Lytic phage strain capable of preventing and controlling tomato bacterial wilt and applications of lytic phage strain

A technology of tomato bacterial wilt and phage, applied in the field of prevention and control of tomato bacterial wilt, can solve the problems of little research and single research, and achieve the effect of strong bactericidal ability

Active Publication Date: 2018-10-12
NANJING AGRICULTURAL UNIVERSITY
View PDF18 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through the unremitting efforts of domestic and foreign medical scientists, phages have played an important role in medicine. However, the research on the prevention and control of soil-borne diseases by phages is relatively single, and most of them are limited to the research on the prevention and control of tobacco bacterial wilt. For the prevention and control of tomato bacterial wilt There is still little research on this aspect, therefore, the use of phage to lyse pathogenic bacteria to prevent and control tomato bacterial wilt has broad application prospects

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Lytic phage strain capable of preventing and controlling tomato bacterial wilt and applications of lytic phage strain
  • Lytic phage strain capable of preventing and controlling tomato bacterial wilt and applications of lytic phage strain
  • Lytic phage strain capable of preventing and controlling tomato bacterial wilt and applications of lytic phage strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Screening of solanacearum obligate phage

[0030] Rhizosphere soil samples of tomato plants were collected from tomato greenhouses in Houcun, Qilin Town, Nanjing City, Jiangsu Province, and a R. solanacearum obligate phage NJ-P3 was screened out by double-layer plate method.

[0031] 1.1 Prepare the required medium:

[0032] NA liquid medium: glucose 10g, peptone 5g, beef extract 3g, yeast powder 0.5g, deionized water 1000mL, pH 7.2-7.4, autoclaved at 115°C for 30min.

[0033] NA semi-solid medium: glucose 10g, peptone 5g, beef extract 3g, yeast powder 0.5g, agar powder 10g, deionized water 1000mL, pH 7.2-7.4, autoclaved at 115°C for 30min.

[0034] NA solid medium: glucose 10g, peptone 5g, beef extract 3g, yeast powder 0.5g, agar powder 20g, deionized water 1000mL, pH 7.2-7.4, autoclaved at 115°C for 30min.

[0035] Ralstonia solanacearum selective medium (SMSA selective medium): add TTC 50mg, crystal violet 50mg, polymyxin 100mg, bacitracin 20mg, chloramph...

Embodiment 2

[0045] Example 2 Determination of Biological Characteristics of Solanacearum Obligate Phage NJ-P3

[0046] 2.1 Phage titer determination

[0047] Using sterile water as a diluent, the purified phage NJ-P3 solution (Example 1) was serially diluted 10 times. Take 10mL of QL-Rs1115 bacteria liquid in the logarithmic growth phase and mix with 150mL NA semi-solid medium cooled to about 30°C, and immediately pour it into the solidified NA solid medium plate to make a double-layer plate, and wait for the upper medium After coagulation, the plates were partitioned, and 20 μL of gradient dilutions (10 1 , 10 2 , 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 ) phage stock solution, cultured upside down in an incubator at 28°C for 24h to 48h, observe and count the formation of phage plaques, and do 3 parallel replicates for each of the above dilution gradients. When calculating the titer of phage, the number of plaques on the selected plate is between 30 and 300, sele...

Embodiment 3

[0061] Embodiment 3 indoor antibacterial test

[0062] Take 50 mL of Ralstonia solanacearum QL-Rs1115 cultured to the logarithmic growth stage, adjust the OD with normal saline 600 to 0.5, sequentially add 190 μL to a sterile 96-well plate, add phage preparations (Example 2) sequentially according to the optimal multiplicity of infection of phages, take no addition of phage preparations as a control (CK), and set 3 parallel experiments for each treatment. repeat. Place in a shaker at 30°C and 170r / min for 36 hours, measure OD with a microplate reader every 4 hours 600 value and record. The antibacterial effect of phage was characterized according to the one-step growth curve of R. solanacearum. Depend on Figure 7It can be seen that after adding phage NJ-P3, R. solanacearum OD 600 The value showed a trend of rising first and then falling until stable, while the OD of R. solanacearum without phage 600 has been showing an upward trend. Compared with CK, phage NJ-P3 had a ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Incubation periodaaaaaaaaaa
Login to view more

Abstract

The invention discloses a lytic phage strain NJ-P3 capable of preventing and controlling tomato bacterial wilt, the classification name of the lytic phage strain NJ-P3 is Podoviridae phage, and the lytic phage strain NJ-P3 is preserved in the China Center for Type Culture Collection (CCTCC) on March 6, 2018, and is assigned with the accession number of CCTCC NO:M 2018099. The invention further discloses applications of the phage NJ-P3 in preventing and treating soil-borne bacterial wilt of tomato and applications of the phage NJ-P3 in preparing phage preparations for preventing and treating soil-borne bacterial wilt of tomato. The phage can realize specific splitting of pathogenic bacteria of bacterial wilt, when the temperature is 20-40 DEG C and the pH is 5-9, the phage has the highest activity and the strongest splitting capacity, the bactericidal ability is shown, the indoor antibacterial test shows that the antibacterial rate achieves 72.4%, and thus the phage can be used for preventing and treating soil-borne bacterial wilt of tomato. When the phage provided by the invention is used for preventing and treating soil-borne bacterial wilt of tomato, the disease prevention rate for the potted plant achieves 60%, and the biocontrol rate for the field achieves 85.19%.

Description

technical field [0001] The invention belongs to the field of prevention and control of tomato bacterial wilt, and relates to a lytic phage NJ-P3 capable of preventing and controlling tomato bacterial wilt and its application. Background technique [0002] Tomato bacterial wilt is a devastating soil-borne bacterial disease caused by Ralstonia solanacearum (referred to as Ralstonia solanacearum), which seriously restricts the development of tomato industry in China. Ralstonia solanacearum enters the tomato root through the elongation zone cells of the plant root tip or the wound of the secondary root growth point, and migrates from the vascular bundle to the aerial part, causing the plant to wilt and die. Due to the strong variability of R. solanacearum in the environment, different types of infecting different plants produce different types of R. solanacearum. In addition, R. solanacearum has a complex transmission route and a wide range of hosts, thus causing problems in pre...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N7/00A01N63/00A01P1/00
CPCA01N63/00C12N7/00C12N2795/00021C12N2795/00051
Inventor 韦中侯玉刚王孝芳徐阳春沈其荣
Owner NANJING AGRICULTURAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap