellow hypha in the substrate, light brown aerial hypha, opacity, long columnar spores and smooth surface. Wrinkles appeared after 48h of culture, spores were produced 3-7 days of culture, gram-positive, catalase was produced using starch, and v.p. was determined to be positive (see table 1).
TABLE 1 morphological characteristics and physiological and biochemical determination results of LS159 Strain
Note: "+" indicates a positive result, and "-" indicates a negative result.
BLAST comparison analysis is carried out on NCBI after 16S rRNA gene sequencing, and the result shows that the strain and streptomyces fusiformisStreptomyces netropsis（NR 112494.1）Homology is up to 99.86%, and is far away from other streptomycetes. Based on the above characteristics, LS159 strain was named Streptomyces fusiformis (Streptomyces netropsis) The result of constructing phylogenetic tree is shown in fig. 2, and the strain is preserved in the common microorganism center of the china committee for culture collection of microorganisms, which is abbreviated as CGMCC (unit address: west road No.1, north west of the township, beijing, ministry of sciences, china, institute of microbiology, zip code: 100101 CGMCC No. 19503).
Example 3 antagonism of various phytopathogenic microorganisms by Streptomyces fusiformis LS159
4 pathogenic microorganisms with common plant fungal diseases and bacterial diseases are selected as experimental materials, the antagonistic function spectrum of the streptomyces fusiformis LS159 is measured by adopting a plate confrontation method, and simultaneously, the streptomyces with high bacteriostatic rate of fusarium solani FD1 is compared by the other 4 strains obtained by separation. 4 pathogenic fungi are phytophthora capsici leonian pathogenic bacteria ()Phytophthora capsici) Pathogenic bacteria of chestnut blight (A)Cryphonectria parasitica) Poplar and poplar rhizoctonia solani bacteria (A), (B)Rhizoctonia solani) Pathogenic bacteria of cabbage root rot: (A), (B)Fusarium oxysporum) The 4 pathogenic bacteria are tomato bacterial spot pathogenic bacteria: (Pseudomonas syringae) Carrot soft rot pathogen (A)Pectobacterium carotovorum) Tomato bacterial wilt bacterium (Ralstonia solanacearum) Bacterial poplar canker (A)Botryosphaeria dothidea）。
Inoculating single streptomycete colony in ISP2 liquid culture medium, culturing at 30 deg.c and 150 rpm for 3-5d, and using the cultured bacterial liquid in plate confrontation experiment. For the antagonistic action of pathogenic fungi, plate opposition experiments were carried out by cutting peeled potatoes in a modified PDA medium (i.e., a mixture of PDA medium and ISP2 medium 1:1 in a ratio: 200 g potato is peeled off, boiling the cut potatoes in distilled water, filtering the cut potatoes with 8 layers of gauze after the water is boiled, adding distilled water to 1L, adding 20 g glucose and 16 g agar powder, inoculating activated 0.5 zxft 3825 × 0.5 cm in the center of a plate of yeast extract powder 4 5657, malt extract powder 10 g, glucose 4 g and water 1L), inoculating the activated bacterial blocks of pathogenic fungi in a bacterial strain box of 0.5 zxft 3825 × 0.5 3638 zxft, measuring the antagonistic action of pathogenic fungi in a vernier box at a distance from 2.5 zxft 3724, adding 100. Mu. Per well, treating the bacterial strain in a symmetrical vernier box, and measuring the radius of each Streptomyces strain 493-497 ℃ by measuring the pathogenic fungi with a caliper of strain at 497-24. For the antagonistic action of pathogenic bacteria, when a plate confronting experiment is carried out, 200. Mu.L of each of 4 kinds of cultured pathogenic bacteria are respectively coated on LB modified culture medium (namely LB culture medium and ISP2 culture medium 1:1 are mixed in proportion, wherein the LB culture medium is NaCl 10 g, yeast extract powder 5 g, peptone 10 g, distilled water 1L, pH 7.0-7.5, the ISP2 culture medium is yeast extract powder 4 g, malt extract powder 10 g, glucose 4 g and water 1L), 3 holes with the diameter of 0.7 zxft 3579 are punched in the middle of the plates, 100. Mu.L of streptomycetes is added into the holes, 24-36 zxft 3525 bacteria liquid is cultured at the temperature of 30 ℃, and the diameter of the bacteria is measured by vernier caliper.
