WO2023246208A1 - Use of bacillus velezensis bv-6 in pest prevention - Google Patents
Use of bacillus velezensis bv-6 in pest prevention Download PDFInfo
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- WO2023246208A1 WO2023246208A1 PCT/CN2023/083575 CN2023083575W WO2023246208A1 WO 2023246208 A1 WO2023246208 A1 WO 2023246208A1 CN 2023083575 W CN2023083575 W CN 2023083575W WO 2023246208 A1 WO2023246208 A1 WO 2023246208A1
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- WIPO (PCT)
- Prior art keywords
- bacillus
- control
- aphid
- aphids
- wettable powder
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- 241000193744 Bacillus amyloliquefaciens Species 0.000 title claims abstract description 36
- 241000607479 Yersinia pestis Species 0.000 title claims abstract description 17
- 230000002265 prevention Effects 0.000 title abstract 6
- 241001124076 Aphididae Species 0.000 claims abstract description 94
- 241001556089 Nilaparvata lugens Species 0.000 claims abstract description 33
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- 239000004563 wettable powder Substances 0.000 claims description 83
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- 238000000855 fermentation Methods 0.000 claims description 37
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- 238000002360 preparation method Methods 0.000 claims description 7
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- 239000005906 Imidacloprid Substances 0.000 description 2
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- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/12—Powders or granules
- A01N25/14—Powders or granules wettable
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P7/00—Arthropodicides
- A01P7/02—Acaricides
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P7/00—Arthropodicides
- A01P7/04—Insecticides
Definitions
- the invention belongs to the technical field of biological control, and specifically relates to the application of Bacillus velezensis Bv-6 in insect control.
- Bacillus veleis can affect plant growth and development in two different ways, directly and indirectly.
- directly promoting plant growth it is usually necessary to provide the plant with a compound synthesized by bacteria or to promote the absorption of nutrients from the environment.
- they can fix nitrogen in the atmosphere and provide minerals such as phosphorus to provide nutrients to the plant; synthesis regulation Enzymes for plant growth and development; synthesize siderophores that can provide iron for plant growth; synthesize a variety of plant hormones—including auxin and cytokinin; release volatile compounds, etc. (Brown, 1974; Davison, 1988; Lambert and Joos, 1989 ; Patten and Glick, 1996).
- B. veleis indirectly promotes plant growth when it can reduce or prevent certain harmful effects of plant pathogenic microorganisms through one or more mechanisms (Glick, 1999).
- chemical control is mainly used to control these diseases, insect pests and mites on crops.
- pathogenic bacteria and aphids have developed varying degrees of resistance to traditional pesticides, there is a high risk of the development of pesticide resistance, resulting in reduced control effects and increased
- long-term use of pesticides will lead to pesticide resistance, pesticide residues, damage to soil ecological balance, environmental pollution and other problems (Biurrun et al., 2010; Liu et al., 2014).
- biopesticides are an inevitable demand for social development (Wu et al., 2009; Zhang et al., 2008) .
- the application of beneficial microorganisms as biopesticides in agriculture has received more and more attention over time.
- Bacillus thuringiensis has a certain control effect on chewing pests such as Lepidoptera and Coleoptera (Bravo et al., 2007; Sanahuja et al., 2011 ).
- Bacillus veleis can induce plants to develop systemic resistance to pathogenic bacteria and fungi, causing massive accumulation of hydrogen peroxide in plants, cell death and callose precipitation in leaves, effectively controlling the growth of various pathogenic bacteria and aphids on plants (Rashid et al., 2019).
- the technical problem to be solved by the present invention is to provide a new option for biological insect control.
- the technical solution of the present invention is the application of Bacillus velezensis Bv-6 in insect control.
- the deposit number of Bv-6 is CCTCC NO:M 20191106.
- the pest control is to control sucking pests.
- sucking pests are brown planthoppers, aphids, Diaphorina citri, Bemisia tabaci or/and mites.
- the aphids are green peach aphid (Myzus persicae), cereal aphid (Rhopalosiphum padi), wheat aphid (Sitobion avenae), melon aphid (Aphis gossypii), pea aphid (Megoura japonica), peach Pink aphid (Hyalopterus arundimis) or rose aphid (Macrosiphum rosirvorum).
- the mite is Panonychus citri or Tetranychus urticae.
- the dosage form of Bacillus veleis Bv-6 is a wettable powder.
- the invention also provides an anti-insect preparation, the main component of which is the fermentation broth or wettable powder of Bacillus velezensis Bv-6.
- the preservation number of Bacillus velezensis Bv-6 is CCTCC NO:M 20191106 .
- the concentration of the wettable powder is 1 ⁇ 10 11 cfu/g.
- the Bacillus velezensis used in the present invention is specifically the Bacillus velezensis Bv-6 strain, which was deposited in the China Typical Culture Collection Center on December 25, 2019 (the address is: Wuhan, China Wuhan University), the collection number is: CCTCC No: M20191106.
- the present invention discloses that Bacillus veleis Bv-6 has a control effect on many pests. Specifically, 500-fold solution of Bacillus veleis Bv-6 wettable powder is effective against Rhopalosiphum padi, Sitobion avenae, Myzus persicae, and Megoura. japonica), cucumber aphid (Aphis gossypii), watermelon aphid (Aphis gossypii), pink peach aphid (Hyalopterus arundimis), rose aphid (Macrosiphum rosirvorum) have 100%, 100%, 98.67%, 100%, 98.17 respectively.
- Bacillus Velez Bv-6 fermentation liquid has 92.9% control effect on the mite citrus spider mite (Panychus citri), Bv-6 wettability
- the 24-hour control effect of 500 times of powder on the mites grapefruit spider mites (Panychus citri) and strawberry spider mites (Tetranychus urticae) was 85.2% and 90.71% respectively.
- the 24-hour control effects of Bv-6 wettable powder 500 times and 100 times on citrus psyllid (Diaphorina citri) were 93.27% and 96.77% respectively.
- Bacillus veleis Bv-6 wettable powder has a control effect of 89% on brown planthoppers.
- the 24-hour control effect of Bv-6 wettable powder 500 times on Bemisia tabaci is 85.86%.
- the present invention provides a new option for biological control of pests.
- Figure 1 shows the control effects of different Bacillus species on green peach aphid. From left to right are Bacillus velezensis Bv-6, Bacillus subtilis Bs-1, Bacillus velezensis Bv-12, Bacillus velezensis Strain Bv-10, Bacillus paraanthracis Bp1, Bacillus paramycoides Bp3, Bacillus paramycoides Bp9, Bacillus paramycoides Bp2, LB medium, red arrows mark dead aphids (most of the green peach aphids treated with Bv-6 fermentation broth turned black, shrank and died).
- Figure 2 shows the control effects of different Bacillus species on the cereal aphid. From left to right are LB medium, Bacillus velezensis Bv-6, Bacillus velezensis Bv-10, Bacillus subtilis subtilis) Bs-1, Bacillus velezensis strain Bv-12, Bacillus paraanthracis Bp1, the red arrow marks the dead aphids (grain cones treated with Bv-6 fermentation broth All aphids turn black, shrivel and die).
- Figure 3 shows the control effect of different concentration dilutions of Bacillus veleis Bv-6 wettable powder on the cereal aphid.
- CK is the shape of the aphid after spraying with clean water (body color is wine red, no death occurred). After spraying with Bv-6 wettable powder, the aphid turned black, shrank and died.
- Figure 4 shows the control effect (2d and 7d) of 500-fold solution of Bacillus veleis Bv-6 wettable powder on wheat aphids (Grain constrictor aphid and Wheat aphid) in the field.
- CK is the form of aphids after spraying with clean water (the number of aphids is large, and only a few die).
- Figure 5 shows the control effect (48h) of 500 times solution of Bacillus veleis Bv-6 wettable powder on pea tail-pruning aphid.
- CK is the shape of the pea aphids after spraying with clean water (the body color is dark green, and no death has occurred). After spraying 500 times of Bv-6 powder, the pea aphids all turned black, shrank and died, and Fall off the plant.
- Figure 6 shows the control effect (24h) of Bacillus veleis Bv-6 wettable powder 500 times on rapeseed.
- CK is the shape of the aphids after spraying with clean water (the body color is dark green, and no death occurred). After spraying 500 times of Bv-6 powder, the rape aphids shrank and died, and fell off from the leaves.
- Figure 7 shows the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder on cucumber aphid (48h).
- the number of aphids on the front and back of the cucumber leaves sprayed with clean water was not reduced, and the cucumber leaves wilted due to aphid damage.
- After spraying with 500 times of Bv-6 powder the number of aphids on the front and back of the cucumber leaves was significantly reduced, and the aphids changed. Black and dead, most of the leaves fall off, and the leaves do not wilt.
- Figure 8 shows the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder on watermelon aphid.
- CK is the shape of aphids after spraying with clean water (body color is dark green or yellow-green, no death occurs). After spraying 500 times of Bv-6 powder, most of the watermelon aphids turned black, shrank and died.
- Figure 9 shows the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder on pink aphid on peach trees.
- CK is the shape of aphids after spraying with clean water (the body color is green, and no death occurs). After spraying 500 times of Bv-6 powder, most of the peach pink aphids turned into dark brown, shriveled and died.
- Figure 10 shows the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder on rose tube aphid.
- CK is the shape of the aphids after spraying with clean water (the body color is green, and no death occurred). After spraying 500 times of Bv-6 powder, most of the rose aphids turned dark brown, shriveled and died.
- Figure 11 shows the control effect of Bacillus veleis Bv-6 fermentation broth on citrus Panonychus mites.
- Figure 12 shows the control effect of Bacillus veleis Bv-6 on strawberry two-spotted spider mite.
- the two-spotted spider mites sprayed with Bv-6 wettable powder shrank and died, while the two-spotted spider mites treated with control (water) were normal and did not die.
- Figure 13 shows the control effect of Bacillus veleis Bv-6 on citrus psyllid.
- the white substance in the control (clear water) treatment picture is the excrement of citrus psyllid.
- Figure 14 shows the control effect of Bacillus veleis Bv-6 on cotton whitefly.
- Figure 15 is a schematic diagram of the control effects of different Bacillus species on rice brown planthopper (48h);
- Picture a from left to right shows the spraying of LB medium, Bacillus paranthracis Bp1, Bacillus subtilis Bs-1, Bacillus velezensis Bv-10, Bacillus velezensis Bv-10, Bacillus velezensis strain (Bacillus velezensis) Bv-12, Bacillus velezensis strain (Bacillus velezensis) Bv-6;
- Picture b is a picture of dead brown planthoppers corresponding to each treatment in picture a, in which the number of dead brown planthoppers after spraying Bacillus is Bv-6>Bv-12>Bs-1>Bv-10>Bp-1>LB.
- Figure 16 is a schematic diagram of the results of the control effect of Bacillus veleis Bv-6 on rice brown planthopper of the present invention (8d);
- Figure 17 is a schematic diagram of the control effect of 500-fold dilution of Bacillus veleis Bv-6 wettable powder on rice brown planthopper (48h);
- brown planthoppers After being treated with a 500-fold dilution of Bacillus veleis Bv-6 wettable powder, brown planthoppers died and turned black. However, the brown planthoppers treated with sterile water control did not die or turn black.
- F The back of the insect
- B The abdomen of the insect.
- the Bacillus velezensis of the present invention is Bacillus velezensis Bv-6, and the deposit number is CCTCC NO: M20191106, which has been disclosed in CN 111254086A.
- the wettable powder preparation method is the same as that of the application, and the concentration of the prepared Bacillus veleis Bv-6 wettable powder is 1 ⁇ 10 11 cfu/g.
- the fermentation medium formula is 3g/L beef extract, 5g/L soy peptone, 20g/L glucose, and pH 7.
- Spray drying indicators inlet air temperature 180°C, outlet air temperature 65°C, pump speed around 3000mL/h (real-time adjustment).
- the preparation method of Bacillus veleis Bv-6 fermentation broth is as follows: take out the Bv-6 strain stored in the laboratory in a -80 degree refrigerator, and streak Bv-6 on an LB solid plate for activation (28 temperature, 24 hours), pick the activated Bv-6 single colony into a 250mL Erlenmeyer flask containing 100mL of LB liquid culture medium with a pH of 7, and shake culture at 28°C and 180rpm for 48h to obtain the BV-6 seed liquid , transfer the seed liquid to a 500mL Erlenmeyer flask containing 250mL of LB liquid culture medium with a pH of 7 according to 5% inoculation amount, and ferment at 28°C and 180rpm for 48h to obtain Bv-6 shake culture fermentation liquid.
- Example 1 Test on the control efficacy of fermentation broths of different Bacillus strains against green peach aphid and cereal aphid
- Green peach aphid (Myzus persicae): Bacillus subtilis Bs-1, Bacillus paraanthracis Bp-1, Bacillus paramycoides strain Bp-2, strain Bp-3, Strain Bp-9, Bacillus velezensis strains Bv-10 and Bv-12, and Bacillus velezensis strain Bv-6 were used to conduct a comparative test on the green peach aphid control effect. After shaking these eight kinds of Bacillus with LB culture medium to obtain fermentation broth, the fermentation broth of each strain was diluted to an OD value of 0.6, and the contact killing activity of the fermentation broth of different strains against green peach aphid was tested. The control was cultured in LB. base treatment.
- Control effect (%) (treatment mortality - control mortality) / (1 - control mortality) ⁇ 100%
- Bacillus veleis strain Bv-6 (97.44%)> Bacillus veleis strain Bv-12 (41.2%)> Bacillus subtilis Bs-1 (27.41%)> Bacillus veleis strain Bv -10(7.41%)>Bacillus paraanthracis Bp1 (4.67%)>Bacillus mycoides Bp2 (4.32%)>Bacillus mycoides Bp3 (1.32%)>Bacillus mycoides Bp9 (1.23%) ( figure 1).
- Rhopalosiphum padi Bacillus subtilis Bs-1, Bacillus paraanthracis Bp-1, Bacillus velezensis strains Bv-10 and Bv-12 , a comparative test on the control effect of green peach aphid with Bacillus velezensis strain Bv-6. After shaking these five kinds of Bacillus with LB medium to obtain fermentation broth, the fermentation broth of each strain was diluted to an OD value of 0.6, and the contact killing activity of the fermentation broth of different strains against the cereal aphid was tested. The control was LB medium treatment. Spray 10 mL of each Bacillus fermentation broth onto wheat leaves containing 30 Aphid graminearum. Each Bacillus strain was repeated three times. After 24 hours, the mortality rate of the Aphid graminearum was calculated to calculate the control effect.
- Control effect (%) (treatment mortality - control mortality) / (1 - control mortality) ⁇ 100%
- Example 2 Test on the control efficacy of Bacillus veleis Bv-6 wettable powder against wheat aphids, broad bean aphids, rapeseed aphids, cucumber aphids, watermelon aphids, peach pink aphids and rose tube aphids
- Control effect (%) (treatment mortality - control mortality) / (1 - control mortality) ⁇ 100%
- Field control of wheat aphids (Rhopalosiphum padi and Sitobion avenae): Use spray method to detect Bacillus veleis Bv-6 wettable powder (1 ⁇ 10 11 cfu/g) 500 For the control effect of twice the liquid on wheat aphids (the cereal aphid and the wheat aphid), weigh 10g of Bv-6 wettable powder, dilute it with 5L of water, and prepare the powder into a 500-fold liquid. In April 2022, two wheat fields (length ⁇ width, 7.6m ⁇ 0.75m) were selected at the experimental base of Huazhong Agricultural University with the same severity of occurrence of wheat aphids (grain aphid and wheat aphid).
