CN114774291B - Trichoderma asperellum 10564 strain and application thereof in biocontrol of tomato fusarium wilt - Google Patents
Trichoderma asperellum 10564 strain and application thereof in biocontrol of tomato fusarium wilt Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/05—Fruit crops, e.g. strawberries, tomatoes or cucumbers
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12N3/00—Spore forming or isolating processes
Abstract
The invention provides a trichoderma asperellum 10564 strain and application thereof in biocontrol of tomato fusarium wilt, and particularly relates to the technical field of biocontrol of plant diseases. The trichoderma asperellum 10564 strain with biocontrol function shows the inhibition effect of the trichoderma asperellum 10564 strain on fusarium oxysporum; the tomato seedlings treated by the mixed spore liquid of the 10564 strain and the fusarium oxysporum show remarkable disease resistance effect on the fusarium oxysporum, and meanwhile, chlorophyll fluorescence related parameters and hormone content in the leaves are measured to find out that the 10564 strain can protect a photosynthetic system II in the tomato seedlings and induce the change of the phytohormone content so as to improve the disease resistance, and the corresponding biocontrol microbial inoculum can be prepared.
Description
Technical Field
The invention belongs to the technical field of biological control of plant diseases, and particularly relates to a trichoderma asperellum 10564 strain and application thereof in biological control of tomato blight.
Background
Trichoderma spp belongs to the phylum Deuteromycotina, class Cellularomyces, order Convergenca, family Convergencaceae. Trichoderma can induce plant system resistance, has competitive action, resistance action and re-mailing action on pathogenic bacteria, and is a fungus with great biocontrol potential.
The current research on trichoderma is mainly focused on trichoderma harzianum (Trichoderma harzianum), trichoderma atroviride (Trichoderma atroviride), trichoderma viride (Trichoderma virens), trichoderma reesei (Trichoderma reesei) and trichoderma longibrachiatum (Trichoderma longbrachiatum), and few studies on trichoderma asperellum strains are known, and the trichoderma asperellum (Trichoderma asperellum) wood-8 strain disclosed in Chinese patent CN107142213A has obvious growth promoting effect, but has less research on tomato blight.
Disclosure of Invention
Accordingly, the invention aims to provide a trichoderma asperellum 10564 strain and application thereof in biocontrol of tomato fusarium wilt, wherein the trichoderma asperellum 10564 strain has an inhibiting effect on fusarium oxysporum, can protect a photosynthetic system II in tomato seedlings and can induce the content change of phytohormone so as to improve disease resistance.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a trichoderma asperellum (Trichoderma asperellum) 10564 strain with a biocontrol function, which is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No.40090.
Preferably, the biocontrol effect comprises inhibition of fusarium oxysporum (Fusarium oxysporum).
The invention also provides application of the 10564 strain in preparing a biocontrol microbial agent.
Preferably, the types of biocontrol agents include conidium eluate and conidium suspension.
The invention provides a preparation method of a conidium eluent of trichoderma asperellum 10564 strain, which comprises the following steps: inoculating Trichoderma asperellum 10564 strain on a PDA plate, and culturing at 25deg.C under illumination for 3d to obtain single colony;
and (3) picking the edge hyphae of the single colony, inoculating the edge hyphae to a PDA flat plate for 7d, adding sterile water, and scraping conidium on the surface of the colony to obtain the conidium eluent.
The invention also provides a preparation method of the conidium suspension of the trichoderma asperellum 10564 strain, which comprises the following steps: trichoderma asperellum 10564 strain is inoculated on a PDA plate for culture, and single colony is obtained;
picking mycelium blocks at the edge of the single colony, and inoculating the mycelium blocks into PDB culture solution for culturing for 3-7 d to obtain a culture;
centrifuging the culture, filtering the supernatant, and collecting filtrate to obtain the conidium suspension.
The invention also provides application of the 10564 strain or the conidium eluent obtained by the preparation method or the conidium suspension obtained by the preparation method in preventing and treating tomato fusarium wilt.
