CN108570433B - Acid-resistant pseudomonas defensins C L P-6 and application thereof - Google Patents
Acid-resistant pseudomonas defensins C L P-6 and application thereof Download PDFInfo
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- CN108570433B CN108570433B CN201810438730.8A CN201810438730A CN108570433B CN 108570433 B CN108570433 B CN 108570433B CN 201810438730 A CN201810438730 A CN 201810438730A CN 108570433 B CN108570433 B CN 108570433B
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Abstract
The invention provides an acid-resistant pseudomonas defensins C L P-6 and application thereof, wherein the strain is preserved in the China general microbiological culture Collection center at 27.10.2016, and the biological preservation number is CGMCC No. 13205. the strain has the advantages of acid resistance, good disease prevention and growth promotion effects and strong stress resistance, has pathogenic bacterium antagonistic activity, has wide tolerance range on soil acidity and wide antibacterial spectrum, has application and development potential in the prevention and control of tobacco bacterial wilt, black shank and brown spot.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to acid-resistant pseudomonas defensins C L P-6 and application thereof.
Background
The tobacco suffers from various fungal and bacterial diseases in the whole growth period, and more than 60 recorded tobacco diseases are recorded in China, wherein the serious diseases comprise nearly 10 of tobacco black shank, bacterial wilt, brown spot, anthracnose, black head drop, powdery mildew and the like. Some pathogens can be harmful in various growth stages of tobacco, rootstock diseases such as tobacco black shank (phytophthora parasitica var. nicotianae) and tobacco bacterial wilt (Ralstonia solanacearum) can be harmful in seedling stage to mature stage of tobacco, tobacco brown spot is serious in later growth stage, and the tobacco is epidemic along with climate of different years, so that the serious yield reduction of tobacco fields seriously affects the yield and quality of tobacco leaves.
Tobacco bacterial wilt, also known as bacterial wilt, is a typical vascular bundle disease caused by ralstonia solanacearum, roots, stems and leaves can be damaged, infected leaves are still green and hung on the stems in the initial stage, and the disease is called bacterial wilt. When the weather is hot, the leaves of the diseased plant are irregularly burnt, the leaves are dry, the stem of the diseased plant with the edge falling off is upright, and dead leaves are hung on the stem; when the soil humidity is high, the roots are soft and sticky, so that the whole plant is withered. The tobacco bacterial wilt mainly damages the root of the tobacco, the most obvious symptom is withering, the withering speed is high, once the tobacco wilt is caused, the whole plant can die, and the tobacco bacterial wilt is a big destructive disease on the tobacco. Tobacco bacterial wilt is the most harmful tobacco bacterial disease in tobacco areas in China, but the disease does not occur in Jilin and Heilongjiang, is commonly generated in Yangtze river basin and southern tobacco areas in China at present, and has a trend towards the north in recent years, and tobacco areas in Shandong, Henan and Liaoning parts also occur, and are seriously harmful in local areas.
Tobacco black shank is a devastating soil-borne disease caused by phytophthora nicotianae (p. parasitica), has high incidence rate and wide distribution range, and causes huge economic loss. In 1950, the disease is found for the first time in Huang-Huai-Yan district in China, and except for sporadic occurrence in the northeast Yan district, all the Yan districts in China occur at present, and the disease is seriously harmful to the production of tobacco. In recent years, as the area of continuous cropping tobacco fields in China is enlarged year by year, the continuous cropping years are increased continuously, and the prevalence of tobacco black shank is increased.
The Alternaria alternate is a leaf spot disease of tobacco leaves in the mature period caused by Alternaria fungus Alternanata, and occurs in each tobacco area in China. In recent years, local tobacco fields in Huang-Huai and northeast tobacco areas become one of destructive diseases, which not only causes incomplete tobacco leaves and grade reduction, but also causes bad taste due to inconsistent internal quality, thereby reducing industrial use value.
At present, a great deal of research work on the prevention and control of tobacco bacterial wilt, blackleg and brown spot, including breeding for disease resistance, agricultural prevention and control, chemical prevention and biological prevention and control, but as the ecological prevention and control microorganisms are one of the most potential means in the green prevention and control of crops, the microorganisms having biological control activity on solanaceae ralstonia mainly come from the endophytic environment of rhizosphere soil and roots of host plants, including Pseudomonas (Pseudomonas), Bacillus (Bacillus), Brevibacillus (Brevibacillus), Paenibacillus (Paenibacillus), Geobacillus (Geobacillus), lysinibacillus (L ysinibacillus), Stenotrophomonas (stenotrophosphas), Acinetobacter (Acinetobacter), Aspergillus (Aspergillus), and the like, and the bacterial strain has a high antagonistic effect on the control of rhizosphere soil diseases, and the bacterial strain of Aspergillus (Aspergillus), and the bacterial strain has been obtained from the domestic and the acidic control of the environment, and the bacterial strain has a high antagonistic effect on the control of the diseases of the Pseudomonas, particularly the acidic soil.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide an acid-resistant disease-preventing growth-promoting biocontrol pseudomonad strain, which is suitable for biological control of tobacco bacterial and fungal diseases under the condition of acid soil and provides a new microbial resource for control of main tobacco root diseases such as bacterial wilt, black shank, brown spot and the like.
In order to achieve the purpose, the invention relates to the following technical scheme:
the invention provides a first aspect of acid-resistant disease-prevention growth-promoting pseudomonas defensins (pseudomonas proteins) C L P-6, which is preserved in China general microbiological culture Collection center (address: No. 3 of No.1 West Chen-Lu of the sunward area, Beijing, China) in 2016 at 10 months and 27 days in 2016, and the biological preservation number of the pseudomonas defensins is CGMCC No. 13205.
In a second aspect of the present invention, there is provided a method for culturing Pseudomonas defensins (Pseudomonas proteins) C L P-6 as described above, comprising culturing Pseudomonas defensins (Pseudomonas proteins) C L P-6 in NB liquid medium under aerobic conditions at 30 ℃ such as shaking culture in a shaker (30 ℃, 150 rpm).
Wherein the formula (g/L) of the NB culture medium comprises beef extract 3.0, peptone 5.0, glucose 2.5, pH7.0 + -0.2;
in a third aspect of the present invention, there is provided a microbial agent comprising a culture of the above-mentioned Pseudomonas defensins (Pseudomonas proteins) C L P-6 or Pseudomonas defensins (Pseudomonas proteins) C L P-6, said microbial agent having the effects of inhibiting tobacco Heptosphaera, Ralstonia solanacearum and Alternaria alternate and promoting tobacco growth.
Preferably, the microbial agent is in the form of wettable powder, water dispersible granules, water suspending agent or dispersible oil suspending agent;
preferably, the microbial agent further comprises an agriculturally and pharmaceutically acceptable auxiliary material, and the agriculturally and pharmaceutically acceptable auxiliary material is selected from one or more of a dispersing agent, a wetting agent, a disintegrating agent, a binder, a defoaming agent, an antifreezing agent, a thickening agent, a filler and a solvent. The invention has no special limitation on the sources of the auxiliary materials acceptable in the agricultural pharmacy, and the like, and generally adopts the products sold in the market.
According to a fourth aspect of the invention, a microbial fertilizer is provided, wherein the microbial fertilizer comprises a culture of the above-mentioned Pseudomonas defensins (Pseudomonas proteins) C L P-6 or Pseudomonas defensins (Pseudomonas proteins) C L P-6.
Preferably, the microbial fertilizer also contains organic matters, total potassium and total nitrogen for providing nutrients.
In a fifth aspect of the invention, the invention provides the application of the Pseudomonas defensins (Pseudomonas proteins) C L P-6, the microbial agent or the microbial fertilizer in the prevention and control of tobacco bacteria and fungal diseases.
Preferably, the tobacco bacterial and fungal diseases include, but are not limited to, tobacco bacterial wilt, tobacco black shank and tobacco brown spot.
In a sixth aspect of the invention, the application of the Pseudomonas defensins (Pseudomonas proteins) C L P-6 and the microbial agent or the microbial fertilizer in promoting the growth of tobacco is provided.
Preferably, the strain, microbial agent or microbial fertilizer is more suitable for functioning in acidic conditions;
further, the acidic condition has a pH of 5.5 to 6.5.
The invention has the beneficial effects that:
the invention screens out a defending pseudomonads (Pseudomonas proteins) C L P-6 from the tobacco healthy plant rhizosphere soil sample of the field where the tobacco bacterial wilt and black shank are mixed by an indoor flat plate opponent method and a greenhouse pot control effect for the first time, the strain has acid resistance, good disease prevention and growth promotion effects and strong stress resistance, has pathogenic bacterium antagonistic activity, wide acidity tolerance range and wide antibacterial spectrum on soil, and has certain application and development potentials in the control of the tobacco bacterial wilt, black shank and red spot.
Drawings
FIG. 1 is a C L P-6 plate colony and thallus morphology map.
FIG. 2 shows the inhibitory effect of C L P-6 sterilized supernatant on Ralstonia solanacearum, wherein the left is the Oxford cup plate diffusion test and the right is the Ralstonia solanacearum treated with C L P-6.
FIG. 3 is a graph showing the degree of destruction of bacterial cells of Ralstonia solanacearum by C L P-6 under an electron microscope.
FIG. 4 is a plate diagram showing the inhibition effect of C L P-6 degerming supernatant on the growth of phytophthora parasitica, wherein the left side is the malformed phytophthora parasitica of the phytophthora parasitica after C L P-6 treatment, and the right side is the normal phytophthora parasitica of the phytophthora parasitica.
FIG. 5 is an optical microscope photograph showing the inhibiting effect of C L P-6 degerming supernatant on the growth of black shank hyphae, wherein the left side is the deformed hyphae of the black shank hyphae treated by C L P-6, and the right side is the normal hyphae of the black shank fungus.
FIG. 6 shows the antagonistic activity of C L P-6 cells against Alternaria alternata.
FIG. 7 is a light microscope photograph showing the effect of C L P-6 on the growth of red star germ hyphae, wherein the right side is abnormal hyphae of red star germ treated with C L P-6, and the left side is normal hyphae of red star germ.
FIG. 8 is a plot of the phosphorus solubilization capacity of the C L P-6 sterilized supernatant.
FIG. 9 is a potassium-releasing map of the C L P-6 sterilized supernatant.