- the control There were a total of 1652 wheat ears in the group, and a total of 1756 wheat ears in the treatment group.
- the test is divided into two treatments.
- Treatment 1 Bv-6 powder 500 times liquid (5L) is evenly sprayed on the wheat ears, leaves and stems of the treatment group;
- Treatment 2 Water (5L) is evenly sprayed on the wheat in the control group. Ears, wheat leaves and stems.
- the mortality of the cereal aphid was counted after 2 days and 7 days respectively, and the control effect was calculated.
- Control effect (%) (treatment mortality - control mortality) / (1 - control mortality) ⁇ 100%
- the control effect of Bv-6 powder 500 times on wheat aphids on 2 days and 7 days was 97% and 99% respectively ( Figure 4).
- Pea aphid (Megoura japonica): The spray method was used to detect the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder (1 ⁇ 10 11 cfu/g) on broad bean aphid (Megoura japonica) , weigh 1g of Bv-6 wettable powder, dilute it with 500mL of water, and prepare a 500-fold solution of the powder.
- broad bean plants with severe pea tail aphid occurrence were selected at the Vegetable Research Institute Base of the Wuhan Academy of Agricultural Sciences for a control test.
- Each broad bean plant in the treatment group was sprayed with 20 mL of Bv-6 powder 500 times, and each broad bean plant in the control group was sprayed Spray 20 mL of clean water, and repeat each treatment three times.
- the population base of pea tail-pruning aphids on each broad bean plant was investigated. 48 hours after spraying, the mortality rate of aphids on each broad bean plant in each treatment was counted to calculate the control effect.
- Control effect (%) (treatment mortality - control mortality) / (1 - control mortality) ⁇ 100%
- the rape aphid green peach aphid (Myzus persicae): Use the spray method to test the control effect of 500 times of Bacillus veleis Bv-6 wettable powder (1 ⁇ 10 11 cfu/g) on the rape aphid. Weigh 1g Bv- 6. Wettable powder, dilute with 500mL of water and prepare 500 times of powder solution. It was conducted in July 2022 in the rapeseed field on the campus of Shihezi University Agricultural College in Xinjiang, and rapeseed plants that were seriously damaged by aphids were selected. The test set up 500 times solution of Bv-6 powder and water control, a total of 2 treatments.
- Insect population reduction rate (number of insects before application - number of insects after application) / number of insects before application * 100%
- Control effect (%) (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) ⁇ 100%
- Cucumber aphid (Aphis gossypii): Use the spray method to detect the control effect of 500-fold solution of Bacillus véleis Bv-6 wettable powder (1 ⁇ 10 11 cfu/g) on cucumber aphid (Aphis gossypii), weigh 10g Bv-6 wettable powder, dilute with 5L water and prepare powder 500 times liquid. It was conducted at the Vegetable Research Institute base of the Wuhan Academy of Agricultural Sciences in May 2022. Cucumber plants with serious melon aphid occurrences in greenhouses were selected. The test equipment was Bv-6 wettable powder 500 times liquid, 70% imidacloprid water dispersant 5000 times liquid and water. Control, 3 treatments in total.
- Control effect (%) (treatment mortality - control mortality) / (1 - control mortality) ⁇ 100%
- Watermelon aphid (Aphis gossypii): Use the spray method to detect the control effect of 500-fold solution of Bacillus véleis Bv-6 wettable powder (1 ⁇ 10 11 cfu/g) on watermelon aphid (Aphis gossypii), weigh 10g of Bv-6 wettable powder is diluted with 5L of water and mixed into 500 times of powder solution. It was conducted at the Horticulture Base of Huazhong Agricultural University in May 2022, and watermelon plants damaged by aphids were selected. The test consisted of 500-fold solution of Bv-6 wettable powder, 5000-fold solution of 10% acetamiprid EC, and water control.
- Insect population reduction rate (number of insects before application - number of insects after application) / number of insects before application ⁇ 100%
- Control effect (%) (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) ⁇ 100%
- the 24h and 48h control effects of Bv-6 wettable powder 500 times on watermelon aphids are 95.01% and 96.42% respectively; the 24h and 48h control effects of 10% acetamiprid EC on watermelon aphids at 24h and 48h are respectively 68.61% and 79.11% ( Figure 8).
- Peach tree pink aphid Hyalopterus arundimis: Use the spray method to detect the control effect of 500-fold solution of Bacillus véleis Bv-6 wettable powder (1 ⁇ 10 11 cfu/g) on peach tree pink aphid, and weigh 5g of Bv-6 wettable powder, dilute with 2500mL of water and prepare 500 times of powder solution. It will be conducted in the peach orchard base of Huazhong Agricultural University in May 2022, and peach tree plants damaged by aphids will be selected. The test consisted of 500-fold solution of Bv-6 wettable powder, 5000-fold solution of 10% acetamiprid EC, and water control.
- Insect population reduction rate (number of insects before application - number of insects after application) / number of insects before application ⁇ 100%
- Control effect (%) (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) ⁇ 100%
- Rose aphid (Macrosiphum rosirvorum): The spray method was used to detect the control effect of 500-fold solution of Bacillus véleis Bv-6 wettable powder (1 ⁇ 10 11 cfu/g) on rose aphid (Macrosiphum rosirvorum). Weigh 1g of Bv-6 wettable powder, dilute it with 500mL of water, and prepare a 500-fold solution of the powder. It will be conducted in the Taoyuan base of Huazhong Agricultural University in May 2022, and rose plants damaged by aphids will be selected. The experiment consisted of 500 times solution of Bv-6 wettable powder, 5000 times solution of 10% acetamiprid EC, and water control.
- Insect population reduction rate (number of insects before application - number of insects after application) / number of insects before application ⁇ 100%
- Control effect (%) (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) ⁇ 100%
- the control effect of Bacillus veleis Bv-6 wettable powder on citrus Panonychus citri The spray method was used to detect the 500 of Bacillus veleis Bv-6 wettable powder (1 ⁇ 10 11 cfu/g) For the control effect of twice the liquid on citrus Panonychus mites, weigh 20g of Bv-6 wettable powder, dilute it with 10L of water, and prepare the powder into 500 times the liquid. It was conducted in the citrus garden at the campus base of Huazhong Agricultural University in May 2022, and grapefruit trees damaged by citrus panonychus mites were selected. The test set up two treatments: 500 times solution of Bv-6 powder and water.
- Each treatment was repeated three times, arranged in random blocks, and two grapefruit trees were sprayed in each plot.
- Reduction rate of live mites (number of live mites before application - number of live mites after application)/number of live mites before application ⁇ 100%
- Control effect (%) (Reduction rate of live mites in the treatment group - Reduction rate of live mites in the control group)/(1 - Reduction rate of live mites in the control group) ⁇ 100%
- the control effects of Bv-6 wettable powder 500 times on citrus Panonychus mites were 87.20%, 90.80% and 88.87% respectively 1d, 2d and 5d after the first spraying, and 1d, 2d and 5d after the second spraying.
- the control effects at 7 days and 7 days were 94.59%, 96.41%, 95.56% and 97.01% respectively (Table 1).
- the control effect of Bacillus veleis Bv-6 wettable powder against two-spotted spider mite (Tetranychus urticae): The spray method was used to detect the 500 of Bacillus veleis Bv-6 wettable powder (1 ⁇ 10 11 cfu/g) For the control effect of twice the liquid on strawberry spider mites (two-spotted spider mites), weigh 1g of Bv-6 wettable powder, dilute it with 500 mL of water, and prepare the powder into 500 times the liquid. The test consisted of 500-fold solution of Bv-6 powder, 1000-fold solution of 10.5% Avi-pyrididab EC, and water control, with a total of 3 treatments, and each treatment was repeated 3 times.
- Reduction rate of live mites (number of live mites before application - number of live mites after application)/number of live mites before application ⁇ 100%
- Control effect (%) (Reduction rate of live mites in the treatment group - Reduction rate of live mites in the control group)/(1 - Reduction rate of live mites in the control group) ⁇ 100%
- the spray method was used to detect the control effect of 500-fold and 100-fold solutions of Bacillus veleis Bv-6 wettable powder (1 ⁇ 10 11 cfu/g) on citrus psyllid. Weighed 1g and 5g of Bv-6 respectively. For wet powder, dilute it with 500mL of water and prepare 500 times powder liquid and 100 times liquid powder. It was conducted in the citrus orchard of the Baiyun Base of the Guangdong Academy of Agricultural Sciences in August 2022, and citrus plants damaged by citrus psyllids were selected.
- the test consisted of 500-fold solution of Bv-6 powder, 100-fold solution of Bv-6 powder, 1,000-fold solution of emamectin and fenfofen, and water control, a total of 4 treatments. Each treatment was repeated three times, arranged in random blocks, and two citrus trees were sprayed in each plot. Select the citrus psyllid-damaged young shoots in the east, south, west, north, and middle directions of the citrus tree. Record the number of citrus psyllid adults and nymphs before applying pesticides and mark them.
- Insect population reduction rate (number of insects before application - number of insects after application) / number of insects before application ⁇ 100%
- Control effect (%) (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) ⁇ 100%
- Insect population reduction rate (number of insects before application - number of insects after application) / number of insects before application ⁇ 100%
- Control effect (%) (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) ⁇ 100%
- Control effect (%) (treatment mortality - control mortality) / (1 - control mortality) * 100%
- the control effect on rice brown planthopper is as follows: Bacillus veleis strain Bv-6 (control effect 71.6%) > Bacillus veleis strain Bv-12 (control effect 25.91%)>Bacillus subtilis Bs-1 (control effect 19.99%)>Bacillus veleis strain Bv-10 (control effect 4.73%)>Bacillus paraanthracis Bp1 (control effect 3.54%) ( Figure 15 ).
- Example 7 In vivo rice pot testing of the control effect of Bacillus veleis Bv-6 fermentation liquid on brown planthopper
- Treatment 1 Each pot of rice seedlings was inoculated with 100 brown planthoppers and then evenly sprayed with 20 mL of Bv-6 shaking culture fermentation broth (OD value 0.6);
- Treatment 2 Each pot of rice seedlings was inoculated with 100 brown planthoppers and then evenly sprayed with 20 mL of LB medium. Each treatment was repeated three times. The rice seedlings were placed in a glass jar and sealed with gauze. After 1, 3, and 8 days of treatment, the number of insects was counted, and the control effect was calculated. After the 8th day of treatment, the survival rates of rice seedlings in the control and Bv-6 treatments were calculated.
- Rice seedling survival rate (%) (number of surviving rice seedlings/total number of rice seedlings)*100%
- Example 8 Control effect test of Bacillus veleis Bv-6 wettable powder on rice brown planthopper
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Abstract
The present invention belongs to the technical field of biological prevention and control, and specifically relates to the use of Bacillus velezensis Bv-6 in pest prevention. The technical solution of the present invention is the use of Bacillus velezensis Bv-6 in pest prevention, wherein Bv-6 has the deposit number of CCTCC NO:M 20191106. The present invention verifies that bacillus velezensis Bv-6 has prevention and control effects on many kinds of pests, and provides a new choice for prevention and control of Nilaparvata lugens, aphids, Diaphorina citri, Bemisiatabaci or/and mites.
Description
本发明属于生物防治技术领域，具体涉及贝莱斯芽孢杆菌Bacillus velezensis Bv-6在防虫中的应用。The invention belongs to the technical field of biological control, and specifically relates to the application of Bacillus velezensis Bv-6 in insect control.
贝莱斯芽孢杆菌可以通过直接和间接两种不同的方式对植物的生长和发育产生影响。一方面，通过直接促进植物生长通常需要为植物提供一种由细菌合成的化合物或促进从环境中吸收营养，例如它们可以固定大气中的氮和提供磷等矿物质为植物提供营养物质；合成调控植物生长发育的酶；合成可以为植物生长提供铁的铁载体；合成多种植物激素—包括生长素和细胞***素；释放挥发性化合物等(Brown,1974；Davison,1988；Lambert and Joos,1989；Patten and Glick,1996)。另一方面，贝莱斯芽孢杆菌可以通过一种或多种机制减少或防止植物病原微生物的某些有害影响时，就会间接促进植物生长(Glick,1999)。Bacillus veleis can affect plant growth and development in two different ways, directly and indirectly. On the one hand, by directly promoting plant growth, it is usually necessary to provide the plant with a compound synthesized by bacteria or to promote the absorption of nutrients from the environment. For example, they can fix nitrogen in the atmosphere and provide minerals such as phosphorus to provide nutrients to the plant; synthesis regulation Enzymes for plant growth and development; synthesize siderophores that can provide iron for plant growth; synthesize a variety of plant hormones—including auxin and cytokinin; release volatile compounds, etc. (Brown, 1974; Davison, 1988; Lambert and Joos, 1989 ; Patten and Glick, 1996). On the other hand, B. veleis indirectly promotes plant growth when it can reduce or prevent certain harmful effects of plant pathogenic microorganisms through one or more mechanisms (Glick, 1999).
农作物正在遭受多种病害，虫害，螨害的威胁，每年造成巨大的损失，这其中包括灰霉病，菌核病，蚜虫，烟粉虱，螨虫等等。目前，针对农作物的这些病虫螨害主要以化学防治为主，但由于病原菌及蚜虫对传统的农药产生了不同程度的抗药性，其对农药抗性发展的高风险，造成防治效果降低、增加了防治的难度，同时长期使用农药会导致病虫害出现抗药性、农药残留、破坏土壤生态平衡、环境污染等问题(Biurrun et al.,2010；刘等，2014)。随着农业和环境的可持续发展，食品安全、环境保护以及农产品质量越来越受到人们的重视，研发生物农药是社会发展必然的需求(Wu et al.,2009；Zhang et al.,2008)。有益微生物作为生物农药在农业中的应用随着时间的推移越来越得到重视。Crops are threatened by a variety of diseases, insect pests, and mites, causing huge losses every year, including gray mold, sclerotinia, aphids, whitefly, mites, etc. At present, chemical control is mainly used to control these diseases, insect pests and mites on crops. However, because pathogenic bacteria and aphids have developed varying degrees of resistance to traditional pesticides, there is a high risk of the development of pesticide resistance, resulting in reduced control effects and increased In addition, long-term use of pesticides will lead to pesticide resistance, pesticide residues, damage to soil ecological balance, environmental pollution and other problems (Biurrun et al., 2010; Liu et al., 2014). With the sustainable development of agriculture and the environment, food safety, environmental protection and the quality of agricultural products have attracted more and more attention. The research and development of biopesticides is an inevitable demand for social development (Wu et al., 2009; Zhang et al., 2008) . The application of beneficial microorganisms as biopesticides in agriculture has received more and more attention over time.
