CN117981773B - Application of verdant polyporus in wheat scab control - Google Patents
Application of verdant polyporus in wheat scab control Download PDFInfo
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- CN117981773B CN117981773B CN202410396655.9A CN202410396655A CN117981773B CN 117981773 B CN117981773 B CN 117981773B CN 202410396655 A CN202410396655 A CN 202410396655A CN 117981773 B CN117981773 B CN 117981773B
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- Agricultural Chemicals And Associated Chemicals (AREA)
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Abstract
The invention belongs to the technical field of microorganisms, and discloses application of verdant trichoderma viride in wheat scab control. The invention knows that the verdant polyporus Clonostachys chloroleuca Cc620,620 can efficiently inhibit the growth of Fusarium graminearum and sexual reproduction, and can be used for developing biocontrol bactericides for preventing and controlling wheat scab.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to application of verdant trichoderma viride in wheat scab control.
Background
Wheat scab (Fusarium head blight, FHB) caused by infection with Fusarium graminearum complex (Fusarium graminearum species complex, FGSC) is a fungal disease that severely jeopardizes wheat production. On one hand, the scab can directly damage the wheat ears, so that kernels are shrunken, and the yield of wheat is seriously affected; on the other hand, the mycotoxin Deoxynivalenol (DON) remaining in the kernel may cause the mycotoxin in the wheat kernel to exceed the standard. Wherein, fusarium graminearum (Fusarium graminearum) is a dominant pathogen causing wheat scab, can be infected in the whole growth period of wheat, and mainly causes seedling blight, stem basal rot, stalk rot and spike rot, wherein the harm of spike rot (scab) is the most serious.
Fusarium graminearum in the form of mycelium or ascus shells overwintering on crop disease residues. In spring of the next year, a large number of mature ascus shells are rapidly generated under proper conditions such as temperature, humidity and the like, and a large number of ascus spores ejected from the apertures of the ascus shells are spread to wheat ears in the flowering stage by means of air and rainwater for infection. Ascospores produced by sexual reproduction of Fusarium graminearum are considered as the primary source of wheat scab.
At present, the prevention and treatment of wheat scab also mainly depend on chemical pesticides, and biological prevention and treatment are important supplementary means.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides application of verdant trichoderma viride in resisting wheat scab pathogenic bacteria.
In order to achieve the aim, the invention provides an application of the verdant mold in preventing and controlling wheat scab, the verruca viridis Clonostachys chloroleuca Cc,620 with the preservation number of CGMCC No. 41029.
Further, the verruca emeraldine comprises the thallus and/or fermentation broth of verruca emeraldine Clonostachys chloroleuca Cc620,620.
Further, the wheat scab pathogen includes fusarium graminearum PH-1.
Further, the fermentation broth of the verruca emeralds can inhibit the saprophytic growth of Fusarium graminearum.
Further, the verruca emeralds is capable of effectively parasitizing Fusarium graminearum.
Further, the emerald is inhibiting formation of fusarium graminearum shells.
In order to better achieve the aim, the invention also provides an agricultural biocontrol microbial inoculum. The agricultural biocontrol microbial inoculum contains verruca viridis, wherein the verruca viridis is verruca viridis Clonostachys chloroleuca Cc620,620 with the preservation number of CGMCC No. 41029.
Compared with the prior art, the technical scheme provided by the invention has the following beneficial effects or advantages.
(1) The verruca viridis Clonostachys chloroleuca Cc620,620 provided by the invention can inhibit Fusarium graminearum with high efficiency.
(2) The verruca delphinii Clonostachys chloroleuca Cc620,620 provided by the invention has extremely strong capability of inhibiting growth and sexual reproduction of fusarium graminearum.
