Enterobacter pseudohormaechei and application thereof
By using the Enterobacter homunculus xWc1 strain as a repellent and antifeedant, the problem of diamondback moth damage to cruciferous vegetables was solved, achieving environmentally friendly pest control and protecting crops and human health.
Patent Information
- Application Number
- CN202511074836.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-01
- Publication Date
- 2025-10-28
AI Technical Summary
Existing technologies are insufficient to effectively control the damage caused by diamondback moth to cruciferous vegetables, and traditional chemical pesticides may be harmful to the environment and human health.
Enterobacter hygroscopicus XWc1 was used as a repellent and antifeedant. The bacterial solution was prepared and sprayed onto crop plants to control agricultural pests by utilizing its antifeedant and repellent effects on diamondback moths.
It provides an environmentally friendly biological control method that significantly reduces the area consumed by diamondback moth and the mortality rate, protects crops from pests, and is safe and non-toxic.
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Figure CN120843361A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of agricultural microbial technology, and in particular relates to the application of Enterobacter cholerae XWc1 as a food repellent for agricultural pests. Background Technology
[0002] Repellents and antifeedants are substances that act on the peripheral nervous system of insects to repel them or stop them from feeding. Their direct purpose is to repel pests and protect humans or crops from harm, rather than to kill them. Using repellents achieves the protective goal while avoiding accidental poisoning and allergic reactions. This is significantly different from insecticides, which act on the central nervous system of insects, and offers advantages such as safety and environmental friendliness.
[0003] The diamondback moth is distributed throughout China. The larvae feed on leaves, damaging cruciferous plants such as cabbage, broccoli, bok choy, mustard greens, cauliflower, Chinese cabbage, rapeseed, and radish. Adults are attracted to light, hiding in concealed areas of plants during the day, and can fly short distances low when startled. In North China, there are 4-6 generations per year; in Hefei, 10-11 generations per year; and in Guangdong, 20 generations per year. The oviposition period can last up to 10 days, with an average of 100-200 eggs per female. The incubation period is 3-11 days. The diamondback moth specifically feeds on cruciferous plants, causing the most serious damage and threat to the production and development of cruciferous vegetables, and is also one of the most difficult major pests to control. Summary of the Invention
[0004] The purpose of this invention is to address the shortcomings of existing technologies by providing an application of Enterobacter cholerae XWc1 as a repellent and antifeedant for agricultural pests.
[0005] To achieve the above objectives, the present invention adopts the following technical solution: A species of Enterobacter quasi-Hodgkinella from the beet armyworm has been named Enterobacter quasi-Hodgkinella. Enterobacter quasihormaechei XWc1, depositary institution: Guangdong Provincial Center for Microbial Culture Collection; depositary address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou; deposit date: April 27, 2023; biological accession number: GDMCC No: 63411.
[0006] The present invention contains Enterobacter quasi-Hodgkin's enterobacter. Enterobacter quasihormaechei XWc1 strain forms round, white colonies with a smooth, glossy surface and a semi-transparent, neatly edged appearance. It occurs singly or in chains. The optimal growth temperature is 35℃, and the optimal pH is 6.0-7.0. It is a facultative anaerobe and does not spore.
[0007] The present invention contains Enterobacter quasi-Hodgkin's enterobacter. Enterobacter quasihormaecheiThe XWc1 strain can utilize d-galactose, d-glucose, maltose, d-mannitol, arabinose, melibiose, d-fructose, d-xylose, trehalose, raffinose, sucrose, d-gentiobiose, L-rhamnose, glycerol, and L-sorbitol.
[0008] The present invention contains Enterobacter quasi-Hodgkin's enterobacter. Enterobacter quasihormaechei The XWc1 strain exhibits a significant antifeedant effect on diamondback moths of all ages.
[0009] Another object of the present invention is to provide a *Enterobacter quasi-Hodgkinella*. Enterobacter quasihormaechei Application of XWc1 strain in the preparation of agricultural pest control agents and its application in the control of agricultural pests.
[0010] Preferably, as a preferred embodiment, the agricultural pest is the diamondback moth.
[0011] Another object of the present invention is to provide a method for controlling agricultural pests, wherein the aforementioned Enterobacter cholerae is used. Enterobacter quasihormaechei After the XWc1 strain is prepared into a bacterial solution, it is sprayed directly onto the crop plants.
[0012] Preferably, the bacterial suspension is prepared as follows: *Enterobacter quasi-Hodgkin's* strain is thawed from -80°C. After the strain thaws into a liquid state, 100 μL of the thawed strain is added to a liquid culture medium cooled to room temperature in a clean bench. The liquid culture medium with added strain is then placed in a constant-temperature shaker set at 30°C and 200 rpm for 16 hours to obtain a culture suspension for expansion. A certain amount of bacterial cells is taken from an LB agar plate inoculated with *Enterobacter quasi-Hodgkin's* and added to a liquid culture medium cooled to room temperature in a clean bench. The liquid culture medium with added bacterial cells is then placed in a constant-temperature shaker set at 30°C and 200 rpm for 16 hours to obtain a bacterial suspension for subsequent experiments.
[0013] Compared with the prior art, the beneficial effects of the present invention are as follows: The present invention provides *Enterobacter quasi-Hodgkinella*. Enterobacter quasihormaechei The XWc1 strain can be used as an environmentally friendly green pesticide to replace environmentally harmful agents, including chemical pesticides, to control agricultural pests such as diamondback moth. It provides a new microbial strain and control strategy for the control of pesticide-treated pests. This strain is harmless to the environment, does not affect human health, and has a very good biological control effect. Attached Figure Description
[0014] Figure 1 The present invention illustrates Enterobacter quasi-Hodgkin's enterobacter of the present invention. Enterobacter quasihormaechei Effects of strain XWc1 on feeding of diamondback moths at different instars. Detailed Implementation
[0015] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. The specific embodiments described below further illustrate the present invention.
[0016] Unless otherwise specified, the reagents and materials used in the embodiments of the present invention can all be obtained through commercial means.
[0017] I. Preparation of bacterial culture of Enterobacter quasihormaechei XWc1 strain The *Enterobacter quasi-Hodgkin's* strain was thawed from -80℃. Once the strain had thawed into a liquid state, 100 μL of the thawed inoculum was added to a liquid culture medium cooled to room temperature in a clean bench. The liquid culture medium with inoculum was then placed in a shaker set at 30℃ and 200 rpm for 16 hours to obtain a culture for expansion. A certain amount of bacterial cells was taken from an LB agar plate inoculated with *Enterobacter quasi-Hodgkin's* and added to a liquid culture medium cooled to room temperature in a clean bench. The liquid culture medium with inoculum was then placed in a shaker set at 30℃ and 200 rpm for 16 hours to obtain a culture for subsequent experiments.
[0018] II. Determination of the antifeedant activity of Enterobacter quasihormaechei XWc1 strain bacterial suspension Non-selective refusal to eat activity assay: Cabbage leaves were cut into leaf discs with a diameter of 60 mm, and the leaves were immersed in OD... 600 =1 Enterobacter cholerae bacterial suspension for 30s. The control group was treated with water immersion, then removed and air-dried at room temperature. After air-drying, the leaves were placed in 60mm diameter petri dishes, with 10 diamondback moth larvae of the same instar in each dish. After 48 hours, the leaf feeding area in each petri dish was calculated. Each treatment was repeated 3 times. The leaf feeding area was calculated using IMAGEJ software.
[0019] Selective feeding refusal activity assay: 1 OD tablet 600Cabbage leaves with a diameter of 30 mm treated with a bacterial solution of 1 were placed in a 120 mm diameter glass dish. A 30 mm diameter cabbage leaf treated with water was also placed in the dish, with the two leaves positioned at opposite ends of the same straight line. After 48 hours, the leaves were removed, and the leaf area consumed was calculated. Each treatment contained 15 larvae. The leaf area consumed was calculated using IMAGEJ software.
