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Endophytic Antagonistic Bacillus zf04 of Phoenix Dancong Tea Tree and Its Application in Biocontrol

A technology of Bacillus and ZF04, which is applied in agricultural biological control, from the endophytic antagonistic bacteria of Fenghuang Dancong tea tree and its application in biological control, can solve the problems that there are not many researches on Fenghuang Dancong tea, and achieve a strong inhibitory effect , the effect of good application prospects

Inactive Publication Date: 2019-03-12
HANSHAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on Fenghuang Dancong tea mainly focuses on the variety identification, heritage diversity analysis, quality chemical characteristics, physiological activity, quality safety and greening process, and there are not many studies on the endophytic bacteria of Fenghuang Dancong tea tree.

Method used

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  • Endophytic Antagonistic Bacillus zf04 of Phoenix Dancong Tea Tree and Its Application in Biocontrol
  • Endophytic Antagonistic Bacillus zf04 of Phoenix Dancong Tea Tree and Its Application in Biocontrol
  • Endophytic Antagonistic Bacillus zf04 of Phoenix Dancong Tea Tree and Its Application in Biocontrol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Isolation, screening and identification of Bacillus sp. ZF04 GDMCC No: 60014

[0039] 1. Isolation and screening of strains

[0040] Bacillus sp. used in the present invention was sampled by Hanshan Teachers College from the root of Fenghuang Dancong tea tree in Fubin Town, Raoping County, Chaozhou, Guangdong Province. Fubin Town, Raoping County, the tree age is 15, 25 and 50 years old, and the planting altitude is 750m. By using the tissue block culture method to isolate the endophytic bacteria in the roots, with different culture temperature, pH value and medium as enrichment conditions, a batch of bacterial strains with good growth were screened out through optimization, and one strain numbered ZF04 was selected. strains.

[0041] Separation steps:

[0042] (1) The separation medium is adopted: the beef extract peptone liquid medium is beef extract 5g, peptone 10g, sodium chloride 5g, distilled water 1000mL, pH 7.0-7.2), and the nutrient agar medium is ...

Embodiment 2

[0057] Example 2: Molecular-level identification of Bacillus sp. ZF04 GDMCC No: 60014

[0058] The DNA of the target strain was extracted using the bacterial DNA genome rapid extraction kit.

[0059] The primers are bacterial universal primers, bacterial 16SrDNA universal primer sequences:

[0060] 27F: 5′-AGAGTTTGATCCTGGCTCAG-3′

[0061] 1492R: 5′-GGTTACCTTGTTACGACTT-3′

[0062] Primer 27F, primer 1492R, and 2×TapPCRMaster mix used in this application were all purchased from Bio-Sanggong (Shanghai) Co., Ltd.

[0063] The 20 μL PCR amplification system used to extract genomic DNA: 1.0 μL of primer 27F and 1492R each, 7.0 μL of 2×TapPCRMaster mix, 1.0 μL of endophytic bacteria DNA mixture, and 10.0 μL of double-distilled water.

[0064] PCR amplification procedure: pre-denaturation at 94°C for 5 min, denaturation at 94°C for 30s, annealing at 55°C for 30s, extension at 72°C for 40s, 30 cycles from denaturation to extension, extension at 72°C for 7 min, and incubation for 4 m...

Embodiment 3

[0068] Example 3: Growth factor of Bacillus sp. ZF04 GDMCC No: 60014

[0069] Table 3: Effects of temperature, pH and NaCl on the growth of strain ZF04

[0070] temperature(℃)

4

10

15

20

25

30

37

40

45

growth situation

-

+

+

++

++

++

+++

+

+

pH

4

5

6

7

8

9

growth situation

-

+

++

+++

+

-

NaCl concentration

5％

6％

7％

8％

9％

10％

15％

growth situation

+++

++

+

+

+

+

-

[0071] Bacillus sp. ZF04 GDMCC No: 60014 was cultured as above, and the culture conditions of the strains were: the culture medium was beef extract peptone liquid medium and nutrient agar medium, culture conditions: pH 7.0, temperature 37 Incubate at ℃ for 20h.

[0072] From Table 3, strain ZF04 is the most suitable growth factor. Through the above results, it is concluded that the optimal culture t...

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Abstract

The invention discloses an endophytic antagonistic Bacillus ZF04 of Fenghuang Dancong tea tree and its application in biocontrol. By sampling from the root of Fenghuang Dancong tea tree, separating, purifying, screening and cultivating, a Bacillus strain with better biocontrol effect is obtained (Bacillus sp.) ZF04 GDMCC No: 60014, the strain has obvious differences with common Bacillus in terms of physiological and biochemical characteristics and molecular level, and is a typical new strain. Leaf blight, black spot fungus of jujube and Rhizoctonia solani had good antibacterial effects, and the mycelial growth inhibition rates were 77.55%, 83.30%, 74.52% and 75.19%. ZF04 inhibited the mycelial growth of Pythium melon and fruit The rate is 31.60%. It is a typical and specific biocontrol bacterium, which has wide application value in biocontrol.

Description

technical field [0001] The invention relates to the technical field of agricultural biological biological control, in particular to the technical field of an endophytic antagonistic bacteria from Fenghuang Dancong tea tree and its application in biological control. Background technique [0002] Endophyte refers to a class of microorganisms that survive in various tissues or intercellular spaces of healthy plants in part or all of their life cycles without causing obvious pathological symptoms to the host. Antagonistic endophytes refer to microorganisms that can produce antagonistic effects. The plant body is a complex micro-ecosystem. Healthy plants contain a large number of endophytic bacteria (Endophytic Bacteria), which have been isolated and identified from more than 50 kinds of plants such as wheat, cotton, rice, peanut, potato, tomato, lemon, and citrus. There are more than 50 genera of endophytic bacteria, and almost all healthy plants contain endophytic bacteria. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00C12R1/07
CPCA01N63/00C12N1/205C12R2001/07
Inventor 傅力蔡丽王忠合王军侯小桢
Owner HANSHAN NORMAL UNIV
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