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Pine endosymbiotic bacterium-new burkholderia cenocepacia NSM-05 and application thereof

A technology of Burkholderia and endophytic bacteria, applied in the application, bacteria, biocide and other directions, can solve the problems of pine dieback and other problems that have not been reported, achieve good effect of field biocontrol test and strong antibacterial effect , the effect of simple cultivation conditions

Inactive Publication Date: 2011-09-21
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, many countries have adopted Bc as raw material to produce biopesticides, but there is no report on the use of Bc to control pine dieback

Method used

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  • Pine endosymbiotic bacterium-new burkholderia cenocepacia NSM-05 and application thereof
  • Pine endosymbiotic bacterium-new burkholderia cenocepacia NSM-05 and application thereof
  • Pine endosymbiotic bacterium-new burkholderia cenocepacia NSM-05 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 Burkholderia neocepacia NSM-05 is to pine blight bacteria ( Sphaeropsis sapinea ) antibacterial effect.

[0036] Use PDA plate culture medium, 25 ℃, constant temperature culture of pine blight bacteria for 3~4 days, use a hole puncher with a diameter of 5mm to get the same quality and same amount of bacteria blocks, inoculate in the center of the PDA plate, and then use NA medium slant The activated Burkholderia neocepacia NSM-05 was streak-inoculated on both sides of the bacterial block 3.5cm away from the pine blight, and the PDA plate only inoculated with the pine blight was used as a control, and each treatment was replicated 3 times , 28 ℃ constant temperature culture for 3 to 5 days, and observe the results when the control covers the entire plate. Test results such as figure 1 The results showed that Burkholderia neocepacia NSM-05 had a strong antibacterial effect on pine shoot fungus, and the average diameter of the inhibition zone (the distance...

Embodiment 2

[0037] Example 2 The antibacterial effect of the metabolites of Burkholderia neocepacia NSM-05 on the pathogen Pine shoot.

[0038] Inoculate Burkholderia cenocepacia NSM-05 in NB culture medium, shake culture at 30°C, 200r / min for 48h, and prepare fermentation broth. Then the fermentation broth was centrifuged at 4° C., 10000 r / min for 20 minutes. Take the supernatant, filter and sterilize through a 0.22 μm microporous bacterial filter, and obtain a sterile fermentation liquid, which is the metabolite of the NSM-05 strain. Sterilize and cool the prepared PDA medium to about 50°C, add 1ml of sterile fermentation broth to every 10ml of the medium, mix well and pour it into a flat plate to make a poisonous plate. 2 days later, the fungus of Pine shoot blight was inoculated, and the PDA medium without adding sterile fermentation broth was used as a control. Incubate at 25°C for 5 days, observe its growth, and calculate the antibacterial rate. Test results such as Figure 9 , ...

Embodiment 3

[0039] Example 3 Field control of Burkholderia neocepacia NSM-05 on pine dieback.

[0040] From May to June 2010, the field control of Burkholderia cepacia NSM-05 against pine dieback was carried out in the pine forest of Nandashan Institute of Forest Pathology, Nanjing Forestry University. The inoculation method of pine dieback fungus: first use a sterile inoculation knife to gently scratch the epidermis of the slash pine branches about 1 cm in length, and then spray the slash pine shoots with the broken liquid of pine dieback fungus. Treatment 1: Root irrigation and inoculation of 100 ml / strain of Burkholderia neocepacia NSM-05 strain fermentation broth (prepared in Example 2) 3 days before inoculation with Pine shoot blight. Treatment 2: Spray and inoculate the fermentation broth of Burkholderia neocepacia NSM-05 strain 3 days before the inoculation of Pine dieback until the pine trees are drenched. Treatment 3: direct inoculation of pine dieback fungus. 20 plants per tre...

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Abstract

The invention discloses a pine endosymbiotic bacterium which is classified and named after a new burkholderia cenocepacia NSM-05 and is stored in the China typical culture collection center with the storage number of CCTCC NO: M2011015 and the storage data of January 11, 2011. The invention also discloses application of the new burkholderia cenocepacia NSM-05 to the preventing and treating pine shoot dieback diseases and promoting the growth of pines. The invention also discloses an antagonistic inoculant for preventing and treating the pine shoot dieback diseases; the antagonistic inoculant is obtained by inoculating NSM-05 bacterium strains in a culture medium. The NSM-05 bacterium strain provided by the invention is the pine endosymbiotic bacterium, has strong bacteriostasis on the pine shoot dieback diseases, has the antagonism effect on multiple woods and crop pathogenic bacteria and road-spectrum antibacterial activity, is safe and non-toxic to people, livestock and plants and has good development and application prospects.

Description

technical field [0001] The invention relates to endophytic bacteria of plants, in particular to a pine endophytic bacterium Burkholderia neocepacia NSM-05 and application thereof. Background technique [0002] Pine dieback (Sphaeropsis sapinea) is one of the most common and widely distributed diseases of coniferous trees worldwide. Among the nearly 30 countries where the disease is distributed in the world, the most affected countries are: New Zealand, Australia, South Africa, the Netherlands, the United States and China. It has a very wide host range and can infect Pinus L., Abies Mill, Larix Mill, Thuja L., Cedrus Trew, More than 60 kinds of coniferous trees belonging to 8 genera including Juniperus L., Picea Dietr. and Pseudotsuga Carr. Although the host range of the pathogen of pine dieback is wide, the susceptibility of different hosts is different. Currently, only slash pine, loblolly pine, masson pine and sylvestris pine are known to cause economic losses. Since 19...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P3/00A01P21/00C12R1/01
Inventor 叶建仁黄洁芳吴小芹袁文婷
Owner NANJING FORESTRY UNIV
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