Application of D-amino acids in inhibition for biofilms of banana bacterial Erwinia carotovora and culture medium thereof

A technology of bacterial biofilm and soft rot pathogens, applied in the direction of microorganism-based methods, bacteria, microorganisms, etc., can solve the problems of death and harm to the banana industry, and achieve the effects of growth inhibition, environmental protection of pesticides, and reduction of use concentration

Active Publication Date: 2015-04-08
PLANT PROTECTION RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The incidence of the disease in the field is 20% to 30%, and it can reach more than 100% in severe cases, and cause the death of the whole plant, which has seriously endangered the banana industry.

Method used

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  • Application of D-amino acids in inhibition for biofilms of banana bacterial Erwinia carotovora and culture medium thereof
  • Application of D-amino acids in inhibition for biofilms of banana bacterial Erwinia carotovora and culture medium thereof
  • Application of D-amino acids in inhibition for biofilms of banana bacterial Erwinia carotovora and culture medium thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: the cultivation of bacteria and the cultivation of bacterial biofilm

[0036]Bacterial strain Dickeya zeae MS1 (Identification of Dickeya zeae as a Causal Agent of Bacterial Soft Rot in Banana in China. Plant Disease, 2014, 98(4): 436-442.) was inoculated with glycerol frozen (-80°C) Streak culture on NA solid medium (beef extract 3g, yeast extract 1g, peptone 5g, glucose 10g, agar 18g, water to 1000mL, pH 7.2-7.4) plate; pick a single colony and culture in liquid NA Base (3g beef extract, 1g yeast extract, 5g peptone, 10g glucose, water to 1000mL, pH 7.2-7.4), 180rpm, 32°C.

[0037] Cultivation of Dickeya zeae MS1 to 10 8 CFU / mL; NA liquid medium to dilute the bacterial solution to 1:1000. Take 2mL of the diluted bacterial solution and add it to a polystyrene 24 microwell plate. Each sample has 3 replicates, and cultured at 32°C. After culturing for 6h, 12h, 24h, 36h, 48h, 60h, 72h, 96h, 144h, and 216h, first extract the planktonic bacteria in a 96-wel...

Embodiment 2

[0040] Embodiment 2: Analysis of the effect of different D-amino acids inhibiting banana bacterial soft rot

[0041] Formulated D-valine, D-methionine, D-tryptophan, D-threonine, D-arginine, D-proline, D-leucine, D-serine, D-histidine , D-aspartic acid, D-glutamic acid, D-glutamine, D-phenylalanine, D-alanine, etc. 100mMol / L storage solution; prepare 10mMol / L of D-tyrosine L storage solution, wherein the D-tyrosine storage solution contains 0.1Mol / L HCl. Prepare series of NA liquid media with D-amino acid concentrations according to different concentration ratios; 500μMol / L (abbreviated as μM) concentration of D-tyrosine NA liquid medium (beef extract 3g, yeast extract 1g, peptone 5g, glucose 10g, water to 1000mL, pH 7.2 ~ 7.4); respectively configure 1 , 3, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 17.5 and 20mMol / L (abbreviated mM) of other D-amino acid NA medium (beef extract 3g, yeast extract 1g, peptone 5g, glucose 10g, water to 1000mL, pH 7.2-7.4).

[0042] Inoculate th...

Embodiment 3

[0047] Embodiment 3: the crystal violet staining analysis of D-amino acid inhibition bacterial biofilm effect

[0048] Inoculate the bacterial strain Dickeya zeae MS1 frozen in glycerol (-80°C) into NA solid medium (beef extract 3g, yeast extract 1g, peptone 5g, glucose 10g, agar 18g, water to 1000mL, pH 7.2-7.4 ) plate, streak culture; pick a single colony and culture in NA liquid medium, 180rpm, 32°C.

[0049] Cultivation of Dickeya zeae MS1 to 10 8 CFU / mL; Prepare 20mM concentration of D-threonine and D-alanine NA liquid medium, 200μM and 420μM concentration of D-tyrosine NA liquid medium, 3mM and 5mM concentration of D-tryptophan NA medium, 5mM and 11mM concentration of D-valine NA medium, 7mM and 15mM concentration of D-methionine NA medium; dilute the bacterial solution to 1:1000 with different concentrations of D-amino acid NA medium, set without D-amino acid bacterial liquid control, and sterile liquid NA medium control. Take 0.2mL of the diluted bacterial solution ...

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Abstract

The invention belongs to the field of crop disease control, and discloses an application of D-amino acids in inhibition for the biofilms of banana bacterial Erwinia carotovora and a culture medium thereof. The D-amino acids screened in the application disclosed by the invention are capable of remarkably and effectively inhibiting the growths of the bacterial biofilms of banana bacterial Erwinia carotovora. According to the application disclosed by the invention, four D-amino acids capable of remarkably inhibiting the growths of the bacterial biofilms of banana bacterial Erwinia carotovora, that is, D-tyrosine, D-valine, D-methionine and D-tryptophan are obtained by screening; and the lowest inhibition concentrations of the four D-amino acids are obtained. The invention further provides a mixing formula of the four D-amino acids, thus the use concentrations of the various D-amino acids are greatly reduced. The invention further provides a D-amino acid culture medium used for the application. On the basis of the contents of the invention, new method and thought can be provided for the development of new control technologies for bacterial Erwinia carotovora; the use effects of control methods such as chemical pesticides and biological pesticides are improved in combination with the use of the D-amino acids.

Description

technical field [0001] The invention belongs to the field of crop disease prevention and control, and particularly relates to an application of D-amino acid in inhibiting bacterial biofilm of banana bacterial soft rot and a culture medium thereof. Background technique [0002] Bananas are mainly grown in tropical and subtropical regions, and bananas have been produced in at least 107 countries; in 2013, bananas were the fourth largest edible plant in the world, second only to rice, wheat and corn. growing trend. However, the crop has been severely damaged by banana bacterial soft rot; since the first report of banana bacterial soft rot caused by Dickeya sp. in mainland China (Lin et al, 2010), the bacterial The expansion and spread of sexual soft rot is rapid, and the disease has been found in the main banana production areas of Guangdong, Hainan, Guangxi, Fujian and other banana production areas. The field incidence of the disease is 20% to 30%, and it can reach more than...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
CPCA01N37/44A01N43/38C12N1/20A01N2300/00
Inventor 张景欣林壁润黄宁沈会芳蒲小明潘群英
Owner PLANT PROTECTION RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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