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Rahnella aquatilis HX2 and application thereof

A technology of aquatic Lahenia and strains, applied in the application, bacteria, biocide and other directions, can solve the problem of inability to prevent and control grape root cancer, and achieve the effect of reducing the incidence rate

Inactive Publication Date: 2007-08-08
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bell reported that the Rahnella aquatili JC577 strain has a plate inhibitory effect on Agrobacterium vine, however it does not prevent grape root cancer

Method used

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  • Rahnella aquatilis HX2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 HX2 strain 16S rDNA analysis

[0023] Using Rahnella aquatilis HX2 genomic DNA as a template, the universal primers 63F 5'-CAGGCCTAACACATGCAAGTC-3' and 1494R 5'-GGYTACCTTGTTACGACTT-3' were amplified with 16S. Amplify in the following PCR amplification reaction system: 50μL amplification system, 10pmol primers, 0.2mM dNTP, 1.5mMMgCl 2 , 2.5 U Taq DNA polymerase (TaKaRa), 1×PCR buffer (TaKaRa), 25ng genomic DNA as template. PCR reaction conditions: denaturation at 94°C for 5 min; denaturation at 94°C for 40 sec, annealing at 55°C for 40 sec, extension at 72°C for 1 min, and a cycle number of 35; extension at 72°C for 10 min. The PCR product was purified by Omega "Gelextraction kit" after 1% agarose gel electrophoresis, connected to pMD18-T Vector, transformed into E.coli DH5α, and sequenced by ABI 3730 DNA sequencer in Invitrogen Company. The nucleic acid sequence is as follows: The list is shown in SEQ ID NO.1. Using the BLAST program in GenBank ( http: / / ww...

Embodiment 2

[0024] Embodiment 2 antibacterial spectrum analysis

[0025] Plate inhibition detection method for plant pathogenic bacteria. The inhibition of HX2 to phytopathogenic bacteria was detected by double-layer culture method. The activated Lahnella aquatica HX2 was formulated into a bacterial suspension (10 9 CFU / mL), draw 10 μL and spot on the PDA medium, one point per plate, culture at 28°C for 48 hours, kill the growing HX2 with 3mL chloroform and its vapor. After 10-12 hours, the cultivated Agrobacterium Bacterial suspensions were prepared from bacterial strains, rice bacterial blight pathogen, cotton bacterial angular spot pathogen, cabbage black rot pathogen, cucumber bacterial angular leaf spot pathogen, tomato canker pathogen, and Bacillus subtilis slant. The final bacterial suspension concentration was determined to be about 10 9 CFU / mL. Take 50 μL of bacterial suspension and add 5 mL of soft YEB medium (sucrose 5 g / L, peptone 5 g / L, beef extract 5 g / L, yeast extract 1...

Embodiment 3

[0043] Example 3 Detection of Effects of Greenhouse Prevention and Control of Root Cancer

[0044] HX2 was inoculated in PDA liquid medium at 28°C and shaken at 150rpm for 28 hours. The tested Agrobacterium was prepared at a concentration of 10 8 The CFU / mL bacterial suspension was mixed with 2 times the volume of HX2 bacterial solution and sterile normal saline, and HX2 bacterial solution and sterile normal saline were inoculated as a control. Use a sterilized inoculation needle to scratch longitudinally on the sunflower stem to form a wound with a length of 0.5 cm. Draw 3 μL of the test solution and inoculate it into the wound, wrap the wound with parafilm, and observe the nodulation after 15 days in the greenhouse. Weigh the tumor weight. There were 30 repetitions for each treatment, and the experiment was repeated three times.

[0045] Control effect (%) = (average weight of control root cancer tumors - average weight of treatment root cancer tumors) ÷ (average weight o...

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Abstract

The invention discloses a new Rahnella aquatilis strain HX2 (CGMCC No.1896), which is characterized by the following: manufacturing antagonistic material and IAA; inhibiting partial pathogenic bacteria (Agrobacterium, Xanthomonas, Clavibacter, Pseudomonas and Bacillus) and partial pathogenic fungus (Fusarium, Alternaria and Botrytis); detecting root cancer based on green house sunflower as indicating plant.

Description

technical field [0001] The present invention relates to a new microbial strain and its application, specifically Lahnella aquatica HX2 and its application in biological control. Background technique [0002] Grape root cancer is common in the world. A typical symptom of grapevine root cancer is the formation of tumors. The diseased part is mainly in the part (root neck) of the trunk close to the ground, and can also occur slightly below the soil surface or on the surface of the grape trellis closer to the ground. Grape root cancer occurs to varying degrees in orchards and nurseries in many provinces and cities in Northeast my country, North China, Northwest China, the Yellow River and the Yangtze River Basin, among which Liaoning, Jilin, Inner Mongolia, Beijing, Hebei, Shanxi, Shandong, Zhejiang and The disease occurred seriously in Shanghai and other places. The incidence rate of some vineyards is as high as 80-90%, and the output loss reaches 30-70%. In severe cases, eve...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02
Inventor 王慧敏陈凡郭岩彬王建辉李金云卢彩鸽
Owner CHINA AGRI UNIV
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