CN116649374A - Pseudomonas aeruginosa QSE-TM7 and application thereof in preparation of biological agent for preventing and treating tomato gray mold - Google Patents
Pseudomonas aeruginosa QSE-TM7 and application thereof in preparation of biological agent for preventing and treating tomato gray mold Download PDFInfo
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Abstract
The invention discloses pseudomonas aeruginosa QSE-TM7 and application thereof in preparation of biological agents for preventing and treating tomato gray mold. The pseudomonas aeruginosa QSE-TM7 is a plant endophyte separated and screened from tomato roots and is classified asPseudomonas aeruginosaThe strain is preserved in China Center for Type Culture Collection (CCTCC) M20231202. The bacterial colony of the pseudomonas aeruginosa QSE-TM7 is round, opaque, smooth in surface and free of wrinkles; the bacterial cells are short rods, are mostly arranged singly, and are red in gram staining. Experiments prove that the pseudomonas aeruginosa QSE-TM7 can obviously inhibit the spore germination, sclerotium germination and hypha growth of the botrytis cinerea, induce the up-regulated expression of plant disease resistance related genes and treat the botrytis cinereaThe occurrence has obvious control effect, can be used for preparing biological agents for controlling the gray mold of tomatoes, and has good market application prospect.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to pseudomonas aeruginosa QSE-TM7 and application thereof in preparation of biological agents for preventing and treating tomato gray mold.
Background
Tomato is widely planted worldwide as an important vegetable and cash crop, and in the planting process, gray mold can infect all organs and tissues of the overground part of the tomato, so that the yield and quality of the tomato are seriously affected; meanwhile, the botrytis cinerea can infect tomato fruits in the links of storage, transportation and retail, and cause great economic loss. In recent years, the cultivation area of tomatoes in a protected area is continuously enlarged, suitable conditions are provided for gray mold generation and epidemic, the diseases spread rapidly, and the disease generation area and severity degree show an ascending trend. At present, the most effective control means for the gray mold of the tomatoes is still chemical control, and the use of chemical pesticides saves huge losses for the tomato planting industry. However, gray mold control becomes extremely difficult due to continuous use of control agents over the years, unreasonable use of chemical pesticides, general multi-drug resistance of gray mold, and the like. In addition, the use of high-toxicity and high-dose traditional chemical pesticides causes serious environmental pollution and ecological balance damage, and pesticide residues affect food safety and endanger human health. Therefore, there is an urgent need for a green control measure in agricultural production to secure and realize the safe, green and sustainable development of agricultural production.
The use of microorganisms (biocontrol bacteria) for controlling plant diseases and insect pests has the advantages of environmental friendliness, biosafety, difficulty in generating drug resistance and the like, and is receiving more and more attention in plant protection application. A large number of microorganisms having biocontrol effect have been isolated from soil, plant rhizosphere and plant tissue, wherein the major group of biocontrol bacteria comprises Bacillus species [ ]Bacillus) Pseudomonas spPseudomonas) Botrytis cinerea (Bulker)Burkholderia) AzotobacterAzotobacter) Agrobacterium(s) aAgrobacterium) And the like, various microorganisms have been developed into biocontrol agents, and are successfully used for controlling plant diseases. Research shows that the biocontrol bacteria can exert biocontrol effect from various aspects, and mainly comprises the following steps: 1) Competing with pathogenic bacteria for nutrients and survival space; 2) Secretion of antibacterial substances inhibits the growth and the virulence of pathogenic bacteria; 3) A re-mailing effect on pathogenic bacteria; 4) Induce plants to develop systemic resistance (induced systemic resistance, ISR). The biological control microorganism with multiple functions is screened from natural environment, and has important application value and potential in the development of biological pesticides and the green control of plant diseases.
Disclosure of Invention
The invention provides pseudomonas aeruginosa QSE-TM7 and application thereof in preparation of biological agents for preventing and treating tomato gray mold. The pseudomonas aeruginosa QSE-TM7 can obviously inhibit the growth of gray mold, induce tomato immune response and has obvious control effect on the occurrence of tomato gray mold.