FIG. 10 is a graph of protease producing capacity (under acidic conditions) of C L P-6 sterilized supernatant.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
The present invention is further illustrated by reference to specific examples, which are intended to be illustrative only and not limiting. If the experimental conditions not specified in the examples are specified, the conditions are generally as usual or as recommended by the reagents company; reagents, consumables and the like used in the following examples are commercially available unless otherwise specified.
In one embodiment of the invention, an acid-resistant disease-preventing growth-promoting Pseudomonas defensins (Pseudomonas protegens) C L P-6 is provided, which is preserved in the China general microbiological culture Collection center (address: No. 3, West Lu No.1, North Cheng, Yang-ward, Beijing, China) in 2016 at 10 months and 27 days of 2016, and has a biological preservation number of CGMCC No. 13205.
In the invention, the strain is finally determined to belong to the Pseudomonas defensins (Pseudomonas proteins) by determining the 16S rDNA gene sequence (shown in SEQ ID NO.1) and the gyrB gene sequence (shown in SEQ ID NO.2) of the strain, combining the morphological characteristics (shown in FIG. 1) and the physiological and biochemical characteristics (shown in the following table) of colonies and thalli.
Physiological and biochemical characteristics of strain C L P-6
Note: positive, Negative, W: weak Positive
In still another embodiment of the present invention, there is provided the method for culturing Pseudomonas defensins (Pseudomonas proteins) C L P-6 as described above, comprising culturing Pseudomonas defensins (Pseudomonas proteins) C L P-6 in NB liquid medium under aerobic conditions at 30 ℃ such as shaking culture in a shaker (30 ℃, 150 rpm).
Wherein the formula (g/L) of the NB culture medium comprises beef extract 3.0, peptone 5.0, glucose 2.5, pH7.0 + -0.2;
in still another embodiment of the present invention, there is provided a microbial agent comprising a culture of Pseudomonas defensins (Pseudomonas proteins) C L P-6 or Pseudomonas defensins (Pseudomonas proteins) C L P-6 as described above, which has the effects of inhibiting Alternaria alternata, Leptosphaeria maculans and Fusarium oxysporum and promoting tobacco growth.
In another specific embodiment of the invention, the formulation of the microbial agent is wettable powder, water dispersible granules, water suspending agent or dispersible oil suspending agent;
preferably, the microbial agent further comprises an agriculturally and pharmaceutically acceptable auxiliary material, and the agriculturally and pharmaceutically acceptable auxiliary material is selected from one or more of a dispersing agent, a wetting agent, a disintegrating agent, a binder, a defoaming agent, an antifreezing agent, a thickening agent, a filler and a solvent. The invention has no special limitation on the sources of the auxiliary materials acceptable in the agricultural pharmacy, and the like, and generally adopts the products sold in the market.
Wherein the dispersant is an anionic dispersant and/or a nonionic dispersant, and can be selected from one or more of sodium lignosulfonate, sodium naphthalene sulfonate formaldehyde condensate, sodium methylene dinaphthalene sulfonate, formaldehyde condensate sulfate, polycarboxylate, alkylphenol polyoxyethylene phosphate and fatty acid polyoxyethylene ester;
the humectant can be one or more selected from sodium dodecyl sulfate, sodium dodecyl benzene sulfonate, fructus Gleditsiae Abnormalis powder, fructus Sapindi Mukouossi powder, tea seed cake powder and nekal BX;
the disintegrant can be one or more selected from bentonite, ammonium sulfate, aluminum chloride, urea, magnesium chloride and glucose;
the binder may be selected from one or more of starch, diatomaceous earth, cyclodextrin, rosin, carboxymethyl cellulose, carboxyethyl cellulose, and carboxymethyl cellulose salts;
the defoaming agent can be one or more of C8-C20 fatty alcohol compounds, C10-C20 saturated fatty acid compounds, epoxidized soybean oil, ethanol, silicone compounds and organic silicone oil;
the antifreeze agent can be selected from one or more of sorbitol, ethylene glycol, polyethylene glycol, propylene glycol, glycerol, urea and sodium chloride;
the thickening agent can be selected from one or more of gelatin, xanthan gum, polyethylene glycol and polyvinyl alcohol;
the filler can be one or more of light calcium carbonate, diatomite, bentonite, attapulgite and white carbon black;
the solvent may be selected from water (preferably deionized water) or methyl oleate;
in another embodiment of the present invention, there is provided a microbial fertilizer, wherein the microbial fertilizer comprises a culture of the above-mentioned Pseudomonas defensins (Pseudomonas proteins) C L P-6 or Pseudomonas defensins (Pseudomonas proteins) C L P-6.
In another embodiment of the present invention, the microbial fertilizer further contains organic matter, total potassium and total nitrogen for providing nutrients.
In another embodiment of the invention, the application of the pseudomonas defensins (pseudomonas proteins) C L P-6, the microbial agent or the microbial fertilizer in the prevention and treatment of tobacco bacteria and fungal diseases is provided.
In yet another embodiment of the present invention, the tobacco bacterial and fungal diseases include, but are not limited to, tobacco bacterial wilt, tobacco black shank and tobacco brown spot.
In another embodiment of the present invention, there is provided the use of Pseudomonas defensins C L P-6, a microbial agent or a microbial fertilizer as described above for promoting the growth of tobacco.
In yet another embodiment of the invention, the strain, microbial agent or microbial fertilizer is more suitable for functioning in acidic conditions;
in yet another embodiment of the present invention, the acidic condition has a pH of 5.5 to 6.5.