各种研究分析了芽孢杆菌对农业病虫螨害的功效，例如苏云金芽孢杆菌对鳞翅目和鞘翅目等咀嚼害虫均具有一定的控制作用(Bravo et al.,2007；Sanahuja et al.,2011)。贝莱斯芽孢杆菌能诱导植物对病原菌和真菌产生***抗性，使植物体内过氧化氢大量积累、细胞死亡和叶片中胼胝质沉淀，有效的控制各类病原菌及蚜虫在植物上的生长(Rashid et al.,2019)。然而，目前少有研究表明芽孢杆菌属对褐飞虱、蚜虫、柑橘木虱、烟粉虱、螨虫等吸食性害虫的杀虫活性。因此，研究这些微生物作为蚜虫、柑橘木虱、烟粉虱、螨虫等吸食性害虫生物防治剂的潜力是有价值的，并有助于发展农业中的环境友好方法。Various studies have analyzed the efficacy of Bacillus against agricultural pests and mites. For example, Bacillus thuringiensis has a certain control effect on chewing pests such as Lepidoptera and Coleoptera (Bravo et al., 2007; Sanahuja et al., 2011 ). Bacillus veleis can induce plants to develop systemic resistance to pathogenic bacteria and fungi, causing massive accumulation of hydrogen peroxide in plants, cell death and callose precipitation in leaves, effectively controlling the growth of various pathogenic bacteria and aphids on plants (Rashid et al., 2019). However, there are few studies showing the insecticidal activity of Bacillus species against sucking pests such as brown planthopper, aphids, citrus psyllid, whitefly, and mites. Therefore, studying the potential of these microorganisms as biocontrol agents of sucking pests such as aphids, citrus psyllid, whitefly, mites, etc. is valuable and can help in the development of environmentally friendly methods in agriculture.
发明内容Contents of the invention
本发明要解决的技术问题是为生物防虫提供一种新选择。The technical problem to be solved by the present invention is to provide a new option for biological insect control.
本发明的技术方案是贝莱斯芽孢杆菌Bacillus velezensis Bv-6在防虫中的应用，Bv-6的保藏号为CCTCC NO:M 20191106。The technical solution of the present invention is the application of Bacillus velezensis Bv-6 in insect control. The deposit number of Bv-6 is CCTCC NO:M 20191106.
进一步的，所述防虫为防治吸食性害虫。Furthermore, the pest control is to control sucking pests.
具体的，所述吸食性害虫为褐飞虱、蚜虫、柑橘木虱(Diaphorina citri)、烟粉虱(Bemisia tabaci)或/和螨虫。
Specifically, the sucking pests are brown planthoppers, aphids, Diaphorina citri, Bemisia tabaci or/and mites.
其中，所述蚜虫为绿桃蚜(Myzus persicae)、禾谷缢管蚜(Rhopalosiphum padi)、麦长管蚜(Sitobion avenae)、瓜蚜(Aphis gossypii)、豌豆修尾蚜(Megoura japonica)、桃粉蚜(Hyalopterus arundimis)或月季长管蚜(Macrosiphum rosirvorum)。Wherein, the aphids are green peach aphid (Myzus persicae), cereal aphid (Rhopalosiphum padi), wheat aphid (Sitobion avenae), melon aphid (Aphis gossypii), pea aphid (Megoura japonica), peach Pink aphid (Hyalopterus arundimis) or rose aphid (Macrosiphum rosirvorum).
其中，所述螨虫为柑橘全爪螨(Panonychus citri)或二斑叶螨(Tetranychus urticae)。Wherein, the mite is Panonychus citri or Tetranychus urticae.
优选的，所述贝莱斯芽孢杆菌Bv-6的剂型为可湿性粉剂。Preferably, the dosage form of Bacillus veleis Bv-6 is a wettable powder.
本发明还提供一种防虫制剂，其主要成分为贝莱斯芽孢杆菌Bv-6的发酵液或可湿性粉剂，所述贝莱斯芽孢杆菌Bacillus velezensis Bv-6的保藏号为CCTCC NO:M 20191106。The invention also provides an anti-insect preparation, the main component of which is the fermentation broth or wettable powder of Bacillus velezensis Bv-6. The preservation number of Bacillus velezensis Bv-6 is CCTCC NO:M 20191106 .
进一步的，所述可湿性粉剂的浓度为1×1011cfu/g。Further, the concentration of the wettable powder is 1×10 11 cfu/g.
本发明所使用的贝莱斯芽孢杆菌具体为贝莱斯芽孢杆菌(Bacillus velezensis)Bv-6菌株，该菌株已于2019年12月25日保存于中国典型培养物保藏中心(地址为：中国武汉武汉大学)，保藏编号为：CCTCC No:M20191106。The Bacillus velezensis used in the present invention is specifically the Bacillus velezensis Bv-6 strain, which was deposited in the China Typical Culture Collection Center on December 25, 2019 (the address is: Wuhan, China Wuhan University), the collection number is: CCTCC No: M20191106.
本发明的有益效果：本发明公开贝莱斯芽孢杆菌Bv-6对诸多的害虫均有防治效果。具体的，贝莱斯芽孢杆菌Bv-6可湿性粉剂500倍液对禾谷缢管蚜(Rhopalosiphum padi)、麦长管蚜Sitobion avenae)、绿桃蚜(Myzus persicae)、豌豆修尾蚜(Megoura japonica)、黄瓜瓜蚜(Aphis gossypii)、西瓜瓜蚜(Aphis gossypii)、桃粉蚜(Hyalopterus arundimis)、月季长管蚜(Macrosiphum rosirvorum)分别具有100％，100％，98.67％，100％，98.17％，96.42％，97.3％，96.02％的防治效果；贝莱斯芽孢杆菌Bv-6发酵液对螨类柑橘红蜘蛛(柑橘全爪螨Panonychus citri)具有92.9％的防治效果，Bv-6可湿性粉剂500倍液对螨类柚子红蜘蛛(柑橘全爪螨Panonychus citri)和草莓红蜘蛛(二斑叶螨Tetranychus urticae)的24h防治效果分别为85.2％，90.71％。Bv-6可湿性粉剂500倍液和100倍液对柑橘木虱(Diaphorina citri)的24h防治效果分别为93.27％，96.77％。贝莱斯芽孢杆菌Bv-6可湿性粉剂对褐飞虱的防治效果达89％。Bv-6可湿性粉剂500倍液对烟粉虱(Bemisia tabaci)的24h防治效果为85.86％。本发明为害虫的生物防治提供了一种新选择。Beneficial effects of the present invention: The present invention discloses that Bacillus veleis Bv-6 has a control effect on many pests. Specifically, 500-fold solution of Bacillus veleis Bv-6 wettable powder is effective against Rhopalosiphum padi, Sitobion avenae, Myzus persicae, and Megoura. japonica), cucumber aphid (Aphis gossypii), watermelon aphid (Aphis gossypii), pink peach aphid (Hyalopterus arundimis), rose aphid (Macrosiphum rosirvorum) have 100%, 100%, 98.67%, 100%, 98.17 respectively. %, 96.42%, 97.3%, 96.02% control effect; Bacillus Velez Bv-6 fermentation liquid has 92.9% control effect on the mite citrus spider mite (Panychus citri), Bv-6 wettability The 24-hour control effect of 500 times of powder on the mites grapefruit spider mites (Panychus citri) and strawberry spider mites (Tetranychus urticae) was 85.2% and 90.71% respectively. The 24-hour control effects of Bv-6 wettable powder 500 times and 100 times on citrus psyllid (Diaphorina citri) were 93.27% and 96.77% respectively. Bacillus veleis Bv-6 wettable powder has a control effect of 89% on brown planthoppers. The 24-hour control effect of Bv-6 wettable powder 500 times on Bemisia tabaci is 85.86%. The present invention provides a new option for biological control of pests.
图1为不同芽孢杆菌对绿桃蚜的防治效果。从左至右依次为贝莱斯芽孢杆菌菌株(Bacillus velezensis)Bv-6，枯草芽孢杆菌(Bacillus subtilis)Bs-1，贝莱斯芽孢杆菌菌株(Bacillus velezensis)Bv-12，贝莱斯芽孢杆菌菌株(Bacillus velezensis)Bv-10，副炭疽芽胞杆菌(Bacillus paranthracis)Bp1，拟蕈状芽胞杆菌(Bacillus paramycoides)Bp3，拟蕈状芽胞杆菌(Bacillus paramycoides)Bp9，拟蕈状芽胞杆菌(Bacillus paramycoides)Bp2，LB培养基，红色箭头所标为死亡蚜虫(经过Bv-6发酵液处理的绿桃蚜大多数都变黑皱缩死亡)。Figure 1 shows the control effects of different Bacillus species on green peach aphid. From left to right are Bacillus velezensis Bv-6, Bacillus subtilis Bs-1, Bacillus velezensis Bv-12, Bacillus velezensis Strain Bv-10, Bacillus paraanthracis Bp1, Bacillus paramycoides Bp3, Bacillus paramycoides Bp9, Bacillus paramycoides Bp2, LB medium, red arrows mark dead aphids (most of the green peach aphids treated with Bv-6 fermentation broth turned black, shrank and died).
图2为不同芽孢杆菌对禾谷缢管蚜的防治效果。从左至右依次为LB培养基，贝莱斯芽孢杆菌菌株(Bacillus velezensis)Bv-6，贝莱斯芽孢杆菌菌株(Bacillus velezensis)Bv-10，枯草芽孢杆菌(Bacillus
subtilis)Bs-1，贝莱斯芽孢杆菌菌株(Bacillus velezensis)Bv-12，副炭疽芽胞杆菌(Bacillus paranthracis)Bp1，红色箭头所标为死亡蚜虫(经过Bv-6发酵液处理的禾谷缢管蚜全部都变黑皱缩死亡)。Figure 2 shows the control effects of different Bacillus species on the cereal aphid. From left to right are LB medium, Bacillus velezensis Bv-6, Bacillus velezensis Bv-10, Bacillus subtilis subtilis) Bs-1, Bacillus velezensis strain Bv-12, Bacillus paraanthracis Bp1, the red arrow marks the dead aphids (grain cones treated with Bv-6 fermentation broth All aphids turn black, shrivel and die).
图3为贝莱斯芽孢杆菌Bv-6可湿性粉剂不同浓度稀释液对禾谷缢管蚜的防治效果。其中CK为喷施清水后蚜虫形态(体色为酒红色，均未发生死亡现象)，经过Bv-6可湿性粉剂喷施过后的禾谷缢管蚜变黑皱缩死亡。Figure 3 shows the control effect of different concentration dilutions of Bacillus veleis Bv-6 wettable powder on the cereal aphid. Among them, CK is the shape of the aphid after spraying with clean water (body color is wine red, no death occurred). After spraying with Bv-6 wettable powder, the aphid turned black, shrank and died.
图4为贝莱斯芽孢杆菌Bv-6可湿性粉剂500倍液对田间小麦蚜虫(禾谷缢管蚜和麦长管蚜)的防治效果(2d和7d)。其中CK为喷施清水后的蚜虫形态(蚜虫数量多，只有极少数发生死亡现象)，经过Bv-6粉剂500倍液喷施处理的活体小麦蚜虫很少，大多数蚜虫死亡变黑皱缩后脱落。Figure 4 shows the control effect (2d and 7d) of 500-fold solution of Bacillus veleis Bv-6 wettable powder on wheat aphids (Grain constrictor aphid and Wheat aphid) in the field. Among them, CK is the form of aphids after spraying with clean water (the number of aphids is large, and only a few die). There are very few living wheat aphids after spraying with 500 times of Bv-6 powder, and most of the aphids die, turn black and shrink. fall off.
图5为贝莱斯芽孢杆菌Bv-6可湿性粉剂500倍液对豌豆修尾蚜的防治效果(48h)。其中CK为喷施清水后豌豆修尾蚜形态(体色为深绿色，均未发生死亡现象)，经过Bv-6粉剂500倍液喷施过后的豌豆修尾蚜全部变黑皱缩死亡，并从植株脱落。Figure 5 shows the control effect (48h) of 500 times solution of Bacillus veleis Bv-6 wettable powder on pea tail-pruning aphid. Among them, CK is the shape of the pea aphids after spraying with clean water (the body color is dark green, and no death has occurred). After spraying 500 times of Bv-6 powder, the pea aphids all turned black, shrank and died, and Fall off the plant.
图6为贝莱斯芽孢杆菌Bv-6可湿性粉剂500倍液对油菜油菜的防治效果(24h)。其中CK为喷施清水后蚜虫形态(体色为深绿色，均未发生死亡现象)，经过Bv-6粉剂500倍液喷施过后的油菜蚜虫皱缩死亡，并从叶片上脱落。Figure 6 shows the control effect (24h) of Bacillus veleis Bv-6 wettable powder 500 times on rapeseed. Among them, CK is the shape of the aphids after spraying with clean water (the body color is dark green, and no death occurred). After spraying 500 times of Bv-6 powder, the rape aphids shrank and died, and fell off from the leaves.
图7为贝莱斯芽孢杆菌Bv-6可湿性粉剂500倍液对黄瓜瓜蚜的防治效果(48h)。喷施清水对照的黄瓜叶片正面及背面蚜虫数量均未减少且黄瓜叶片经蚜虫危害出现萎蔫现象，经过Bv-6粉剂500倍液喷施后的黄瓜叶片正背面蚜虫数量显著减少，蚜虫虫体变黑死亡，大多数从叶片脱落，叶片未发生萎蔫现象。Figure 7 shows the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder on cucumber aphid (48h). The number of aphids on the front and back of the cucumber leaves sprayed with clean water was not reduced, and the cucumber leaves wilted due to aphid damage. After spraying with 500 times of Bv-6 powder, the number of aphids on the front and back of the cucumber leaves was significantly reduced, and the aphids changed. Black and dead, most of the leaves fall off, and the leaves do not wilt.
图8为贝莱斯芽孢杆菌Bv-6可湿性粉剂500倍液对西瓜瓜蚜的防治效果。其中CK为喷施清水后蚜虫形态(体色为深绿色或黄绿色，均未发生死亡现象)，经过Bv-6粉剂500倍液喷施过后的西瓜瓜蚜大部分都变黑皱缩死亡。Figure 8 shows the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder on watermelon aphid. Among them, CK is the shape of aphids after spraying with clean water (body color is dark green or yellow-green, no death occurs). After spraying 500 times of Bv-6 powder, most of the watermelon aphids turned black, shrank and died.
图9为贝莱斯芽孢杆菌Bv-6可湿性粉剂500倍液对桃树桃粉蚜的防治效果。其中CK为喷施清水后蚜虫形态(体色为绿色，均未发生死亡现象)，经过Bv-6粉剂500倍液喷施过后的桃粉蚜大部分都变成黑褐色皱缩死亡。Figure 9 shows the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder on pink aphid on peach trees. Among them, CK is the shape of aphids after spraying with clean water (the body color is green, and no death occurs). After spraying 500 times of Bv-6 powder, most of the peach pink aphids turned into dark brown, shriveled and died.
图10为贝莱斯芽孢杆菌Bv-6可湿性粉剂500倍液对月季长管蚜的防治效果。其中CK为喷施清水后蚜虫形态(体色为绿色，均未发生死亡现象)，经过Bv-6粉剂500倍液喷施过后的月季长管蚜大部分都变成黑褐色皱缩死亡。Figure 10 shows the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder on rose tube aphid. Among them, CK is the shape of the aphids after spraying with clean water (the body color is green, and no death occurred). After spraying 500 times of Bv-6 powder, most of the rose aphids turned dark brown, shriveled and died.
图11为贝莱斯芽孢杆菌Bv-6发酵液对柑橘全爪螨防治效果。经过Bv-6发酵液处理后的柑橘全爪螨虫体死亡并皱缩，经过LB培养基处理的柑橘全爪螨并未出现死亡现象。Figure 11 shows the control effect of Bacillus veleis Bv-6 fermentation broth on citrus Panonychus mites. The citrus panonychus mites that were treated with Bv-6 fermentation liquid died and shrank, but the citrus panonychus mites that were treated with LB medium did not die.
