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Pseudomonas syringae, screening method thereof and application to kill nematode

A technology of Pseudomonas and cloves, applied in the field of Pseudomonas syringae and its screening and application in killing nematodes, achieving remarkable beneficial technical effects

Inactive Publication Date: 2014-09-10
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The bacterial strains in the Pseudomonas genus reported above all have the ability to control nematodes, but so far, there is no report that Pseudomonas syringae in the Pseudomonas genus can kill nematodes

Method used

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  • Pseudomonas syringae, screening method thereof and application to kill nematode
  • Pseudomonas syringae, screening method thereof and application to kill nematode
  • Pseudomonas syringae, screening method thereof and application to kill nematode

Examples

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Embodiment 1

[0030] Example 1: is an example of Pseudomonas syringae of the present invention. Taxonomically named Pseudomonas syringae MB03 ( Pseudomonas syringae MB03), the preservation date is: April 2, 2014, the preservation unit is: China Type Culture Collection Center, address: Wuhan University, Luojia Mountain, Wuchang City, Wuhan City, the preservation center preservation number: CCTCC NO: M 2014114; the clove The morphological and physiological and biochemical characteristics of Pseudomonas MB03 include: the colony protrudes on LB solid medium, is milky white, and the bacterial body is rod-shaped under the microscope, which is a Gram-negative bacterium.

[0031] The components of the LB solid medium are: 10 g sodium chloride, 10 g peptone, 5 g yeast powder, 15 g agar, distilled water to 1000 mL, pH 7.0-7.2.

[0032] Such as image 3 Shown is the colony morphology of Pseudomonas syringae MB03 cultured on LB solid medium for 24 hours, which is round, milky white, smooth, with n...

Embodiment 2

[0034] Example 2: It is an embodiment of a screening method for Pseudomonas syringae of the present invention. Specifically, the screening method for Pseudomonas syringae MB03 comprises the following steps:

[0035] A. Collect the plant tissues with obvious frostbite in the open-air vegetation of Shizishan Mountain, Wuhan City, Hubei Province, and use them to isolate the raw material of Pseudomonas syringae MB03;

[0036] B. Mince the collected plant tissue, then homogenize it in a glass homogenizer, then add sterile water to a shaker at 200 r / min at room temperature and soak for 1 hour, then take 100 μL of soaking solution Spread on NA solid medium after 10-fold serial dilution, and culture in an incubator at 28°C for 48 hours;

[0037] C. Pick the grown single colonies and separate and purify them by streaking, then use morphological characteristics observation, physiological and biochemical characteristics measurement, colony in situ hybridization and nucleic acid inform...

Embodiment 3

[0039] Embodiment 3, It is an embodiment of the application of Pseudomonas syringae in killing nematodes in the present invention. In this embodiment, Pseudomonas syringae MB03 cells are used to kill Caenorhabditis elegans and Meloidogyne incognita. The preparation method of Pseudomonas syringae MB03 cells is as follows: Streak inoculation of Pseudomonas syringae MB03 strains on LB solid medium for cultivation, the temperature is 28 °C, and the cultivation time is 12 h; then from the above solid medium Pick a single colony of Pseudomonas syringae MB03 and put it into a PA bottle with 5 mL of LB liquid medium, and heat it at 28 °C, 160 r min -1 Cultivate for 10 h; then expand the cultivation of Pseudomonas syringae, that is, transfer the inoculation amount of 1% by volume of the activated Pseudomonas syringae MB03 strain in the above-mentioned PA bottle to the medium containing 200 mL LB liquid medium. 28 ℃, 160 r·min in the Erlenmeyer flask -1 Extended culture for 24h; then...

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Abstract

The invention relates to a pseudomonas syringae, a screening method thereof and application to kill nematode. The classification name of the pseudomonas syringae is pseudomonas syringae MB03, colonies has protuberances and show milk white, the thalline is in the shape of a rod through microscopic examination and belongs to a gram-negative bacterium. The screening method comprises: acquiring frozen plant tissue, chopping and culturing in a medium, and finally picking growing single colony, and performing separation purification through streaking. The application to kill nematode means that the thalline, a fermentation supernatant, a cell disruption solution, or extracellular crude protein and / or intracellular crude protein of pseudomonas syringae MB03 subjected to ammonium sulfate precipitation is used to kill caenorhabditis elegans and meloidogyne incognita. The advantages comprise that pseudomonas syringae is low in production cost and free of pollution to environment, and has relatively killing effect on caenorhabditis elegans and meloidogyne incognita. The factor, killing nematode, in pseudomonas syringae is protein, so that pseudomonas syringae is sutiable for molecular gene operation, and has great value on microbe control resource and mechanism research.

Description

technical field [0001] The present invention relates to a kind of Pseudomonas syringae and its screening method and application in killing nematodes, and the classification of Pseudomonas syringae is named as Pseudomonas syringae MB03 ( Pseudomonas syringae MB03), its application in killing nematodes is especially suitable for killing Caenorhabditis elegans ( Caenorhabditis elegans ) and Meloidogyne incognita ( Meloidogyne incognita ), belonging to the technical field of microbial prevention and control of agricultural diseases. Background technique [0002] Agriculture is the basis of human survival. However, diseases, their wide distribution and variety, have caused losses of more than 100 billion U.S. dollars to crop yields worldwide every year. Among them, root-knot nematodes ( Meloidogyne spp. ) accounted for approximately US$77 billion in losses, so the need for root-knot nematode control is self-evident. At present, chemical control is more commonly used in the...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/04A01N63/02A01P5/00C12R1/38
Inventor 李林秦旭孙孝文顾桐张震
Owner HUAZHONG AGRI UNIV
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