Preferably, the method comprises dipping roots by using the conidium eluent before transplanting tomato seedlings;
and/or after transplanting the tomato seedlings, root irrigation is carried out by utilizing the conidium suspension.
The invention also provides a method for preventing and treating tomato leaf blight, which comprises the following steps: before transplanting tomato seedlings, dipping roots of the seedling roots by using the conidium eluent obtained by the preparation method.
The invention also provides a method for protecting a tomato seedling photosynthetic system II and improving the content of resistance-related hormone, which comprises the following steps: after the tomato seedlings are transplanted, the conidium suspension obtained by the preparation method is used for root irrigation.
The beneficial effects are that: the invention provides a trichoderma asperellum 10564 strain with biocontrol function, which is used for carrying out a plate counter experiment on the trichoderma asperellum 10564 strain and fusarium oxysporum, and shows the inhibition effect of the trichoderma asperellum 10564 strain on the fusarium oxysporum; the tomato seedlings treated by the mixed spore liquid of the 10564 strain and the fusarium oxysporum show remarkable disease resistance effect on the fusarium oxysporum, and meanwhile, chlorophyll fluorescence related parameters and hormone content in the leaves are measured to find out that the 10564 strain can protect a photosynthetic system II in the tomato seedlings and induce the change of phytohormone content so as to improve disease resistance.
Biological preservation information
Trichoderma asperellum (Trichoderma asperellum) 10564 is stored in China general microbiological culture Collection center (China general microbiological culture Collection center), and the specific storage address is China national academy of sciences of China, including No. 3, no. 1, of North Chen West Lu in the Korean area of Beijing, and the storage time is 2022, 03, 04 days, and the storage number is CGMCC No.40090.
Drawings
FIG. 1 shows the inhibition of growth of Fusarium oxysporum by Trichoderma asperellum 10564 strain and Fusarium oxysporum co-cultured on PDA medium for 7 days;
FIG. 2 shows the phenotype of tomato seedlings after the bacterial strain spore eluent of Trichoderma asperellum 10564 is used for root soaking, wherein control in the figure represents clear water treatment, F.oxy represents Fusarium oxysporum treatment, 564& F.oxy represents mixed treatment of Fusarium oxysporum and 10564, 564 represents treatment by using the bacterial strain 10564, and the following is the same;
FIG. 3 shows plant disease index after soaking tomato seedlings with Trichoderma asperellum 10564 strain spore eluent;
FIG. 4 shows the incidence of plant disease after the tomato seedlings are rooted with Trichoderma asperellum 10564 strain spore eluate;
FIG. 5 shows that after the spore suspension of Trichoderma asperellum 10564 strain is used for root soaking of tomato seedlings, the chlorophyll fluorescence related parameters in tomato leaves are changed, so that the stress resistance of the tomato is enhanced;
FIG. 6 shows the change in jasmonic acid content of tomato plants after rooting tomato seedlings with Trichoderma asperellum 10564 strain spore suspension.
Detailed Description
The invention provides a trichoderma asperellum (Trichoderma asperellum) 10564 strain with a biocontrol effect, wherein the 10564 strain is preserved with the preservation number of CTCCSJ-W-AW10564. The 10564 strain is separated from the beach wetland, and the strain is identified by molecular level, and the result is trichoderma asperellum (Trichoderma asperellum).
The trichoderma asperellum 10564 is cultured on a PDA culture medium for one week, can generate a large amount of aerial hyphae and generate dark green conidium clusters, and is beneficial to the rapid growth and spore production of the trichoderma asperellum by culturing at the temperature of 25-28 ℃ under illumination.
