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Biocontrol agent for preventing and curing bacterial wilt and club roots of plant as well as preparation method and application thereof

A bio-control preparation and plant technology, which are applied in the field of bio-control preparations for preventing and controlling plant bacterial wilt and clubroot and their preparation, can solve the problems that restrict the sustainable and stable development of oilseed and vegetable industries to increase farmers' income, lack high-efficiency and low-residue chemical agents, The research is not comprehensive and systematic, etc., to achieve the effect of vigorous growth and metabolism of bacteria, promoting the effect of increasing production and simple process

Inactive Publication Date: 2012-10-31
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clubroot is an important soil-borne disease on cruciferous crops. It is caused by Plasmodiophora brassicae infection. The damage area is about 60 million mu, with an average yield loss of 20% to 30%. Yield loss of more than 60%, or even no harvest, causes economic loss of about 1 billion yuan in rapeseed and 20 billion yuan in vegetable economic loss every year, which greatly restricts the sustainable and stable development of the oilseed and vegetable industry and the increase of farmers' income
However, for the prevention and control of clubroot, there is still a lack of high-efficiency, low-toxicity, low-residue, and low-cost chemical agents; in terms of agricultural control, the research is not comprehensive and systematic, and there are many disadvantages

Method used

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  • Biocontrol agent for preventing and curing bacterial wilt and club roots of plant as well as preparation method and application thereof
  • Biocontrol agent for preventing and curing bacterial wilt and club roots of plant as well as preparation method and application thereof
  • Biocontrol agent for preventing and curing bacterial wilt and club roots of plant as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 prepares seed solution

[0031] The preserved Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) ZJUB2008 (preserved in the China Center for Type Culture Collection, the preservation number is CCTCC NO: M209145) was streaked on the NA agar plate by the three-section line method, and cultured at 30°C 24h; Inoculate a single colony of Bacillus amyloliquefaciens ZJUB2008 into LB liquid medium, shake and culture in a shaker at 30°C and 160r / min for 12h, and make a seed liquid, the concentration of bacteria in the seed liquid is 10 8 CFU / ml.

Embodiment 2

[0032] The optimization of embodiment 2 fermentation medium

[0033] Five media component factors including peptone, beef extract, glucose, yeast powder and NaCl were selected, and each factor was set to 4 levels. Use L 16 (4 5 ) orthogonal table, random arrangement factors and matching levels (see Table 1).

[0034] According to the orthogonal test design table (seeing table 3), respectively configure the culture medium, about pH 7.0; v / v); at 30° C., 160 r / min shaker shaking culture for 24 hours. Each treatment has 3 repetitions.

[0035] The antibacterial activity was determined by the agar diffusion method: the fermented liquid was added to the agar plate containing R. solanacearum, and after culturing in an incubator at 30° C. for 48 hours, the diameter of the inhibition zone of each treatment was measured.

[0036] The results showed that the components of the culture medium, peptone, beef extract, glucose, yeast powder, and NaCl, all had an effect on the antibacter...

Embodiment 3

[0046] The optimization of the initial pH of embodiment 3 medium

[0047] The initial pH of culture medium is set 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0 eight gradients, inoculate the bacillus amyloliquefaciens ZJUB2008 seed liquid of embodiment 1 in the culture medium after embodiment 2 optimization, inoculum size 0.5% (v / v); at 30°C, 160r / min shaker shaking culture for 24h; measure the size of the inhibition zone and the number of bacteria.

[0048] see results figure 1 , when the initial pH is 7.5, the antibacterial activity of Bacillus amyloliquefaciens ZJUB2008 is the largest; when the initial pH is 8.0, the number of bacteria of Bacillus amyloliquefaciens ZJUB2008 is the largest. Considering the growth of the bacteria and the production of antibacterial substances, and focusing on the antibacterial activity, the initial pH of the medium most suitable for the fermentation of Bacillus amyloliquefaciens ZJUB2008 was determined to be 7.5.

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Abstract

The invention discloses a biocontrol agent for preventing and curing bacterial wilt and club roots of a plant as well as preparation method and application thereof. The preparation method of the biocontrol agent comprises the following steps: inoculating activated bacillus amyloliquefaciens ZJUB2008 into a culture solution, and fermenting and culturing at 25-45 degree C for 6-120 hours, wherein the culture solution comprises the following components by taking the volume of 1L as the basis: 3-15g of peptone, 0-3g of beef extract, 0-5g of glucose, 0-5g of yeast powder, 0-8g of NaCl, and the balance of water; and the pH value of the culture solution is 5.5-9.0. According to the invention, the bacillus amyloliquefaciens ZJUB2008 is used to prepare the biocontrol agent on / under the appropriate fermentation medium and culture conditions by means of microbial fermentation; in addition, the prepared biocontrol agent has a better prevention effect to different chemotyped ralstonia and plasmodiophora, can remarkably lowering the occurrence rate and severity of the bacterial wilt and the club roots, and is suitable for large-scale production.

Description

technical field [0001] The invention relates to the field of plant disease prevention and control, in particular to a biocontrol agent for preventing and treating bacterial wilt and clubroot, a preparation method and application thereof. Background technique [0002] Bacterial wilt of crops is a systemic and devastating soil-borne disease caused by Ralstonia solanacearum, commonly known as "plant blast". R. solanacearum has a wide range of hosts and can infect hundreds of plant species in 44 families. At present, the prevention and control of bacterial wilt is a major problem recognized in the world. It is widely distributed all over the world, and it occurs seriously in southern provinces and cities in my country, especially in Zhejiang. In recent years, not only Solanaceae and melons have been damaged, but also large-scale Mulberry bacterial wilt has caused serious losses to sericulture production. [0003] my country began to carry out research on biological control agen...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P1/00A01P3/00C12R1/07
Inventor 周青谢关林苏婷刘宝平朱勃李斌
Owner ZHEJIANG UNIV
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