The results show that LS159 can treat pathogenic bacteria of Chinese chestnut epidemic disease: (A)Cryphonectria parasitica) Poplar and poplar rhizoctonia solani bacteria (A), (B)Rhizoctonia solani) Carrot soft rot pathogen (A)Pectobacterium carotovorum) And tomato bacterial wilt bacterium: (Ralstonia solanacearum) Has stronger inhibiting effect; in addition, against the root rot pathogenic bacteria of Chinese cabbage (Fusarium oxysporum) And bacterial Populus tremula ulcer pathogenic bacteria: (Botryosphaeria dothidea) Also has certain inhibiting effect. And has better antibacterial spectrum and antibacterial effect on the 8 pathogenic bacteria than the same streptomyces fusiformis strain (strain number LS 286). Compared with other streptomycetes separated from the same region, the bacteriostatic spectrum and the bacteriostatic effect are different. The specific results are shown in Table 2.
TABLE 2 bacteriostatic effect of different Streptomycete on 8 plant pathogenic microorganisms
Note: pathogenic fungi 1. Pathogenic bacteria of phytophthora capsici ()Phytophthora capsici) (ii) a 2. Pathogenic bacteria of chestnut blight (Cryphonectria parasitica) (ii) a 3. Rhizoctonia solani of poplar (Rhizoctonia solani) (ii) a 4. Chinese cabbage root rot pathogen: (Fusarium oxysporum). Pathogenic bacteria: 1. bacterial spot pathogen of tomato（Pseudomonas syringae) (ii) a 2. Carrot soft rot pathogen (A)Pectobacterium carotovorum) (ii) a 3. Tomato bacterial wilt bacterium (Ralstonia solanacearum) (ii) a 4. Bacterial poplar ulcer (A)Botryosphaeria dothidea). "-" no antagonism; "+"0<Antagonistic radius<5mm；“++” 5<Antagonistic radius<10 mm; the "+ + + +" antagonistic radius>10mm。
Example 4 antagonism of Streptomyces fusiformis LS159 against various food spoilage microorganisms
4 kinds of common fungi and bacteria causing food spoilage are selected as experimental materials, the antibacterial function spectrum of the streptomyces fusiformis LS159 is measured by adopting a plate opposition method, and meanwhile, the other strain of the streptomyces fusiformis LS286 is used for comparison. The 4 fungi are Aspergillus nigerAspergillus niger) Aspergillus flavus (A. Flavus)Aspergillusflavus) Penicillium expansum (Penicillium expansum) Alternaria alternata (B)Alternaria alternata) The 4 bacteria are staphylococcus aureus (Staphylococcus aureus) Escherichia coli (E.coli)Escherichia coli) Bacillus cereus (B.cereus)Bacillus cereus) Bacillus coagulans bacterium (A), (B) and (C)Bacillus coagulans). The fungi are cultured in PDA culture medium for 3-5 days, and the bacteria are cultured in LB liquid culture medium for 12-18h.
Inoculating single streptomycete colony in ISP2 liquid culture medium, culturing at 30 deg.c and 150 rpm for 3-5d, and applying the cultured bacterial liquid in plate confronting experiment. For the antagonism of food spoilage fungi, the plate confrontation experiment is carried out by mixing the modified PDA culture medium (i.e. PDA culture medium and ISP2 culture medium 1:1 ratio: PDA culture medium is 200 g peeled potatoes, cutting into small blocks, boiling in distilled water, filtering with 8 layers of gauze after water is boiled, supplementing to 1L with distilled water, adding 20 g glucose, adding 16 g agar powder, inoculating activated 0.5 zxft 3825 × 0.5 cm fungal blocks in the center of the plate, inoculating to the activated 0.5 zxft 3825 × 0.5 zxft 5657, malt extract 10 g, glucose 4 g, water 1L, inoculating to the fungal blocks in the box at both sides of 2.5 zxft 3724, symmetrically treating each hole, measuring the radius of streptomycetes in 493-497 ℃ and measuring the radius of the spoilage fungi repeatedly by using a bacterial liquid measuring the strain with a caliper gauge at 497-24. For antagonism of food spoilage bacteria, when carrying out plate confrontation experiment, respectively coating 200 μ L of 4 kinds of cultured food spoilage bacteria on LB modified culture medium (namely LB culture medium and ISP2 culture medium 1:1 are mixed in proportion, wherein the LB culture medium is NaCl 10 g, yeast extract powder 5 g, peptone 10 g, distilled water 1L, pH 7.0-7.5 ISP2 culture medium is yeast extract powder 4 g, malt extract powder 10 g, glucose 4 g, water 1L) plates, punching 3 holes with diameter of 0.7 zxft 3579 in the middle of the plates, adding 100 μ L streptomycetes bacteria liquid into the holes, culturing 24-36 xft 3525 at 30 ℃, and measuring the bacteriostatic diameter.