The biocontrol effect of the invention preferably comprises inhibition of fusarium oxysporum (Fusarium oxysporum), and in the embodiment, the experiment using a flat plate for the counter verifies that the trichoderma asperellum 10564 strain can inhibit the growth of fusarium oxysporum, and the embodiment comprises the following steps: the trichoderma asperellum 10564 strain can occupy more growth space and nutrition of a flat plate in the same time, can generate resistant substances to obviously inhibit the growth of fusarium oxysporum hyphae, and forms a bacteriostasis ring between the trichoderma asperellum 10564 strain and the fusarium oxysporum hyphae, so that the 10564 strain can be applied to the preparation of biocontrol microbial agents.
The invention also provides application of the 10564 strain in preparing a biocontrol microbial agent.
Types of biocontrol agents according to the invention preferably include conidium eluate and conidium suspension.
The invention provides a preparation method of a conidium eluent of trichoderma asperellum 10564 strain, which comprises the following steps: inoculating Trichoderma asperellum 10564 strain on a PDA plate, and culturing at 25deg.C under illumination for 3d to obtain single colony;
and (3) picking the edge hyphae of the single colony, inoculating the edge hyphae to a PDA flat plate for 7d, adding sterile water, and scraping conidium on the surface of the colony to obtain the conidium eluent.
Trichoderma asperellum 10564 strain is inoculated on a PDA plate for culture to obtain single colony, wherein the temperature of the culture is preferably 25 ℃ and the time is preferably 3d. The composition of the PDA plate is not particularly limited, and the PDA plate is preferably prepared by using a conventional plate culture medium in the art. The formulation of the PDA plate used in the examples preferably comprises: 200g of potato, 20g of glucose, 10g of agar powder and 1000ml of distilled water are supplemented. In the preparation of the PDA flat plate, the invention preferably comprises the steps of slicing the peeled potatoes after washing, adding water, boiling for about half an hour, filtering the potatoes by using gauze, adding glucose, adding agar powder into water, stirring uniformly, adding the agar powder into the solution, adding water for constant volume, stirring, dissolving, packaging and sterilizing.
After obtaining single colony, the invention picks up the edge hypha of the single colony, inoculates the single colony to a PDA flat plate, cultures the single colony for 7d under the illumination condition at 25 ℃, and scrapes conidium on the surface of the colony after adding sterile water to obtain the conidiumAnd (5) sub-eluent. According to the invention, the edge hyphae of the single colony are preferably selected and inoculated on a new PDA plate for culture, the temperature of the culture is preferably 25 ℃, the time is preferably 7d, then water is added into a culture dish containing the PDA plate, conidia on the surface of the colony are scraped, and the conidia eluent is obtained after uniform mixing. The invention preferably further comprises counting after obtaining said conidial eluate, e.g. by counting under a microscope using a blood cell counting plate, and adjusting the concentration of the conidial eluate to 1x10 7 Each of the above-mentioned components/ml was used as a working fluid.
The invention also provides a preparation method of the conidium suspension of the trichoderma asperellum 10564 strain, which comprises the following steps: trichoderma asperellum 10564 strain is inoculated on a PDA plate for culture, and single colony is obtained;
picking mycelium blocks at the edge of the single colony, and inoculating the mycelium blocks into PDB culture solution for culturing for 3-7 d to obtain a culture;
centrifuging the culture, filtering the supernatant, and collecting filtrate to obtain the conidium suspension.
Trichoderma asperellum 10564 strain is inoculated on a PDA flat plate for culture, and single bacterial colony is obtained. The cultivation according to the invention preferably comprises cultivation under light conditions at 25℃for 3 days, whereby single colonies can be produced.
After obtaining single colony, the invention picks up mycelium blocks at the edge of the single colony and inoculates the mycelium blocks into PDB culture solution for 3-7 d to obtain a culture. The formula of the PDB culture solution is completely different from the formula of the PDA plate except that the PDB culture solution does not contain agar. The culture according to the present invention preferably comprises shaking culture at a temperature of 25℃and 150r/min for 7 days.