The invention is further illustrated by the following examples, which are not to be construed as limiting the invention thereto.
Example 1C L P-6 and its fermentation broth for the antagonism of tobacco bacterial wilt and phytophthora parasitica antagonism activity of its fermentation broth: antagonism activity of puerto tobacco plants in Cili county, Zhang Nanjing, NA medium was mixed with appropriate amount of acidic buffer (0.2 mol/L sodium acetate and 0.3 mol/L acetic acid in a volume ratio of 1: 9) to prepare NA medium with pH of 5.5, 6.0, 6.5, 7.0 and 7.5 respectively, pH difference of bacteriostatic activity of C L P-6 strain fermentation broth under different acid conditions was determined by oxford cup method, C L P-6 was inoculated on NA plate for 48h at 30 ℃, then plate stand-off method was used, i.e. black shin cake (diameter of 5mm) and red spot are inoculated on oat and potato dextrose broth (plate) plate, C L P-6 was inoculated on agar plate, pH of agar plate for 30 ℃ for culturing for 48h, C L P-6 was inoculated on beef broth, pH of agar plate was used for culturing, after observation of bacterial cake (diameter of bacterial wilt, pH of the bacterial cake) and pH of the supernatant, the supernatant was inoculated on agar plate, the supernatant of agar, the supernatant of the supernatant, the supernatant was taken, and the supernatant was taken after observation, the supernatant was taken, and was added to obtain the supernatant was added, and the supernatant was added.
As can be seen from tables 1 and 2, in the range of acidity (pH is more than or equal to 5.5 and less than or equal to 6.5), the C L P-6 has strong inhibition effect on bacterial wilt and phytophthora parasitica and is higher than neutral or alkaline conditions, which indicates that the acidic conditions are favorable for the bacterial strain to exert the optimal inhibition effect, so that the bacterial strain has great potential when being applied to biological control of bacterial wilt and phytophthora parasitica under the acidic soil conditions.
TABLE 1 bacteriostatic effect of acid-resistant antagonistic strain C L P-6 on Ralstonia solanacearum
Note: the letters after the same column of data represent significant differences (p.ltoreq.0.01).
TABLE 2 comparison of antagonistic Activity of acid resistant antagonistic Strain C L P-6 Strain in acidic and neutral conditions
Note: the letters after the same column of data represent significant differences (p.ltoreq.0.05).
Example 2C L P-6 and its fermentation broth are used to destroy Ralstonia solanacearum thallus by inoculating C L P-6 on NA plate at 30 deg.C for 48h, then using plate opposition method to inhibit Ralstonia solanacearum by Oxfordii cup plate diffusion method, inoculating C L P-6 in beef extract peptone liquid culture solution (NB), shaking at 30 deg.C and 150rpm for 2 days, taking appropriate amount of fermentation broth and sterile supernatant (filtered by bacterial filter), placing in Oxfordii cup on beef extract peptone medium plate (NA) containing Ralstonia solanacearum, standing for 2 days, observing the inhibition of C L P-6 on the above pathogenic bacteria, observing the inhibition effect of C L P-6 thallus, fermentation broth and sterile supernatant on Ralstonia solanacearum from indoor plate (see FIG. 2).
The antagonistic activity of antibacterial substances secreted by C L P-6 strain on the destructive action of bacterial body of Ralstonia solanacearum is obtained by inoculating activated C L P-6 into NB culture medium, performing shake culture at 30 deg.C for 48 hr to obtain fermentation liquor, collecting the supernatant after removing bacterial bodyMixing the clear solution with a suspension of Ralstonia solanacearum (concentration 2 × 10)8cfu/ml), the blank control is equal amount of sterile water and bacterial suspension for mixing treatment, the treatment is carried out for 24 hours, the diluted mixed solution after each treatment is observed under a scanning electron microscope, the result shows that bacteriostatic substances secreted by the C L P-6 strain have stronger destructive effect on the ralstonia solanacearum thallus, so that the rod-shaped thallus is deformed, the content is leaked, and the ralstonia solanacearum rod-shaped thallus of the sterile water control is normal (see figure 3).
Example 3C L P-6 and its fermentation broth inhibiting the growth of Digitalis retziae hypha by inoculating C L P-6 to NA plate at 30 deg.C for 48h, inoculating Digitalis retziae to oat plate, picking C L P-6 with inoculating loop, drawing line with the bacterial cake as center at equal distance, culturing at 30 deg.C for 5d, picking edge hypha of Digitalis cake under microscope, the result shows that the edge hypha of Digitalis cake treated by C L P-6 has spherical cytoplasm in hypha cell aggregation and breaks away from cell membrane and cell wall, thus affecting its normal growth (see FIG. 4).
Example 4 inhibition of Alternaria alternata and its effect on hyphal growth by the thallus of C L P-6 Strain:
the red star disease bacteria are used as target bacteria, namely C L P-6 strains are inoculated on an NA plate and cultured for 48 hours at the temperature of 30 ℃, then a plate opposition method is adopted, namely the red star disease bacteria are inoculated on a PDA plate, C L P-6 strains are picked by an inoculating loop and are scribed in parallel at equal intervals on two sides with a bacterial cake as the center, the red star disease bacteria are cultured for 5 days at the temperature of 30 ℃, then hyphae at the edge of the red star disease bacterial cake are picked and are examined under a microscope, the result shows that the growth of the red star disease bacterial colony treated by C L P-6 is inhibited, the growth of the red star disease bacterial colony adjacent to the C L P-6 strains is slow, the bacterial colony is in long and narrow development (figure 6), and compared with the normal hyphae, the inhibition effect of the hyphae treated by the C L P-6 red star disease bacterial hyphae is shown as malformation, the deformation and swelling of hyphae among divisions, the dispersion or aggregation of the inclusions, the growth is not vigorous, and the growth is.
Example 5C L P-6 and its fermentation broth have phosphorus-dissolving effect, acid-resistant antagonistic strain culture broth is measured by molybdenum-antimony colorimetry to determine its water-soluble phosphorus content, determine its phosphorus-dissolving capacity, the acid-resistant antagonistic strain standard bacterial liquid is inoculated to the phosphorus-dissolving culture medium with pH of 5.5 and 7.0 respectively according to 1%, shaking culture is carried out at 28 deg.C and 170r/min for 5d, the cultured fermentation broth is put into a centrifuge at 4000r/min, centrifuged for 20min, the supernatant is poured out and fixed to 50m L, the precipitate is ground in a mortar with 1g of quartz sand for 10min and washed down with distilled water, the ground liquid is centrifuged in a centrifuge at 4000r/min for two times, the precipitate is discarded, the supernatant is similarly fixed to 50m L, the two centrifuged supernatants are combined together, the test solution is measured for the water-soluble phosphorus in the strain fermentation broth, and the phosphorus-dissolving capacity is determined, each treatment is repeated for 3 times.
5ml of the supernatant was taken and 0.5 mol/L NaHCO was added3Adding distilled water 35m L into the solution 10m L, sucking the molybdenum antimony color-developing reagent 5.0m L with a pipette, shaking up, standing at room temperature for 30min, performing color comparison at 720nm wavelength, and adjusting OD of the non-inoculated phosphorus-dissolving culture medium720And (5) determining the absorbance of the supernatant of the strain to be detected, and comparing the absorbance with a standard curve to obtain the content of soluble phosphorus in the supernatant of the strain.
And (3) selecting activated C L P-6 from an inoculating loop, inoculating the inoculating loop on a phosphorus dissolving culture medium, culturing at 30 ℃ for 48h, and observing whether a halo is generated around a C L P-6 colony or not, wherein the halo is generated if the halo is generated, so that the result is positive, the result shows that a halo with a certain width is generated around a C L P-6 colony (as shown in figure 8), and the strain has a certain phosphorus dissolving capacity.
As shown in FIG. 5 and Table 3, the difference in the water-soluble phosphorus content in the C L P-6 culture solution under acidic-neutral conditions is significant, and the water-soluble phosphorus content is 46.2 mg/L under acidic conditions of pH5.5 and 21.1 mg/L under neutral conditions of pH7.0, so that the phosphorus-dissolving capacity of C L P-6 under acidic conditions (pH 5.5) is stronger than that under neutral conditions (pH 7.0).
TABLE 3 content of water-soluble phosphorus in acid-neutral medium by acid-resistant antagonistic strain C L P-6
Note: the letters after the same column of data represent significant differences (p.ltoreq.0.05).
Example 6C L P-6 and its fermentation broth have potassium-solubilizing effect, using sodium tetraphenylborate method to measure the water-soluble potassium content in the acid-resistant antagonistic strain culture broth, determining the potassium-solubilizing ability of the strain, inoculating 1% of the acid-resistant antagonistic bacteria standard bacterial liquid into potassium-solubilizing culture medium with pH of 5.5 and 7.0, shaking at 28 ℃ and 170r/min for 5d, placing the cultured fermentation broth into a centrifuge at 4000r/min, centrifuging for 20min, pouring out the supernatant and fixing the volume to 50m L, adding 1g of quartz sand into the precipitate, grinding for 10min in a mortar, washing with distilled water, placing the grinding liquid into a centrifuge at 4000r/min, centrifuging for 20min twice, discarding the precipitate, fixing the supernatant at the same volume of 50m L, combining the two centrifuged supernatants together, measuring the water-soluble potassium in the strain fermentation broth, determining the potassium-solubilizing ability, and repeating the steps 3 times.
Adding 5.0m L of the centrifugal solution into a colorimetric tube, adding 1m L formaldehyde-EDTA masking agent, shaking up, sucking 1.0m L of sodium tetraphenylborate solution by a pipette, adding into the colorimetric tube, shaking up, standing at room temperature for 15min, shaking up again, carrying out colorimetric analysis under the condition of 420nm wavelength, and adjusting OD of a potassium-dissolving culture medium without inoculation420And (5) determining the absorbance of the supernatant of the strain to be detected, and comparing the absorbance with a standard curve to obtain the content of the water-soluble potassium in the supernatant of the strain.
The activated C L P-6 point is selected from an inoculating loop and inoculated on a potassium-releasing culture medium, the inoculating loop is cultured for 48 hours at the temperature of 30 ℃, whether a halo is generated around a C L P-6 bacterial colony or not is observed, if the halo is generated, the halo is positive, the result shows that (shown in figure 10), a halo with a certain width is generated around a C L P-6 bacterial colony, the bacterial strain has certain potassium-releasing capacity, the potassium-releasing bacteria can convert ineffective potassium in soil into effective potassium and increase potassium elements in the soil, the tobacco is a potassium-favored crop, and if the C L P-6 is applied to the tobacco, sufficient potassium nutrition is provided for the tobacco plant, so that the inoculating loop has a great application value in the aspect of improving the quality and the yield of the tobacco.