In order to achieve the aim of the invention, the invention is realized by adopting the following technical scheme:
the invention provides a pseudomonas aeruginosa QSE-TM7 for preventing and treating tomato gray mold, which is classified as pseudomonas aeruginosaPseudomonas aeruginosaThe strain is preserved in China Center for Type Culture Collection (CCTCC) M20231202.
Furthermore, the bacterial colony of the pseudomonas aeruginosa QSE-TM7 is round, opaque, smooth in surface and free of wrinkles; no growth at 4 ℃ and growth at 28 ℃ and 42 ℃; QSE-TM7 cells are short rods, are arranged singly, and are red in gram staining.
Further, the Pseudomonas aeruginosa QSE-TM7 is extracted from a greenhouse tomato plant.
Further, the nucleotide sequence of the 16S rDNA of the pseudomonas aeruginosa QSE-TM7 is shown as SEQ ID No: 1.
Further, the pseudomonas aeruginosa QSE-TM7rpoDThe partial nucleotide sequence of the gene is shown in SEQ ID No: 2.
Furthermore, the invention also provides application of the pseudomonas aeruginosa QSE-TM7 in preparing biological agents for preventing and treating tomato gray mold.
Further, the Pseudomonas aeruginosa QSE-TM7 can inhibit hypha growth, spore germination and sclerotium germination of the Botrytis cinerea.
Further, the concentration of the pseudomonas aeruginosa QSE-TM7 for inhibiting the growth of the mycelium and the germination of spores of the griseomyces is 1-15% (v/v).
Further, the optimal concentration of Pseudomonas aeruginosa QSE-TM7 to inhibit the growth of the mycelial hyphae and spore germination was 15% (v/v).
Further, the concentration of the pseudomonas aeruginosa QSE-TM7 inhibiting the germination of the gray mold and the gray mold sclerotium is 15% (v/v).
Furthermore, the pseudomonas aeruginosa QSE-TM7 can induce the expression of tomato leaf disease resistance related genes.
Further, the genes comprise tomato disease resistance related genes SlPR1a, slPR1b, slPR2 and SlPR5.
Furthermore, the pseudomonas aeruginosa QSE-TM7 has remarkable control effect on tomato gray mold.
Further, when the pseudomonas aeruginosa QSE-TM7 is applied, the pseudomonas aeruginosa QSE-TM7 fermentation broth is prepared.
Furthermore, the pseudomonas aeruginosa QSE-TM7 fermentation broth can be used alone or prepared into biological agents with other materials.
Further, the biological agent comprises pseudomonas aeruginosa QSE-TM7 fermentation broth.
Further, the preparation method of the pseudomonas aeruginosa QSE-TM7 fermentation broth comprises the following steps: inoculating pseudomonas aeruginosa QSE-TM7 into a TM liquid culture medium, shake culturing for 4 days at 25 ℃ and 180rpm, centrifuging the culture solution at 10000rpm, and filtering the supernatant by a 0.45 μm sterile filter to obtain pseudomonas aeruginosa QSE-TM7 fermentation broth without thalli.
Further, the added volume ratio of the pseudomonas aeruginosa QSE-TM7 fermentation broth in the biological preparation is 1-15%.
Further, the use method of the biological agent comprises the following steps: the prepared solution is uniformly sprayed on the whole tomato plant.
Compared with the prior art, the invention has the following advantages and beneficial effects: the pseudomonas aeruginosa QSE-TM7 is extracted from greenhouse tomato plants, has remarkable inhibition effect on the gray mold, can inhibit the growth of gray mold hyphae, and can inhibit the spore germination and sclerotium germination of the gray mold, so that the growth of the tomato gray mold on the plants is inhibited from multiple aspects; in addition, the pseudomonas aeruginosa QSE-TM7 can induce the expression of tomato disease resistance related genes, so that the resistance of tomatoes to gray mold can be enhanced, and therefore, the pseudomonas aeruginosa QSE-TM7 has dual functions in the control of tomato gray mold. The pseudomonas aeruginosa QSE-TM7 is prepared into strain fermentation broth, can be used singly or prepared into biological agents for preventing and treating the gray mold of tomatoes with other materials, and the biological agents do not contain thalli, are simple to use, have good effects, are safe and reliable, have long storage period, and therefore have good market application prospects.
