Compound microbial agent for preventing and treating areca yellows as well as preparation method and application of compound microbial agent

Abstract

The invention discloses a compound microbial agent for preventing and treating areca yellows, the active ingredient of the microbial agent is a functional biological flora, and the functional biological flora comprises rhizosphere colonization guard bacteria, pathogen parasitic attack bacteria, antibiotic producing bacteria, plant growth-promoting bacteria and host original flora which are obtained through coupling symbiotic culture, the invention relates to the technical field of agricultural biotechnology and plant disease prevention and control, internal functions of flora are clearly divided, a three-dimensional prevention and control network is formed through the synergistic effect of multiple ways of ecological niche competition, pathogen parasitism, antibacterial substance secretion, plant growth promotion and the like, the effect is obviously superior to that of a single fungicide, and the cost is low. Through coupled culture with an original flora of a rhizosphere host of the betel nut, the colonization competitiveness and durability of the functional flora in a target environment are remarkably improved, the stability of the control effect is ensured, pathogens are only directly inhibited, the tree vigor is enhanced through a growth promoting effect, and the disease resistance of the betel nut is improved.

Description

Technical Field

[0001] This invention relates to the fields of agricultural biotechnology and plant disease control technology, specifically to a compound microbial agent for controlling areca nut yellowing disease, its preparation method, and its application. Background Technology

[0002] Areca nut yellowing disease is a devastating phytoplasmic disease, often referred to as the "cancer" of the areca nut industry. Its main symptoms include yellowing leaves, shrunken canopy, reduced fruit yield and quality, and ultimately, plant death. The disease spreads rapidly and is extremely difficult to control. Currently, there are no effective treatments available in production, and control relies mainly on removing diseased plants to manage the spread of the pathogen. This not only causes significant economic losses but also fails to fundamentally curb the disease's spread. Chemical pesticides have minimal effect on phytoplasmic diseases, and long-term use leads to pesticide residues, environmental pollution, and the development of pesticide resistance in pathogens.

[0003] In recent years, biocontrol using beneficial microorganisms has been considered a promising green control strategy. However, existing microbial agents are mostly single-species or simple compound formulations, generally suffering from limited functionality, poor colonization in complex soil environments, and weak synergistic effects with native microorganisms. Consequently, their control efficacy against areca nut yellowing disease, a systemic disease, is unstable and unsatisfactory. Therefore, there is an urgent need to develop a compound microbial preparation that can specifically target areca nut yellowing disease, utilize multi-species synergy, and is highly adapted to the areca nut rhizosphere environment, along with its application method. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a compound microbial agent for the prevention and control of areca nut yellowing disease, its preparation method, and its application, solving the problems of unstable and unsatisfactory prevention and control effects of existing technologies on areca nut yellowing disease.

[0005] To achieve the above objectives, the present invention is implemented through the following technical solution: a compound microbial agent for preventing and treating areca nut yellowing disease, wherein the active ingredient of the agent is a functional biological community, which includes rhizosphere colonizing and protecting bacteria, pathogenic parasitic and attacking bacteria, antibiotic-producing bacteria, plant growth-promoting bacteria, and host original microbiota obtained through coupled symbiotic culture.

[0006] Preferably, based on the total viable count of the functional microbial community, its microbial composition includes: Rhizosphere colonization and protection bacteria, with a content of 50%-60%; The content of pathogenic parasitic attacking bacteria is 20%-30%; Antibiotic-producing bacteria, with a content of 10%-15%; Plant growth-promoting bacteria, with a content of 5%-10%; The original host flora, its content is 1%-5%.

[0007] Preferably, the rhizosphere colonizing and protecting bacteria include Bacillus subtilis and / or Bacillus amyloliquefaciens; the pathogenic parasitic attacking bacteria include Trichoderma harzianum and / or Trichoderma viride; the antibiotic producing bacteria include Streptomyces jingyangensis and / or Streptomyces griseus; and the plant growth promoting bacteria include Bacillus mucilaginosus and / or Bacillus megaterium.

[0008] Preferably, the original host microbiota originates from the rhizosphere soil of a healthy areca nut plantation without a history of areca nut yellowing disease.

[0009] Preferably, the total viable count of the microbial agent is not less than 2 billion / gram.

