Method for promoting growth of beauveria bassiana hyphae
By adding arginine to the culture medium of leucorrhizalis, the rapid growth of mycelium is promoted, the problem of inefficient pest control in the prior art is solved, and the efficiency of pest control is improved.
Patent Information
- Application Number
- CN202510562676.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-30
- Publication Date
- 2025-07-22
AI Technical Summary
The prior art is difficult to effectively promote the rapid growth of the mycelium of Cyclopsia leukobassium in insects, resulting in inefficient pest control.
By preparing a culture medium containing 8mM or 10mM arginine, combined with PDA medium, it promotes the culture of the spores of the white coccidioida under specific conditions to achieve rapid growth of the mycelium.
It significantly promotes the growth of the mycelium of Cyperus leucorrhea, improves the efficiency of pest control, and shortens the time of pest death.
Smart Images
Figure CN120349955A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of agricultural microorganisms, and specifically relates to a method for promoting the mycelial growth of Beauveria bassiana. Background Art
[0002] Beauveria bassiana is an entomopathogenic fungus that can parasitize more than 700 insects and mites in 15 orders such as Lepidoptera and Hymenoptera, including common agricultural and forestry pests such as corn borers, pine caterpillars, grubs, and thrips. It is widely used for the control of pests on barley, vegetables (such as eggplants and tomatoes), fruit trees (such as apples and lychees), and forest trees (such as pine trees and poplar trees). The infection of insects by Beauveria bassiana mostly starts with the attachment of aerial conidia to the insect body surface. Then, the conidia germinate to produce germ tubes by means of nutrients and moisture on the host body surface, and at the same time secrete protease, chitinase, and lipase. Under the action of mechanical pressure and enzyme degradation at the hyphal tip, the insect cuticle is damaged, and the hyphae enter the insect hemocoel. Then, the hyphae absorb the water and nutrients of the host to grow rapidly and produce blastospores. The blastospores continue to grow hyphae to invade various organs of the insect, gradually filling the entire hemocoel, and at the same time secrete various toxins such as beauvericin and oosporein to block the insect immune system, damage the insect nerve conduction pathway, cause changes in the physical and chemical properties of the insect blood, metabolic disorders, and ultimately die due to the inability to carry out normal life activities. In view of the process of Beauveria bassiana infecting insects, by promoting the growth and extension of hyphae in the insect body, the death of pests can be accelerated and the loss of plants damaged by them can be reduced. Summary of the Invention
[0003] To solve the above technical problems, the present invention provides a method for promoting the mycelial growth of Beauveria bassiana, including the steps of:
[0004] S1: Prepare a PDA medium and a medium containing arginine;
[0005] S2: Prepare a Beauveria bassiana spore suspension. Heat and melt the PDA medium obtained in step S1, place it to cool to 50 °C, pour it into a petri dish. After solidification, inoculate the Beauveria bassiana spores at the center of the plate medium and culture it under the conditions of a temperature of 27 ± 1 °C and a relative humidity of 70 - 80%.
[0006] Further, in step S1, the preparation steps of the PDA medium are: Weigh 8.2 g of potato dextrose agar medium (PDA) powder into a conical flask, add 200 ml of ultrapure water, heat it to melt in a microwave oven, dispense it into 250 ml conical flasks, wrap it with a sealing film and put it into an autoclave, sterilize it at 121 °C for 20 min, take it out and cool it for standby.
[0007] Further, in step S1, the preparation steps of the arginine-containing medium are as follows: Weigh 8.2 g of potato dextrose agar medium (PDA) powder into a conical flask, add 200 ml of ultrapure water, heat it in a microwave oven until melted, weigh 0.27872 g of arginine or 0.03484 g of arginine, add it to the above 200 ml of PDA solution, wrap it with a sealing film and put it into an autoclave, sterilize it at 121 °C for 20 min, take it out and cool it for later use.
[0008] In summary, the present invention has the following beneficial effects compared with the prior art:
[0009] By adding 8 mM or 10 mM arginine, the present invention achieves the purpose of rapid growth of Beauveria bassiana hyphae. Description of the Drawings
[0010] The drawings described herein are used to provide a further understanding of the present invention, and constitute a part of the present invention. The schematic embodiments of the present invention and their descriptions are used to explain the present invention, and do not constitute an improper limitation to the present invention. In the drawings:
[0011] Figure 1 are the colonies of Beauveria bassiana cultured at different arginine concentrations;
[0012] Figure 2 is the effect of 10 mM arginine on the hyphal growth of Beauveria bassiana. Detailed Embodiments
[0013] It should be noted that, without conflict, the embodiments in the present invention and the features in the embodiments can be combined with each other. The present invention will be described in detail below with reference to the drawings and in conjunction with the embodiments.