Drawings
FIG. 1 is a colony morphology of the Pseudomonas aeruginosa QSE-TM7 on TM medium.
FIG. 2 shows the growth of Pseudomonas aeruginosa QSE-TM7 under different temperature conditions.
FIG. 3 is a pattern and color under a microscope after gram staining of the Pseudomonas aeruginosa QSE-TM7.
FIG. 4 is a phylogenetic tree of the Pseudomonas aeruginosa QSE-TM7.
FIG. 5 shows that the Pseudomonas aeruginosa QSE-TM7 fermentation broth inhibits the growth of the mycelial of Botrytis cinerea on the PDA plate, the abscissa shows the concentration of the fermentation broth containing the QSE-TM7 strain in the PDA plate, and the ordinate shows the inhibition rate of the mycelial growth of Botrytis cinerea.
FIG. 6 shows that the Pseudomonas aeruginosa QSE-TM7 fermentation broth inhibits the germination of the spores of Botrytis cinerea, the abscissa shows the concentration of the fermentation broth containing the QSE-TM7 strain in the spore liquid, and the ordinate shows the germination rate of the spores of Botrytis cinerea.
FIG. 7 shows that the Pseudomonas aeruginosa QSE-TM7 broth inhibited the germination of Gramineae on PDA plates, with 15% QSE-TM7 strain broth concentration, and CK indicated the absence of QSE-TM7 strain broth.
FIG. 8 shows the relative expression level of the tomato leaf disease resistance related gene induced by the Pseudomonas aeruginosa QSE-TM7 fermentation broth, wherein the abscissa indicates the sampling time after the treatment of the QSE-TM7 fermentation broth, the ordinate indicates the relative expression level before the treatment, and the tomato action gene is used as an internal reference.
FIG. 9 shows the effect of the Pseudomonas aeruginosa QSE-TM7 fermentation broth on controlling tomato gray mold in vitro, wherein the whole leaf is treated with the QSE-TM7 fermentation broth, inoculated with a Botrytis cinerea cake, and the diameter of the lesion is measured after inoculation for 2 days, and CK represents the use of H 2 O replaces QSE-TM7 strain broth to treat the leaves.
FIG. 10 shows the in vivo experimental effect of the Pseudomonas aeruginosa QSE-TM7 fermentation broth on controlling tomato gray mold, spraying the mold spore liquid inoculated with the whole plant after the treatment of the QSE-TM7 fermentation broth, and observing the disease condition of the tomato plant after 7 days.
Description of the embodiments
The technical scheme of the invention is further described in detail below with reference to the attached drawings and specific embodiments. The experimental methods, in which specific conditions are not noted in the following examples, are generally conducted under conventional conditions or under conditions recommended by the manufacturer.
Example 1
1. Isolation and screening of QSE-TM7 Strain
2019. Collecting 5 parts of tomato roots in a green house in Qingdao city for 21 days in 12 months, cleaning soil on the root surfaces, sterilizing with 0.1% sodium hypochlorite for 3min, sterilizing with 70% ethanol for 1min, and cleaning with sterile water for three times; grinding the juice in a sterile mortar, and continuously and gradually diluting the juice with sterile water; 100. Mu.l of each of the tissue suspensions diluted 100-fold and 1000-fold was uniformly spread on a TM plate (peptone 10g, acid hydrolyzed casein 1g, glucose 5g, agar powder 15g, and water 1L, autoclaved at 121℃for 20 min) with a spreader, and incubated in an incubator at 28℃for 24-48h.
