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Acinetobacter guillouiae PxCG3 strain of plutella xylostella (L) and application of acinetobacter guillouiae PxCG3 strain

A technology of diamondback moth and bacterial strains, applied in the field of agricultural microorganisms

Active Publication Date: 2021-02-05
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, patent CN109402008A discloses a strain of Acinetobacter TAT1-6A with indole degrading ability and its application. The indole degrading ability measurement results of said strain show that the removal effect of indole is good, and it can effectively degrade indole in feces. , the highest degradation rate can reach 84.45%, showing high application potential; CN109161511A discloses a kind of Acinetobacter inducing tomato resistance to M. The use of nematodes can induce tomato to produce resistance to root-knot nematode incognita during seed germination and growth; the above-mentioned patents all involve related functional research on Acinetobacter, but the control of Acinetobacter on Lepidoptera pests and the The effect of Bt toxin on insecticidal activity is still rarely reported

Method used

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  • Acinetobacter guillouiae PxCG3 strain of plutella xylostella (L) and application of acinetobacter guillouiae PxCG3 strain
  • Acinetobacter guillouiae PxCG3 strain of plutella xylostella (L) and application of acinetobacter guillouiae PxCG3 strain
  • Acinetobacter guillouiae PxCG3 strain of plutella xylostella (L) and application of acinetobacter guillouiae PxCG3 strain

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Isolation and cultivation of the bacterial strain PxCG3 of embodiment 1

[0024] (1) Preparation of selective separation medium:

[0025] LB medium (10g peptone, 5g yeast extract, 10g NaCl, 15g agar, dissolved in 1L sterile ddH2O water, adjust pH to 7.0);

[0026] Nutrient agar medium NA (3g beef extract, 10g peptone, 5g NaCl, 15g agar, dissolved in 1L sterile ddH2O, adjust pH to 7.0).

[0027] (2) Dissect and plate: Dissect the 3rd instar larvae of Plutella xylostella in an ultra-clean bench under sterile conditions and take the midgut contents, centrifuge to get the supernatant and sediment, dilute 5 concentration gradients, and dilute 10 -4 ~10 -5 After doubling, NA plates were respectively coated, placed in a constant temperature incubator at 30°C, and observed every 24 hours;

[0028] (3) Continuous purification culture and taking pictures: After a single colony grows in the selective medium, first pick a single colony according to the color, size and shape of t...

Embodiment 2

[0029] Example 2 Identification and Phylogenetic Analysis of Bacterial Strain PxCG3

[0030] 1. Traditional biological identification

[0031] (1) Colony Morphological Characteristics

[0032] From the morphological point of view, PxCG3 bacteria form round gray-white colonies with neat edges on the LB plate. Gram staining is sometimes not easy to decolorize. . This bacterium is obligate to aerobic, and the optimum growth temperature is 35°C; the nutrient requirements are not high, and it grows well on ordinary culture medium;

[0033] (2) Determination of physiological and biochemical characteristics of Acinetobacter guilin

[0034] Bacterial physiological and biochemical assays were carried out with reference to the method in the "Common Bacterial System Identification Manual", using bacterial micro-biochemical reaction tubes. The test results of available carbon sources are shown in Table 1. Galactose, lactose, sorbitol, rhamnose, arabinose, xylose, starch, melibiose, D-...

Embodiment 3

[0043] Example 3 The synergistic effect of PxCG3 strain on Bt

[0044] 1. Removal of intestinal bacteria in diamondback moth

[0045] Prepare antibiotic solution: 1mg / mL ciprofloxacin; 1mg / mL levofloxacin; 2mg / mL metronidazole;

[0046] The method of eliminating intestinal bacteria of diamondback moth and feeding antibiotics. Specific method: Collect diamondback moth egg cards and disinfect: collect egg cards produced during the peak oviposition period of diamondback moth, first place them in 5‰ sodium hypochlorite disinfectant for 10 minutes, and then soak them in clean water twice, each time for 5 minutes. Minutes, finally blotted with absorbent paper and placed in the insect box, after 24 hours, add feed to feed;

[0047] Antibiotic feeding treatment: take 4g of diamondback moth artificial feed in a sterile petri dish, then add ciprofloxacin, levofloxacin and metronidazole antibiotic solution prepared above 333 μL respectively, mix well and place in the insect culture box...

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Abstract

The invention discloses an acinetobacter guillouiae PxCG3 strain of plutella xylostella (L) and application of the acinetobacter guillouiae PxCG3 strain. The strain is preserved in guangdong microbialculture collection center (GDMCC) on August 11, 2020, and the preservation number of the strain is GDMCC No: 61134. Researches show that the acinetobacter guillouiae PxCG3 strain has the effects of improving the insecticidal activity of Bt toxin and synergizing Cry1Ac prototoxin to quickly kill the plutella xylostella (L); and the acinetobacter guillouiae PxCG3 strain can be used as a novel biological prevention and control bacterium, is used for prevention and control of cruciferous vegetable pests, and has extremely good biological prevention and control potential and application prospects.

Description

technical field [0001] The invention relates to the technical field of agricultural microorganisms, and more specifically relates to a strain of Acinetobacter guillouiae PxCG3 of diamondback moth and application thereof. Background technique [0002] Diamondback moth Plutella xylostella (L) is one of the most serious insect pests against cruciferous crops, usually causing severe economic losses to many important vegetables such as cabbage, broccoli and cauliflower (Talekar and Shelton, 1993). Plutella xylostella has developed severe resistance to all insecticides used to control Lepidoptera pests and Bacillus thuringiensis (Bt), making its control increasingly difficult (Talekar and Shelton, 1993; Baxter et al., 2005 ). Therefore, it is necessary to find a method that can solve or alleviate the resistance of diamondback moth to Bt, thereby improving the insecticidal activity of Bt protein. [0003] There are different types and quantities of insect symbiotic bacteria distr...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/20A01N63/50A01N63/23A01P7/04C12R1/01
CPCA01N63/20A01N63/50A01N63/23C12R2001/01C12N1/205Y02A50/30
Inventor 金丰良许小霞李树忠付东冉
Owner SOUTH CHINA AGRI UNIV
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