图12为贝莱斯芽孢杆菌Bv-6对草莓二斑叶螨防治效果。经过Bv-6可湿性粉剂喷施过的二斑叶螨虫体皱缩死亡，对照(清水)处理的二斑叶螨虫体正常并未出现死亡现象。Figure 12 shows the control effect of Bacillus veleis Bv-6 on strawberry two-spotted spider mite. The two-spotted spider mites sprayed with Bv-6 wettable powder shrank and died, while the two-spotted spider mites treated with control (water) were normal and did not die.
图13为贝莱斯芽孢杆菌Bv-6对柑橘木虱的防治效果。经过Bv-6可湿性粉剂喷施过的柑橘木虱虫体
干瘪死亡，大多数从叶片脱落，对照(清水)处理图中的白色物质是柑橘木虱的***物。Figure 13 shows the control effect of Bacillus veleis Bv-6 on citrus psyllid. Citrus psyllids sprayed with Bv-6 wettable powder They shriveled and died, and most of them fell off the leaves. The white substance in the control (clear water) treatment picture is the excrement of citrus psyllid.
图14为贝莱斯芽孢杆菌Bv-6对棉花烟粉虱的防治效果。经过Bv-6可湿性粉剂喷施过的烟粉虱死亡，对照(清水)处理的烟粉虱正常并未出现死亡现象。Figure 14 shows the control effect of Bacillus veleis Bv-6 on cotton whitefly. The whiteflies sprayed with Bv-6 wettable powder died, and the whiteflies treated with control (water) did not die normally.
图15为不同芽孢杆菌对水稻褐飞虱的防治效果结果示意图(48h)；Figure 15 is a schematic diagram of the control effects of different Bacillus species on rice brown planthopper (48h);
a图从左到右依次为喷施LB培养基，副炭疽芽胞杆菌(Bacillus paranthracis)Bp1，枯草芽孢杆菌(Bacillus subtilis)Bs-1，贝莱斯芽孢杆菌菌株(Bacillus velezensis)Bv-10，贝莱斯芽孢杆菌菌株(Bacillus velezensis)Bv-12，贝莱斯芽孢杆菌菌株(Bacillus velezensis)Bv-6；Picture a from left to right shows the spraying of LB medium, Bacillus paranthracis Bp1, Bacillus subtilis Bs-1, Bacillus velezensis Bv-10, Bacillus velezensis Bv-10, Bacillus velezensis strain (Bacillus velezensis) Bv-12, Bacillus velezensis strain (Bacillus velezensis) Bv-6;
b图为对应a图各个处理死亡褐飞虱的图片，其中喷施芽孢杆菌后褐飞虱的死亡数量Bv-6>Bv-12>Bs-1>Bv-10>Bp-1>LB。Picture b is a picture of dead brown planthoppers corresponding to each treatment in picture a, in which the number of dead brown planthoppers after spraying Bacillus is Bv-6>Bv-12>Bs-1>Bv-10>Bp-1>LB.
图16为本发明贝莱斯芽孢杆菌Bv-6对水稻褐飞虱的防治效果结果示意图(8d)；Figure 16 is a schematic diagram of the results of the control effect of Bacillus veleis Bv-6 on rice brown planthopper of the present invention (8d);
其中经过Bv-6发酵液喷施后的水稻植株上的褐飞虱虫口数量显著减少，水稻植株生长状况良好，并未发生枯萎现象，经过LB培养基喷施的CK对照处理，褐飞虱的虫口数量并未出现减退，水稻植株受褐飞虱危害后出现枯萎死亡现象。Among them, the number of brown planthoppers on the rice plants after spraying with Bv-6 fermentation broth was significantly reduced. The rice plants grew in good condition and did not wither. After the CK control treatment sprayed with LB medium, the number of brown planthoppers did not increase. The occurrence of rice plants has declined, and rice plants have withered and died after being damaged by brown planthoppers.
图17为贝莱斯芽孢杆菌Bv-6可湿性粉剂500倍稀释液对水稻褐飞虱的防治效果结果示意图(48h)；Figure 17 is a schematic diagram of the control effect of 500-fold dilution of Bacillus veleis Bv-6 wettable powder on rice brown planthopper (48h);
经过贝莱斯芽孢杆菌Bv-6可湿性粉剂500倍稀释液处理过后，褐飞虱出现死亡和虫体变黑现象。而无菌水对照处理的褐飞虱并未出现死亡及虫体变黑现象。F：虫子背部，B：虫子腹部。After being treated with a 500-fold dilution of Bacillus veleis Bv-6 wettable powder, brown planthoppers died and turned black. However, the brown planthoppers treated with sterile water control did not die or turn black. F: The back of the insect, B: The abdomen of the insect.
本发明所述的贝莱斯芽孢杆菌为贝莱斯芽孢杆菌(Bacillus velezensis)Bv-6，保藏编号为CCTCC NO:M20191106，已在CN 111254086A中公开，本发明使用的贝莱斯芽孢杆菌Bv-6可湿性粉剂制备方法同该申请，制备得到的贝莱斯芽孢杆菌Bv-6可湿性粉剂浓度为1×1011cfu/g。The Bacillus velezensis of the present invention is Bacillus velezensis Bv-6, and the deposit number is CCTCC NO: M20191106, which has been disclosed in CN 111254086A. The Bacillus velezensis Bv-6 used in the present invention 6. The wettable powder preparation method is the same as that of the application, and the concentration of the prepared Bacillus veleis Bv-6 wettable powder is 1×10 11 cfu/g.
发酵液制备：Fermentation broth preparation:
将-80℃甘油中保存的贝莱斯芽抱杆菌Bv-6划线接种于固体NA培养基上，在温度为30℃恒温箱中培养24h，然后挑取单菌落接种于100mL的液体NB培养基中，在30℃、180rpm条件下振荡培养24h，作为贝莱斯芽孢杆菌Bv-6的10L发酵罐种子液。Streak-inoculate Bacillus veleis Bv-6 stored in -80°C glycerol on solid NA medium, culture it in an incubator at 30°C for 24 hours, and then pick a single colony and inoculate it into 100 mL of liquid NB for culture In the base, it was shaken and cultured at 30°C and 180 rpm for 24 hours to prepare the 10L fermentation tank seed liquid of Bacillus veleis Bv-6.
发酵培养基配方为牛肉浸膏3g/L、大豆蛋白胨5g/L、葡萄糖20g/L、pH7。The fermentation medium formula is 3g/L beef extract, 5g/L soy peptone, 20g/L glucose, and pH 7.
将上述发酵培养基6升在10L发酵罐中121℃下高压蒸汽灭菌0.5h，灭菌后降温至40℃立即接种1×107cfu/mL浓度的贝莱斯芽孢杆菌种子液300mL(5％接种量)，培养温度30℃、转速300r/min、气压0.05MPa、通气量0.4(V/V·min)，培养72h，得到贝莱斯芽孢杆菌Bv-6菌株发酵液，该发酵液的有效菌浓度是1×1010cfu/mL。6 liters of the above fermentation culture medium were sterilized by high-pressure steam at 121°C for 0.5h in a 10L fermentation tank. After sterilization, the temperature was lowered to 40 °C and immediately inoculated with 300mL (5 % inoculation amount), the culture temperature is 30°C, the rotation speed is 300r/min, the air pressure is 0.05MPa, and the ventilation volume is 0.4 (V/V·min). Cultivate for 72h to obtain the fermentation liquid of Bacillus veleis Bv-6 strain. The fermentation liquid is The effective bacterial concentration is 1×10 10 cfu/mL.
可湿性粉剂的制备：Preparation of wettable powder:
上述得到的贝莱斯芽孢杆菌Bv-6的发酵液中，以1L发酵液加入100g可湿性淀粉，搅拌均匀，经过喷雾干燥机喷雾干燥得到母粉。To the fermentation broth of Bacillus veleis Bv-6 obtained above, add 100g of wettable starch to 1L of the fermentation broth, stir evenly, and spray-dry with a spray dryer to obtain a mother powder.
喷雾干燥指标：进风温度180℃，出风温度65℃，进泵速度3000mL/h左右(实时调节)。Spray drying indicators: inlet air temperature 180℃, outlet air temperature 65℃, pump speed around 3000mL/h (real-time adjustment).
将收集得到的母粉按照以下配方比例进行配制：母粉10g，丁基萘磺酸钠5g，木质素磺酸钙5g，高
岭土80g，混匀即得可湿性粉剂。Prepare the collected mother powder according to the following formula proportions: 10g mother powder, 5g sodium butylnaphthalene sulfonate, 5g calcium lignosulfonate, high 80g of ling clay, mix well to get wettable powder.
将上述可湿性粉剂样品，用无菌水稀释配至106-1010倍，取其0.1mL均匀涂布在NA平板上，于28℃培养。48h后，观察并记录单菌落数量，重复3次。经检测贝莱斯芽孢杆菌Bv-6可湿性粉剂浓度为1×1011cfu/g，本发明或简称该可湿性粉剂为粉剂。Dilute the above wettable powder sample with sterile water to 10 6 -10 10 times, take 0.1 mL and spread it evenly on the NA plate, and incubate at 28°C. After 48 hours, observe and record the number of single colonies and repeat three times. It is detected that the concentration of Bacillus veleis Bv-6 wettable powder is 1×10 11 cfu/g, and the wettable powder may be referred to as powder in the present invention.
下述实施例中贝莱斯芽孢杆菌Bv-6发酵液的制备方法为：在-80度冰箱中取出实验室保存的Bv-6菌株，将Bv-6划线于LB固体平板上活化(28度，24小时)，挑取活化好的Bv-6单菌落挑取到装有100mL pH为7的LB液体培养基的250mL锥形瓶中，28℃，180rpm摇培48h获得BV-6种子液，按5％接种量将种子液转接到盛有250mL pH为7的LB液体培养基的500mL锥形瓶中，在温度28℃，180rpm发酵48h得到Bv-6摇培发酵液。In the following examples, the preparation method of Bacillus veleis Bv-6 fermentation broth is as follows: take out the Bv-6 strain stored in the laboratory in a -80 degree refrigerator, and streak Bv-6 on an LB solid plate for activation (28 temperature, 24 hours), pick the activated Bv-6 single colony into a 250mL Erlenmeyer flask containing 100mL of LB liquid culture medium with a pH of 7, and shake culture at 28℃ and 180rpm for 48h to obtain the BV-6 seed liquid , transfer the seed liquid to a 500mL Erlenmeyer flask containing 250mL of LB liquid culture medium with a pH of 7 according to 5% inoculation amount, and ferment at 28°C and 180rpm for 48h to obtain Bv-6 shake culture fermentation liquid.
实施例1不同芽孢杆菌菌株发酵液对绿桃蚜和禾谷缢管蚜的防效试验Example 1 Test on the control efficacy of fermentation broths of different Bacillus strains against green peach aphid and cereal aphid
绿桃蚜(Myzus persicae)：枯草芽孢杆菌(Bacillus subtilis)Bs-1、副炭疽芽胞杆菌(Bacillus paranthracis)Bp-1、拟蕈状芽孢杆菌(Bacillus paramycoides)菌株Bp-2、菌株Bp-3、菌株Bp-9、贝莱斯芽孢杆菌(Bacillus velezensis)菌株Bv-10和Bv-12，与贝莱斯芽孢杆菌(Bacillus velezensis)菌株Bv-6做绿桃蚜防效对比试验。分别将这八种芽孢杆菌用LB培养基摇配得到发酵液后，分别将各种菌株的发酵液稀释至OD值为0.6，检测不同菌株发酵液对绿桃蚜的触杀活性，对照采用LB培养基处理。每种芽孢杆菌发酵液喷施10mL到含有绿桃蚜(30只)的油菜叶片上，每种芽孢杆菌三个重复，48小时后统计绿桃蚜的死亡率，计算防治效果。Green peach aphid (Myzus persicae): Bacillus subtilis Bs-1, Bacillus paraanthracis Bp-1, Bacillus paramycoides strain Bp-2, strain Bp-3, Strain Bp-9, Bacillus velezensis strains Bv-10 and Bv-12, and Bacillus velezensis strain Bv-6 were used to conduct a comparative test on the green peach aphid control effect. After shaking these eight kinds of Bacillus with LB culture medium to obtain fermentation broth, the fermentation broth of each strain was diluted to an OD value of 0.6, and the contact killing activity of the fermentation broth of different strains against green peach aphid was tested. The control was cultured in LB. base treatment. Spray 10 mL of each Bacillus fermentation broth onto rape leaves containing green peach aphids (30 individuals). Each Bacillus strain was repeated three times. After 48 hours, the mortality rate of green peach aphids was calculated to calculate the control effect.
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)×100％Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) × 100%
防治效果如下：贝莱斯芽孢杆菌菌株Bv-6(97.44％)>贝莱斯芽孢杆菌菌株Bv-12(41.2％)>枯草芽孢杆菌Bs-1(27.41％)>贝莱斯芽孢杆菌菌株Bv-10(7.41％)>副炭疽芽胞杆菌Bp1(4.67％)>拟蕈状芽胞杆菌Bp2(4.32％)>拟蕈状芽胞杆菌Bp3(1.32％)>拟蕈状芽胞杆菌Bp9(1.23％)(图1)。The control effects are as follows: Bacillus veleis strain Bv-6 (97.44%)> Bacillus veleis strain Bv-12 (41.2%)> Bacillus subtilis Bs-1 (27.41%)> Bacillus veleis strain Bv -10(7.41%)>Bacillus paraanthracis Bp1 (4.67%)>Bacillus mycoides Bp2 (4.32%)>Bacillus mycoides Bp3 (1.32%)>Bacillus mycoides Bp9 (1.23%) ( figure 1).
禾谷缢管蚜(Rhopalosiphum padi)：枯草芽孢杆菌(Bacillus subtilis)Bs-1、副炭疽芽胞杆菌(Bacillus paranthracis)Bp-1、贝莱斯芽孢杆菌(Bacillus velezensis)菌株Bv-10和Bv-12，与贝莱斯芽孢杆菌(Bacillus velezensis)菌株Bv-6做绿桃蚜防效对比试验。分别将这五种芽孢杆菌用LB培养基摇配得到发酵液后，分别将各种菌株的发酵液稀释至OD值为0.6，检测不同菌株发酵液对禾谷缢管蚜的触杀活性，对照采用LB培养基处理。每种芽孢杆菌发酵液喷施10mL到含有禾谷缢管蚜(30只)的小麦叶片上，每种芽孢杆菌三个重复，24小时后统计禾谷缢管蚜的死亡率，计算防治效果。Rhopalosiphum padi: Bacillus subtilis Bs-1, Bacillus paraanthracis Bp-1, Bacillus velezensis strains Bv-10 and Bv-12 , a comparative test on the control effect of green peach aphid with Bacillus velezensis strain Bv-6. After shaking these five kinds of Bacillus with LB medium to obtain fermentation broth, the fermentation broth of each strain was diluted to an OD value of 0.6, and the contact killing activity of the fermentation broth of different strains against the cereal aphid was tested. The control was LB medium treatment. Spray 10 mL of each Bacillus fermentation broth onto wheat leaves containing 30 Aphid graminearum. Each Bacillus strain was repeated three times. After 24 hours, the mortality rate of the Aphid graminearum was calculated to calculate the control effect.