[0010] The preparation method of the compound microbial agent includes the following steps: (1) Activate and ferment the rhizosphere colonization and protection bacteria, pathogenic parasitic attack bacteria, antibiotic producing bacteria and plant growth promoting bacteria at high density to obtain high concentration bacterial solutions of single strains; (2) The host original microbial community was extracted from the rhizosphere soil of a healthy areca nut plantation to obtain a suspension of the host original microbial community; (3) Mix the various single strain bacterial solutions obtained in step (1) in proportion, and then mix them with the host original bacterial suspension obtained in step (2) for coupled symbiotic culture to obtain a composite functional bacterial community. (4) The composite functional microbial community is mixed with the carrier to prepare a microbial agent.

[0011] Preferably, in step (3), the conditions for the coupled symbiotic culture are: temperature 26-30℃, pH 6.0-7.0, and aeration culture for 60-96 hours.

[0012] The application of the compound microbial agent in the prevention and control of areca nut yellowing disease includes at least one of the following methods: soil treatment, root irrigation, or foliar spraying.

[0013] Preferably, the soil treatment involves applying 20-30 kg of the microbial agent per acre; the root irrigation involves diluting the microbial agent 500-800 times and applying 10-15 liters to each areca nut tree. Beneficial effects

[0014] This invention provides a compound microbial agent for preventing and treating areca nut yellowing disease, its preparation method, and its application, which have the following beneficial effects: Synergistic control through multiple mechanisms: The microbial community has a clear division of functions and works synergistically through multiple pathways such as competing for ecological niches, parasitizing pathogens, secreting antimicrobial substances, and promoting plant growth, forming a three-dimensional control network with significantly better effects than single microbial agents; Strong environmental adaptability: Through coupled culture with the original microbial community of the areca rhizosphere host, the colonization competitiveness and persistence of the functional microbial community in the target environment are significantly improved, ensuring the stability of the control effect; It addresses both the symptoms and the root cause: it not only directly inhibits pathogens, but also enhances tree vigor through growth promotion, improves the disease resistance of areca palms themselves, and improves the overall rhizosphere microecological environment, fundamentally reducing the risk of disease. Green and safe: It is based entirely on biological control, with no chemical pesticide residues, is environmentally friendly, and meets the requirements for agricultural product quality and safety and sustainable agricultural development. Detailed Implementation

[0015] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0016] This invention provides a technical solution: a compound microbial agent for preventing and treating areca nut yellowing disease, wherein the active ingredient of the agent is a functional biological community, which includes rhizosphere colonizing and protecting bacteria, pathogenic parasitic and attacking bacteria, antibiotic-producing bacteria, plant growth-promoting bacteria, and host original microbiota obtained through coupled symbiotic culture.

[0017] This embodiment is further configured such that, based on the total viable number of functional microbial communities, the composition of the microbial species includes: Rhizosphere colonization and protection bacteria, with a content of 50%-60%; The content of pathogenic parasitic attacking bacteria is 20%-30%; Antibiotic-producing bacteria, with a content of 10%-15%; Plant growth-promoting bacteria, with a content of 5%-10%; The original host flora, its content is 1%-5%; [Table 1] Comparison of bacterial strain composition and measured viable count In this embodiment, the rhizosphere colonizing and protecting bacteria include Bacillus subtilis and / or Bacillus amyloliquefaciens; the pathogenic parasitic attacking bacteria include Trichoderma harzianum and / or Trichoderma viride; the antibiotic producing bacteria include Streptomyces jingyangensis and / or Streptomyces griseus; and the plant growth promoting bacteria include Bacillus gelatinosa and / or Bacillus megaterium.

[0018] In this embodiment, the original host microbiota is further configured to originate from the rhizosphere soil of a healthy areca nut plantation without a history of areca nut yellowing disease.

[0019] In this embodiment, the total viable count of the microbial agent is not less than 2 billion / gram; [Table 2] Verification of Total Viable Bacteria Count Testing items Measured value Claim threshold in conclusion Total viable bacteria count 2.23 billion / gram ≥2 billion / gram ✓ Qualified Moisture 4.1 % ≤5% (implicit) ✓ Qualified granularity 90% passing through 100 mesh 80-200 mesh ✓ Qualified The preparation method of the compound microbial agent includes the following steps: (1) Activate and ferment the rhizosphere colonization and protection bacteria, pathogenic parasitic attack bacteria, antibiotic producing bacteria and plant growth promoting bacteria at high density to obtain high concentration bacterial solutions of single strains; (2) The host original microbial community was extracted from the rhizosphere soil of a healthy areca nut plantation to obtain a suspension of the host original microbial community; (3) Mix the various single strain bacterial solutions obtained in step (1) in proportion, and then mix them with the host original bacterial suspension obtained in step (2) for coupled symbiotic culture to obtain a composite functional bacterial community. (4) The composite functional microbial community is mixed with the carrier to prepare a microbial agent.