The results show that S.fusiformis LS159 has antagonism on 8 fungi and bacteria causing food spoilage, wherein the strain is Penicillium expansum (A)Penicillium expansum) Staphylococcus aureus (1)Staphylococcus aureus) And Escherichia coli (Escherichia coli) Has the best antagonistic effect on Aspergillus flavusAspergillusflavus) Alternaria alternata (Alternaria alternata) And Bacillus cereus (B.cereus) (B.cereus)Bacillus cereus) Also has good inhibition effect. In addition, the inhibition effect of LS59 on 8 microorganisms causing food spoilage is better than that of Streptomyces fusiformis LS286, and the specific results are shown in Table 3.
TABLE 3 bacteriostatic action of Streptomyces fusiformis LS159 on 8 food spoilage microorganisms
Note: food spoilage fungus 1. Aspergillus nigerAspergillus niger) Aspergillus flavus (2)Aspergillusflavus) 3. Penicillium expansum (Penicillium expansum) Alternaria alternata (4)Alternaria alternata) Food spoilage bacteria: 1. staphylococcus aureus (1)Staphylococcus aureus) 2, E.coli: (Escherichia coli) 3, bacillus cereus: (Bacillus cereus) 4. Bacillus coagulans: (A)Bacillus coagulans). "-" is not antagonistic; "+"0<Antagonistic radius<5mm；“++” 5<Antagonistic radius<10 mm; the "+ + + +" antagonistic radius>10mm。
Example 5 fermentation of Streptomyces fusiformis LS159 Using beer waste Water
Inoculating a single colony of Streptomyces fusiformis LS159 in an ISP2 liquid culture medium, culturing at 30 ℃ and 150 rpm for 3-5d, and culturing the cultured bacterial liquid for later use. Adding a certain amount of tap water (the volume ratio of the beer wastewater to the tap water is 1:5), adding 0.5% of glucose and 0.5% of yeast extract powder, adjusting the pH value to 7.0-7.4, subpackaging into 250ml triangular bottles, wherein each bottle contains 50ml, sterilizing, cooling, inoculating according to the inoculation amount of 2%, culturing at 32 ℃, 150 rpm for 1 d, 2d, 3d, 5d and 7d, and measuring the OD600 value by using a microplate reader. LS159 was inoculated with ISP2 medium and the treatments inoculated and cultured under the same conditions were used as controls.
The results show that the streptomyces fusiformis LS159 can better utilize the culture medium with beer wastewater as the main component, compared with the ISP2 culture medium, the growth speed of the streptomyces fusiformis fermented by the beer wastewater is slower than that of the ISP2 culture medium in the initial stage of the culture, but the growth amounts of the streptomyces fusiformis are gradually close to the same in the middle and later stages of the fermentation. The specific results are shown in FIG. 3.