After obtaining the culture, the invention centrifugates the culture, filters the centrifugate supernatant, and collects the filtrate to obtain the conidium suspension. The centrifugation according to the invention is 8000rpm. The invention filters the centrifuged supernatant, preferably comprising a two-stage filtration, first by a filter paper filtration, and the primary filtrate obtained is filtered again by a bacterial filter to obtain a conidium suspension.
The invention also provides application of the 10564 strain or the conidium eluent obtained by the preparation method or the conidium suspension obtained by the preparation method in preventing and treating tomato fusarium wilt.
The application of the invention preferably comprises dipping roots with the conidium eluent before transplanting tomato seedlings; and/or after transplanting the tomato seedlings, root irrigation is carried out by utilizing the conidium suspension.
The invention also provides a method for preventing and treating tomato leaf blight, which comprises the following steps: before transplanting tomato seedlings, dipping roots of the seedling roots by using the conidium eluent obtained by the preparation method.
In the invention, when the root dipping is carried out, the root dipping is preferably carried out by using the working solution of the conidium eluent, and the root dipping time is preferably 15-20 min. The invention preferably further comprises the steps of taking out tomato seedlings growing for 3 weeks from the plug tray, washing the roots with clean water, and then carrying out root dipping. The root dipping agent can inhibit fusarium oxysporum parasitized in the soil of the root and the rhizosphere of the tomato, and effectively prevent and treat the fusarium wilt of the leaf of the tomato.
The invention also provides a method for protecting a tomato seedling photosynthetic system II and improving the content of resistance-related hormone, which comprises the following steps: after transplanting tomato seedlings, the concentration of the conidium suspension obtained by the preparation method is adjusted to be 1x10 7 Root irrigation was performed at a concentration of each ml. The root irrigation of the invention is preferably carried out 3 times at 3 weeks of transplanting, each time is spaced for 3 days, and each time is 20ml.
The root irrigation method can improve the wilt resistance of tomato plants by protecting the photosynthesis system II of tomato seedlings from being damaged, and is specifically characterized in that: chlorophyll fluorescence related parameters in leaves of tomato seedlings treated by spore liquid of Trichoderma asperellum 10564 strain all show a tendency of promoting photosynthesis of tomatoes; the root irrigation can also improve the content of the hormone related to the resistance of the tomatoes, and the content of jasmonic acid in the tomato seedlings is obviously improved.
The Trichoderma asperellum 10564 strain and the application thereof in biocontrol of tomato blight provided by the invention are described in detail below with reference to examples, but they are not to be construed as limiting the scope of the invention.
Example 1
Separation and purification of trichoderma asperellum 10564
Retrieving soil containing trichoderma from the field, naturally airing the collected soil sample, crushing the soil sample, and sieving the crushed soil sample. 10g of soil sample is weighed and placed in a flask containing 90mL of sterile water, and is fully oscillated by a vortex machine, 10mL of suspension is sucked and diluted in 90mL of sterile water from the diluted suspension, and the like, and the volume fraction is diluted to be 10 -3 、10 -4 、10 -5 . 0.1mL of the suspension is sucked from the soil suspension with each content by a micropipette and dispersed and dripped on a Ma Dingshi-Bengalhon plate, then the mixture is uniformly coated by a sterilized triangular glass rod, and after standing for 20min, the mixture is inversely cultured in a constant temperature incubator at 25 ℃, and the colony is immediately purified after being formed.
Identification of Trichoderma species
Trichoderma is cultured on a PDA plate for 3d, a puncher with the diameter of 0.5cm is used for punching a bacterial cake, the bacterial cake is placed on the PDA plate, and is cultured for 5d under the condition of 25 ℃ and illumination, and the shape, the color and the like of bacterial colonies are observed. And observing branching conditions of the conidiophores of the trichoderma and the forms of the conidiophores under a microscope, and carrying out species identification by combining culture characters and referring to a trichoderma population classification system of Rifai and Bissett.