Picking single colony growing on TM plate, inoculating on Botrytis cinerea PDA plate (potato 200g is cut into pieces and boiled in boiling water for 15min, three layers of gauze are filtered, supernatant is obtained, glucose 20g, agar powder 15g, water is added to 1L, and high pressure sterilization is carried out at 121deg.C for 20 min), placing in a 25 deg.C incubator for opposite growth experiment, culturing for 2-3d, and observing that Botrytis cinerea mycelium growth is inhibited. Selecting bacterial strain with good effect of inhibiting growth of gray mold, picking bacterial colony into 100 μl of sterile water, sucking into bacterial suspension with a pipetting gun, continuously gradient diluting with sterile water, respectively taking 100 μl of 10-4 times and 10-5 times of diluted solution, coating onto TM plate, and culturing in a 28 ℃ incubator for 24h to obtain single bacterial colony. And selecting single bacterial colony, and verifying the inhibiting effect of the purified bacterial strain on the gray mold through a plate counter experiment to obtain the single bacterial colony bacterial strain with good inhibiting effect on the gray mold.
The purified strain was inoculated into a TM liquid medium (peptone 10g, acid hydrolyzed casein 1g, glucose 5g, water to 1L, autoclaving at 121℃for 20 min), shake-cultured at 28℃and 180rpm for 4 days, centrifuged at 10000rpm, and the supernatant was filtered through a 0.45 μm sterile filter to obtain a bacterial-free fermentation broth. On the gray mold PDA plate which has been cultivated for 24 hours, placing oxford cup at a position 4cm away from the edge of the bacterial colony, sucking 200 μl of fermentation liquor, adding the fermentation liquor into the oxford cup, continuously cultivating for 2-3d in a 25 ℃ incubator, observing the condition that growth of gray mold hypha is inhibited, obtaining a strain with strong inhibition effect on growth of gray mold hypha by the fermentation liquor, and naming the strain as QSE-TM7.
2. Classification and identification of QSE-TM7 Strain
1. Morphological and physiological biochemical identification:
the bacterial colony formed by the QSE-TM7 strain after 24 hours of growth on the TM culture medium is round, opaque, smooth and wrinkle-free (figure 1); no growth at 4 ℃ and growth at 28 ℃ and 42 ℃ (fig. 2); the cells were red after gram staining, were gram-negative, were short-rod-shaped, and were arranged singly (FIG. 3). Preliminary judgment of the QSE-TM7 Strain as PseudomonasPseudomonas）。
2. Molecular identification:
extracting genome DNA of QSE-TM7 strain, and using the genome DNA as a template to carry out PCR amplification by using a specific primer; specific genes and primers amplified are shown in Table 1;
TABLE 1 amplified genes and primers used
The PCR reaction system is as follows: 5×TransStart R FastPfuBuffer 10. Mu.l; 10 mM dNTP mix 1. Mu.l; 2 μl of template; 1 μl of primer-F (10 μM); primer-R (10. Mu.M) 1. Mu.l;TransStart R FastPfu DNA polymerase 1 μl；ddH 2 O 34 μl；Total 50 μl。
the PCR reaction procedure was as follows: pre-denaturation at 95 ℃ for 5 min; denaturation at 95℃of 20s; annealing at 54 ℃ for 30 s; extending at 72 ℃ by 40 s; extending at 72 ℃ for 5 min;32 cycles; maintained at 4 ℃.
And (5) cutting and recycling after the PCR product is subjected to gel running, and sending the recycled product to a sequencing company for sequencing. 16S rDNA and of the strain QSE-TM7, sequencedrpoD The fragment sequences of the genes are shown as SEQ ID No. 1 and SEQ ID No. 2. Comparing the sequence obtained by sequencing with the sequence in NCBI database, respectively, the 16S rDNA sequence of the strain QSE-TM7 to be detected andrpoDgene fragment sequence and pseudomonas aeruginosaPseudomonas aeruginosaHas the highest sequence homology. Based on sequence alignment of 2 different gene fragments, a phylogenetic tree (FIG. 4) was constructed, and the strain QSE-TM7 was identified as Pseudomonas aeruginosa by comprehensive analysisPseudomonas aeruginosa。
The pseudomonas aeruginosa QSE-TM7 screened by the invention is subjected to strain preservation, and the preservation unit is as follows: china center for type culture Collection; address: chinese university of Wuhan; preservation date: 2023, 7, 6; pseudomonas aeruginosaPseudomonas aeruginosaThe preservation number of QSE-TM7 is CCTCC M20231202.