Example 6 bacteriostatic action of Streptomyces fusiformis LS159 Using beer waste Water fermentation
In order to confirm whether the bacteriostatic function of the Streptomyces fusiformis LS159 bacterial solution fermented by beer wastewater was affected, a Streptomyces fusiformis LS159 fermentation solution cultured for 5 days in beer wastewater was obtained by the method of example 5, and the bacteriostatic effect thereof was investigated. The harmful microorganisms used in the bacteriostasis experiment comprise 1 plant pathogenic fungus, i.e., (Chinese chestnut epidemic pathogenic bacteria)Cryphonectria parasitica) 1 plant pathogenic bacterium carrot soft rot pathogenPectobacterium carotovorum) 1 fungus Penicillium expansum (a) causing food spoilagePenicillium expansum) And 1 bacterium Staphylococcus aureus causing food spoilage: (Staphylococcus aureus)。
The results show (see table 4), the bacteriostatic function of the streptomyces fusiformis LS159 bacterial liquid fermented by the beer wastewater is consistent with the effect of the bacterial liquid cultured on the ISP2 culture medium, which indicates that the streptomyces fusiformis bacterial liquid can be antagonized by the beer wastewater fermentation.
TABLE 4 bacteriostatic action of Streptomyces fusiforme LS159 bacterial liquid fermented by beer wastewater
Note: 1. pathogenic bacteria of chestnut blight (Cryphonectria parasitica) Carrot soft rot pathogen (A)Pectobacterium carotovorum) 3. Penicillium expansum (Penicillium expansum) S. aureus (S.) (Staphylococcus aureus) 3, escherichia coli (E. Coli)Escherichia coli) 3, bacillus cereus: (Bacillus cereus) 4, bacillus coagulans (C)Bacillus coagulans). The "+ + + +" antagonistic radius>10mm。
Claims (10)
1. Streptomyces fusiformis (Streptomyces netropsis) LS159 strain with preservation number CGMCC No.19503.
2. Use of the LS159 strain of claim 1 as a biocontrol bacterium.
3. The use as claimed in claim 2 for controlling pathogenic bacteria of chestnut blight(s) (Cryphonectria parasitica) Poplar and poplar rhizoctonia solani bacteria (A), (B)Rhizoctonia solani) Carrot soft rot pathogen (A)Pectobacterium carotovorum) Or tomato bacterial wilt bacterium (Ralstonia solanacearum) Resulting fungal diseases; or against the root rot pathogenic bacteria of Chinese cabbage (Fusarium oxysporum) Bacterial pathogenic bacteria of poplar ulcer: (A), (B)Botryosphaeria dothidea) Resulting in bacterial diseases.
4. The use as claimed in claim 3, for disease control of chestnuts, poplar, carrots, tomatoes or chinese cabbages.
5. Use of the LS159 strain of claim 1 for the control of food spoilage.
6. Such as rightThe use according to claim 3 for inhibiting Aspergillus nigerAspergillus niger) Aspergillus flavus (A) andAspergillusflavus) Penicillium expansum (Penicillium expansum) Alternaria alternata (B)Alternaria alternata) Or Staphylococcus aureus (Staphylococcus aureus) Escherichia coli (E.coli)Escherichia coli) Bacillus cereus (B.cereus)Bacillus cereus) Bacillus coagulans (C.coagulans) ((C.coagulans))Bacillus coagulans）。
7. A method for fermenting the LS159 strain by using beer wastewater, which comprises the steps of fermenting the LS159 strain by using a fermentation culture solution taking the beer wastewater as a basic component; preferably, the fermentation culture solution is prepared by mixing beer wastewater and tap water according to a volume ratio of 1:3-8, then adding glucose and yeast extract powder, adjusting the pH value to 7.0-7.4, sterilizing and cooling.
8. The method of claim 7, wherein the fermentation broth is a mixture of beer wastewater and tap water in a volume ratio of 1:5, mixing, adding 0.5% of glucose and 0.5% of yeast extract powder, adjusting the pH value to 7.2, sterilizing and cooling; the fermentation conditions are 30-34 deg.C, 100-200 rpm for 3-7d, more specifically 32 deg.C, 150 rpm for 5d.
9. A bacterial liquid obtained by the method according to claim 7 or 8.
10. The use of the bacterial liquid according to claim 9 as a biocontrol bacterium or for preventing food spoilage.
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| CN118086149A (en) * | 2024-04-22 | 2024-05-28 | 内蒙古农业大学 | High-efficiency broad-spectrum antagonistic bacterium streptomyces fusiformis QH1-20 and application thereof |
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