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)×100％Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) × 100%
防治效果如下：贝莱斯芽孢杆菌菌株Bv-6(100％)>枯草芽孢杆菌Bs-1(69.9％)>贝莱斯芽孢杆菌菌株Bv-10(46.87％)>贝莱斯芽孢杆菌菌株Bv-12(34.5％)>副炭疽芽胞杆菌Bp1(0％)(图2)。The control effects are as follows: Bacillus veleis strain Bv-6 (100%)>Bacillus subtilis Bs-1 (69.9%)>Bacillus veleis strain Bv-10 (46.87%)>Bacillus veleis strain Bv -12 (34.5%) > Bacillus paraanthracis Bp1 (0%) (Fig. 2).
实施例2贝莱斯芽孢杆菌Bv-6可湿性粉剂对小麦蚜虫、蚕豆蚜虫、油菜蚜虫、黄瓜蚜虫、西瓜瓜蚜、桃树桃粉蚜和月季长管蚜的防效试验Example 2 Test on the control efficacy of Bacillus veleis Bv-6 wettable powder against wheat aphids, broad bean aphids, rapeseed aphids, cucumber aphids, watermelon aphids, peach pink aphids and rose tube aphids
室内不同浓度Bv-6可湿性粉剂对禾谷缢管蚜(Rhopalosiphum padi)的防效：采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)的不同浓度稀释液对禾谷缢管蚜的防效，称取1g Bv-6可湿性粉剂，用1000mL清水稀释，配置成粉剂1000倍液。然后再用清水分别稀释成粉剂2000倍液、4000倍液、
6000倍液、8000倍液和10000倍液。取两周龄的小麦幼苗，每盆小麦幼苗接种500只三龄禾谷缢管蚜(每盆50株小麦幼苗)，将不同浓度的贝莱斯芽孢杆菌Bv-6可湿性粉剂稀释液和清水分别喷施含有禾谷缢管蚜的小麦幼苗(5mL/盆)，每个处理重复三盆小麦。24小时后统计禾谷缢管蚜的死亡率，计算防治效果。Indoor control effect of different concentrations of Bv-6 wettable powder on Rhopalosiphum padi: Spray method was used to detect different concentrations of Bv-6 wettable powder of Bacillus véleis (1×10 11 cfu/g) For the control effect of the diluent on the cereal aphid, weigh 1g of Bv-6 wettable powder, dilute it with 1000mL of water, and prepare a 1000-fold solution of the powder. Then dilute it with water into powder 2000 times liquid, 4000 times liquid, 6000 times liquid, 8000 times liquid and 10000 times liquid. Take two-week-old wheat seedlings and inoculate each pot with 500 three-year-old cereal aphids (50 wheat seedlings per pot). Mix different concentrations of Bacillus velevis Bv-6 wettable powder diluents and water. Wheat seedlings containing Aphis graminearum (5 mL/pot) were sprayed separately, and each treatment was repeated in three pots of wheat. After 24 hours, the mortality rate of the cereal aphid was counted and the control effect was calculated.
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)×100％Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) × 100%
防治效果如下：Bv-6粉剂1000倍液(100％)>Bv-6粉剂2000倍液(100％)>Bv-6粉剂4000倍液(96.07％)>Bv-6粉剂6000倍液(91.4％)>Bv-6粉剂8000倍液(52.27％)>Bv-6粉剂10000倍液(12.27％)(图3)。The control effect is as follows: Bv-6 powder 1000 times liquid (100%)>Bv-6 powder 2000 times liquid (100%)>Bv-6 powder 4000 times liquid (96.07%)>Bv-6 powder 6000 times liquid (91.4%) )>Bv-6 powder 8000 times liquid (52.27%)>Bv-6 powder 10000 times liquid (12.27%) (Figure 3).
田间防治小麦蚜虫(禾谷缢管蚜(Rhopalosiphum padi)和麦长管蚜(Sitobion avenae))：采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)500倍液对小麦蚜虫(禾谷缢管蚜和麦长管蚜)的防效，称取10g Bv-6可湿性粉剂，用5L清水稀释，配置成粉剂500倍液。于2022年4月在华中农业大学校内试验基地选取两块小麦蚜虫(禾谷缢管蚜和麦长管蚜)发生严重程度一致的小麦地(长×宽，7.6m×0.75m)，其中对照组总计1652株麦穗，处理组总计1756株麦穗。试验分为两个处理，处理一：Bv-6粉剂500倍液(5L)均匀喷施处理组小麦麦穗，麦叶及茎秆；处理二：清水(5L)均匀喷施对照组的小麦麦穗，麦叶及茎秆。分别于2d和7d后统计禾谷缢管蚜的死亡率，计算防治效果。
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)×100％
Bv-6粉剂500倍液对小麦蚜虫的2d和7d的防治效果分别为97％和99％(图4)。Field control of wheat aphids (Rhopalosiphum padi and Sitobion avenae): Use spray method to detect Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) 500 For the control effect of twice the liquid on wheat aphids (the cereal aphid and the wheat aphid), weigh 10g of Bv-6 wettable powder, dilute it with 5L of water, and prepare the powder into a 500-fold liquid. In April 2022, two wheat fields (length × width, 7.6m × 0.75m) were selected at the experimental base of Huazhong Agricultural University with the same severity of occurrence of wheat aphids (grain aphid and wheat aphid). Among them, the control There were a total of 1652 wheat ears in the group, and a total of 1756 wheat ears in the treatment group. The test is divided into two treatments. Treatment 1: Bv-6 powder 500 times liquid (5L) is evenly sprayed on the wheat ears, leaves and stems of the treatment group; Treatment 2: Water (5L) is evenly sprayed on the wheat in the control group. Ears, wheat leaves and stems. The mortality of the cereal aphid was counted after 2 days and 7 days respectively, and the control effect was calculated.
Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) × 100%
The control effect of Bv-6 powder 500 times on wheat aphids on 2 days and 7 days was 97% and 99% respectively (Figure 4).
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)×100％
Bv-6粉剂500倍液对小麦蚜虫的2d和7d的防治效果分别为97％和99％(图4)。Field control of wheat aphids (Rhopalosiphum padi and Sitobion avenae): Use spray method to detect Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) 500 For the control effect of twice the liquid on wheat aphids (the cereal aphid and the wheat aphid), weigh 10g of Bv-6 wettable powder, dilute it with 5L of water, and prepare the powder into a 500-fold liquid. In April 2022, two wheat fields (length × width, 7.6m × 0.75m) were selected at the experimental base of Huazhong Agricultural University with the same severity of occurrence of wheat aphids (grain aphid and wheat aphid). Among them, the control There were a total of 1652 wheat ears in the group, and a total of 1756 wheat ears in the treatment group. The test is divided into two treatments. Treatment 1: Bv-6 powder 500 times liquid (5L) is evenly sprayed on the wheat ears, leaves and stems of the treatment group; Treatment 2: Water (5L) is evenly sprayed on the wheat in the control group. Ears, wheat leaves and stems. The mortality of the cereal aphid was counted after 2 days and 7 days respectively, and the control effect was calculated.
Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) × 100%
The control effect of Bv-6 powder 500 times on wheat aphids on 2 days and 7 days was 97% and 99% respectively (Figure 4).
豌豆修尾蚜(Megoura japonica)：采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)500倍液对蚕豆蚜虫(豌豆修尾蚜Megoura japonica)的防效，称取1g Bv-6可湿性粉剂，用500mL清水稀释，配置成粉剂500倍液。于2022年4月在武汉市农业科学院蔬菜研究所基地选取豌豆修尾蚜发生严重的蚕豆植株进行防治试验，处理组对每株蚕豆喷施Bv-6粉剂500倍液20mL，对照组每株蚕豆喷施清水20mL，每个处理各三个重复。喷施前调查每株蚕豆上豌豆修尾蚜的虫口基数，喷药48小时后统计各处理每株蚕豆上蚜虫的死亡率，计算防治效果。Pea aphid (Megoura japonica): The spray method was used to detect the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) on broad bean aphid (Megoura japonica) , weigh 1g of Bv-6 wettable powder, dilute it with 500mL of water, and prepare a 500-fold solution of the powder. In April 2022, broad bean plants with severe pea tail aphid occurrence were selected at the Vegetable Research Institute Base of the Wuhan Academy of Agricultural Sciences for a control test. Each broad bean plant in the treatment group was sprayed with 20 mL of Bv-6 powder 500 times, and each broad bean plant in the control group was sprayed Spray 20 mL of clean water, and repeat each treatment three times. Before spraying, the population base of pea tail-pruning aphids on each broad bean plant was investigated. 48 hours after spraying, the mortality rate of aphids on each broad bean plant in each treatment was counted to calculate the control effect.
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)×100％Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) × 100%
48小时后经过Bv-6粉剂500倍液喷施后的豌豆修尾蚜全部变黑死亡，防治效果达到100％(图5)。After 48 hours, all the pea aphids that were sprayed with 500 times of Bv-6 powder turned black and died, and the control effect reached 100% (Figure 5).
油菜蚜虫绿桃蚜(Myzus persicae)：采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)的500倍液对油菜蚜虫的防效，称取1g Bv-6可湿性粉剂，用500mL清水稀释，配置成粉剂500倍液。于2022年7月在新疆石河子大学农学院校内基地油菜田进行，选取蚜虫危害严重的油菜植株。试验设Bv-6粉剂500倍液和清水对照，共2个处理。每个处理挑选3株蚜虫危害严重的油菜植株，施药前记录蚜虫数量，并做标记。采用手动喷雾器对油菜叶片正反面均匀喷施，至叶片有少量药液下滴为止。喷药24h后调查各个叶片上蚜虫的活虫数，计算虫口减退率和防治效果。
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数*100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％The rape aphid green peach aphid (Myzus persicae): Use the spray method to test the control effect of 500 times of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) on the rape aphid. Weigh 1g Bv- 6. Wettable powder, dilute with 500mL of water and prepare 500 times of powder solution. It was conducted in July 2022 in the rapeseed field on the campus of Shihezi University Agricultural College in Xinjiang, and rapeseed plants that were seriously damaged by aphids were selected. The test set up 500 times solution of Bv-6 powder and water control, a total of 2 treatments. For each treatment, 3 rapeseed plants with serious aphid damage were selected, and the number of aphids was recorded and marked before application. Use a manual sprayer to spray evenly on the front and back of the rape leaves until a small amount of liquid drops on the leaves. 24 hours after spraying, the number of live aphids on each leaf was investigated, and the population reduction rate and control effect were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application * 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数*100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％The rape aphid green peach aphid (Myzus persicae): Use the spray method to test the control effect of 500 times of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) on the rape aphid. Weigh 1g Bv- 6. Wettable powder, dilute with 500mL of water and prepare 500 times of powder solution. It was conducted in July 2022 in the rapeseed field on the campus of Shihezi University Agricultural College in Xinjiang, and rapeseed plants that were seriously damaged by aphids were selected. The test set up 500 times solution of Bv-6 powder and water control, a total of 2 treatments. For each treatment, 3 rapeseed plants with serious aphid damage were selected, and the number of aphids was recorded and marked before application. Use a manual sprayer to spray evenly on the front and back of the rape leaves until a small amount of liquid drops on the leaves. 24 hours after spraying, the number of live aphids on each leaf was investigated, and the population reduction rate and control effect were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application * 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
Bv-6可湿性粉剂500倍液对油菜蚜虫的24h防治效果为98.67％(图6)。The 24-hour control effect of Bv-6 wettable powder 500 times on rapeseed aphids was 98.67% (Figure 6).
黄瓜瓜蚜(Aphis gossypii)：采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)的500倍液对黄瓜瓜蚜(Aphis gossypii)的防效，称取10g Bv-6可湿性粉剂，用5L清水稀释，配置成粉剂500
倍液。于2022年5月在武汉市农业科学院蔬菜研究所基地进行，选取大棚内瓜蚜发生严重的黄瓜植株，试验设Bv-6可湿性粉剂500倍液、70％吡虫啉水分散剂5000倍液和清水对照，共3个处理。每个处理3次重复。施药前记录黄瓜叶片蚜虫数量，采用手动喷雾器对黄瓜叶片正反面均匀喷施，至叶片有少量药液下滴为止。喷药48h后调查各个叶片上蚜虫的活虫数，计算虫口减退率和防治效果。Cucumber aphid (Aphis gossypii): Use the spray method to detect the control effect of 500-fold solution of Bacillus véleis Bv-6 wettable powder (1×10 11 cfu/g) on cucumber aphid (Aphis gossypii), weigh 10g Bv-6 wettable powder, dilute with 5L water and prepare powder 500 times liquid. It was conducted at the Vegetable Research Institute base of the Wuhan Academy of Agricultural Sciences in May 2022. Cucumber plants with serious melon aphid occurrences in greenhouses were selected. The test equipment was Bv-6 wettable powder 500 times liquid, 70% imidacloprid water dispersant 5000 times liquid and water. Control, 3 treatments in total. Each treatment was repeated 3 times. Before applying pesticide, record the number of aphids on cucumber leaves, and use a manual sprayer to spray evenly on the front and back of the cucumber leaves until a small amount of pesticide drops on the leaves. 48 hours after spraying, the number of live aphids on each leaf was investigated, and the population reduction rate and control effect were calculated.
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)×100％Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) × 100%
Bv-6可湿性粉剂500倍液对黄瓜蚜虫的48h防治效果为98.17％，70％吡虫啉水分散剂5000倍液对黄瓜蚜虫的48h防治效果为99.37％(图7)。The 48h control effect of Bv-6 wettable powder 500 times on cucumber aphids is 98.17%, and the 48h control effect of 70% imidacloprid water dispersant on cucumber aphids is 99.37% (Figure 7).
西瓜瓜蚜(Aphis gossypii)：采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)的500倍液对西瓜瓜蚜(Aphis gossypii)的防效，称取10g Bv-6可湿性粉剂，用5L清水稀释，配置成粉剂500倍液。于2022年5月在华中农业大学校内园艺基进行，选取蚜虫危害的西瓜植株。试验设Bv-6可湿性粉剂500倍液、10％啶虫脒乳油5000倍液和清水对照，共3个处理，每个处理3次重复。每个处理挑选3株蚜虫危害严重的西瓜植株，施药前记录蚜虫数量，并做标记。采用手动喷雾器对西瓜叶片正反面均匀喷施，至叶片有少量药液下滴为止。喷药24h和48h后调查各个叶片上蚜虫的活虫数，计算虫口减退率和防治效果。
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数×100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％Watermelon aphid (Aphis gossypii): Use the spray method to detect the control effect of 500-fold solution of Bacillus véleis Bv-6 wettable powder (1×10 11 cfu/g) on watermelon aphid (Aphis gossypii), weigh 10g of Bv-6 wettable powder is diluted with 5L of water and mixed into 500 times of powder solution. It was conducted at the Horticulture Base of Huazhong Agricultural University in May 2022, and watermelon plants damaged by aphids were selected. The test consisted of 500-fold solution of Bv-6 wettable powder, 5000-fold solution of 10% acetamiprid EC, and water control. There were 3 treatments in total, and each treatment was repeated 3 times. For each treatment, 3 watermelon plants with severe aphid damage were selected, and the number of aphids was recorded and marked before application. Use a manual sprayer to spray evenly on the front and back of the watermelon leaves until a small amount of liquid drops on the leaves. 24h and 48h after spraying, the number of live aphids on each leaf was investigated, and the population reduction rate and control effect were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application × 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数×100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％Watermelon aphid (Aphis gossypii): Use the spray method to detect the control effect of 500-fold solution of Bacillus véleis Bv-6 wettable powder (1×10 11 cfu/g) on watermelon aphid (Aphis gossypii), weigh 10g of Bv-6 wettable powder is diluted with 5L of water and mixed into 500 times of powder solution. It was conducted at the Horticulture Base of Huazhong Agricultural University in May 2022, and watermelon plants damaged by aphids were selected. The test consisted of 500-fold solution of Bv-6 wettable powder, 5000-fold solution of 10% acetamiprid EC, and water control. There were 3 treatments in total, and each treatment was repeated 3 times. For each treatment, 3 watermelon plants with severe aphid damage were selected, and the number of aphids was recorded and marked before application. Use a manual sprayer to spray evenly on the front and back of the watermelon leaves until a small amount of liquid drops on the leaves. 24h and 48h after spraying, the number of live aphids on each leaf was investigated, and the population reduction rate and control effect were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application × 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
Bv-6可湿性粉剂500倍液对西瓜瓜蚜的24h、48h的防治效果分别为95.01％和96.42％；10％啶虫脒乳油5000倍液对西瓜瓜蚜的24h、48h的防治效果分别为68.61％和79.11％(图8)。The 24h and 48h control effects of Bv-6 wettable powder 500 times on watermelon aphids are 95.01% and 96.42% respectively; the 24h and 48h control effects of 10% acetamiprid EC on watermelon aphids at 24h and 48h are respectively 68.61% and 79.11% (Figure 8).