Wherein:
1. preparation of trichoderma asperellum 10564 spore eluent:
1) Inoculating the trichoderma asperellum 10564 strain on a PDA culture medium for culture to obtain trichoderma asperellum 10564 strain colonies;
2) Inoculating the bacterial colony edge hypha of the trichoderma asperellum 10564 strain obtained in the step 1) onto a PDA (personal digital assistant) culture dish for culture;
3) Adding sterile water into the culture dish in the step 2), scraping and washing conidium on the surface of the bacterial colony, and uniformly mixing to obtain the conidium eluent of the trichoderma asperellum 10564 strain.
4) Counting the conidium eluate of step 3) under microscope by using a blood cell counting plate, and adjusting the concentration of the conidium eluate to be1x10 7 And each ml.
2. Preparation of a conidium suspension of Trichoderma asperellum 10564 strain:
1) Inoculating the trichoderma asperellum 10564 strain on a PDA culture medium for culture to obtain trichoderma asperellum 10564 strain colonies;
2) Inoculating mycelium blocks at the edge of the trichoderma asperellum 10564 colony obtained in the step 1) into PDB culture solution for culture to obtain a culture;
3) Centrifuging the culture obtained in the step 2), filtering the centrifugated supernatant by filter paper, and filtering the obtained primary filtrate again by a bacterial filter to obtain filtrate.
Example 2
Inhibition of trichoderma asperellum 10564 strain on fusarium oxysporum
Activating the stored 10564 strain and Fusarium oxysporum on a PDA plate respectively, punching holes on a PDA culture medium plate by using a puncher in an ultra-clean workbench to obtain a bacterial cake with the length of 5 multiplied by 5mm, transferring the two bacteria to a new PDA plate, and culturing for 7 days at room temperature.
As shown in FIG. 1, the Trichoderma asperellum 10564 strain can occupy more growth space and nutrition of the plate in the same time, can generate resistant substances to remarkably inhibit the growth of fusarium oxysporum hyphae, and forms a bacteriostasis ring between the two.
Example 3
Prevention and treatment effect of trichoderma asperellum 10564 strain on tomato fusarium wilt
Removing tomato seedlings growing for 3 weeks from the tray, washing the roots with clear water, soaking in clear water, respectively, and soaking in Trichoderma asperellum 10564 strain spore eluate (1 x 10) 7 Individual/ml), fusarium oxysporum spore eluate (1 x 10) 7 Each ml), trichoderma asperellum 10564 strain and Fusarium oxysporum spore mixed eluent (1 x 10) 7 Each ml) is planted in a plug for 15-20 min, and each treatment is carried out for 24 plants. After treatment, tomato shows phenotype difference in seedling stage, as shown in figure 2, the mixed spore liquid treated group and the single trichoderma treated group have strong tomato seedling growth, and the disease resistance of plants is obviously enhanced.
The preparation method of fusarium oxysporum spore eluent comprises the following steps:
inoculating fusarium oxysporum on a PDA culture medium, and culturing for 3d at 25 ℃ to obtain fusarium oxysporum colonies;
inoculating fusarium oxysporum edge hyphae on a PDA culture dish for culturing for 7d;
adding sterile water into the culture dish, scraping and washing conidium on the surface of the bacterial colony, and uniformly mixing to obtain fusarium oxysporum conidium eluent.
Counting the obtained conidium eluate under microscope by using a blood cell counting plate, and adjusting the concentration of the conidium eluate to 1×10 7 And each ml.
The preparation steps of the mixed eluent of trichoderma asperellum 10564 and fusarium oxysporum are the same as the above, so that the concentration of the two strains is 1x10 7 And each ml.
The tomato plants are classified into 4 grades according to the disease state:
level 0: the plants grow strongly, the leaves are naturally stretched, green and bright, and the stems are thick and strong;
stage 1: the plant stands upright, the leaf yellowing and wilting area is less than 50% of the total leaf area, and the stem base is brown;
2 stages: the plant leaf yellowing and wilting area is larger than 50% of the total leaf area, the top leaf is yellow and shrunken, and the stem is withered and shrunken;
3 stages: the plants are dwarfed and withered, the leaves are withered and curled and shrunken, only growing points survive, and stems wilt;
4 stages: plants wilt to death.