As shown in FIG. 9 and Table 4, the acid-resistant antagonism of the content of water-soluble potassium in the culture solution of C L P-6 under the acidic-neutral condition is not significant, the content of water-soluble potassium in the culture solution is 3.2 mg/L under the acidic condition of pH5.5, and the content of water-soluble potassium in the culture solution is 2.9 mg/L under the neutral condition of pH7.0, so that the potassium-dissolving capacity of C L P-6 under the acidic condition (pH 5.5) is equivalent to that under the neutral condition (pH 7.0).
TABLE 4C L P-6 Water-soluble Potassium content in acidic-neutral Medium
Note: the letters after the same column of data represent significant differences (p.ltoreq.0.05).
Example 7C L P-6 and its fermentation broth have protease secretion ability by inoculating standard bacterial suspension into NB with pH of 5.5 and 7.0 at 1% under aseptic condition, repeating each treatment for 3 times, centrifuging at 28 deg.C and 170r/min for 3d, 10000r/min for 10min, collecting 1m L supernatant as enzyme solution, placing in 3 tubes, preheating in 37 deg.C water bath for 2min, adding 1m L preheated casein solution into tubes, mixing at 37 deg.C for 30min, adding 2ml 0.4 mol/L trichloroacetic acid solution into tubes, mixing at 37 deg.C for 30min, centrifuging at 10000g for 10min, collecting 1m L supernatant, adding 5m L0.4.4 mol/L Na into tubes2CO3Mixing the solution and 1m L Folin phenol reagent, developing in 40 deg.C water bath for 20min, and simultaneously setting blank control, i.e. adding trichloroacetic acid solution into enzyme solution, then adding casein solution, and measuring absorbance at 680nm wavelength after color development.
As shown in FIG. 10 and Table 5, the results of the experiments revealed that the bacterial enzyme activity of C L P-6 in neutral-acidic medium, C L P-6 in acidic medium with pH5.5, was 49.89U/m L, and the bacterial enzyme activity of C L P-6 in neutral medium with pH7.0 was 38.15U/m L, so that the acidic conditions were suitable for the secretion of protease from C L P-6.
TABLE 5 acid-resistant antagonistic strain C L P-6 enzyme-producing ability (acid condition pH5.5)
Note: the letters after the same column of data represent significant differences (p.ltoreq.0.05).
Example 8C L P-6 and its fermentation broth have phosphatase secretion ability the phosphatase activity determination is based on phosphomolybdic blue colorimetry, a standard bacterial suspension is inoculated in Pikovskaya liquid medium with pH5.5 and 7.0 according to 1% under aseptic condition, each treatment is repeated for 3 times, the solution is oscillated at 28 ℃ and 170r/min for 3d, centrifuged at 10000r/min for 10min, 1m L supernatant is diluted 10 times with distilled water, the diluted fermentation broth is accurately sucked, a blank Pikovskaya liquid medium which is not inoculated after sterilization is used as a control, the absorbance value is determined, and the phosphorus content in the sample solution is checked on a standard curve.
The test results are shown in tables 5 and 6, the difference of the C L P-6 phosphatase activity in the acidic-neutral culture solution is obvious, the C L P-6 phosphatase activity is 5.12U/m L under the acidic condition of pH5.5, and the C L P-6 phosphatase activity is 8.27U/m L under the neutral condition of pH7.0, which indicates that the phosphatase producing capability of C L P-6 is lower than that of the neutral condition (pH7.0) under the weakly acidic condition (pH 5.5).
TABLE 6 acid-resistant antagonistic Strain C L P-6 phosphatase Activity in acid-neutral Medium
Note: the letters after the same column of data represent significant differences (p.ltoreq.0.01).
Example 9C L P-6 and the growth-promoting effect of fermentation broth on tobacco plants, the experiment was conducted in a nutrient tray planting mode in natural soil and the seedlings were treated by root soaking by preparing a substrate from field soil, farmyard manure and vermiculite in a ratio of 7:2:1, filling natural soil in a nursery tray, carefully digging out 3-4 tobacco seedlings with true leaves, gently shaking off the soil attached to the roots, and soaking in a pre-selected standard biocontrol bacteria suspension (concentration about 10) to be tested8cfu/m L), taking out after 40min, transplanting into a seedling tray, filling roots with standard bacterial suspension for 1 time at an interval of 7d, 5m L/plant, treating 10 plants with clear water as a control, repeating for three times every time, performing moisture preservation culture at 30 ℃ in a greenhouse, treating 20d after the last time, randomly selecting 10 tobacco seedlings, carefully digging out the whole seedlings, washing away soil at the root, measuring the plant height, the fresh weight of the whole plant, the fresh weight of the root and other indexes, drying to constant weight at 180 ℃, measuring the dry weight and the dry weight of the root, and promoting growth, wherein the dry weight and the dry weight of the root are measuredTest results (see tables 7 and 8) show that when tobacco seedlings planted in natural soil are inoculated with C L P-6, tobacco growth can be obviously promoted, the fresh weight and dry weight of the whole tobacco seedlings are obviously increased, the leaves are dark green, the leaves are thick and large, in the natural bacterial soil, all indexes of the tobacco seedlings processed by C L P-6 are higher than those of a control, compared with CK, the strain processed by C L P-6 can effectively promote the tobacco growth, the plant height, the fresh weight, the dry weight and root dry weight of the whole tobacco seedlings and the growth difference of the root dry weight are obvious, the tobacco plants processed by C L P-6 have obvious growth promoting effects on the plant height, the fresh weight, the dry weight and the root dry weight of the whole tobacco seedlings, the growth promoting effects are respectively increased by 31.2%, 40.0%, 70.6% and 45.2%, and the growth promoting effect of the strain is extremely strong, and the C L P-6 is safe for the tobacco.