[0020] Selective antifeeding activity assay using potted plants: The treatment group used cabbage leaves that had grown to the five true leaf stage, with added OD... 600 The entire plant was treated with a solution of *Enterobacter quasi-Horacetamella* (=1). The control group was treated with sterile water from potted cabbage plants grown at the same time. Both groups were sprayed evenly onto the entire plant at 9:00 AM on days 1, 3, and 5, and observations were made at 4:00 PM. On day 7, the final treatment was performed at 9:00 AM, spraying only the undersides of the leaves. Leaf morphology and wilting were observed during this period. After treatment on day 7, the air-dried cabbage plants from both the treatment and control groups, whose growth and development were not significantly affected, were placed in 60*60*60cm nylon mesh cages. One cabbage plant from each group and one from the control group were placed at each diagonal end of each cage. Simultaneously, 50 third-instar diamondback moth larvae were placed in the center of each cage. Each treatment was repeated three times. Three standard leaves were selected from each plant, and observations were made every 24 hours. Feeding on the standard leaves was recorded after 72 hours, and the feeding area was calculated using IMAGEJ.
[0021] OD 600 Table 1 shows the non-selective antifeeding activity of Enterobacter quasi-Horaceae bacterial suspension at a concentration of 1 against diamondback moths of all ages. As can be seen from Table 1, Enterobacter quasi-Horaceae has a significant antifeeding effect on diamondback moths of all ages, and data analysis shows significant differences between the treatment group and the control group.
[0022]
[0023] The selective antifeeding activity of Enterobacter quasi-Hodgkin's bacterial solution against third-instar diamondback moth was determined by the leaf immersion method, as shown in Table 2. The results of selective antifeeding activity showed that Enterobacter quasi-Hodgkin's bacterial solution had antifeeding activity against third-instar diamondback moth larvae, with an antifeeding rate of 72.1%.
[0024]
[0025] The selective antifeeding activity of Enterobacter quasi-Hodgkin's bacterial suspension against third-instar diamondback moth larvae was determined by pot method, as shown in Table 3. The results of selective antifeeding activity showed that Enterobacter quasi-Hodgkin's bacterial suspension had antifeeding activity against third-instar diamondback moth larvae, with an antifeeding rate of 83.8%.
[0026]
[0027] III. Avoidance Test Cut the cabbage leaves into uniform sizes of 60mm in diameter, and place the leaves from the treatment group into the OD. 600 Immerse the leaves in a 1% solution of *Enterobacter hygroscopicus* for 30 seconds, then air-dry at room temperature. Place the air-dried leaves into disposable petri dishes with a diameter of 60 mm, and use cotton plugs to lift the dishes and place them in the center of a 120 mm diameter glass dish. Apply petroleum jelly around the glass dish to prevent larvae from crawling out. Leaves from the control group were immersed in water for 30 seconds, air-dried at room temperature, and then placed in disposable petri dishes with a diameter of 60 mm, serving as the no-food-attracting group. Ten third-instar diamondback moth larvae were placed in each group, and each treatment was repeated three times. Observations were made at 24 h and 48 h. At 48 h, the number of larvae that completely crawled out of the 60 mm diameter petri dish and landed in the glass dish, as well as the number of dead larvae, were recorded.
[0028] Similarly, the treated leaf discs were placed in disposable petri dishes. A leaf treated with water was placed in the center of the glass dish under a cotton plug to serve as a food attraction group. Ten third-instar diamondback moth larvae were placed in each group, and each treatment was repeated three times. Observations were made at 24 h and 48 h. At 48 h, the number of insects that completely crawled out of the 60 mm diameter petri dish and landed on the glass dish, as well as the number of dead larvae, were recorded.
[0029] The experimental results are shown in Table 4. As can be seen from Table 4, when the Caesarium cholerae of the present invention was used to conduct a repellency test on the third instar diamondback moth, the mortality rate of the treatment group was 100%, which was significantly higher than that of the control group.
[0030]
Claims
1. A strain of Enterobacter quasi-Hodgkinella XWc1, characterized in that, This strain was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 27, 2023, with the accession number GDMCC NO:63411.
2. The application of *Enterobacter cholerae* as described in claim 1 in the preparation of agricultural pest control agents.
3. An agricultural pest control agent, characterized in that, It contains the Enterobacter quasi-Hodgkin's XWc1 strain and / or its bacterial culture as described in claim 1.
4. The agricultural pest control agent according to claim 3, characterized in that, The bacterial solution concentration was OD600=1.
5. The application according to claim 2, characterized in that, Protecting crops from pests, the agricultural pest referred to is the diamondback moth.
6. A method for controlling agricultural pests, characterized in that, The Enterobacter quasi-Hodgkin's XWc1 strain described in claim 1 was prepared into a bacterial solution and then sprayed directly onto the plants.