Example 2: inhibition of growth and development of Botrytis cinerea by QSE-TM7 fermentation liquor
Inoculating the pseudomonas aeruginosa QSE-TM7 into a TM liquid culture medium, shake culturing for 4 days at 28 ℃ and 180rpm, centrifuging the culture solution at 10000rpm, and filtering the supernatant by a 0.45 mu m sterile filter to obtain QSE-TM7 fermentation broth without QSE-TM7 thalli.
1. Inhibition of growth of Botrytis cinerea mycelium by QSE-TM7 fermentation broth
QSE-TM7 fermentation brothPDA plates containing different concentrations of QSE-TM7 broth were prepared by adding 1%,2%,3%,6%,9%,12%,15% to PDA medium, and 4 plates of each of the concentration of CK and 6 different QSE-TM7 broth were used as Controls (CK) without QSE-TM7 broth added. The center of the plate was inoculated with the mold spore liquid (3. Mu.l, 10) 6 cfu), culturing for 3d in a 25 ℃ incubator, measuring the colony diameters (a and b) of the gray mold on the flat plate by adopting a crisscross method, calculating the colony area according to an elliptic area formula (S=pi ab/4), and calculating the inhibition rates of QSE-TM7 fermentation broths with different concentrations on the growth of gray mold hyphae according to the area and a formula 1.
（1）
Wherein,,mean value of the areas of the gray mold colonies in the control group, [ about ]>The average value of the areas of the gray mold colonies in the treated group to which the QSE-TM7 fermentation broth was added was shown.
As shown in FIG. 5, the 6-concentration QSE-TM7 fermentation liquor has inhibition effect on the growth of the mycelial of the Botrytis cinerea, and the inhibition rate of the mycelial growth is obviously improved along with the increase of the concentration, and the inhibition rate is 79.3% when the concentration of the QSE-TM7 fermentation liquor is 15%.
2. Inhibition of Botrytis cinerea spore germination by QSE-TM7 fermentation broth
Collecting Botrytis cinerea spores with 1/10 concentration of YEPD medium (formula: yeast extract 10g, peptone 20g, glucose 20g, water to 1L, and autoclaving at 121deg.C for 20 min), and adjusting to concentration of 10 4 cfu. To the spore suspension, QSE-TM7 broth at final concentrations of 3%,9% and 15% was added as treatment group, equal volume of TM medium was added as CK group, 20. Mu.l spores were pipetted onto glass slides, placed in a cassette for humidification, and incubated in an incubator at 25 ℃. When the spore germination in the CK group exceeds 90%, counting the spore germination number in the treatment group and the CK groupAmount of the components. 5 slides were counted per treatment, about 100 spores per slide were counted, and the average of the number of spores germinated was plotted.
As shown in FIG. 6, the average germination rate of the spore suspension in the CK group was 99.5% when the spore liquid was cultured for about 6 hours, and the germination rates of the spores of the QSE-TM7 fermentation broths containing 3%,9% and 15% were 97.8%,93.6% and 0%, respectively, which indicated that the concentration of the QSE-TM7 fermentation broths reached 15%, and that the spore suspension had a strong inhibitory effect on the germination of the Botrytis cinerea spores.
3. Inhibition of Botrytis cinerea sclerotium germination by QSE-TM7 fermentation broth
PDA plates containing 15% QSE-TM7 broth were prepared and inoculated with the surface inverted (side of the sclerotium-producing plate near the medium facing upward) to remove residual airborne hyphae from the surface, with PDA plates without QSE-TM7 broth as controls. The PDA plate was incubated in an incubator at 25℃for 2 days, and the germination of sclerotium on the PDA plate was observed.