TABLE 7 growth promotion of acid-resistant antagonistic strain C L P-6 in soil at pH5.5
Note: the letters after the same column of data represent significant differences (p.ltoreq.0.05).
TABLE 8 acid-tolerant antagonistic strain C L P-6 in soil at normal pH (7.1) growth promoting action
Note: the letters after the same column of data represent significant differences (p.ltoreq.0.05).
In conclusion, the strain of the Pseudomonas defensins (Pseudomonas proteins) C L P-6 enriches the types and application range of the Pseudomonas defensins, shows strong growth capability and stable activity in different culture media, and has further research and development values.
According to the invention, a strain of Pseudomonas defensins (C L P-6) is screened out for the first time through indoor plate culture and living greenhouse control, the strain is separated from acidic tobacco root soil, has antagonistic activity on pathogenic strains of tobacco bacterial wilt, black shank and red star disease, and has the capacities of acid resistance, phosphorus dissolution, potassium dissolution, protease secretion, phosphatase and the like, and the C L P-6 and the microbial agent thereof can effectively prevent and control tobacco soil-borne diseases under continuous cropping or acidic soil conditions, are novel high-quality microbial pesticide resources, and have wide application prospects.
It should be noted that the above examples are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the examples given, those skilled in the art can modify the technical solution of the present invention as needed or equivalent substitutions without departing from the spirit and scope of the technical solution of the present invention.
SEQUENCE LISTING
<110> tobacco institute of Chinese academy of agricultural sciences
<120> acid-resistant pseudomonas defensins C L P-6 and application thereof
<130>
<160>1
<170>PatentIn version 3.5
<210>1
<211>1381
<212>DNA
<213> Pseudomonas protegens 16s rDNA
<400>1
cggcagcacg ggtacttgta cctggtggcg agcggcggac gggtgagtaa tgcctaggaa 60
tctgcctagt agtgggggat aacgtccgga aacgggcgct aataccgcat acgtcctacg 120
ggagaaagtg ggggatcttc ggacctcacg ctattagatg agcctaggtc ggattagcta 180
gttggtgagg taatggctca ccaaggcgac gatccgtaac tggtctgaga ggatgatcag 240
tcacactgga actgagacac ggtccagact cctacgggag gcagcagtgg ggaatattgg 300
acaatgggcg aaagcctgat ccagccatgc cgcgtgtgtg aagaaggtct tcggattgta 360
aagcacttta agttgggagg aagggcagtt acctaatacg tgattgtttt gacgttaccg 420
acagaataag caccggctaa ctctgtgcca gcagccgcgg taatacagag ggtgcaagcg 480
ttaatcggaa ttactgggcg taaagcgcgc gtaggtggtt tgttaagttg gatgtgaaag 540
ccccgggctc aacctgggaa ctgcatccaa aactggcaag ctagagtatg gtagagggtg 600
gtggaatttc ctgtgtagcg gtgaaatgcg tagatatagg aaggaacacc agtggcgaag 660
gcgaccacct ggactgatac tgacactgag gtgcgaaagc gtggggagca aacaggatta 720
gataccctgg tagtccacgc cgtaaacgat gtcaactagc cgttgggagc cttgagctct 780
tagtggcgca gctaacgcat taagttgacc gcctggggag tacggccgca aggttaaaac 840
tcaaatgaat tgacgggggc ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac 900
gcgaagaacc ttaccaggcc ttgacatcca atgaactttc tagagataga ttggtgcctt 960
cgggaacatt gagacaggtg ctgcatggct gtcgtcagct cgtgtcgtga gatgttgggt 1020
taagtcccgt aacgagcgca acccttgtcc ttagttacca gcacgttatg gtgggcactc 1080
taaggagact gccggtgaca aaccggagga aggtggggat gacgtcaagt catcatggcc 1140
cttacggcct gggctacaca cgtgctacaa tggtcggtac aaagggttgc caagccgcga 1200
ggtggagcta atcccataaa accgatcgta gtccggatcg cagtctgcaa ctcgactgcg 1260
tgaagtcgga atcgctagta atcgcgaatc agaatgtcgc ggtgaatacg ttcccgggcc 1320
ttgtacacac cgcccgtcac accatgggag tgggttgcac cagaagtagc tagtctaacc 1380
t 1381
<210>2
<211>652
<212>DNA
<213> Pseudomonas protegens gyrB
<400>2
ggtagtgaac gccctgtccg aggagctgat cctcaccgtg cgccgtagcg gcaagatctg 60
ggaacagacc tatgtccacg gtgttccgca agagcggatg aaaatcgttg gtgacagcga 120
aaccaccggt acccagatcc acttcaagcc ttcggctgaa accttcaaga acatccactt 180
cagctgggac gtcctggcca agcggatccg tgaactgtcc ttcctcaact ccggtgtcgg 240
catcgtcctc aaggacgagc gcagcggcaa ggaagagctg ttcaagtacg aaggcggcct 300
gcgggcattc gttgaatacc tgaacaccaa caagactgcg gtcaaccagg tgttccactt 360
caacatccag cgtgaagacg gcatcggcgt ggaaatcgcc ttgcagtgga acgacagctt 420
caacgagaac ctgttgtgct tcaccaacaa cattcctcag cgcgacggcg gtacccacct 480
ggtgggcttc cgttcggccc tgacccgtaa cctgaacaac tacatcgagc aggaaggcct 540
ggccaagaaa cacaaggtcg ccaccactgg tgacgacgct cgtgaaggcc tgaccgcgat 600
catctcggtg aaggtgccgg atccgaagtt cagctcccag accaaggaca ag 652
Claims (11)
1. An acid-resistant disease-preventing growth-promoting Pseudomonas defensins (Pseudomonas protegens) C L P-6, which is preserved in China general microbiological culture Collection center (CGMCC) at 10-27 th in 2016, and the biological preservation number is CGMCC No. 13205.
