Lysobacter enzymogenes and application thereof

An enzyme-producing lysobacteria, acting technology, applied in the application, bacteria, fungicides and other directions, can solve the problems of complex production process, resource shortage, industrialization degree and application scope limitation, and achieve simple nutritional requirements and fast reproduction. Effect

Active Publication Date: 2017-08-29
四川金珠生态农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Fungi are used as bio-control bacteria, mainly using fungal spore preparations, the production process is complicated, the production cost is high, and its industrialization degree and application range are limited to a certain extent
Streptomyces plays a biocontrol role mainly through the production of antibiotics, but due to the shortage of resources and the limitation of the mechanism of action, this kind of biocontrol agents are also restricted by the cost of research and development and practical application

Method used

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  • Lysobacter enzymogenes and application thereof
  • Lysobacter enzymogenes and application thereof
  • Lysobacter enzymogenes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: Isolation and identification of bacteria

[0033] 1. Isolation of bacteria

[0034] LE16 was isolated from farmland soil in Yuxi City, Yunnan Province. During the long period of flue-cured tobacco boom in 2016, the rhizosphere soil of flue-cured tobacco was collected, and the microorganisms in the soil were separated by the dilution plate coating method. Weigh 1.0 g of soil and place it in a 250 mL Erlenmeyer flask filled with 100 mL of sterile water, shake it in a shaker at 28 °C and 150 r / min for 1 h to prepare a soil suspension. Then, the soil suspension was diluted into different concentration gradients on the aseptic operating table, and the dilution factor was 10. -2 ~10 -7 (i.e. 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 and 10 -7 ). Use a sterilized pipette gun to draw 0.5mL to dilute 10 -2 ~10 -7times the soil suspension on the beef extract peptone solid medium and spread evenly, and each dilution was repeated 5 times. After the operation was ...

Embodiment 2

[0037] Example 2: Identification of antagonistic bacterial strains

[0038] 1. Morphological observation of bacterial colonies

[0039] The above-mentioned bacterial strains with the strongest antagonistic effect were inoculated on beef extract peptone medium, LB medium, TSA medium and 10% TSA medium, cultured at 32°C for 12 hours, and the colony morphology was observed. Such as figure 1 For the colony morphology of LE16 in the present invention on 4 different culture media, on beef extract peptone culture medium, LB culture medium and TSA culture medium, the colony is light yellow, the thalline is moist and shiny, the surface is prominent, and it is mucus-like. Not easy to provoke; on 10% TSA medium, the colony is milky white, the surface is flat, and it is not easy to provoke. Cultivated on various media, the strains all produced unpleasant odors.

[0040] 2. Bacterial Gram staining reaction

[0041] (1) Smear: Take a clean glass slide, drop a small drop of sterile disti...

Embodiment 3

[0060] Example 3: 16S rDNA sequencing

[0061] First, the DNA of LE16 was extracted with the TaKaRaAgarose Gel DNApurification Bacterial Genomic DNA Extraction Kit, which was used as a template for 16S rDNA amplification. The bacterial universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACCTTGTTACGACTT-3') were used for PCR amplification. PCR reaction system 20μL:

[0062] 10×Ex Taqbuffer 2.0μL; 2.5Mm DNTP Mixture 1.6μL; 5pmol / μL primer 27F 0.8μL; 5pmol / μL primer 1492R 0.8μL; template DNA 0.5μL; 5u / μL Ex Taq DNA polymerase 0.2μL; ddH 2 O 14.1 μL. The PCR amplification program was: 95°C pre-denaturation for 5 min, 95°C denaturation for 30 s, 55°C annealing for 30 s, 72°C extension for 1 min 30 s, 24 cycles, and finally 72°C extension for 10 min. The PCR reaction product was detected by 1% agarose coagulation electrophoresis. Compared with DL2000Marker, the fragment size was correct, and it was the target fragment. See Figure 5 . After the target fragment was...

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Abstract

The invention relates to a biocontrol strain LE16 and application thereof. The biocontrol strain LE16 is characterized in that the strain has a nucleotide sequence in LE16 16S rDNA sequence table and is Lysobacter enzymogenes, belonging to the family Xanthomonadaceae and the genus Lysobacter, and preserved with a preservation number of CGMCC No. 14215. The Lysobacter enzymogenes LE16 has the advantages of high propagating speed, simple nutrition requirements, good environmental adaptability, high rhizosphere soil planting capacity and the like. The strain can self-dissolve during liquid culture, and fermentation broth prepared provides great antagonistic effect for various plant pathogenic fungi and can effectively control black shank and powdery mildew of tobacco. The strain has great potential of application in controlling plant fungal diseases.

Description

technical field [0001] The invention relates to an enzyme-producing lysobacterium, and also relates to the application of the fermented liquid of the bacterium in broad-spectrum and high-efficiency antagonism against plant pathogenic fungi, belonging to the field of agricultural biocontrol microorganisms. Background technique [0002] 70% to 80% of plant diseases are caused by pathogenic fungi. Fungal diseases have always been one of the main reasons for crop yield reduction. Globally, crop diseases caused by fungi directly cause hundreds of billions of dollars in economic losses every year. Fungal diseases not only cause crop yield and quality reduction, but also produce toxic metabolites during the infection process, which poses a great threat to the safety of agricultural products. At present, crop diseases caused by fungi have become a global food safety problem. [0003] At present, the prevention and control of fungal diseases mainly depends on the use of chemical pest...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/01
CPCA01N63/10C12N1/205C12R2001/01
Inventor 袁玲陈丹梅黄建国杨红军夏志林赵建段玉琪
Owner 四川金珠生态农业科技有限公司
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