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Biological agent for preventing and treating verticillium dahliae, preparation and application method thereof

A technology of cotinus fusarium wilt and biological preparations, applied in biochemical equipment and methods, botany equipment and methods, methods based on microorganisms, etc., can solve the problems of poor control effect of cotinus cotinus wilt and achieve good application prospects

Active Publication Date: 2021-04-09
济南市园林绿化工程质量与安全中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of this, the object of the present invention is to solve the problem that existing medicines have poor control effects on Cotinus Fusarium Wilt, and provide a biological agent for preventing and treating Cotinus Cotinus Fusarium Wilt and its preparation and application method

Method used

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  • Biological agent for preventing and treating verticillium dahliae, preparation and application method thereof
  • Biological agent for preventing and treating verticillium dahliae, preparation and application method thereof
  • Biological agent for preventing and treating verticillium dahliae, preparation and application method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1: Isolation and identification of bacteria

[0017] 1.1 Isolation of bacteria

[0018] (1) In the ultra-clean workbench, put the newly collected branches of Cotinus cotinus wilt in 100 mL of sterile water, shake and rinse for 30 minutes, and suspend them sufficiently to make a bacterial suspension.

[0019] (2) After gradiently diluting the bacterial suspension with sterile water, take 100uL from each concentration of bacterial suspension dilution and spread it on the LB medium plate, place it at 30°C for 48 hours, and then pick it out on the plate Streak and purify single colonies with different shapes, sizes, and colors on LB plates, and number them.

[0020] (3) Take the purified strains with different numbers and culture them in LB liquid medium at 30°C and 180rpm / min for 24 hours, then take 2mL of fermentation broth and mix them into 15mL of PDA medium, pour the plate, after solidification, inoculate in the center of the plate The inhibitory co-cultur...

Embodiment 2

[0026] Embodiment 2: the antagonism of methylotrophic bacillus KNK-1 to Verticillium dahliae (co-cultivation method)

[0027] (1) After the purified strains with different numbers were cultured in LB liquid medium at 30°C and 180rpm / min for 24 hours, 2mL of fermentation broth was mixed into 15mL of PDA medium, poured onto the plate, solidified, and set aside.

[0028] (2) Use a hole puncher with a diameter of 2.5 cm to punch holes on the edge of the cultivated Verticillium dahliae colony to obtain bacterial flakes, which are set aside.

[0029] (3) Use a sterile picking needle to pick the above-mentioned bacterial slices into the center position of the PDA plate in (1), and set the PDA plate only inoculated with Verticillium dahliae bacterial slices as a control group at the same time, cultivate at 30°C for 4-6 days, Observe the growth colony speed and colony radius of Verticillium dahliae. Three replicates were set up for each, and the results are shown in Table 1.

[0030]...

Embodiment 3

[0033] Example 3: Detection of the control effect of KNK-1 on Cotinus cotinus wilt

[0034] (1) Inoculate KNK-1 into LB culture medium, and culture at 180rpm / min at 30°C for 24h to obtain seed solution with a concentration of about 10 9 cfu / mL;

[0035] (2) Strain KNK-1 was fermented with 60 liters of mechanically stirred stainless steel fermenters with six consecutive tanks: the fermentation medium was molasses medium (1 L): 100 mL of molasses, 1 g of sodium chloride, and 3 g of urea. The liquid loading volume is 40L, the inoculum size is 6%, the temperature is 30°C, the stirring speed is 300rpm / min, the ventilation rate is 3L / min, the fermentation time is 2 days, and the biomass reaches 2.10×10 10 cfu / mL;

[0036] (3) Preparation of finished powder: add carrier diatomaceous earth and starch to the fermented KNK-1 fermentation liquid according to the saturated adsorption amount, stir once every 1 hour to make the carrier fully adsorb the bacteria, dry at low temperature aft...

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Abstract

The invention discloses a biological agent for preventing and treating verticillium dahliae, preparation and application method thereof. The preservation unit of bacillus methylotrophicus is China General Microbiological Culture Collection Center; the address is No.3, Yard 1, West Beichen Road, Chaoyang District, Beijing; the preservation date is November 30, 2018; the preservation number of the strain is CGMCC NO: 16842. The bacillus methylotrophicus provided by the invention not only can efficiently inhibit verticillium dahliae during plate culture, but also has a prevention and treatment effect as high as 66.2% in a field experiment.

Description

technical field [0001] The invention relates to the technical field of biological medicines, in particular to a biological preparation for preventing and treating Cotinus cotinus wilt and its preparation and application methods. Background technique [0002] Cotinus coggygria (Cotinus coggygria) belongs to the Sumac family Sumac genus. Its autumn leaves are bright red after frost. It is widely distributed in Northwest my country, North China, Southwest China, Zhejiang, Anhui and other places. It has great ornamental and economic value. Since the 1980s, Cotinus cotinus in Beijing, Qingzhou, Jinan and other places in Shandong have suffered from different degrees of fusarium wilt. The diseased plants in the Cotinus cotinus forest in Xiangshan, Beijing are 0.11%-46.2%, and the diseased plants are scattered or distributed in patches , the light ones will affect the red leaf landscape, and the severe ones will die soon. As a destructive forest disease, it has caused serious harm ...

Claims

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Application Information

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IPC IPC(8): A01N63/22A01P3/00C12N1/20C12R1/07
CPCA01N63/22C12N1/20
Inventor 杜欣韩梅珍张倩段伟张秀明康建美张新建周红姿范素素
Owner 济南市园林绿化工程质量与安全中心
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