Calculating disease index according to disease grade: disease index = Σ (number of disease plants at each stage×corresponding disease stage)/(total number of disease stages×highest disease stage) ×100. The disease index is used as the inhibition effect of the conidium eluent of the trichoderma asperellum 10564 strain on the conidium eluent of the fusarium oxysporum. As shown in fig. 3 and table 1.
TABLE 1 inhibition effect of different treatment groups on Fusarium oxysporum spore eluate
Calculating the morbidity (at 2-4 levels, namely judging as a tomato plant with wilt): incidence% = number of plants treated per total number of plants treated x 100. The morbidity is taken as the control effect of the conidium eluent of the trichoderma asperellum 10564 strain on the fusarium oxysporum to cause tomato fusarium wilt. As shown in fig. 4 and table 2.
TABLE 2 control effects of different treatment groups on tomato blight
Example 4
Trichoderma asperellum 10564 strain improves chlorophyll fluorescence content of tomato leaves so as to improve stress resistance of tomato plants
Tomato seedlings growing for 3 weeks are planted in the field, and clear water, trichoderma asperellum 10564 spore suspension, fusarium oxysporum suspension, and mixed suspension of trichoderma asperellum 10564 and fusarium oxysporum spores prepared in example 1 are irrigated to the root of each seedling respectively. Root irrigation is performed once every 3 days, and 3 times are continuous. Sampling after phenotype difference occurs in the four treatments, picking leaves at the same part of the tomato, carrying out dark treatment for 15min, and measuring chlorophyll fluorescence related parameters Fv/Fm, ETR= [ (Fm ' -F) Fm ' ] ×PFD, qP= (Fm ' -Fs)/Fv ' and NPQ= (Fm-Fm ')/Fm ' =Fm/Fm ' -1 in the samples by using a fluorometer PAM series.
Fv/Fm reflects the potential maximum photosynthetic capacity (i.e., photosynthetic efficiency) of a plant;
ETR is apparent photosynthetic electron transfer rate;
qP is fluorescence quenching caused by photosynthesis and reflects the level of photosynthesis activity;
NPQ reflects the ability of a plant to dissipate excess light energy as heat, i.e., photoprotection.
The preparation method of fusarium oxysporum spore suspension comprises the following steps:
inoculating fusarium oxysporum to PDA culture at 25 ℃ under dark condition for 3d to obtain fusarium oxysporum colonies;
inoculating mycelia at the edge of Fusarium oxysporum into a PDB culture solution for culturing to obtain a culture, wherein the culture condition is that shaking culture is carried out for 5 days under the conditions that the temperature is 25 ℃ and 150 r/min;
centrifuging the obtained culture, filtering the supernatant with filter paper to obtain spore suspension, and adjusting concentration to 1×10 with blood cell counting plate 7 And each ml.
A method of mixing spore suspensions comprising the steps of:
inoculating Trichoderma asperellum 10564 strain to PDA culture at 25deg.C under illumination for 3 days to obtain Trichoderma asperellum 10564 strain colony;
inoculating mycelium blocks at the edge of the obtained trichoderma asperellum 10564 colony into PDB culture solution for culturing to obtain a culture, wherein the culture condition is that shaking culture is carried out for 5 days under the conditions that the temperature is 25 ℃ and 150 r/min;
centrifuging the obtained culture, filtering the supernatant with filter paper to obtain spore suspension, and adjusting concentration to 1×10 with blood cell counting plate 7 And each ml.
Will be 2x10 7 Fusarium oxysporum spore suspension at a concentration of 2X10 per ml 7 Uniformly mixing the trichoderma asperellum 10564 spore suspension with the same volume per ml to obtain the mixed spore suspension of the trichoderma asperellum 10564 spore suspension.