(3) The verruca viridis Clonostachys chloroleuca Cc620,620 provided by the invention can solve the problem of controlling primary infection bacteria sources in wheat scab control, and by spraying the verruca viridis Clonostachys chloroleuca Cc620,620 provided by the invention on straws, not only can the growth of fusarium graminearum on the straws be inhibited, but also the formation of ascus shells and ascus spores on the straws can be inhibited, so that the primary infection bacteria sources are effectively reduced.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required to be used in the description of the embodiments or the prior art will be briefly described below, it being obvious that the drawings in the description are only some embodiments of the invention, and that other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a plate diagram of a culture medium for antagonizing Fusarium graminearum from a sterile fermentation broth of Saprolegnia emeraldii Clonostachys chloroleuca Cc620,620 in accordance with the present invention. CK in fig. 1 is a control group. T1 in FIG. 1 is the treatment group.
FIG. 2 is a graph showing the results of antagonizing Fusarium graminearum colony diameters in a sterile fermentation broth of Sauropus turcz Clonostachys chloroleuca Cc620,620 in accordance with the present invention. CK in fig. 2 is a control group. T1 in fig. 2 is a treatment group.
FIG. 3 is a graph showing the parasitic effects of the Severe viridis Clonostachys chloroleuca Cc620,620 described in the present invention on Fusarium graminearum on carrot medium. CK in fig. 3 is a control group. T1 in fig. 3 is the treatment group.
Fig. 4 is a diagram showing the formation of fusarium graminearum shells inhibited by the verruca emeralds Clonostachys chloroleuca Cc620,620 in wheat and corn stalks. CK in fig. 4 is a control group. T1 and T2 in FIG. 4 are treatment groups.
Detailed Description
The following describes the technical aspects of the present invention with reference to examples, but the present invention is not limited to the following examples.
The experimental methods and the detection methods in each embodiment are conventional methods unless otherwise specified; the reagents and materials are commercially available unless otherwise specified.
The invention provides an application of verdant trichoderma viride in wheat scab control. Wherein the verruca viridis comprises thalli and/or fermentation liquor of the verruca viridis Clonostachys chloroleuca Cc620,620. The pathogenic bacteria of wheat scab comprise fusarium graminearum (Fusarium graminearum). In the following examples, the verruca emeraldine is verruca emeraldine Clonostachys chloroleuca Cc620,620 with a preservation number of CGMCC No. 41029. The verdant Sauropus oligosporus Clonostachys chloroleuca Cc620 is preserved in China general microbiological culture Collection center (China Committee for culture Collection) at a preservation address of North Star, west Lu 1, no. 3, china academy of sciences of China, which is the Korean area of Beijing. For a detailed description of the calicheamicin emeraldrich see patent application 202410330190.7.
The invention is further illustrated by the following examples.
Example 1
This example describes a bacterial fermentation broth antagonism of Fusarium graminearum biological assay of Sauropus delbrueckii Clonostachys chloroleuca Cc620,620.
1. Preparation of Endosporium emeraldine Clonostachys chloroleuca Cc620,620 spore suspension
Verruca emeraldine Clonostachys chloroleuca Cc620,620 was inoculated on PDA medium with a diameter of 60mm and cultured for 10d at 25 ℃. To each medium was added 3mL of Tween-20 at a volume fraction of 0.1%, the surface conidia were scraped with a coater, filtered with 4 layers of sterile filter cloth, and the spore concentration was adjusted to 1X 10 6 spores/mL with Tween-20 at a volume fraction of 0.1%.
The PDA medium composition: 200g of potato, 20g of glucose, 15-20 g of agar and 1000mL of water.
The preparation method of the Tween-20 with the concentration of 0.1 percent comprises the following steps: adding distilled water into the stock solution of 100 mu LTween-20 to 100mL, mixing uniformly, and sterilizing at 121 ℃ for 20min by moist heat.
2. Preparation of aseptic fermentation liquor of verruca emeraldine Clonostachys chloroleuca Cc620,620
The Sauropus turquoise Clonostachys chloroleuca Cc620 was cultured on PDA medium for 10d, and 5 bacterial cakes with a diameter of 5mm were punched out by a puncher and transferred into 150mL Erlenmeyer flasks containing 50mL PDB. After shaking culture at a constant temperature of 175rpm for 9 to 10 days at 25℃the resulting culture was filtered through 0.45 μm and 0.25 μm bacterial filters, respectively, to obtain sterile fermentation broths.
