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Plutella xylostella (L) Enterococcus mundtii PxG1 bacterial strain and application thereof

A technology of Enterococcus monsonii and cabbage, applied in the field of EnterococcusPxG1 strain of diamondback moth

Active Publication Date: 2021-01-05
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned patents all involve the relevant functional research of intestinal symbiotic Enterococcus monsonii, but there are few reports about its influence on the activity of Enterococcus monsonii on insecticides such as Bt protein

Method used

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  • Plutella xylostella (L) Enterococcus mundtii PxG1 bacterial strain and application thereof
  • Plutella xylostella (L) Enterococcus mundtii PxG1 bacterial strain and application thereof
  • Plutella xylostella (L) Enterococcus mundtii PxG1 bacterial strain and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Isolation and cultivation of embodiment 1 bacterial strain PxG1

[0026] (1) Preparation of selective separation medium:

[0027] LB medium (10g peptone, 5g yeast extract, 10g NaCl, 15g agar, dissolved in 1L sterile ddHO 2 O water, adjust pH to 7.0);

[0028] Enterococcus isolation medium EC (3g beef extract powder, 17g tryptone, 5g yeast powder, 10g ox gall powder, 5g sodium chloride, 1g aescin, 0.5g ferric ammonium citrate, 0.25g sodium azide, 1g Sodium citrate, 13.5 g agar, dissolved in 1 L sterile ddHO 2 O, adjust the pH to 7.0).

[0029] (2) Dissect and plate: Dissect the 3rd instar larvae of Plutella xylostella in an ultra-clean bench under aseptic 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, ECs were coated on selective plates, placed in a constant temperature incubator at 30°C, and observed every 24 hours;

[0030] (3) Continuous purificati...

Embodiment 2

[0031] Example 2 Identification and Phylogenetic Analysis of Bacterial Strain PxG1

[0032] 1. Traditional biological identification

[0033] (1) Colony Morphological Characteristics

[0034]Morphologically, PxG1 bacteria are round or oval yellow colonies with a smooth and shiny surface, Gram-positive cocci arranged in chains. Generally, the chain is long in liquid medium and short in solid medium, without flagella and spores, and is an aerobic or facultative anaerobic bacteria. Like Streptococcus, this bacterium also has high nutritional requirements and grows well on medium containing serum.

[0035] (2) Determination of physiological and biochemical characteristics of Enterococcus

[0036] Physiological and biochemical measurements of bacteria were carried out with reference to the method in the "Common Bacterial System Identification Manual", using bacterial micro-biochemical reaction tubes. The test results for available carbon sources are shown in Table 1, for galact...

Embodiment 3

[0045] The eclosion rate of embodiment 3 PxG1 bacterial strains to diamondback moth

[0046] The preserved glycerol bacterium PxG1 was taken out and transferred to LB liquid medium at a ratio of 1:1000 for overnight activation to obtain seed bacteria, and the seed bacteria were transferred to fresh medium at a ratio of 1:100 for about 6 hours. After cultivation, the bacterial liquid was centrifuged to enrich the bacterial cells, ddH 2 O Repeated washing 3 times to remove residual medium, washed with ddH 2 O was diluted to OD600=1.0 and stored for later use.

[0047] The third-instar diamondback moth with the same healthy development period was selected. Weigh 4g of diamondback moth artificial feed in a sterile petri dish, then add 1mL of the above-mentioned bacterium solution, mix well and place it in the insect rearing box, then change the feed every day until eclosion is observed, and the control group is fed with normal feed. The result is as image 3 ,Depend on image...

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Abstract

The invention discloses a Plutella xylostella (L) Enterococcus mundtii PxG1 bacterial strain and application thereof. The bacterial strain is collected in the GuangDong Microbial Culture Collection Center (GDMCC) on June 29th, 2020, and a culture collection number is GDMCC No: 61067. Research indicates that the eclosion rate of the Plutella xylostella (L) fed with the Enterococcus mundtii PxG1 bacterial strain is obviously lowered, and simultaneously indicates that the Enterococcus mundtii PxG1 bacterial strain has a function for improving Bt toxin insecticidal activity and carrying out synergy on Cry1Ac original toxins to quickly kill the Plutella xylostella (L), and the Enterococcus mundtii PxG1 bacterial strain can be used as a novel biological prevention and control bacterium, is usedfor preventing and controlling brassicaceae vegetable pests, and has good biological prevention and treatment potential and a good application prospect.

Description

technical field [0001] The invention relates to the technical field of agricultural microorganisms, and more specifically relates to a Enterococcus PxG1 strain of diamondback moth and its application. 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 distributed in various ti...

Claims

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Application Information

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