桃树桃粉蚜(Hyalopterus arundimis)：采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)的500倍液对桃树桃粉蚜的防效，称取5g Bv-6可湿性粉剂，用2500mL清水稀释，配置成粉剂500倍液。于2022年5月在华中农业大学校内桃园基地进行，选取蚜虫危害的桃树植株。试验设Bv-6可湿性粉剂500倍液、10％啶虫脒乳油5000倍液和清水对照，共3个处理，每个处理3次重复。在桃树的东、南、西、北、中5个方位选定桃粉蚜危害枝条，施药前记录桃粉蚜数量，并做标记。采用背负式电动喷雾器对整棵桃树叶片正反面均匀喷雾，使枝条树叶充分接触药液，至叶片有少量药液下滴为止。喷药24h后调查各个处理嫩梢上桃粉蚜的活虫数，统计各处理桃粉蚜存活虫总数，并计算虫口减退率和防治效果。
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数×100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％Peach tree pink aphid (Hyalopterus arundimis): Use the spray method to detect the control effect of 500-fold solution of Bacillus véleis Bv-6 wettable powder (1×10 11 cfu/g) on peach tree pink aphid, and weigh 5g of Bv-6 wettable powder, dilute with 2500mL of water and prepare 500 times of powder solution. It will be conducted in the peach orchard base of Huazhong Agricultural University in May 2022, and peach tree plants damaged by aphids will be selected. The test consisted of 500-fold solution of Bv-6 wettable powder, 5000-fold solution of 10% acetamiprid EC, and water control. There were 3 treatments in total, and each treatment was repeated 3 times. Select the branches damaged by pink aphids in the east, south, west, north and middle directions of the peach tree. Record the number of pink aphids and mark them before applying pesticides. Use a knapsack electric sprayer to evenly spray the front and back of the entire peach tree leaves, so that the branches and leaves are fully exposed to the liquid until a small amount of liquid drips from the leaves. 24 hours after spraying, the number of live pink aphids on the young shoots of each treatment was investigated, the total number of surviving pink aphids in each treatment was counted, and the population reduction rate and control effect were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application × 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数×100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％Peach tree pink aphid (Hyalopterus arundimis): Use the spray method to detect the control effect of 500-fold solution of Bacillus véleis Bv-6 wettable powder (1×10 11 cfu/g) on peach tree pink aphid, and weigh 5g of Bv-6 wettable powder, dilute with 2500mL of water and prepare 500 times of powder solution. It will be conducted in the peach orchard base of Huazhong Agricultural University in May 2022, and peach tree plants damaged by aphids will be selected. The test consisted of 500-fold solution of Bv-6 wettable powder, 5000-fold solution of 10% acetamiprid EC, and water control. There were 3 treatments in total, and each treatment was repeated 3 times. Select the branches damaged by pink aphids in the east, south, west, north and middle directions of the peach tree. Record the number of pink aphids and mark them before applying pesticides. Use a knapsack electric sprayer to evenly spray the front and back of the entire peach tree leaves, so that the branches and leaves are fully exposed to the liquid until a small amount of liquid drips from the leaves. 24 hours after spraying, the number of live pink aphids on the young shoots of each treatment was investigated, the total number of surviving pink aphids in each treatment was counted, and the population reduction rate and control effect were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application × 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
Bv-6可湿性粉剂500倍液对桃粉蚜的24h防治效果为97.3％，10％啶虫脒乳油5000倍液对桃粉蚜的24h防治效果为97.4％(图9)。The 24-hour control effect of Bv-6 wettable powder 500 times on peach pink aphid is 97.3%, and the 24-hour control effect of 10% acetamiprid EC on 5000 times solution of peach pink aphid is 97.4% (Figure 9).
月季长管蚜(Macrosiphum rosirvorum)：采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)的500倍液对月季长管蚜(Macrosiphum rosirvorum)的防效，称取1g Bv-6可湿性粉剂，用500mL清水稀释，配置成粉剂500倍液。于2022年5月在华中农业大学校内桃园基地进行，选取蚜虫危害的月季植株。试验设Bv-6可湿性粉剂500倍液、10％啶虫脒乳油5000倍液和清水对照，共3个处理，每个处理选取3个月季枝条。施药前每个枝条统计5片叶片的蚜虫数量，并做标记。采用手动喷雾器对月季枝条喷雾，叶片正反面均匀喷施，使枝条树叶充分接触药液，至叶片有少量药液下滴为止。喷药24h后调查各个处理
叶片上月季长管蚜的活虫数，统计各处理月季长管蚜存活虫总数，并计算虫口减退率和防治效果。
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数×100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％Rose aphid (Macrosiphum rosirvorum): The spray method was used to detect the control effect of 500-fold solution of Bacillus véleis Bv-6 wettable powder (1×10 11 cfu/g) on rose aphid (Macrosiphum rosirvorum). Weigh 1g of Bv-6 wettable powder, dilute it with 500mL of water, and prepare a 500-fold solution of the powder. It will be conducted in the Taoyuan base of Huazhong Agricultural University in May 2022, and rose plants damaged by aphids will be selected. The experiment consisted of 500 times solution of Bv-6 wettable powder, 5000 times solution of 10% acetamiprid EC, and water control. There were 3 treatments in total, and 3 rose branches were selected for each treatment. Before applying pesticides, count the number of aphids on 5 leaves of each branch and mark them. Use a manual sprayer to spray the rose branches, evenly spraying the front and back of the leaves, so that the branches and leaves are fully exposed to the liquid until a small amount of liquid drips from the leaves. Investigate each treatment 24 hours after spraying The number of living rose aphids on the leaves was counted. The total number of surviving rose aphids in each treatment was counted, and the population reduction rate and control effect were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application × 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数×100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％Rose aphid (Macrosiphum rosirvorum): The spray method was used to detect the control effect of 500-fold solution of Bacillus véleis Bv-6 wettable powder (1×10 11 cfu/g) on rose aphid (Macrosiphum rosirvorum). Weigh 1g of Bv-6 wettable powder, dilute it with 500mL of water, and prepare a 500-fold solution of the powder. It will be conducted in the Taoyuan base of Huazhong Agricultural University in May 2022, and rose plants damaged by aphids will be selected. The experiment consisted of 500 times solution of Bv-6 wettable powder, 5000 times solution of 10% acetamiprid EC, and water control. There were 3 treatments in total, and 3 rose branches were selected for each treatment. Before applying pesticides, count the number of aphids on 5 leaves of each branch and mark them. Use a manual sprayer to spray the rose branches, evenly spraying the front and back of the leaves, so that the branches and leaves are fully exposed to the liquid until a small amount of liquid drips from the leaves. Investigate each treatment 24 hours after spraying The number of living rose aphids on the leaves was counted. The total number of surviving rose aphids in each treatment was counted, and the population reduction rate and control effect were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application × 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
Bv-6可湿性粉剂500倍液对月季长管蚜的24h防治效果为96.02％，10％啶虫脒乳油5000倍液对桃粉蚜的24h防治效果为98.04％(图10)。The 24-hour control effect of Bv-6 wettable powder 500 times on rose tube aphid is 96.02%, and the 24-hour control effect of 10% acetamiprid EC on peach pink aphid is 98.04% (Figure 10).
实施例3贝莱斯芽孢杆菌Bv-6对作物螨害的防效试验Example 3 Test on the control effect of Bacillus veleis Bv-6 on crop mite damage
贝莱斯芽孢杆菌Bv-6发酵液对柑橘全爪螨(Panonychus citri)的防效：检测贝莱斯芽孢杆菌Bv-6的LB摇培发酵液对柑橘全爪螨的触杀活性，对照采用LB液体培养基处理。Bv-6发酵液喷施10mL到含有柑橘全爪螨的柑橘叶片上，叶片正反面都喷，每个试验每个处理三个重复，48小时后用放大镜检查结果。用毛笔轻轻触碰螨虫，以螨足不动者为死亡。统计柑橘全爪螨的死亡率，计算防治效果。
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)×100％
48小时后BV-6发酵液对柑橘全爪螨防治效果为92.9％(图11)。Control effect of Bacillus veleis Bv-6 fermentation broth on citrus Panonychus citri: The contact killing activity of LB shaking culture fermentation broth of B. veleis Bv-6 against citrus Panonychus citri was tested, and LB was used as the control Liquid media handling. Spray 10 mL of Bv-6 fermentation broth onto citrus leaves containing Panonychus citrus mites, spraying both the front and back of the leaves. Each test was repeated three times for each treatment, and the results were checked with a magnifying glass after 48 hours. Use a brush to gently touch the mites. If the mites do not move, they will die. The mortality rate of Panonychus citrus mites was counted and the control effect was calculated.
Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) × 100%
After 48 hours, the control effect of BV-6 fermentation broth on citrus Panonychus mites was 92.9% (Figure 11).
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)×100％
48小时后BV-6发酵液对柑橘全爪螨防治效果为92.9％(图11)。Control effect of Bacillus veleis Bv-6 fermentation broth on citrus Panonychus citri: The contact killing activity of LB shaking culture fermentation broth of B. veleis Bv-6 against citrus Panonychus citri was tested, and LB was used as the control Liquid media handling. Spray 10 mL of Bv-6 fermentation broth onto citrus leaves containing Panonychus citrus mites, spraying both the front and back of the leaves. Each test was repeated three times for each treatment, and the results were checked with a magnifying glass after 48 hours. Use a brush to gently touch the mites. If the mites do not move, they will die. The mortality rate of Panonychus citrus mites was counted and the control effect was calculated.
Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) × 100%
After 48 hours, the control effect of BV-6 fermentation broth on citrus Panonychus mites was 92.9% (Figure 11).
贝莱斯芽孢杆菌Bv-6可湿性粉剂对柑橘全爪螨(Panonychus citri)的防效：采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)的500倍液对柑橘全爪螨的防效，称取20g Bv-6可湿性粉剂，用10L清水稀释，配置成粉剂500倍液。于2022年5月在华中农业大学校内基地柑橘园进行，选取柑橘全爪螨危害的柚子树。试验设Bv-6粉剂500倍液和清水对照两个处理。每个处理3次重复，随机区组排列，每个小区施药2株柚子树。在柚子树的东、南、西、北、中5个方位选定柑橘全爪螨危害嫩梢，施药前记录柑橘全爪螨数量，并做标记。采用手动喷雾器对树冠嫩梢喷雾，叶片正反面均匀喷施，使嫩叶嫩梢充分接触药液，至叶片有少量药液下滴为止。喷药后1d、2d和5d用放大镜检查各个处理叶片上柑橘全爪螨的活螨数，用毛笔轻轻触碰螨虫，以螨足不动者为死亡。第一次喷药5d后，再进行第二次喷药，第二次喷药后1d、2d、5d和7d后调查统计活螨数，计算活螨减退率和防治效果。
活螨减退率＝(施药前活螨数-施药后活螨数)/施药前活螨数×100％
防治效果(％)＝(处理组活螨减退率-对照组活螨减退率)/(1-对照组活螨减退率)×100％The control effect of Bacillus veleis Bv-6 wettable powder on citrus Panonychus citri: The spray method was used to detect the 500 of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) For the control effect of twice the liquid on citrus Panonychus mites, weigh 20g of Bv-6 wettable powder, dilute it with 10L of water, and prepare the powder into 500 times the liquid. It was conducted in the citrus garden at the campus base of Huazhong Agricultural University in May 2022, and grapefruit trees damaged by citrus panonychus mites were selected. The test set up two treatments: 500 times solution of Bv-6 powder and water. Each treatment was repeated three times, arranged in random blocks, and two grapefruit trees were sprayed in each plot. Select the citrus panonychus mites to harm the young shoots in the east, south, west, north, and middle directions of the grapefruit tree. Record the number of citrus panonychus mites before applying pesticides and mark them. Use a manual sprayer to spray the young tips of the crown of the tree, and spray evenly on the front and back of the leaves so that the young leaves and tips are fully exposed to the liquid until a small amount of liquid drips from the leaves. 1d, 2d and 5d after spraying, use a magnifying glass to check the number of live citrus Panonychus mites on the leaves of each treatment, touch the mites gently with a brush, and mites that cannot move their legs are considered dead. 5 days after the first spraying, the second spraying was carried out. The number of live mites was investigated and counted 1d, 2d, 5d and 7d after the second spraying, and the reduction rate of live mites and the control effect were calculated.
Reduction rate of live mites = (number of live mites before application - number of live mites after application)/number of live mites before application × 100%
Control effect (%) = (Reduction rate of live mites in the treatment group - Reduction rate of live mites in the control group)/(1 - Reduction rate of live mites in the control group) × 100%
活螨减退率＝(施药前活螨数-施药后活螨数)/施药前活螨数×100％
防治效果(％)＝(处理组活螨减退率-对照组活螨减退率)/(1-对照组活螨减退率)×100％The control effect of Bacillus veleis Bv-6 wettable powder on citrus Panonychus citri: The spray method was used to detect the 500 of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) For the control effect of twice the liquid on citrus Panonychus mites, weigh 20g of Bv-6 wettable powder, dilute it with 10L of water, and prepare the powder into 500 times the liquid. It was conducted in the citrus garden at the campus base of Huazhong Agricultural University in May 2022, and grapefruit trees damaged by citrus panonychus mites were selected. The test set up two treatments: 500 times solution of Bv-6 powder and water. Each treatment was repeated three times, arranged in random blocks, and two grapefruit trees were sprayed in each plot. Select the citrus panonychus mites to harm the young shoots in the east, south, west, north, and middle directions of the grapefruit tree. Record the number of citrus panonychus mites before applying pesticides and mark them. Use a manual sprayer to spray the young tips of the crown of the tree, and spray evenly on the front and back of the leaves so that the young leaves and tips are fully exposed to the liquid until a small amount of liquid drips from the leaves. 1d, 2d and 5d after spraying, use a magnifying glass to check the number of live citrus Panonychus mites on the leaves of each treatment, touch the mites gently with a brush, and mites that cannot move their legs are considered dead. 5 days after the first spraying, the second spraying was carried out. The number of live mites was investigated and counted 1d, 2d, 5d and 7d after the second spraying, and the reduction rate of live mites and the control effect were calculated.
