Method of fast and efficiently screening antagonistic bacteria from compost

A technology of bacteria and composting, applied in the field of agricultural biotechnology and biological control, can solve the problems of accelerating the development and application of antagonistic bacteria, heavy workload of antagonistic bacteria, and reducing screening workload, so as to speed up development and utilization, shorten screening time, The effect of reducing workload

Inactive Publication Date: 2013-04-03
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to use the traditional dilution plate coating method to screen antagonistic bacteria from compost with a large workload and low efficiency. By establishing an efficient and fast method for screening antag

Method used

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  • Method of fast and efficiently screening antagonistic bacteria from compost
  • Method of fast and efficiently screening antagonistic bacteria from compost
  • Method of fast and efficiently screening antagonistic bacteria from compost

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0033] Example 1, Primary Screening of Antagonistic Bacteria in Vinegar Grains Compost

[0034] (1) Prepare solid potato glucose medium (potato 200 g / L, glucose 20 g / L, agar 20 g / L).

[0035] (2) Weigh 10 g of vinegar lees compost, add it into a 500 ml Erlenmeyer flask filled with 90 ml of sterile water, shake it fully, place the Erlenmeyer flask in a shaker at 37°C at 200 r / min, and shake the flask for 30 min .

[0036] (3) Use a pipette gun to draw 5ml of the vinegar lees suspension prepared in the above steps and add it to a 150ml triangular flask containing 45ml of sterile water to make 10 -2 compost suspension, mix well. Then prepare 10 in turn -3 , 10 -4 , 10 -5 , 10 -6series of compost dilutions.

[0037] （ 4) Use a pipette gun to draw 100 μl of the above series of compost dilutions, drop them on the potato dextrose agar medium, and then spread them evenly with a coating stick.

[0038] (5) After the plate of potato dextrose agar medium coated with compost su...

Embodiment 2

[0040] Embodiment 2, the purification of target bacterium

[0041] After the potato dextrose agar plate described in Example 1 was grown in the incubator for 7 days, the mycelium of a large amount of Fusarium oxysporum cucumber-specific pathogenic bacteria was overgrown on the plate, and at 10 -5 and 10 -6 Obvious inhibition zone can be seen on the plate (see figure 1 ). Use an inoculation loop to pick the bacteria in different inhibition zones and mark them on the solid yeast extract-peptone medium (yeast extract 5 g / l, peptone 10 g / l, sodium chloride 10 g / l). Cultivate in an incubator at 30°C for one day. After a single colony grows on the yeast extract-peptone medium, observe the shape of the colonies in each plate, and purify the impure strains by streaking. until the bacterial colonies in each Petri dish have the same shape, size and color (see figure 2 ). Finally, a total of 34 purified bacterial single colonies were picked.

Embodiment 3

[0042] Embodiment 3, re-screening of antagonistic bacteria

[0043] Pick the target bacterium that 34 strains screened out in embodiment 2 are cultivated in solid yeast extract peptone with a toothpick, inoculate in the liquid medium containing 50 ml yeast extract peptone (yeast extract 5 g / l, peptone 10 g / l, Sodium chloride 10 g / l) in a 150 ml Erlenmeyer flask, shake at 30 °C and 200 r / min to logarithmic phase. In the center of the potato dextrose agar medium plate, transfer the Fusarium oxysporum specialized type strains of cucumber punched with a hole puncher (diameter 5 cm), around the place about 2.5 cm away from the central pathogenic bacteria (four points equidistant), and respectively spot 0.5 μl of the bacterial solution in the above logarithmic phase. Cultivate in an incubator at 30°C for about 7 days. By observing the presence or absence of the inhibition zone, a total of 8 antagonistic bacteria were screened, and the antagonistic effect of some antagonistic bac...

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Abstract

The invention discloses a method of fast and efficiently screening antagonistic bacteria from compost. The method comprises the steps of: firstly coating compost suspension on a flat plate, then directly coating pathogenic bacteria liquid on the flat plate so as to enable target bacteria and pathogenic bacteria to be directly confronted, conducting ruling and purification on the mixed target bacteria from an inhibition zone after the inhibition zone is formed, purifying and then picking single target bacterium to conduct the confrontation of the flat plate, thereby screening out the antagonistic bacteria. According to the method, the cost is low, the workload is greatly reduced, the time for screening the antagonistic bacteria can be shortened, and the screening efficiency is improved. According to the method, the development and the utilization of the biocontrol bacterium can be quickened, the development of biological control and sustainable agriculture in China can be promoted, and practical application values are important.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology and biological control, in particular to a method for quickly and efficiently screening antagonistic bacteria from compost. Background technique [0002] Soil-borne diseases are extremely serious plant diseases. Extensive application of chemical fertilizers, continuous cropping of soil, and improper management are all likely to cause outbreaks of soil-borne diseases. Pathogens cause plant death by invading plant roots or stems, causing major economic losses. According to the statistics of the Food and Agriculture Organization of the United Nations, the annual loss of production due to plant diseases is 10% to 15% of the total production. Therefore, the research on the prevention and control of soil-borne diseases has always been a hot and difficult topic. Although the traditional chemical pesticide control can solve the problem of large-scale disease in the intensive planting of agricul...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/07C12R1/22
Inventor 李萍萍林英司春灿杜道林赵青松朱咏莉
Owner JIANGSU UNIV
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