3. Antagonism of the sterile fermentation broth against Fusarium graminearum test
Control group (CK): PDA medium.
Treatment group (T1): sterile fermentation broth of Sauropus turcz Clonostachys chloroleuca Cc620 was mixed with 2 x PDA medium at 1: mixing uniformly in proportion of 1.
20ML of the corresponding culture medium was poured into a culture dish with a diameter of 90mm, fusarium graminearum PH-1 bacterial cake was inoculated at the center of the plate, each treatment was repeated 3 times, and after 3d of culture at 25 ℃, photographing and measurement were performed, and experimental results are shown in FIG. 1 and FIG. 2.
Colony diameters were measured by the crisscross method, and the average value thereof was taken. And calculating the hypha growth inhibition rate according to a formula.
Colony diameter (mm) =average value of three diameters-5.0 (cake diameter)
Hypha growth inhibition = (control colony diameter-treated colony diameter bacteria)/control colony diameter x 100%.
As shown in fig. 1 and 2, the colony diameter of fusarium graminearum in the treated group was significantly smaller than that of the control group. The colony diameter of T1 is calculated to be 29.2+/-1.0 mm, and the hypha growth inhibition rate is 54.31 +/-1.43%. The sterile fermentation broth of the verruca emeraldine Clonostachys chloroleuca Cc620,620 can obviously inhibit the growth of Fusarium graminearum.
Example 2
This example describes the test of the husk of Fusarium emeraldine Clonostachys chloroleuca Cc620,620 parasitic to Fusarium graminearum.
Fusarium graminearum PH-1 is inoculated on a carrot culture medium flat plate with the diameter of 60mm and placed in a 25 ℃ incubator to be cultured for 5-6 d. And (5) when the hyphae grow fully.
The preparation method of the carrot culture medium comprises the following steps: 250g of fresh carrot is peeled and cut into small pieces, 600mL of distilled water is added, and the fresh carrot is sterilized by moist heat at 121 ℃ for 10min. Crushing carrot in a crusher, adding 20g of agar and supplementing water to 1L, and finally carrying out damp-heat sterilization at 121 ℃ for 35min for later use.
Control group (CK): 600 μl of Tween-20 solution with a volume fraction of 0.1% was added dropwise, and the mycelia were pressed.
Treatment group 1 (T1): 600. Mu.L of spore suspension of the strain Cuiyusa Clonostachys chloroleuca Cc620 prepared with Tween-20 at a volume fraction of 0.1% and having a concentration of 1X 10 6 pieces/mL was added, and the mixture was uniformly dropped on the surface of the mycelia to wet the mycelia entirely.
The mycelia of Fusarium graminearum PH-1 are gently poured onto the carrot culture medium by a sterilizing medicine spoon, and the redundant mycelia are removed by a toothpick. Placing in a black light lamp incubator with a light-dark alternation period of 12h to 12h for culturing for 7d at 25 ℃.3 replicates were set for each group, and the number of ascus shells and the parasitic rate of the ascus shells parasitized by the verdant polyporus were counted after 8-10 days and photographed. The test results are shown in FIG. 3.
As shown in FIG. 3, the left side of the graph in T1 shows the results of culturing in carrot culture medium plates, the middle graph shows the photographed results of magnifying the carrot culture medium plates under a stereoscopic vision by 11.2 times, and the right side of the graph shows the photographed results of magnifying the carrot culture medium plates under a fluorescence microscope by 20 times. The results indicate that verruca emeraldine Clonostachys chloroleuca Cc620,620 is capable of parasitizing fusarium graminearum shells. The average parasitic ascal amount per 100 ascal in the T1 group was 76.67±3.51, and the parasitic ratio was 76.67%. The verruca emeraldine Clonostachys chloroleuca Cc620,620 is shown to be effective in parasitizing Fusarium graminearum.
