Application of bacillus wurrensis preparation in prevention and control of acer truncatum tortoise shell disease
By using Bacillus Vares preparation, combined with biomass charcoal and specific auxiliary materials, the prevention and treatment problems of Yuanbao Maple Turtle shell disease are solved, and efficient biological control effects are achieved, which is significantly better than chemical methods and provides a brand new bio-drug agent .
Patent Information
- Application Number
- CN202411218600.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-02
- Publication Date
- 2024-12-13
AI Technical Summary
The existing technology cannot effectively prevent and treat Yuanbao Maple Turtle shell disease. Chemical control methods have drug resistance and environmental pollution problems, and there is a lack of microbial control methods for this disease.
Bacillus Valez preparations, including Bacillus Valez fermentation broth, biomass charcoal as carriers and peptone, yeast extract, dihydrogen phosphate, and sodium alginate as auxiliary materials, were sprayed on the trunk of Yuanbao maple to prevent the turtle shell sick.
Bacillus Vares preparation significantly improved its prevention effect on Yuanbao Maple Turtle shell disease, reaching 98.3%, better than the commonly used chemical preparation triazolone, and reduced Bacillus death by providing sufficient nutrients and enhancing adhesion. The rate of prevention and control has been significantly improved.
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Abstract
Description
Technical Field
[0001] The invention relates to the field of biological control, and in particular to application of a Varese bacillus preparation in preventing and controlling Acer truncatum tortoise shell disease. Background Art
[0002] Acer truncatum Bunge, also known as flat maple, is a deciduous tree of the genus Acer in the family Aceraceae. Acer truncatum Bunge is 8-10m tall, with longitudinal fissures in the bark; single leaves, opposite leaves, 5 main veins, palm-shaped; petioles 3-5cm long, umbels terminal, yellow-green flowers; flowering in May, fruiting in September. The bark of Acer truncatum Bunge is grayish yellow, with shallow longitudinal fissures, and the twigs are grayish yellow (twigs of one-year-old trees are tender green), smooth and hairless. Acer truncatum is tolerant to partial shade, cold, relatively wind-resistant, and intolerant to dry heat and strong sunlight. The cultivation of Acer truncatum needs attention. It can be planted in late autumn, early winter, and spring. It is better to plant after the soil thaws in spring. Acer truncatum Bunge will face the threat of various diseases during its growth, mainly including maple bubble disease, maple tortoise shell disease, smut, and powdery mildew.
[0003] Maple tortoise shell disease is a common bark disease of Acer truncatum. After the disease occurs, smooth or compressed depressions will form on the bark surface of Acer truncatum, which looks like a tortoise shell, so it is named maple tortoise shell disease. At present, the cause of maple tortoise shell disease is still unclear, but after infection with the disease, the wood tissue of Acer truncatum will be destroyed, and the trunk of Acer truncatum will also be broken, eventually leading to the death of Acer truncatum.
[0004] At present, the main method of preventing and controlling maple shell disease is chemical prevention and control, such as using triadimefon for prevention and control during the peak period of the disease. Due to the singleness of its chemical control drugs, the prevention and control effect of triadimefon on maple shell disease has been seriously reduced. Biological control has become the mainstream prevention and control method because of its advantages of not easily producing drug resistance, no pollution to the environment and good prevention effect. However, there is currently no microorganism that can target maple shell disease. Therefore, it is very necessary to find a microorganism that can effectively prevent and control maple shell disease. Summary of the invention
[0005] The purpose of the present invention is to provide an application of a Bacillus Varese preparation in preventing and controlling the tortoise shell disease of Acer truncatum to solve the problems existing in the above-mentioned prior art. The present invention firstly discovers that Bacillus Varese can effectively prevent and control the tortoise shell disease of Acer truncatum.
[0006] To achieve the above object, the present invention provides the following solutions:
[0007] The invention provides application of Bacillus velezensis in preventing and treating Acer truncatum tortoise shell disease.
[0008] The invention provides a biological preparation for preventing and treating Acer truncatum tortoise shell disease. The biological preparation comprises a fermentation liquid of Bacillus varese, a carrier and auxiliary materials.
[0009] Preferably, the mass ratio of the Bacillus Varezius fermentation broth, the carrier and the auxiliary material is (3-5):2:1.
[0010] Preferably, the carrier is biochar.
[0011] Preferably, the auxiliary materials are peptone, yeast extract, dihydrogen phosphate and sodium alginate.
[0012] Preferably, the mass ratio of the peptone, the yeast extract, the dihydrogen phosphate and the sodium alginate is 2:1:1:1.
[0013] The present invention provides a method for preparing the above-mentioned biological agent, comprising the following steps:
[0014] inoculating the Varese Bacillus into a fermentation medium for fermentation culture to obtain a Varese Bacillus fermentation liquid;
[0015] The biological preparation is obtained by mixing the Varese Bacillus fermentation liquid, the carrier and the auxiliary material.
[0016] Preferably, the fermentation medium is LB medium; the fermentation culture condition parameters are: 35°C, 150rpm, fermentation culture for 48-72h;
[0017] The mass ratio of the Varese Bacillus fermentation broth, the carrier and the auxiliary material is (3-5):2:1.
[0018] The present invention provides application of the above biological preparation in preventing and treating Acer truncatum tortoise shell disease.
[0019] The invention provides a method for preventing and treating Acer truncatum tortoise shell disease, comprising the following steps:
[0020] The above biological preparations were sprayed on the trunks of Acer truncatum trees.
[0021] The present invention discloses the following technical effects:
[0022] The present invention first discovered that Bacillus Varese can effectively prevent and treat the tortoise shell disease of Acer truncatum. Through verification by the examples of the present invention, it can be seen that the prevention effect of Bacillus Varese on the tortoise shell disease of Acer truncatum reaches 98.3%, which is significantly better than the commonly used chemical agent triadimefon. It can be seen that Bacillus Varese can be used for the prevention and treatment of the tortoise shell disease of Acer truncatum.
[0023] At the same time, the present invention also provides a biological agent for preventing and treating the tortoise shell disease of Acer truncatum, which comprises a fermentation liquid of Bacillus Varese, a carrier and an auxiliary material. Using biochar as a carrier enhances the control effect of Bacillus Varese; while using peptone, yeast extract, dihydrogen phosphate and sodium alginate as auxiliary materials, sufficient nutrients can be provided for Bacillus Varese, ensuring the reproduction of Bacillus Varese, reducing the mortality rate of Bacillus Varese, and thus ensuring the control effect of Bacillus Varese on the tortoise shell disease of Acer truncatum. The present invention provides a new biocontrol agent for preventing and treating the tortoise shell disease of Acer truncatum, and it has a wide source, simple preparation and broad application prospects. DETAILED DESCRIPTION
[0024] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.
[0025] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.
[0026] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.
[0027] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.
[0028] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.
[0029] Unless otherwise specified, the components and culture medium used in the present invention are all purchased routinely by those skilled in the art.
[0030] Example 1
[0031] The activated Bacillus Varese FZB42 (the strain was disclosed in "Research on the Antagonism of Bacillus Varese FZB42 against Two Tree Pathogenic Fungi and Its Mechanism", and promised to be released to the public for 20 years) was inoculated into LB medium for fermentation to obtain Bacillus Varese fermentation liquid. The effective viable bacteria count of Bacillus Varese in the fermentation liquid was 3.4×10 8 CFU / mL; the inoculation volume was 5% of the volume of the LB medium, the fermentation time was 54h, the temperature was 35°C, and the rotation speed was 150rpm;
[0032] The fermentation broth of Bacillus varese, the carrier (biomass carbon) and the auxiliary materials (peptone, yeast extract, dihydrogen phosphate and sodium alginate) are mixed in a mass ratio of 3:2:1 to obtain a biological preparation for preventing and treating the tortoise shell disease of Acer truncatum, wherein the mass ratio of peptone, yeast extract, dihydrogen phosphate and sodium alginate is 2:1:1:1.