[0014] It should be noted that the terms used herein are only for describing specific embodiments, and are not intended to limit the exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form can also include the plural form. In addition, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components and / or their combinations.
[0015] Unless otherwise specifically stated, the relative arrangements of components and steps, numerical expressions, and numerical values set forth in these embodiments do not limit the scope of the present invention. At the same time, it should be understood that for the sake of description, the dimensions of the various parts shown in the drawings are not drawn in actual proportional relationships. Technologies, methods, and devices known to those of ordinary skill in the relevant art may not be discussed in detail, but where appropriate, the technologies, methods, and devices should be regarded as part of the authorization specification. In all the examples shown and discussed here, any specific value should be construed as merely exemplary and not as a limitation. Therefore, other examples of the exemplary embodiments may have different values. It should be noted that similar reference numerals and letters denote similar items in the following drawings, and thus, once an item is defined in one drawing, it does not need to be further discussed in subsequent drawings.
[0016] See Figure 1 As shown, the present invention provides a method for increasing the sporulation amount of Beauveria bassiana, comprising the steps of:
[0017] S1: Prepare a PDA medium and a medium containing arginine.
[0018] Prepare the PDA medium: Weigh 8.2 g of potato dextrose agar medium (PDA) powder into a conical flask, add 200 ml of ultrapure water, heat it in a microwave oven until melted, dispense it into 250-ml conical flasks, wrap it with a sealing film, and put it into an autoclave. Sterilize it at 121 °C for 20 min, take it out and cool it for later use.
[0019] Prepare the medium containing arginine: Weigh 8.2 g of potato dextrose agar medium (PDA) powder into a conical flask, add 200 ml of ultrapure water, heat it in a microwave oven until melted, weigh 0.27872 g of arginine or 0.03484 g of arginine, add it to the above 200-ml PDA solution, wrap it with a sealing film, and put it into an autoclave. Sterilize it at 121 °C for 20 min, take it out and cool it for later use.
[0020] S2: Culture Beauveria bassiana. Heat and melt the above PDA medium and the medium containing arginine, let it cool to about 50 °C and pour it into a petri dish. After solidification, inoculate a 10 4 spores / ml Beauveria bassiana spore suspension at the center of the plate medium, and culture it under the conditions of a temperature of (27 ± 1) °C and a relative humidity of 70-80%.
[0021] After inoculation, the growth of Beauveria bassiana colonies and hyphae can be seen Figure 1 . From Figure 1It can be seen that on the 15th day after inoculation, white colonies were visible on both the control group and the medium treated with arginine, and the hyphae were dense; the colonies on the medium containing 8 mM and 10 mM arginine were significantly larger than those in the control group. This indicates that the addition of 8 mM and 10 mM arginine promoted the hyphal growth of Beauveria bassiana. Judging from the colony size, the promoting effect of 10 mM arginine on hyphal growth was stronger.
[0022] The colony diameters were measured, and the results showed that from the 5th day, the colony diameter of the 10 mM arginine group was significantly larger than that of the control group (p > 0.05). This indicates that the promotion of hyphal growth by the addition of 10 mM arginine mainly occurred after 3 days of inoculation.
[0023] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention can have various modifications and changes. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.
Claims
1. A method for promoting the mycelial growth of Beauveria bassiana, characterized in that, Including the steps: S1: Prepare PDA medium and arginine-containing medium; S2: Prepare Beauveria bassiana spore suspension. Heat and melt the PDA medium obtained in step S1, place it to cool to 50 °C, pour it into a petri dish, and after solidification, inoculate the Beauveria bassiana spores at the center of the plate medium, and culture it under the conditions of a temperature of 27 ± 1 °C and a relative humidity of 70 - 80%.
2. The method for promoting the mycelial growth of Beauveria bassiana according to claim 1, wherein In step S1, the preparation steps of the PDA medium are as follows: Weigh 8.2 g of potato dextrose agar medium (PDA) powder into a conical flask, add 200 ml of ultrapure water, heat it to melt in a microwave oven, dispense it into 250 ml conical flasks, wrap it with a sealing film and put it into an autoclave, sterilize it at 121 °C for 20 min, take it out and cool it for standby.
3. The method for promoting the mycelial growth of Beauveria bassiana according to claim 1, characterized in that, In step S1, the preparation steps of the arginine-containing medium are as follows: Weigh 8.2 g of potato dextrose agar medium (PDA) powder into a conical flask, add 200 ml of ultrapure water, heat it to melt in a microwave oven, weigh 0.27872 g of arginine or 0.03484 g of arginine, add it to the above 200 ml PDA solution, wrap it with a sealing film and put it into an autoclave, sterilize it at 121 °C for 20 min, take it out and cool it for standby.