Reduction rate of live mites = (number of live mites before application - number of live mites after application)/number of live mites before application × 100%
Control effect (%) = (Reduction rate of live mites in the treatment group - Reduction rate of live mites in the control group)/(1 - Reduction rate of live mites in the control group) × 100%
Bv-6可湿性粉剂500倍液对柑橘全爪螨第一次喷施后1d、2d和5d的防治效果分别为87.20％，90.80％和88.87％，第二次喷施后1d、2d、5d和7d的防治效果分别为94.59％，96.41％，95.56％和97.01％，(表1)。The control effects of Bv-6 wettable powder 500 times on citrus Panonychus mites were 87.20%, 90.80% and 88.87% respectively 1d, 2d and 5d after the first spraying, and 1d, 2d and 5d after the second spraying. The control effects at 7 days and 7 days were 94.59%, 96.41%, 95.56% and 97.01% respectively (Table 1).
表1 Bv-6可湿性粉剂500倍液对柑橘全爪螨的防效
Table 1 The control effect of Bv-6 wettable powder 500 times on citrus Panonychus mites
Table 1 The control effect of Bv-6 wettable powder 500 times on citrus Panonychus mites
贝莱斯芽孢杆菌Bv-6可湿性粉剂对二斑叶螨(Tetranychus urticae)的防效：采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)的500倍液对草莓叶螨(二斑叶螨)的防效，称取1g Bv-6可湿性粉剂，用500mL清水稀释，配置成粉剂500倍液。试验设Bv-6粉剂500倍液、10.5％阿维·哒螨灵乳油1000倍液、以及清水对照，共3个处理，每个处理3次重复。施药前每个草莓枝条统计3片叶片的二斑螨虫数量，并做标记。采用手动喷雾器对草莓叶片喷雾，叶片正反面均匀喷施，使叶片充分接触药液，至叶片有少量药液下滴为止。喷药24h和48h后用放大镜检查各个处理叶片上二斑叶螨的活螨数，用毛笔轻轻触碰螨虫，以螨足不动者为死亡。计算活螨减退率和防治效果。
活螨减退率＝(施药前活螨数-施药后活螨数)/施药前活螨数×100％
防治效果(％)＝(处理组活螨减退率-对照组活螨减退率)/(1-对照组活螨减退率)×100％The control effect of Bacillus veleis Bv-6 wettable powder against two-spotted spider mite (Tetranychus urticae): The spray method was used to detect the 500 of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) For the control effect of twice the liquid on strawberry spider mites (two-spotted spider mites), weigh 1g of Bv-6 wettable powder, dilute it with 500 mL of water, and prepare the powder into 500 times the liquid. The test consisted of 500-fold solution of Bv-6 powder, 1000-fold solution of 10.5% Avi-pyrididab EC, and water control, with a total of 3 treatments, and each treatment was repeated 3 times. Before application, count the number of two-spotted mites on 3 leaves of each strawberry branch and mark them. Use a manual sprayer to spray the strawberry leaves, evenly spraying the front and back of the leaves, so that the leaves are fully exposed to the liquid until a small amount of liquid drips from the leaves. 24h and 48h after spraying, use a magnifying glass to check the number of living two-spotted spider mites on the leaves of each treatment. Use a brush to gently touch the mites. The mites that cannot move their legs are considered dead. Calculate the reduction rate of live mites and the control effect.
Reduction rate of live mites = (number of live mites before application - number of live mites after application)/number of live mites before application × 100%
Control effect (%) = (Reduction rate of live mites in the treatment group - Reduction rate of live mites in the control group)/(1 - Reduction rate of live mites in the control group) × 100%
活螨减退率＝(施药前活螨数-施药后活螨数)/施药前活螨数×100％
防治效果(％)＝(处理组活螨减退率-对照组活螨减退率)/(1-对照组活螨减退率)×100％The control effect of Bacillus veleis Bv-6 wettable powder against two-spotted spider mite (Tetranychus urticae): The spray method was used to detect the 500 of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) For the control effect of twice the liquid on strawberry spider mites (two-spotted spider mites), weigh 1g of Bv-6 wettable powder, dilute it with 500 mL of water, and prepare the powder into 500 times the liquid. The test consisted of 500-fold solution of Bv-6 powder, 1000-fold solution of 10.5% Avi-pyrididab EC, and water control, with a total of 3 treatments, and each treatment was repeated 3 times. Before application, count the number of two-spotted mites on 3 leaves of each strawberry branch and mark them. Use a manual sprayer to spray the strawberry leaves, evenly spraying the front and back of the leaves, so that the leaves are fully exposed to the liquid until a small amount of liquid drips from the leaves. 24h and 48h after spraying, use a magnifying glass to check the number of living two-spotted spider mites on the leaves of each treatment. Use a brush to gently touch the mites. The mites that cannot move their legs are considered dead. Calculate the reduction rate of live mites and the control effect.
Reduction rate of live mites = (number of live mites before application - number of live mites after application)/number of live mites before application × 100%
Control effect (%) = (Reduction rate of live mites in the treatment group - Reduction rate of live mites in the control group)/(1 - Reduction rate of live mites in the control group) × 100%
Bv-6可湿性粉剂500倍液对二斑叶螨的24h和48h防治效果分别为90.71％，93.4％(图12、表2)。The 24h and 48h control effects of Bv-6 wettable powder 500 times on two-spotted spider mites were 90.71% and 93.4% respectively (Figure 12, Table 2).
表2 Bv-6可湿性粉剂对二斑叶螨的防效
Table 2 Control effectiveness of Bv-6 wettable powder against two-spotted spider mites
Table 2 Control effectiveness of Bv-6 wettable powder against two-spotted spider mites
实施例4贝莱斯芽孢杆菌Bv-6可湿性粉剂对柑橘木虱(Diaphorina citri)的防效试验Example 4 Control effect test of Bacillus veleis Bv-6 wettable powder on citrus psyllid (Diaphorina citri)
采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)的500倍液和100倍液对柑橘木虱的防效，分别称取1g和5g Bv-6可湿性粉剂，用500mL清水稀释，配置成粉剂500倍液和100倍液。于2022年8月在广东省农业科学院白云基地柑橘园进行，选取柑橘木虱危害的柑橘植株。试验设Bv-6粉剂500倍液、Bv-6粉剂100倍液、甲维·虫螨腈1000倍液、以及清水对照，共4个处理。每个处理3次重复，随机区组排列，每个小区施药2株柑橘树。在柑橘树的东、南、西、北、中5个方位选定柑橘木虱危害嫩梢，施药前记录柑橘木虱成虫和若虫数量，并做标记。采用手动喷雾器对树冠嫩梢喷雾，叶片正反面均匀喷施，使嫩叶嫩梢充分接触药液，至叶片有少量药液下滴为止。喷药24h后调查各个处理嫩梢上柑橘木虱的活虫数，统计各处理柑橘木虱存活虫总数，并计算虫口减退率和防治效果。
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数×100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％The spray method was used to detect the control effect of 500-fold and 100-fold solutions of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) on citrus psyllid. Weighed 1g and 5g of Bv-6 respectively. For wet powder, dilute it with 500mL of water and prepare 500 times powder liquid and 100 times liquid powder. It was conducted in the citrus orchard of the Baiyun Base of the Guangdong Academy of Agricultural Sciences in August 2022, and citrus plants damaged by citrus psyllids were selected. The test consisted of 500-fold solution of Bv-6 powder, 100-fold solution of Bv-6 powder, 1,000-fold solution of emamectin and fenfofen, and water control, a total of 4 treatments. Each treatment was repeated three times, arranged in random blocks, and two citrus trees were sprayed in each plot. Select the citrus psyllid-damaged young shoots in the east, south, west, north, and middle directions of the citrus tree. Record the number of citrus psyllid adults and nymphs before applying pesticides and mark them. Use a manual sprayer to spray the young tips of the crown of the tree, and spray evenly on the front and back of the leaves so that the young leaves and tips are fully exposed to the liquid until a small amount of liquid drips from the leaves. 24 hours after spraying, the number of live citrus psyllids on the young shoots of each treatment was investigated, the total number of surviving citrus psyllids in each treatment was counted, and the population reduction rate and control effect were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application × 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数×100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％The spray method was used to detect the control effect of 500-fold and 100-fold solutions of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) on citrus psyllid. Weighed 1g and 5g of Bv-6 respectively. For wet powder, dilute it with 500mL of water and prepare 500 times powder liquid and 100 times liquid powder. It was conducted in the citrus orchard of the Baiyun Base of the Guangdong Academy of Agricultural Sciences in August 2022, and citrus plants damaged by citrus psyllids were selected. The test consisted of 500-fold solution of Bv-6 powder, 100-fold solution of Bv-6 powder, 1,000-fold solution of emamectin and fenfofen, and water control, a total of 4 treatments. Each treatment was repeated three times, arranged in random blocks, and two citrus trees were sprayed in each plot. Select the citrus psyllid-damaged young shoots in the east, south, west, north, and middle directions of the citrus tree. Record the number of citrus psyllid adults and nymphs before applying pesticides and mark them. Use a manual sprayer to spray the young tips of the crown of the tree, and spray evenly on the front and back of the leaves so that the young leaves and tips are fully exposed to the liquid until a small amount of liquid drips from the leaves. 24 hours after spraying, the number of live citrus psyllids on the young shoots of each treatment was investigated, the total number of surviving citrus psyllids in each treatment was counted, and the population reduction rate and control effect were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application × 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
Bv-6可湿性粉剂500倍液和100倍液对柑橘木虱的24h防治效果分别为93.27％，96.77％(图13、表3)。The 24-hour control effects of Bv-6 wettable powder 500 times and 100 times on citrus psyllid were 93.27% and 96.77% respectively (Figure 13, Table 3).
表3 Bv-6可湿性粉剂对柑橘木虱若虫的田间防效
Table 3 Field control effectiveness of Bv-6 wettable powder against citrus psyllid nymphs
Table 3 Field control effectiveness of Bv-6 wettable powder against citrus psyllid nymphs
实施例5贝莱斯芽孢杆菌Bv-6可湿性粉剂对烟粉虱(Bemisia tabaci)的防效试验Example 5 Control effect test of Bacillus veleis Bv-6 wettable powder on Bemisia tabaci
采用喷雾法检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)的500倍液对烟粉虱的防效，称取1g Bv-6可湿性粉剂，用500mL清水稀释，配置成粉剂500倍液。于2022年9月在华中农业大学校内棉花实验田进行，选取烟粉虱危害严重的棉花植株。试验设Bv-6粉剂500倍液、22％氟啶虫胺腈1000倍液、以及清水对照，共3个处理。每个处理随机选取三株棉花植株，施药前记录棉花植株烟粉虱数量，并做标记。采用喷雾器对整株棉花叶片喷雾，叶片正反面均匀喷施，使棉叶正反两面充分接触药液，至叶片有少量药液下滴为止。喷药24h后调查各个处理棉花叶片活虫数，计算烟粉虱虫口减退率和防治效果。
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数×100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％Use the spray method to test the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) on Bemisia tabaci. Weigh 1g of Bv-6 wettable powder and dilute it with 500mL of water. , configured into powder 500 times liquid. It was conducted in the cotton experimental field of Huazhong Agricultural University in September 2022, and cotton plants seriously damaged by whitefly were selected. The test consisted of 500-fold solution of Bv-6 powder, 1000-fold solution of 22% sulfoxaflor, and water control, a total of 3 treatments. Three cotton plants were randomly selected from each treatment, and the number of whiteflies tabaci on the cotton plants was recorded and marked before spraying. Use a sprayer to spray the entire cotton leaves, and spray evenly on the front and back of the leaves, so that both sides of the cotton leaves are fully exposed to the liquid until a small amount of liquid drips from the leaves. 24 hours after spraying, the number of live insects on cotton leaves in each treatment was investigated, and the population reduction rate and control effect of Bemisia tabaci were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application × 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
虫口减退率＝(施药前虫数-施药后虫数)/施药前虫数×100％
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口减退率)×100％Use the spray method to test the control effect of 500 times solution of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) on Bemisia tabaci. Weigh 1g of Bv-6 wettable powder and dilute it with 500mL of water. , configured into powder 500 times liquid. It was conducted in the cotton experimental field of Huazhong Agricultural University in September 2022, and cotton plants seriously damaged by whitefly were selected. The test consisted of 500-fold solution of Bv-6 powder, 1000-fold solution of 22% sulfoxaflor, and water control, a total of 3 treatments. Three cotton plants were randomly selected from each treatment, and the number of whiteflies tabaci on the cotton plants was recorded and marked before spraying. Use a sprayer to spray the entire cotton leaves, and spray evenly on the front and back of the leaves, so that both sides of the cotton leaves are fully exposed to the liquid until a small amount of liquid drips from the leaves. 24 hours after spraying, the number of live insects on cotton leaves in each treatment was investigated, and the population reduction rate and control effect of Bemisia tabaci were calculated.
Insect population reduction rate = (number of insects before application - number of insects after application) / number of insects before application × 100%
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) × 100%
Bv-6可湿性粉剂500倍液对棉花烟粉虱24h的防治效果为85.86％(图14，表4)。The control effect of Bv-6 wettable powder 500 times on cotton whitefly in 24 hours was 85.86% (Figure 14, Table 4).
表4 Bv-6可湿性粉剂对棉花烟粉虱的田间防效
Table 4 Field control effectiveness of Bv-6 wettable powder against cotton whitefly
Table 4 Field control effectiveness of Bv-6 wettable powder against cotton whitefly
实施例6不同芽孢杆菌菌株对水稻褐飞虱的防效试验Example 6 Test on the control efficacy of different Bacillus strains against rice brown planthopper
枯草芽孢杆菌(Bacillus subtilis)Bs1、副炭疽芽胞杆菌(Bacillus paranthracis)Bp1、贝莱斯芽孢杆菌(Bacillus velezensis)菌株Bv-10和Bv-12，与贝莱斯芽孢杆菌(Bacillus velezensis)菌株Bv-6做水稻褐飞虱防效对比试验。分别检测Bs1，Bp1，Bv-10，Bv-12和Bv-6这五种芽孢杆菌的LB摇培发酵液(OD＝0.6)对水稻褐飞虱的触杀活性，对照采用LB培养基处理。每种芽孢杆菌发酵液喷施10mL到含有水稻褐飞虱(30只)的塑料杯内，每种芽孢杆菌三个重复，48小时后统计水稻褐飞虱的死亡率，计算防治效果。
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)*100％
对水稻褐飞虱的防治效果如下：贝莱斯芽孢杆菌菌株Bv-6(防效71.6％)>贝莱斯芽孢杆菌菌株Bv-12
(防效25.91％)>枯草芽孢杆菌Bs-1(防效19.99％)>贝莱斯芽孢杆菌菌株Bv-10(防效4.73％)>副炭疽芽胞杆菌Bp1(防效3.54％)(图15)。Bacillus subtilis Bs1, Bacillus paraanthracis Bp1, Bacillus velezensis strains Bv-10 and Bv-12, and Bacillus velezensis strain Bv- 6. Conduct a comparative test on the control effectiveness of rice brown planthopper. The contact activity of LB shake culture fermentation broth (OD=0.6) of Bacillus Bs1, Bp1, Bv-10, Bv-12 and Bv-6 against rice brown planthopper was tested respectively, and the control was treated with LB medium. Spray 10 mL of each Bacillus fermentation liquid into a plastic cup containing 30 rice brown planthoppers. Each type of Bacillus was repeated three times. After 48 hours, the mortality of rice brown planthoppers was counted and the control effect was calculated.
Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) * 100%
The control effect on rice brown planthopper is as follows: Bacillus veleis strain Bv-6 (control effect 71.6%) > Bacillus veleis strain Bv-12
(control effect 25.91%)>Bacillus subtilis Bs-1 (control effect 19.99%)>Bacillus veleis strain Bv-10 (control effect 4.73%)>Bacillus paraanthracis Bp1 (control effect 3.54%) (Figure 15 ).
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)*100％
对水稻褐飞虱的防治效果如下：贝莱斯芽孢杆菌菌株Bv-6(防效71.6％)>贝莱斯芽孢杆菌菌株Bv-12
(防效25.91％)>枯草芽孢杆菌Bs-1(防效19.99％)>贝莱斯芽孢杆菌菌株Bv-10(防效4.73％)>副炭疽芽胞杆菌Bp1(防效3.54％)(图15)。Bacillus subtilis Bs1, Bacillus paraanthracis Bp1, Bacillus velezensis strains Bv-10 and Bv-12, and Bacillus velezensis strain Bv- 6. Conduct a comparative test on the control effectiveness of rice brown planthopper. The contact activity of LB shake culture fermentation broth (OD=0.6) of Bacillus Bs1, Bp1, Bv-10, Bv-12 and Bv-6 against rice brown planthopper was tested respectively, and the control was treated with LB medium. Spray 10 mL of each Bacillus fermentation liquid into a plastic cup containing 30 rice brown planthoppers. Each type of Bacillus was repeated three times. After 48 hours, the mortality of rice brown planthoppers was counted and the control effect was calculated.
Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) * 100%
The control effect on rice brown planthopper is as follows: Bacillus veleis strain Bv-6 (control effect 71.6%) > Bacillus veleis strain Bv-12
(control effect 25.91%)>Bacillus subtilis Bs-1 (control effect 19.99%)>Bacillus veleis strain Bv-10 (control effect 4.73%)>Bacillus paraanthracis Bp1 (control effect 3.54%) (Figure 15 ).
实施例7活体水稻盆栽检测贝莱斯芽孢杆菌Bv-6发酵液对褐飞虱的防治效果Example 7 In vivo rice pot testing of the control effect of Bacillus veleis Bv-6 fermentation liquid on brown planthopper
取一周龄水稻幼苗六盆(每盆30株)，试验分为两个处理，处理一：每盆水稻幼苗接种100头褐飞虱后均匀喷施Bv-6摇培发酵液(OD值0.6)20mL；处理二：每盆水稻幼苗接种100头褐飞虱后均匀喷施LB培养基20mL。每个处理三个重复，水稻幼苗放置玻璃缸后用纱布封好，处理1d，3d，8d后统计虫体数量，并计算防治效果。在处理的第8d后分别统计对照和Bv-6处理的水稻幼苗存活率。
虫口减退率(％)＝(防治前的活虫数-防治后的活虫数)/防治前的活虫数*100
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口率)*100％Six pots of one-week-old rice seedlings (30 plants per pot) were taken. The experiment was divided into two treatments. Treatment 1: Each pot of rice seedlings was inoculated with 100 brown planthoppers and then evenly sprayed with 20 mL of Bv-6 shaking culture fermentation broth (OD value 0.6); Treatment 2: Each pot of rice seedlings was inoculated with 100 brown planthoppers and then evenly sprayed with 20 mL of LB medium. Each treatment was repeated three times. The rice seedlings were placed in a glass jar and sealed with gauze. After 1, 3, and 8 days of treatment, the number of insects was counted, and the control effect was calculated. After the 8th day of treatment, the survival rates of rice seedlings in the control and Bv-6 treatments were calculated.
Insect population reduction rate (%) = (number of live insects before control - number of live insects after control)/number of live insects before control * 100
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) * 100%
虫口减退率(％)＝(防治前的活虫数-防治后的活虫数)/防治前的活虫数*100
防治效果(％)＝(处理组虫口减退率-对照组虫口减退率)/(1-对照组虫口率)*100％Six pots of one-week-old rice seedlings (30 plants per pot) were taken. The experiment was divided into two treatments. Treatment 1: Each pot of rice seedlings was inoculated with 100 brown planthoppers and then evenly sprayed with 20 mL of Bv-6 shaking culture fermentation broth (OD value 0.6); Treatment 2: Each pot of rice seedlings was inoculated with 100 brown planthoppers and then evenly sprayed with 20 mL of LB medium. Each treatment was repeated three times. The rice seedlings were placed in a glass jar and sealed with gauze. After 1, 3, and 8 days of treatment, the number of insects was counted, and the control effect was calculated. After the 8th day of treatment, the survival rates of rice seedlings in the control and Bv-6 treatments were calculated.
Insect population reduction rate (%) = (number of live insects before control - number of live insects after control)/number of live insects before control * 100
Control effect (%) = (Insect population reduction rate in the treatment group - Insect population reduction rate in the control group) / (1 - Insect population reduction rate in the control group) * 100%
水稻幼苗存活率(％)＝(存活水稻幼苗株数/总水稻幼苗株数)*100％Rice seedling survival rate (%) = (number of surviving rice seedlings/total number of rice seedlings)*100%
Bv-6喷施后的水稻植株上的褐飞虱虫口数量显著减少，水稻植株生长状况良好，并未发生枯萎现象，
经过LB培养基喷施的CK处理，褐飞虱的虫口数量并未出现减退，水稻植株受褐飞虱危害后出现枯萎现象。经过Bv-6发酵液处理1d后对水稻褐飞虱的防效为64.69％，3d后的防效为70.21％，8d后的防效为76.27％。在处理的第8d对照组水稻幼苗存活率为15.56％，经过Bv-6处理的水稻幼苗存活率为100％，在处理的第10d对照组水稻幼苗全部枯死(存活率为0％)，经过Bv-6处理的水稻幼苗存活率为100％(图16)。After Bv-6 was sprayed, the number of brown planthoppers on the rice plants was significantly reduced, and the rice plants grew in good condition without wilting. After the CK treatment sprayed on LB medium, the number of brown planthoppers did not decrease, and the rice plants withered after being damaged by the brown planthoppers. The control effect against rice brown planthopper after treatment with Bv-6 fermentation broth for 1 day was 64.69%, the control effect after 3 days was 70.21%, and the control effect after 8 days was 76.27%. On the 8th day of treatment, the survival rate of rice seedlings in the control group was 15.56%. The survival rate of rice seedlings treated with Bv-6 was 100%. On the 10th day of treatment, all rice seedlings in the control group died (survival rate was 0%). After Bv-6 treatment, the survival rate of rice seedlings was 100%. The survival rate of rice seedlings treated with -6 was 100% (Figure 16).
实施例8贝莱斯芽孢杆菌Bv-6可湿性粉剂对水稻褐飞虱的防效试验Example 8 Control effect test of Bacillus veleis Bv-6 wettable powder on rice brown planthopper
检测贝莱斯芽孢杆菌Bv-6可湿性粉剂(1×1011cfu/g)的500倍稀释液对水稻褐飞虱的防效，称取1g Bv-6可湿性粉剂用500mL无菌水稀释配置成粉剂500倍稀释液，对照采用无菌水处理。取一周龄水稻幼苗六盆(每盆30株)，试验分为两个处理，处理一：每盆水稻幼苗接种30头褐飞虱后均匀喷施Bv-6可湿性粉剂500倍稀释液20mL；处理二：每盆水稻幼苗接种100头褐飞虱后均匀喷施无菌水20mL。每个处理三个重复，水稻幼苗放置玻璃缸后用纱布封好，分别在24h，48h，72h后统计各个处理死亡率并计算防治效果。
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)*100％To test the 500-fold dilution of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) against rice brown planthopper, weigh 1g of Bv-6 wettable powder and dilute it with 500mL of sterile water to prepare The powder was diluted 500 times, and the control was treated with sterile water. Six pots of one-week-old rice seedlings (30 plants per pot) were taken. The test was divided into two treatments. Treatment one: each pot of rice seedlings was inoculated with 30 brown planthoppers and then evenly sprayed with 20 mL of 500-fold dilution of Bv-6 wettable powder; treatment two : Each pot of rice seedlings is inoculated with 100 brown planthoppers and then sprayed evenly with 20 mL of sterile water. Each treatment was repeated three times. The rice seedlings were placed in glass jars and sealed with gauze. The mortality of each treatment was calculated after 24h, 48h, and 72h respectively, and the control effect was calculated.
Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) * 100%
防治效果(％)＝(处理死亡率-对照死亡率)/(1-对照死亡率)*100％To test the 500-fold dilution of Bacillus veleis Bv-6 wettable powder (1×10 11 cfu/g) against rice brown planthopper, weigh 1g of Bv-6 wettable powder and dilute it with 500mL of sterile water to prepare The powder was diluted 500 times, and the control was treated with sterile water. Six pots of one-week-old rice seedlings (30 plants per pot) were taken. The test was divided into two treatments. Treatment one: each pot of rice seedlings was inoculated with 30 brown planthoppers and then evenly sprayed with 20 mL of 500-fold dilution of Bv-6 wettable powder; treatment two : Each pot of rice seedlings is inoculated with 100 brown planthoppers and then sprayed evenly with 20 mL of sterile water. Each treatment was repeated three times. The rice seedlings were placed in glass jars and sealed with gauze. The mortality of each treatment was calculated after 24h, 48h, and 72h respectively, and the control effect was calculated.
Control effect (%) = (treatment mortality - control mortality) / (1 - control mortality) * 100%
Bv-6可湿性粉剂500倍稀释液对褐飞虱的24h，48h和72h防治效果分别为76.84％，86.77％，89.46％(图17)。
The 24h, 48h and 72h control effects of the 500-fold dilution of Bv-6 wettable powder on brown planthoppers were 76.84%, 86.77% and 89.46% respectively (Figure 17).
Claims (8)
- 贝莱斯芽孢杆菌Bacillus velezensis Bv-6在防虫中的应用，其特征在于，所述贝莱斯芽孢杆菌Bacillus velezensis Bv-6的保藏号为CCTCC NO:M 20191106。The application of Bacillus velezensis Bv-6 in insect control is characterized in that the deposit number of Bacillus velezensis Bv-6 is CCTCC NO:M 20191106.
- 如权利要求1所述应用，其特征在于，所述防虫为防治吸食性害虫。The application according to claim 1, characterized in that the insect control is to control sucking pests.
- 如权利要求2所述应用，其特征在于，所述吸食性害虫为褐飞虱、蚜虫、柑橘木虱(Diaphorina citri)、烟粉虱(Bemisia tabaci)或/和螨虫。Application as claimed in claim 2, characterized in that the sucking pests are brown planthoppers, aphids, citrus psyllids (Diaphorina citri), whitefly (Bemisia tabaci) or/and mites.
- 如权利要求3所述应用，其特征在于，所述蚜虫为绿桃蚜(Myzus persicae)、禾谷缢管蚜(Rhopalosiphum padi)、麦长管蚜(Sitobion avenae)、瓜蚜(Aphis gossypii)、豌豆修尾蚜(Megoura japonica)、桃粉蚜(Hyalopterus arundimis)或月季长管蚜(Macrosiphum rosirvorum)。Application as claimed in claim 3, characterized in that the aphids are green peach aphid (Myzus persicae), cereal aphid (Rhopalosiphum padi), wheat aphid (Sitobion avenae), melon aphid (Aphis gossypii), Pea aphid (Megoura japonica), pink peach aphid (Hyalopterus arundimis) or rose aphid (Macrosiphum rosirvorum).
- 如权利要求3所述应用，其特征在于，所述螨虫为柑橘全爪螨(Panonychus citri)或二斑叶螨(Tetranychus urticae)。Application as claimed in claim 3, characterized in that the mite is Panonychus citri or Tetranychus urticae.
- 如权利要求1所述应用，其特征在于，所述的贝莱斯芽孢杆菌(Bacillus velezensis)Bv-6的剂型为可湿性粉剂。Application as claimed in claim 1, characterized in that the dosage form of Bacillus velezensis Bv-6 is a wettable powder.
- 一种防虫制剂，其特征在于，其主要成分为贝莱斯芽孢杆菌Bv-6的发酵液或可湿性粉剂，所述贝莱斯芽孢杆菌Bacillus velezensis Bv-6的保藏号为CCTCC NO:M 20191106。An anti-insect preparation, characterized in that its main component is the fermentation broth or wettable powder of Bacillus velezensis Bv-6, and the preservation number of Bacillus velezensis Bv-6 is CCTCC NO:M 20191106 .
- 如权利要求7所述防虫制剂，其特征在于，所述可湿性粉剂的浓度为1×1011cfu/g。 The anti-insect preparation according to claim 7, wherein the concentration of the wettable powder is 1×10 11 cfu/g.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20180020676A1 (en) * | 2014-12-29 | 2018-01-25 | Fmc Corporation | Bacillus velezensis rti301 compositions and methods of use for benefiting plant growth and treating plant disease |
| CN111254086A (en) * | 2019-12-27 | 2020-06-09 | 华中农业大学 | Bacillus belgii and application thereof in biocontrol |
| CN114410498A (en) * | 2021-09-24 | 2022-04-29 | 成都特普生物科技股份有限公司 | Microbial preparation, preparation method and application thereof |
| CN115005234A (en) * | 2022-06-24 | 2022-09-06 | 华中农业大学 | Application of Bacillus beilesiensis Bv-6 in preventing and treating brown planthopper |
| CN115399338A (en) * | 2022-09-19 | 2022-11-29 | 华中农业大学 | Application of Bacillus velezensis Bv-6 in insect prevention |
-
2023
- 2023-03-24 WO PCT/CN2023/083575 patent/WO2023246208A1/en unknown
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20180020676A1 (en) * | 2014-12-29 | 2018-01-25 | Fmc Corporation | Bacillus velezensis rti301 compositions and methods of use for benefiting plant growth and treating plant disease |
| CN111254086A (en) * | 2019-12-27 | 2020-06-09 | 华中农业大学 | Bacillus belgii and application thereof in biocontrol |
| CN114410498A (en) * | 2021-09-24 | 2022-04-29 | 成都特普生物科技股份有限公司 | Microbial preparation, preparation method and application thereof |
| CN115005234A (en) * | 2022-06-24 | 2022-09-06 | 华中农业大学 | Application of Bacillus beilesiensis Bv-6 in preventing and treating brown planthopper |
| CN115399338A (en) * | 2022-09-19 | 2022-11-29 | 华中农业大学 | Application of Bacillus velezensis Bv-6 in insect prevention |
Non-Patent Citations (1)
| Title |
|---|
| CAI GAOLEI; ZHANG FAN; OUYANG YOUXIANG; ZHAO CHANGSONG; PENG XUANHE; JIANG AIMING: "Research Progress on Bacillus Velezensis", NORTHERN HORTICULTURE, no. 12, 30 June 2018 (2018-06-30), pages 162 - 167, XP009551475, ISSN: 1001-0009 * |
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