Example 3
This example describes the inhibition of the formation of aschersonia aleyrodis Clonostachys chloroleuca Cc620,620 on wheat and corn stover.
Fresh wheat stalks and corn stalks were cut into 4cm long sections, soaked with 75% alcohol for 2min and rinsed three times with sterile water, followed by wiping off the surface moisture with sterile filter paper.
Preparation of the suspension of the spores of Saurosporine Clonostachys chloroleuca Cc620,620 was carried out as in example 1.
The preparation of Fusarium graminearum PH-1 spore suspension was the same as in example 1, except that the concentration of Fusarium graminearum PH-1 spore suspension was 2X 10 5 pieces/mL.
Control group (CK): the fresh wheat stalks and the corn stalks are respectively placed in 20mL of fusarium graminearum PH-1 spore suspension, 25 ℃ and 90rpm, and after shaking culture for 1h, the fresh wheat stalks and the corn stalks are taken out and placed in a sterile culture dish for culture for 8h.
Treatment group 1 (T1): placing the fresh wheat stalks in 20mL of the emerald Clonostachys chloroleuca Cc spore suspension, shaking at 25 ℃ and 90rpm for 1 hour, taking out, placing in a sterile culture dish, placing in 20mL of the fusarium graminearum PH-1 spore suspension after 48 hours, shaking at 25 ℃ and 90rpm for 1 hour, taking out, and placing in the sterile culture dish for culturing for 8 hours; the corn stalks are treated as fresh wheat stalks.
Treatment group 2 (T2): placing the fresh wheat stalks in 20mL of fusarium graminearum PH-1 spore suspension at 25 ℃ and 90rpm, shaking and culturing for 1h, taking out, placing in a sterile culture dish, placing in 20mL of fusarium graminearum Clonostachys chloroleuca Cc spore suspension with the concentration of 1X 10 6 pieces/mL after 48h, shaking and culturing for 1h at 25 ℃ and 90rpm, taking out, and placing in the sterile culture dish for culturing for 8h; the corn stalks are treated as fresh wheat stalks.
Equal amount (2 g) of sterilized vermiculite is paved on a culture dish with the diameter of 60mm, 3mL of sterile water is added for moisturizing, then treated fresh wheat and corn straw are put into the culture dish, and the result is counted after outdoor culture for 6 weeks. The experimental results are shown in FIG. 4.
As can be seen from fig. 4, the number of ascal shells on fresh wheat stalks and corn stalks of the treatment groups 1,2 is significantly lower than that of the control group. The verdant polyporus Clonostachys chloroleuca Cc,620 has strong colonization ability on wheat and corn stalks, application of verruca emeraldsis Clonostachys chloroleuca Cc620 either before or after fusarium graminearum inoculation significantly inhibits fusarium graminearum capsule shell formation.
As described above, the basic principles, main features and advantages of the present invention are better described. The above examples and description are merely illustrative of preferred embodiments of the present invention, and the present invention is not limited to the above examples, and various changes and modifications to the technical solution of the present invention should be made by those skilled in the art without departing from the spirit and scope of the present invention, and all the changes and modifications fall within the scope of the present invention defined by the present invention.
Claims (4)
1. The application of the verruca emeraldine in preventing and controlling wheat scab is characterized in that the verruca emeraldine is verruca emeraldine Clonostachys chloroleuca Cc620,620, and the preservation number is CGMCC No.41029;
The verruca viridis is applied to crop straws, and the primary infection bacterial source of wheat scab is reduced by inhibiting the growth of Fusarium graminearum and sexual reproduction.
2. The use according to claim 1, wherein the fermentation broth of verruca delphinii inhibits growth of fusarium graminearum.
3. The use according to claim 1, characterized in that said verruca emeralda parasitica is of fusarium graminearum.
4. The use according to claim 1, wherein the verruca emeraldine inhibits the formation of fusarium graminearum shells.
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