Bacillus atrophaeus having poisoning and controlling effects on potato beetles

A technology of Bacillus atrophaeus and potato beetle, which is applied in the direction of bacteria, insecticides, and microbial-based methods, can solve problems such as unseen research reports, achieve strong pathogenicity and control effects, alleviate the resulting resistance, and potential The effect of developing application value and commercial promotion benefits

Inactive Publication Date: 2014-11-26
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the pathogenic effect and biocontrol application of Bacillus atr...

Method used

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  • Bacillus atrophaeus having poisoning and controlling effects on potato beetles
  • Bacillus atrophaeus having poisoning and controlling effects on potato beetles
  • Bacillus atrophaeus having poisoning and controlling effects on potato beetles

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Example 1: Isolation, purification and screening scheme of Bacillus atrophaeus

[0031] Isolation and culture of Bacillus atrophaeus : Use Luria-Bertani medium (LB), its formula is tryptone 1%, yeast powder 0.5%, NaCl 1%, agar 2%, tap water 1000mL, pH7.0. Heat and melt the weighed agar; separately weigh yeast powder, tryptone and NaCl, dissolve in a small amount of hot water, add and mix well, then add water to make up to 1000 mL. The pH was adjusted with NaOH. After dispensing into test tubes or Erlenmeyer flasks, sterilize at 121°C for 25 minutes. Heat and melt before use, pour into sterilized petri dishes or test tubes to make flat or slant medium for strain isolation and culture or strain preservation.

[0032] Isolation culture and purification : Collect dead potato beetle samples under natural conditions, use sterile tweezers to take the dead beetles and immerse them in 70% alcohol for about 2 seconds. After the surface of the beetles is wet, move them int...

Embodiment 2

[0036] Embodiment 2: the cultivation of bacillus atrophaeus and inoculation condition experimental scheme

[0037] temperature test : Inoculate fresh strains (inoculated and cultured for 12~24h) on LB solid culture plates, and inoculate those with higher temperature (above 37°C) in liquid medium, respectively set at 4°C, 20°C, 30°C, and 37°C , 41°C, 45°C and 65°C were initially treated at 8 temperatures, each treatment was repeated 3 times, and cultured in a water bath above 37°C. Place them in an incubator and a constant temperature water bath with the required temperature set in advance for cultivation, and observe the growth situation day by day. After selecting the temperature range, conduct further temperature tests, and set different treatments according to the preliminary test results. Therefore, the optimal growth temperature of Bacillus atrophaeus was determined.

[0038] pH test: LB liquid medium was used in the experiment, and 11 pH treatments were set up. ...

Embodiment 3

[0042] Embodiment 3: Indoor pathogenicity test of germs : Wash the activated bacterial colonies into the Erlenmeyer flask with sterile water, fully shake them evenly, and make proper dilution to make spore suspension, the concentration is 10 8 cells / mL and above orders of magnitude. Place the 1st and 2nd instar larvae of potato beetle in a sterilized petri dish with a diameter of 15 cm (the petri dish is lined with a layer of absorbent paper, 10 per dish), and use a pipette to draw the prepared spore suspension and spray it evenly on the Around the body wall of the worm body, it is advisable to have full contact. When the body wall of the insect body is relatively dry and the absorbent paper is relatively dry, add a certain amount of clean and fresh potato leaves, then cover with gauze, and place it at room temperature to allow it to grow. In addition, the test insects were sprayed with sterilized water as a control. Three replicates were set up for each strain treatment a...

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Abstract

Bacillus atrophaeus CPB072 having poisoning and controlling effects on potato beetles is separated from natural environment, can grow and massively produce spores under conventional culture conditions, an optimum culture medium is a Luria-Bertani culturing medium, an optimum temperature is 30DEG C, and an optimal pH value is 7.0; and an optimum condition of fungal infection pathogenicity is that the inoculation concentration is more than 10<8>/mL. The above bacterium has strong pathogenic poisoning effects on potato beetle larvae, has strong insecticidal activity on lepidopteran insects, can be processed through a bacterium fermentation technology to produce a safe and environmentally-friendly bacterial pesticide without pollution or residual, can also clone other insecticidal functional genes for research and development of transgenic beetle-resistant potato, and has latent commercial development and application values.

Description

technical field [0001] The invention belongs to the technical field of biological control of agricultural pests, and in particular relates to a strain of Bacillus atrophaeus which can kill and control potato beetles. Background technique [0002] potato beetle ( L. decemlineata , English abbreviation CPB) is one of the top ten destructive pests recognized internationally, and it is also one of the major quarantine objects and important invasive alien species in my country. The insect feeds on adults and larvae, and the most preferred host is the cultivated potato ( Solanum tuberosum ) and eggplant ( S. melongena ), often eating up the whole potato leaves, causing serious damage and yield loss. According to literature reports and damage surveys of potato beetle-occurring areas in my country, the yield loss due to potato beetle damage is generally 20-50%, and in severe cases it can reach more than 80%, or even result in extinction. At the same time, CPB can also spread p...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01P7/04C12R1/07
Inventor 陆慧慧谭万忠罗华东郭文超
Owner SOUTHWEST UNIVERSITY
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