As shown in FIG. 5 and Table 3, trichoderma inhibits Fusarium oxysporum activity, prevents tomato seedling from feeling ill, protects plant photosynthetic system II from damage, and improves leaf photosynthetic efficiency.
TABLE 3 influence of different treatment groups on photosynthesis
Example 5
Trichoderma asperellum 10564 strain induces the change of the content of jasmonic acid which is a hormone related to the resistance of tomato plants, and improves the disease resistance of tomatoes
Tomato seedlings growing for 3 weeks are planted in the field, and clear water, trichoderma asperellum 10564 spore suspension, fusarium oxysporum suspension, and mixed suspension of trichoderma asperellum 10564 and fusarium oxysporum spores prepared in example 1 are irrigated to the root of each seedling respectively. Root irrigation is performed once every 3 days, and 3 times are continuous. Sampling after phenotype difference occurs in the four treatments, and picking leaves at the same part of the tomato to determine the jasmonic acid content.
The results are shown in fig. 6 and table 4, the trichoderma regulates the change of resistance hormone in plants, induces the defense reaction of tomato seedlings and improves the disease resistance.
TABLE 4 Effect of different treatment groups on disease resistance of tomato
| CK | F. | 564&F.oxy | 564 | |
| 14.94 | 14.96 | 31.34 | 34.64 | |
| 19.44 | 12.34 | 32.43 | 33.60 | |
| 14.03 | 12.49 | 31.05 | 30.41 | |
| 16.28 | 11.62 | 32.12 | 29.75 |
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (1)
1. A method for protecting photosynthetic system II of tomato seedlings and increasing resistance-related hormone content, comprising the steps of: after tomato seedling transplanting, root irrigation is carried out by utilizing a conidium suspension of 564 strain of trichoderma asperellum (Trichoderma asperellum);
a method for preparing a conidium suspension of trichoderma asperellum 564 strain, comprising the steps of: inoculating Trichoderma asperellum 564 strain on a PDA plate for culture to obtain single colony; the preservation number of the trichoderma asperellum 564 strain is CGMCC No.40090;
picking mycelium blocks at the edge of the single colony, and inoculating the mycelium blocks into PDB culture solution for culturing for 3-7 d to obtain a culture;
centrifuging the culture, filtering the supernatant, and collecting filtrate to obtain the conidium suspension.
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| ES2188385B1 (en) * | 2001-06-26 | 2004-12-01 | Universidad De Barcelona | SUBSTRATES FOR BIOLOGICAL CONTROL OF PLANT DISEASES. |
| RU2629776C1 (en) * | 2016-11-23 | 2017-09-04 | федеральное государственное автономное образовательное учреждение высшего образования "Казанский (Приволжский) федеральный университет" (ФГАОУ ВО КФУ) | Method for obtaining compost suppressive with respect to fusarium oxysporum pathogene of plant fusarium wilt |
| CN107142213B (en) * | 2017-05-25 | 2021-02-19 | 金正大生态工程集团股份有限公司 | Trichoderma asperellum with growth promoting effect and culture method and application thereof |
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| CN113025501B (en) * | 2021-04-01 | 2022-05-31 | 山东农业大学 | Multifunctional trichoderma asperellum and application thereof |
| CN113788886B (en) * | 2021-08-23 | 2022-11-08 | 广东省农业科学院蔬菜研究所 | Application of cucumber photosynthetic system II oxygen evolution enhancement protein CsPSII-OEEP in resistance to melon epidemic diseases |
| CN114258923B (en) * | 2021-12-30 | 2022-10-14 | 南京工业大学 | Application of trichoderma asperellum microbial agent in prevention and treatment of crop diseases |
| CN114410481B (en) * | 2021-12-30 | 2023-09-15 | 广州市土根旺生物科技有限公司 | Trichoderma asperellum and application thereof |
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