Rice bacterial brown streak germ phage and application thereof

A brown streak pathogen, bacterial technology, applied in the field of rice bacterial brown streak pathogen phage, can solve the problem of no phage, etc., and achieve a strong cracking effect

Active Publication Date: 2015-05-20
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] So far, there are no related reports on phages cap...

Method used

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  • Rice bacterial brown streak germ phage and application thereof
  • Rice bacterial brown streak germ phage and application thereof
  • Rice bacterial brown streak germ phage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Isolation of Rice Bacterial Brown Streak Phage

[0030] (1) Isolation of rice bacterial brown streak pathogen

[0031] ① After soaking and grinding the rice disease species, let it stand for a period of time, take the supernatant for gradient dilution and spread on the plate, and place the plate upside down in a 30°C incubator for cultivation;

[0032] ② After culturing for 48 hours, a single colony of suspected rice bacterial brown streak pathogen was picked and purified;

[0033] ③The isolated strains were tested for tobacco allergy and rice pathogenicity, and the strains that were all positive were taken, and the genomic DNA was extracted for 16S rDNA sequencing (see SEQ ID No.1 for the sequencing results).

[0034] Thus, a rice bacterial brown streak pathogen (Aaa RS-2) was isolated and verified, and the bacteriophage was isolated using this strain as a host bacterium.

[0035] (2) Isolation of rice bacterial brown streak pathogen phage

[0036] ① Soak ...

Embodiment 2

[0040] Example 2 Determination of biological properties of rice bacterial brown streak pathogen phage

[0041] (1) Determination of phage titer

[0042] After the last purification, evenly distributed and uniformly sized phage plaques were obtained. Prick a single phage plaque with a tip and soak the tip in 1ml of SM liquid medium at 4°C overnight. Take 0.1 phage plaque the next day. Inoculate 5ml of LB liquid medium with 0.1ml of supernatant and 0.1ml of host bacteria solution, incubate at 30°C for 6 hours, then centrifuge and filter, make 10-fold gradient dilution with SM solution, and finally dilute to 10 -8 , absorb 0.1ml of the last three dilutions of phage dilution and mix with 0.1ml of host bacteria, incubate at 30°C for 20min and spread double-layer agar plates. Observe the number of plaques and calculate the phage titer according to the following formula:

[0043] Phage titer = number of plaques × dilution factor / sampling volume;

[0044] Three parallel samples a...

Embodiment 3

[0051] Example 3 Bacteriophages are used to control rice bacterial brown streak disease

[0052] (1) Under in vitro conditions, the lysis curve of phages against the rice bacterial brown streak pathogen

[0053] The host bacterium Aaa RS-2 was cultivated to the pre-logarithmic phase, and 2ml of the culture was added to 5ml of LB liquid medium; among them, 20μl of phage (2×10 7 pfu / ml), the control group was added with 20 μl LB liquid medium, and cultured in a shaker. Samples were taken at each time point in a 96-well plate for OD value determination, and the lysis curve was drawn. The lysis curve was as follows: Figure 4 .

[0054] Such as Figure 4 As shown, the rice bacterial brown streak pathogen phage significantly reduces the number and growth rate of host bacteria, and can be used to prevent and control rice bacterial brown streak pathogen.

[0055] (2) Control effect of phage on rice bacterial brown streak disease under living conditions

[0056] Soak the seeds in...

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PUM

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Abstract

The invention discloses a rice bacterial brown streak germ phage and an application thereof. The classification term of the phage is rice bacterial brown streak germ phage AP1 (Acidovorax avenae subsp.Avenae phage AP1), and the preservation number is CCTCC M 2014599. The application comprises an application of the rice bacterial brown streak germ phage in preparation of a phage preparation used for inhibiting rice bacterial brown streak germs, an application for inhibiting the rice bacterial brown streak germs and an application of preventing and treating the rice bacterial brown streak germs. The rice bacterial brown streak germ phage disclosed by the invention can specifically crack the rice bacterial brown streak germ, the burst size is about 160PFU/infected cells, and the cracking performance is stronger, so that the rice bacterial brown streak germ phage can be used for preventing and treating rice bacterial brown streak caused by the rice bacterial brown streak germs.

Description

technical field [0001] The invention belongs to the technical field of prevention and treatment of bacterial brown streak disease of rice, and in particular relates to a phage of bacterial brown streak disease of rice and an application thereof. Background technique [0002] Rice is the main food crop and economic crop in my country, and is the staple food of one-third of the human beings in the world. Rice bacterial brown streak pathogen (Acidovorax avenae subsp. avenae) is a bacterium of the genus Pseudomonas, which mainly overwinters on rice seeds and straw, and invades and causes damage through stomata and wounds of rice plants. The main symptoms caused by the pathogen are brown streaks on the leaf sheaths along the veins on the rice seedlings, which can cause the heart and leaves to die in severe cases; early ear deformities and false fruit rate increase on rice seedlings. [0003] Rice brown streak disease is one of the most important bacterial diseases in rice produc...

Claims

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Application Information

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IPC IPC(8): C12N7/00A01N63/00A01P1/00C12R1/92
Inventor 李斌王丽罗金燕张阳杨樱子方雨诗王晓璇葛梦禹
Owner ZHEJIANG UNIV
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