[0033] Example 2
[0034] The steps are the same as those in Example 1, except that the fermentation liquid of Bacillus varezii, the carrier and the auxiliary materials are mixed in a mass ratio of 4:2:1.
[0035] Example 3
[0036] The steps are the same as those in Example 1, except that the fermentation liquid of Bacillus varezii, the carrier and the auxiliary materials are mixed in a mass ratio of 5:2:1.
[0037] Comparative Example 1
[0038] The steps are the same as those in Example 3, except that the fermentation broth of Bacillus varezii is replaced with Bacillus velez TCS001, which has an inhibitory effect on a variety of plant pathogenic fungi and is disclosed in "Identification and antifungal activity of Bacillus velez TCS001 from marine habitats".
[0039] Comparative Example 2
[0040] The steps are the same as those in Example 3, except that vermiculite is used as the carrier.
[0041] Comparative Example 3
[0042] The steps are the same as those in Example 3, except that LB solid culture medium powder is used as a carrier.
[0043] Comparative Example 4
[0044] The steps are the same as those in Example 3, except that the auxiliary materials are maltodextrin, sodium dodecyl sulfate, dihydrogen phosphate and sodium alginate; and the mass ratio of maltodextrin, sodium dodecyl sulfate, dihydrogen phosphate and sodium alginate is 2:1:1:1.
[0045] Comparative Example 5
[0046] Triadimefon was used as a control agent.
[0047] Comparative Example 6
[0048] The steps are the same as those in Example 1, except that the fermentation liquid of Bacillus varezii, the carrier and the auxiliary materials are mixed in a mass ratio of 2:2:1.
[0049] Comparative Example 7
[0050] The steps are the same as those in Example 1, except that the fermentation liquid of Bacillus varezii, the carrier and the auxiliary materials are mixed in a mass ratio of 7:2:1.
[0051] Experimental example
[0052] 1. Experimental methods
[0053] Acer truncatum forest that had suffered from maple shell disease in the previous year was selected for the experiment. The experiment was divided into 11 groups. Groups 1-10 were sprayed with the biological preparations or control agents for preventing and controlling the maple shell disease prepared in Examples 1-3 and Comparative Examples 1-7, respectively. Specifically, the biological preparations or control agents for preventing and controlling the maple shell disease were diluted 500 times and then sprayed on the trunks of the maple until droplets were left on the surface of the trunks. The spraying period was the leaf germination period, and the spraying was performed once every 15 days, for a total of 4 times. The experimental area of each treatment was 100m 2 , each treatment was repeated 3 times; group 11 was the water control. All experimental plots adopted the same management measures, including appropriate water and fertilizer management, pruning and cleaning, and conventional methods for comprehensive disease prevention and control except maple tortoise shell disease.
[0054] 2. Data investigation
[0055] Five months after the experiment, the incidence rate was counted, and the disease rate was recorded in units of plants. According to the disease grading standards, the number of diseased plants with root rot at each level was counted, and the disease index was calculated. The specific grading standards are shown in Table 1, and the survival rate of Acer truncatum in each treatment was counted. The calculation formulas for the disease index, incidence rate, and survival rate are as follows:
[0056] Disease index = Σ(number of diseased plants at each level × relative level value) / (total number of plants surveyed × highest level representative value) × 100%;
[0057] Incidence rate (%) = number of diseased plants / total number of plants investigated × 100%;
[0058] Survival rate (%) = number of surviving plants / total number of surveyed plants × 100%.
[0059] Table 1 Maple shell disease classification standard
[0060]
[0061]
[0062] 3 Experimental results
[0063] The survey results of the morbidity, disease index and survival rate of different treatments are shown in Table 2.
