Pseudomonas for bacterial disease control and application

A technology for Pseudomonas and disease control, applied in the biological field, can solve the problems of lack of control means and unsatisfactory control effect, and achieve the effect of ensuring soil quality, excellent control effect, and reducing residues

Inactive Publication Date: 2017-07-25
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The special disease cycle and infection route of R. solanacearum cause the spread and infectivity of the disease to be extremely strong, and there is a lack of effective control methods in agricultural production.
Although chemical agents can inhibit the occurrence of diseases to a certain extent, it is difficult to form a sustained effect on R. solanacearum, and the control effect is not ideal

Method used

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  • Pseudomonas for bacterial disease control and application
  • Pseudomonas for bacterial disease control and application
  • Pseudomonas for bacterial disease control and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] (1) Isolation and identification of Pseudomonas A1

[0017] Soil samples were collected from a peanut planting site in Quanjiao County, Anhui Province, inoculated into enriched medium at an inoculum size of 3%, and propagated at 30°C for 12 hours; 2 mL of the supernatant was transferred to 10 mL of fresh NB medium Cultivate at 30°C for 24h. The enriched culture is plated with a gradient concentration, and a single colony of the bacterial culture in the culture medium is isolated, and one of the single colonies is the Pseudomonas strain of the present invention, named A1.

[0018] 16S rRNA universal primers (16SF: 5′-AGAGTTTGATCATGGCTCA-3′; 16SR: 5′-GGTACCTTGTTACGACTT-3′) were designed, and the genome of the isolated strain AH was extracted as a template to amplify the 16S rRNA sequence of the strain. The PCR amplification conditions were: 95°C / 5 min+(94°C / 45 sec+52°C / 30 sec+72°C / 1min30 sec)×33 cycles+72°C / 10min. The PCR product was sequenced and compared with the GenB...

Embodiment 2

[0020] Pseudomonas After sp. A1 was streak cultured on NB and LB solid media, there was no obvious polysaccharide production, and the colony color was dark brown. Because it grows relatively well on NB solid plates, the color is slightly darker. The Pseudomonas fluorescens cultured at the same time was off-white in NB and LB. It is precisely because of the special colony growth color of the isolated strain that it has attracted attention ( figure 2 A). In minimal medium MMX, the isolated strain could not use galactose, mannitol or aditol as the sole carbon source, but it had the ability to assimilate fructan, fructose, inositol and trehalose ( figure 2 B). The strains grew fastest on MMX medium containing fructose and fructans, probably the most efficient carbon source. Pseudomonas fluorescens in parallel experiments could utilize galactose, mannitol or aditol in addition to fructan, fructose, inositol and trehalose, indicating that the isolated strain was not exactly...

Embodiment 3

[0022] (1) Antibacterial effect of Pseudomonas on several plant pathogenic bacteria

[0023] Liquid cultivation of phytopathogenic bacteria in rich media: Xanthomonas oryzae ( Xanthomonas oryzae pv. oryzae ), Rice streak disease ( Xanthomonas oryzae pv. oryzicola ), Canker sorrel citrus ( Xanthomonas citri subsp. citri ) and Ralstonia solanacearum ( Ralstonia solanacearum ), adjust the OD600 value of the cultured bacterial solution to 1.0. Before preparing the plate, add it to the NA medium at a ratio of 1:100, shake well and make a plate with bacteria. Add 2 μl of Pseudomonas solution (OD 600 =1.0); after static culture at 28°C for 48 hours, observe the size of the inhibition zone ( image 3 ). The results showed that the A1 strain had obvious inhibitory effects on the four tested strains.

[0024] (2) Control effect of Pseudomonas A1 on tomato bacterial wilt

[0025] At the 5-6 leaf stage, a knife wound was made at 1 cm from the root of the tomato plant,...

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Abstract

The invention belongs to the technical field of biology and provides a pseudomonas for bacterial disease control and an application. A microorganism with bacteriostatic activity on phytopathogenic bacteria is separated from peanut foot rot soil; and a pseudomonas bacterium is determined according to 16S rRNA and a whole genome sequence. By adopting a bacteria solution root-irrigation method, tomato bacterial wilt can be effectively inhibited. The pseudomonas can be used for green biological control of the phytopathogenic bacteria.

Description

technical field [0001] This patent belongs to the field of biotechnology, and relates to the isolation of a pseudomonas strain that has antibacterial effects on a variety of plant pathogenic bacteria from the soil, and the effective control and application of R. solanacearum in tomato. Background technique [0002] In the natural ecology of the field, crops are threatened by a variety of pathogens, and creating broad-spectrum and efficient new pesticides is an effective strategy for disease control. However, the comprehensive control technology of diseases in my country's agricultural production needs to be improved, and there is still a lot of room for improvement, mainly because (1) the commonly used fungicides are highly specific, and they are often "press the gourd to float the scoop", and the prevalence of crops is high. The disease broke out alternately every other year; (2) The spraying of fungicides, mainly chemical pesticides, caused serious soil environmental pollut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01P1/00C12R1/38
CPCC12N1/205C12R2001/38
Inventor 邹华松孙东玲范晓静卓涛户勋
Owner FUJIAN AGRI & FORESTRY UNIV
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