The sclerotium germination results are shown in FIG. 7, and after 2d of culture, the sclerotium inoculated on the PDA plate without QSE-TM7 fermentation broth all germinated, and white mycelium was produced around the sclerotium contact medium, and colonies were formed; sclerotium inoculated on PDA plate containing 15% QSE-TM7 broth had no hyphae produced near the medium and no colonies formed. The result shows that the QSE-TM7 fermentation broth has a strong inhibition effect on the mycelium generated by the germination of the sclerotium of the Botrytis cinerea.
Example 3: QSE-TM7 fermentation liquid induces expression of tomato leaf disease resistance related genes
Tomato seedlings of 2 weeks old were uniformly sprayed with the QSE-TM7 fermentation broth prepared in example 2, treated and cultured in a greenhouse, and tomato leaves were taken at 4h,8h,12h,16h and 20h after the treatment, respectively, and immediately taken as 0h after the spraying. 3 seedlings are respectively taken at each time point, 6 leaves are respectively taken for each seedling, 18 leaves are mixed and quickly frozen by liquid nitrogen, and the obtained mixture is placed in a refrigerator at the temperature of minus 80 ℃ for preservation. After all materials are sampled, grinding each leaf to powder in liquid nitrogen, taking a proper amount of powder, extracting RNA of the tomato leaf by using a plant RNA extraction kit, carrying out reverse transcription by using a reverse transcription kit to obtain cDNA, and detecting the relative expression levels of tomato disease-resistant related genes SlPR1a, slPR1b, slPR2 and SlPR5 by using fluorescence quantitative PCR (qRT-PCR). The primers used for qRT-PCR are shown in Table 2.
TABLE 2 qRT-PCR primers
The qRT-PCR reaction system is as follows: 2 XqRT-PCR Mix 10. Mu.l; 2 μl of cDNA template was added after 10-fold dilution; 1 μl of primer-F (10 μM); primer-R (10. Mu.M) 1. Mu.l; ddH 2 O 6 μl；Total 20 μl。
The qRT-PCR reaction procedure was as follows: pre-denaturation at 95 ℃ for 20s; denaturation at 95℃5 s; annealing at 58 ℃ for 20s; extension at 72 DEG C
20s; 40 cycles.
The results of qRT-PCR were analyzed with tomato Actin (SlActin) gene expression as an internal control and 0h as a control, and the results of gene relative expression were as shown in FIG. 8, wherein QSE-TM7 fermentation broth treatment induced significant upregulation of expression of 4 genes, with the highest relative levels at 12h (12 hpi) of treatment. The results show that QSE-TM7 fermentation broth treatment induces rapid response of tomato leaf disease resistance related genes.
Example 4: application of QSE-TM7 fermentation liquor in preventing and treating gray mold of tomatoes
1. In vitro experiment of QSE-TM7 fermentation liquor for preventing and treating gray mold of tomatoes
Selecting tomato seedlings with uniform growth vigor and 4 weeks old, uniformly spraying the whole plant with QSE-TM7 fermentation liquor to obtain a treatment group, treating the whole plant with water to obtain a control group (CK), and respectively taking 18 leaves of the control group and the treatment group, and placing the 18 leaves on wet filter paper. The patties were removed from the edges of the 3d culture plate with a 4mm diameter punch and inoculated onto the treated tomato leaves. Moisturizing for 60 hours, and observing the infection condition of the gray mold; the diameter from the center of the bacterial cake to the edge of the infection spot is measured by a crisscross method, the oval area is calculated, meanwhile, the occupied area of the bacterial cake (with the diameter of 4 mm) is subtracted, and the residual area (ring shape) is used as the area of the infection spot. Statistical analysis was performed on the area of the infested plaques in the treated and control groups.
As shown in fig. 9, the leaf area of the infection spots in the treatment group sprayed with QSE-TM7 fermentation broth was significantly smaller than that in the control group; the inhibition of the ash mold infestation by the QSE-TM7 fermentation broth was calculated to be 71.6% according to equation 2. The experiment was performed in three biological replicates with substantially identical results.
（2）
Wherein the method comprises the steps ofMean value of the areas of the gray mold infested spots in the control group is shown, and +.>Mean gray mold infested spot area in the treatment group is shown.