[0020] In this embodiment, the conditions for the coupled symbiotic culture in step (3) are: temperature 26-30℃, pH 6.0-7.0, and aeration culture for 60-96 hours. Table 3. Comparison of Coupled Culture Parameters Parameter Items Scope of claims Example setting values Compliance temperature 26-30℃ 28℃ ✓ pH 6.0-7.0 6.5 ✓ Ventilation time 60-96 h 72 h ✓ Host microbial community volume ratio 1-5% 3% ✓ The application of the compound microbial agent in the prevention and control of areca nut yellowing disease includes at least one of the following methods: soil treatment, root irrigation, or foliar spraying.

[0021] In this embodiment, the soil treatment involves applying 20-30 kg of the microbial agent per acre; the root irrigation involves diluting the microbial agent 500-800 times and applying 10-15 liters to each areca nut tree.

[0022]

Table 4

Table 5

[0023] It should be noted that, in this document, relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, the phrase "comprising an element defined as..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0024] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A complex microbial agent for controlling yellowing disease of Areca catechu, characterized by, The active ingredient of the microbial agent is a functional microbial flora, which comprises rhizosphere colonization guardian bacteria, pathogenic parasitic attack bacteria, antibiotic-producing bacteria, plant growth-promoting bacteria and host original flora obtained by coupling symbiotic culture.

2. The complex microbial agent according to claim 1, characterized by, The strain composition of the functional microbial flora includes, based on the total number of viable bacteria: rhizosphere colonization guardian bacteria, accounting for 50%-60%; pathogenic parasitic attack bacteria, accounting for 20%-30%; antibiotic-producing bacteria, accounting for 10%-15%; plant growth-promoting bacteria, accounting for 5%-10%; host original flora, accounting for 1%-5%.

3. The complex microbial agent according to claim 2, characterized by, The rhizosphere colonization guardian bacteria include Bacillus subtilis and / or Bacillus amyloliquefaciens; the pathogenic parasitic attack bacteria include Trichoderma harzianum and / or Trichoderma viride; the antibiotic-producing bacteria include Streptomyces jingyangensis and / or Streptomyces griseus; and the plant growth-promoting bacteria include Bacillus mycoides and / or Bacillus megaterium.

4. The complex microbial agent according to any one of claims 1 to 3, characterized in that, The host original flora is derived from the rhizosphere soil of a healthy areca garden without areca yellowing disease history.

5. The complex microbial agent according to any one of claims 1 to 4, characterized in that, The total number of viable bacteria of the microbial agent is not less than 2.0 billion per gram.

6. A method for preparing the complex microbial agent according to any one of claims 1 to 5, characterized by, The method comprises the following steps: (1) activating and high-density fermenting the strains of rhizosphere colonization guardian bacteria, pathogenic parasitic attack bacteria, antibiotic-producing bacteria and plant growth-promoting bacteria respectively to obtain high-concentration bacterial liquid of single strain; (2) extracting host original flora from the rhizosphere soil of a healthy areca garden to obtain host original flora suspension; (3) mixing the high-concentration bacterial liquid of single strain obtained in step (1) in proportion, and then mixing with the host original flora suspension obtained in step (2) to perform coupling symbiotic culture to obtain a compound functional flora; (4) mixing the compound functional flora with a carrier to prepare a microbial agent.

7. The method of claim 6, wherein, In step (3), the coupling symbiotic culture is performed under the following conditions: temperature 26-30℃, pH 6.0-7.0, and aeration culture for 60-96 hours.

8. Application of the compound microbial agent according to any one of claims 1-5 in the prevention and treatment of areca yellowing disease, wherein the application mode comprises at least one of soil treatment, root irrigation or foliar spraying.

9. Use according to claim 8, characterized in that, The soil treatment is to apply 20-30 kg of the microbial agent per mu; and the root irrigation is to dilute the microbial agent by 500-800 times and then apply 10-15 liters per areca tree.