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Method for efficiently and directionally screening biocontrol bacteria in soil-borne diseases of crops

A soil-borne disease, directional screening technology, applied in microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve problems such as poor colonization ability of antagonistic bacteria, and achieve the goal of overcoming poor colonization ability, strong pertinence, The effect of avoiding waste of human and financial resources

Active Publication Date: 2010-08-18
南京科绿丰科技有限公司
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In view of the problem that the antagonistic bacteria screened in the existing bio-control bacteria screening methods do not necessarily have a good control effect in the field, that is, the selected antagonistic bacteria have poor colonization ability, the purpose of the present invention is to provide a directional and efficient screening of crops Methods of Biocontrol of Soilborne Diseases

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  • Method for efficiently and directionally screening biocontrol bacteria in soil-borne diseases of crops
  • Method for efficiently and directionally screening biocontrol bacteria in soil-borne diseases of crops

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Screening of Antagonistic Bacteria of Cotton Verticillium Wilt by Plate Confrontation Method

[0038] Specifically include the following steps:

[0039] (1) Strain activation: 115 strains, such as the bacteria NCD-2 strain preserved at low temperature (preservation number: CGMCC1019, see patent number: ZL200310109619.8), were isolated from crop field soil bacteria (Plant Protection of Hebei Academy of Agriculture and Forestry Sciences). The Institute collects and isolates from the whole country, mainly in Hebei Province. Partial bacterial strains are shown in Table 1) on NA plate culture medium at 37 DEG C of activation culture, pick single bacterial colony on NA slant culture medium and multiply at 37 DEG C; NA plate medium composition and proportion: beef extract 0.5%, peptone 1.0%, NaCl 0.5%, agar powder 1.5%, the rest is water, pH7.4.

[0040] (2) pathogenic bacteria cultivation: the cotton Verticillium dahliae Kleb strain (Verticillium dahliae Kleb) VD-1...

Embodiment 2

[0048] Example 2 Detection test of the biofilm ability of antagonizing bacteria

[0049] Proceed as follows:

[0050] (1) Strain activation: the 17 strains of antagonistic bacteria screened in Example 1 were respectively activated on the NA plate medium, and a single colony was picked and multiplied on the NA slant medium;

[0051] (2) Bacterial strain liquid culture: 17 strains of antagonistic bacterial strains and 168 bacterial strains after picking up multiplication with sterile bamboo sticks are inoculated in DSM culture medium respectively (its composition and weight percent are: beef extract 1.0%, KCl 0.12 %, MgSO 4 .7H 2 O 0.00144%, Ca(NO 3 ) 2 0.00492%, MnCl 2 0.0000378%, FeSO 4 0.0000456%, the rest is water) and cultured overnight at 37°C. The next day, it was diluted with freshly prepared DSM medium at a ratio of 1:1000. Transfer 0.5ml to a new 2.0ml centrifuge tube, and culture at 37°C for 48h.

[0052] (3) Crystal violet staining: Add 550 μl of 0.1-0.3% ...

Embodiment 3

[0054] Example 3 Calcium Phytate Hydrolysis Ability Determination Test

[0055] The specific method is as follows:

[0056] (1) Strain activation: the numbered NCD-2 and C-39, NCD-25, C-148, C-94, DMT-36, and C-28 bacterial strains screened out in the embodiment 2 of cryopreservation were respectively placed in Activate on NA plate medium, pick a single colony and multiply on NA slant medium;

[0057] (2) the above-mentioned 7 strains of bacterial strains for testing are connected to the phytase detection medium with sterilized toothpicks respectively (its composition and weight percentage are: sucrose 0.2%, KCl 0.2% g, MnSO 4 0.6%, NH 4 NO 3 0.4%, calcium phytate 8.0%, MgSO 4 0.7%, Fe2(SO 4 ) 3 0.7%, agar powder 4.0%, all the other are water; pH7.0), after 37 ℃ of cultivations for 5 days, it is found that only the NCD-2 bacterial strain has a large and transparent digestion circle on the phytase detection medium ( figure 2 ). It shows that the NCD-2 strain can sec...

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Abstract

The invention discloses a method for efficiently and directionally screening biocontrol bacteria in soil-borne diseases of crops, belonging to the field of biological control. The method of the invention comprises the following steps: firstly, selecting antagonistic bacteria of pathogenic bacteria by a plate confrontation cultural method; then screening the antagonistic bacteria with capacity of generating a biological film by a crystal violet dyeing method; and then further screening the hydrolysis capacity of calcium phytate. The method of the invention has strong pertinence, simplicity, high efficiency and low cost, and avoids the waste of manpower and financial resources caused by too many greenhouse tests and field tests.

Description

technical field [0001] The invention belongs to the field of agricultural biological control, and in particular relates to a method for efficiently and directional screening biocontrol bacteria for soil-borne diseases of crops. Background technique [0002] Soil-borne diseases are plant diseases that are difficult to control. The traditional control methods are mainly to cultivate disease-resistant varieties and apply chemical pesticides. However, due to the diversity of soil-borne diseases and the limitation of resource sources of disease-resistant varieties, the application of breeding disease-resistant varieties is limited; and the use of chemical pesticides is generally not effective, and will cause pesticide residues and environmental pollution problems. Therefore, new solutions need to be found. Biological control has attracted more and more attention in the control of plant pests and diseases due to its advantages of non-toxic, harmless, non-polluting, not easy to pr...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/01
Inventor 马平李社增鹿秀云郭庆港李宝庆
Owner 南京科绿丰科技有限公司
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