2. Potting experiment for preventing and treating gray mold of tomatoes by using QSE-TM7 fermentation liquor
Selecting 4-week-old potted tomato seedlings with consistent growth vigor, uniformly spraying the whole plants with QSE-TM7 fermentation liquor to serve as a treatment group, treating with water to serve as a control group (CK), and naturally airing for 12h. The gray mold spore liquid (10) 6 cfu) are uniformly sprayed on plant leaves, and are placed in an illumination incubator with the humidity of more than 85% for continuous cultivation. After 7 days of incubation, the onset of disease was observed in the control and treatment groups.
As a result, as shown in fig. 10, the number of leaves of tomato plants treated with QSE-TM7 fermentation broth was significantly smaller than that of the control group, and the growth state of the whole plants was significantly better than that of the control group.
In combination with all the results, the QSE-TM7 fermentation liquor has remarkable direct inhibition effect on the gray mold, can induce the tomato leaves to generate immune response, and further can improve the resistance of the tomatoes to the gray mold, so that the QSE-TM7 fermentation liquor has a good control effect on the gray mold of the tomatoes, and can be developed into a biological agent for effectively controlling the gray mold of the tomatoes.
The above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be apparent to one skilled in the art that modifications may be made to the technical solutions described in the foregoing embodiments, or equivalents may be substituted for some of the technical features thereof; such modifications and substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Claims (10)
1. Pseudomonas aeruginosa QSE-TM7 is characterized by being classified as Pseudomonas aeruginosaPseudomonas aeruginosaThe strain is preserved in China Center for Type Culture Collection (CCTCC) M20231202.
2. The pseudomonas aeruginosa QSE-TM7 according to claim 1, wherein the pseudomonas aeruginosa QSE-TM7 colony is round, opaque, smooth in surface and wrinkle-free; no growth at 4 ℃ and growth at 28 ℃ and 42 ℃; QSE-TM7 cells are short rods, are arranged singly, and are red in gram staining.
3. Use of pseudomonas aeruginosa QSE-TM7 according to claim 1 for preparing biological agents for controlling tomato gray mold.
4. The use according to claim 3, wherein the pseudomonas aeruginosa QSE-TM7 is capable of significantly inhibiting mycelial growth, spore germination and sclerotium germination of gray mold.
5. The use according to claim 3, wherein the pseudomonas aeruginosa QSE-TM7 is capable of inducing expression of tomato leaf disease resistance related genes.
6. The use according to claim 5, wherein said tomato disease-resistance related genes comprise SlPR1a, slPR1b, slPR2 and SlPR5.
7. The use according to claim 3, wherein the biological agent comprises a pseudomonas aeruginosa QSE-TM7 fermentation broth.
8. The use according to claim 7, wherein the preparation method of the pseudomonas aeruginosa QSE-TM7 fermentation broth comprises: inoculating pseudomonas aeruginosa QSE-TM7 into TM liquid culture medium, shake culturing, centrifuging culture solution, filtering supernatant by sterile filter, and obtaining pseudomonas aeruginosa QSE-TM7 fermentation broth without thallus.
9. The use according to claim 7, wherein the pseudomonas aeruginosa QSE-TM7 fermentation broth is added in a biological preparation at a volume ratio of 1-15%.
10. The use according to claim 3, wherein the biological agent is used in the following way: uniformly spraying on the whole tomato plants.
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| WO2017178529A1 (en) * | 2016-04-12 | 2017-10-19 | Bioscienz Holding B.V. | Pseudomonas strains and consortia thereof for use in protection against plant diseases |
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| US20220232834A1 (en) * | 2021-01-28 | 2022-07-28 | T3 Bioscience, LLC | Pseudomonas chlororaphis species and its use in the control of diseases caused by bacteria and fungi |
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| WO2017178529A1 (en) * | 2016-04-12 | 2017-10-19 | Bioscienz Holding B.V. | Pseudomonas strains and consortia thereof for use in protection against plant diseases |
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