[0064] Table 2 Statistics on the incidence, disease index and survival rate of Acer truncatum shell disease
[0065] Grouping Incidence (%) Disease index Survival rate (%) Example 1 5.7 3.43 98.7 Example 2 4.8 2.55 99 Example 3 3.1 1.24 100 Comparative Example 1 34.2 7.31 43.2 Comparative Example 2 25.6 5.29 40.4 Comparative Example 3 19.5 4.22 57.8 Comparative Example 4 17.6 3.95 60.5 Comparative Example 5 10.7 5.88 89.3 Comparative Example 6 7.2 3.17 90.8 Comparative Example 7 7.5 3.36 91.3 Clear water control 58.9 11.39 39.2
[0066] As can be seen from Table 2, the biological preparations for preventing and controlling the tortoise shell disease of Acer truncatum prepared in Examples 1-3 of the present invention can significantly control the occurrence and progression of the tortoise shell disease of Acer truncatum, reduce the incidence and disease index of the tortoise shell disease of Acer truncatum, and improve the survival rate of planted Acer truncatum, which is significantly better than the commonly used fungicide triadimefon.
[0067] Combined with Comparative Example 1, it can be found that although Bacillus Velezii TCS001 also has an inhibitory effect on a variety of plant pathogenic fungi, it cannot effectively prevent and treat the tortoise shell disease of Acer truncatum. Combined with Comparative Examples 2-4, it can be seen that replacing other carriers or excipients with the same effect will also reduce the control effect of Bacillus Varezii on the tortoise shell disease of Acer truncatum. In order to further prove the effect of Bacillus Varezii, it was compared with the control effect of the chemical agent triadimefon. The control effect and survival rate of Bacillus Varezii are much higher than those of triadimefon, and it can also effectively reduce the disease index. Moreover, in order to study whether the dosage of the fermentation liquid of Bacillus Varese has an effect on the control effect, the present invention further set up comparative examples 6 and 7. The results show that the dosage of the fermentation liquid of Bacillus Varese also has a great influence on the control effect. The small dosage of the fermentation liquid of Bacillus Varese may be due to the fact that the number of Bacillus Varese cannot occupy an advantage, resulting in a reduced effect; the dosage of the fermentation liquid of Bacillus Varese may be too much, and the carrier and the auxiliary materials may not be able to effectively attach the Bacillus Varese, and cannot meet the normal growth of the Bacillus Varese, thereby resulting in a reduced control effect.
[0068] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.
Claims
1. Application of Bacillus velezensis in the prevention and treatment of Acer truncatum tortoise shell disease.
2. A biological agent for preventing and treating Acer truncatum tortoise shell disease, characterized in that: The biological preparation comprises Bacillus Varese fermentation liquid, a carrier and auxiliary materials.
3. The biological agent according to claim 2, characterized in that The mass ratio of the Varese Bacillus fermentation broth, the carrier and the auxiliary material is (3-5):2:
1.
4. The biological agent according to claim 3, characterized in that The carrier is biochar.
5. The biological agent according to claim 3, characterized in that The auxiliary materials are peptone, yeast extract, dihydrogen phosphate and sodium alginate.
6. The biological agent according to claim 5, characterized in that The mass ratio of the peptone, the yeast extract, the dihydrogen phosphate and the sodium alginate is 2:1:1:
1.
7. The method for preparing a biological agent according to any one of claims 2 to 6, characterized in that: The following steps are involved: inoculating the Varese Bacillus into a fermentation medium for fermentation culture to obtain a Varese Bacillus fermentation liquid; The biological preparation is obtained by mixing the Varese Bacillus fermentation liquid, the carrier and the auxiliary material.
8. The preparation method according to claim 7, characterized in that: The fermentation medium is LB medium; the fermentation culture condition parameters are: 35°C, 150rpm, fermentation culture for 48-72h; The mass ratio of the Varese Bacillus fermentation broth, the carrier and the auxiliary material is (3-5):2:
1.
9. Use of the biological agent according to any one of claims 2 to 6 in preventing and treating Acer truncatum tortoise shell disease.
10. A method for preventing and treating Acer truncatum tortoise shell disease, characterized in that: The following steps are involved: The biological agent according to any one of claims 2 to 6 is sprayed on the trunk of the Acer truncatum tree.