2. The method for culturing Pseudomonas defensive (Pseudomonas proteins) C L P-6 as claimed in claim 1, wherein the Pseudomonas defensive (Pseudomonas proteins) C L P-6 is cultured in NB liquid medium.
3. The method for culturing Pseudomonas defensins (Pseudomonas proteins) C L P-6 as claimed in claim 2, wherein the culture condition is aerobic culture at 30 ℃.
4. A microbial agent comprising a culture of Pseudomonas defensive (Pseudomonas proteins) C L P-6 of claim 1 or Pseudomonas defensive (Pseudomonas proteins) C L P-6 of claim 1.
5. The microbial agent according to claim 4, wherein the microbial agent is in the form of wettable powder, water dispersible granule, water suspension or dispersible oil suspension.
6. A microbial fertilizer, comprising a culture of Pseudomonas defensive (Pseudomonas proteins) C L P-6 of claim 1 or Pseudomonas defensive (Pseudomonas proteins) C L P-6 of claim 1.
7. The microbial fertilizer according to claim 6, wherein the microbial fertilizer further comprises organic matter, total potassium and total nitrogen.
8. Use of the pseudomonads defensins (Pseudomonas proteins) C L P-6 of claim 1, the microbial agent of claim 4 or 5, the microbial fertilizer of claim 6 or 7 for the control of tobacco bacterial wilt, tobacco black shank and tobacco brown spot.
9. Use of the pseudomonads defensins (Pseudomonas proteins) C L P-6 of claim 1, the microbial agent of claim 4 or 5, the microbial fertilizer of claim 6 or 7 for promoting the growth of tobacco.
10. Use according to claim 8 or 9, wherein the application environment is an acidic condition.
11. The use according to claim 10, wherein the acidic conditions have a pH of 5.5 to 6.5.
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| CN109182213B (en) * | 2018-10-16 | 2021-07-09 | 云南省烟草公司普洱市公司 | Sphingomonas X1-8 in tobacco planting soil and application thereof in prevention and treatment of tobacco black shank |
| CN110643538B (en) * | 2019-10-15 | 2021-04-27 | 浙江大学 | Pseudomonas fluorescens XY2F4 and application thereof in preventing and treating crop verticillium wilt |
| CN111778183B (en) * | 2020-06-30 | 2021-11-30 | 中国农业科学院烟草研究所 | Acidophilic nitrogen-producing pseudomonas strain and application thereof |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1347981A (en) * | 2000-10-11 | 2002-05-08 | 常州兰陵制药有限公司 | Pseudomonas stutzeri Zh9944 and its screening method and bacterial wilt preventing and controlling microbial prepn |
| CN105543133A (en) * | 2015-12-31 | 2016-05-04 | 山东农业大学 | Separation and application of pseudomonas with significant antagonism to plant pathogenic bacteria |
| CN106916764A (en) * | 2017-02-15 | 2017-07-04 | 中国农业科学院烟草研究所 | One plant of acid proof South Korea pseudomonad CLP 7 and its application |
| CN106978373A (en) * | 2017-05-09 | 2017-07-25 | 福建农林大学 | One plant is used for pseudomonad and the application that bacteriosis is prevented and treated |
-
2018
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1347981A (en) * | 2000-10-11 | 2002-05-08 | 常州兰陵制药有限公司 | Pseudomonas stutzeri Zh9944 and its screening method and bacterial wilt preventing and controlling microbial prepn |
| CN105543133A (en) * | 2015-12-31 | 2016-05-04 | 山东农业大学 | Separation and application of pseudomonas with significant antagonism to plant pathogenic bacteria |
| CN106916764A (en) * | 2017-02-15 | 2017-07-04 | 中国农业科学院烟草研究所 | One plant of acid proof South Korea pseudomonad CLP 7 and its application |
| CN106978373A (en) * | 2017-05-09 | 2017-07-25 | 福建农林大学 | One plant is used for pseudomonad and the application that bacteriosis is prevented and treated |
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