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New application of bacillus pumilus HR10 in prevention and treatment of forest diseases

A technology of Bacillus pumilus and bacteria agent, applied in the field of biological control, can solve the problems of unseen strain application reports, achieve environmental friendly side effects, and reduce the effect of use

Active Publication Date: 2020-09-18
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there is no report on the application of this strain in the prevention and treatment of tree branch diseases

Method used

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  • New application of bacillus pumilus HR10 in prevention and treatment of forest diseases
  • New application of bacillus pumilus HR10 in prevention and treatment of forest diseases
  • New application of bacillus pumilus HR10 in prevention and treatment of forest diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Determination of the antagonistic effect of Bacillus pumilus HR10 on S. sapinea mycelial growth and spore germination

[0027] Inoculate B.pumilusHR10 on Luria-Bertani (LB) liquid medium, culture at 28°C with shaking at 200r / min until logarithmic growth phase; inoculate S.sapinea on potato dextrose agar (PDA) medium, at 25°C , cultivated in the dark for 5 days for later use.

[0028] 1. Determination of the antagonistic effect of B. pumilus HR10 on the growth of S. sapinea mycelium

[0029] S. sapinea and B. pumilus HR10 were cultured on PDA medium for 5 days, and the size of the inhibition zone was observed. The specific method is as follows:

[0030] B.pumilus HR10 preserved at low temperature was streak cultured in LB plate medium (recipe: 10g / L sodium chloride + 10g / L tryptone + 5g / L yeast extract powder + 15g / L agar) and placed at 28°C Cultivate in the dark in an incubator for 24 hours, and use a sterile inoculation loop to pick a single colony of the ...

Embodiment 2

[0035] Example 2: Determination of the influence of HR10 bacterial strain on S. sapinea cell membrane permeability and cell content

[0036] The growth of pathogenic bacteria is inseparable from the physiological metabolism of cells, and the normal physiological metabolism of cells depends on the integrity of cell membranes and the normal synthesis of cell contents. Relative conductivity, nucleic acid extravasation and malondialdehyde (MDA) content are important indicators to measure cell membrane permeability. Higher relative conductivity means more leakage of electrolytes in the cell membrane and more severe cell damage. In order to investigate whether the HR10 strain inhibits the growth of S. sapinea hyphae and spore germination, whether it achieves the purpose of antagonism by affecting the physiological metabolism of pathogenic bacteria cells, it is necessary to investigate the effect of HR10 strain on S. sapinea cell membrane permeability and cell content. Impact.

[0...

Embodiment 3

[0042] Embodiment 3: Investigate the influence of HR10 bacterial strain on S. sapinea infecting pine needles

[0043] Centrifuge the bacillus pumilus HR10 cultured for 24 hours at 10,000 r / min for 10 minutes, collect the bacteria, wash with sterile water and dilute to 1×10 8 After cfu / mL, spray 2mL evenly on the pine tip of Masson pine, use sterile water as the blank control, spray S. sapinea spore suspension 3 days later, and after another 3 days, cut the needles and leaves of the pine tip for fixation, disinfection and drying , gold spraying and other treatments, the growth status of S. sapinea was observed with an environmental scanning electron microscope ((Quanta 200, USA)).

[0044] Under normal circumstances, S. sapinea hyphae can directly invade through the stomata on needles ( Figure 4 A), the morphology of the spores is plump and can normally germinate mycelium ( Figure 4 B); and a large number of HR10 strains can be seen in and around the stomata of needles spra...

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Abstract

The invention discloses new application of bacillus pumilus HR10 in prevention and treatment of forest diseases, and belongs to the technical field of biological prevention and treatment. A further test on the rhizosphere growth-promoting bacterium bacillus pumilus HR10 obtained in the earlier stage of the laboratory finds that the rhizosphere growth-promoting bacterium bacillus pumilus HR10 has an antagonistic effect on S. salina of pine wilt tip pathogenic bacterium; 1) HR10 strain fermentation broth can cause S. salina vegetative body mycelium deformity, spore germination delay and myceliumgrowth capability inhibition, and can destroy normal synthesis of pathogenic bacteria cell membranes and cell inclusions; and 2) the HR10 strain can be colonized on pine trees and gathered around coniferous stomata to prevent pathogenic bacteria hyphae from invading, and the hyphae are thin and weak. In a greenhouse test, the prevention and treatment effect of the bacillus pumilus HR10 on the pine wilt reaches up to 90%. Therefore, the bacillus pumilus HR10 has a good application prospect in prevention and treatment of pine shoot blight.

Description

technical field [0001] The invention belongs to the technical field of biological control, and more specifically relates to a new application of bacillus pumilus HR10 in preventing and controlling forest diseases. Background technique [0002] Pine is the most important building tree species in forests in the northern hemisphere. Its products are widely used in construction, papermaking, food, industrial supplies, and medicine. Therefore, pine has extensive and important ecological and economic values. Pine dieback is a worldwide distribution of pine diseases caused by the pathogenic fungus Sphaeropsis sapinea. Pine tree damage is mainly manifested as dead shoots or trunk ulcers, fat flow and root and neck rot. Because the disease has many kinds of harmful trees, a large area of ​​occurrence, and rapid development, the pine plantations in many countries have suffered serious damage and heavy losses. At present, the prevention and treatment of pine dieback mainly depends on ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N63/22A01P3/00C12N1/20C12R1/07
CPCA01N63/22C12N1/20
Inventor 吴小芹戴云王亚会叶建仁
Owner NANJING FORESTRY UNIV
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