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Method for promoting stable germination of Ophiostoma fimbriatum conidia and obtaining optimum observation effect

A technology of conidia and black spot bacteria, which is applied in the direction of microbial-based methods, biochemical equipment and methods, and microbial measurement/inspection, which can solve the problems of no reports, achieve simplified and efficient operations, and improve efficiency and accuracy , highly operable effect

Pending Publication Date: 2017-03-15
INST OF PLANT PROTECTION HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on how to ensure the germination rate of conidia of the black spot fungus that infects sweet potato is more than 90%, maintain stable germination and obtain the best observation effect.

Method used

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  • Method for promoting stable germination of Ophiostoma fimbriatum conidia and obtaining optimum observation effect
  • Method for promoting stable germination of Ophiostoma fimbriatum conidia and obtaining optimum observation effect
  • Method for promoting stable germination of Ophiostoma fimbriatum conidia and obtaining optimum observation effect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The preparation of embodiment 1 experimental material

[0027] Sweet potato black spot strain: sweet potato long beak shell (Ceratocystis fimbriata) CFSC-01, isolated from Sichuan diseased potato pieces, purified, identified and preserved; preservation number: CGMCC No.3.18030, preservation date: September 26, 2016; Preservation Unit: General Microbiology Center of China Microbiological Culture Collection Management Committee, Preservation Address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0028] P+S medium: Peel and cut potatoes and sweet potatoes into pieces, weigh 20g of potato pieces and 100g of sweet potato pieces, mix them, add water and boil until rotten (the amount of water added this time should not exceed 1L), filter, add glucose 0.02g to the filtrate, agar Powder 13g, add water to make up to 1L.

[0029] Sweet potato leaf tissue fluid: Grind and centrifuge 1g of sweet potato leaf tissue with 2ml of water, take the supernatant, add w...

Embodiment 2

[0030] Example 2 The relationship between conidia germination rate and germ tube length and time under interstitial fluid stimulation

[0031] Pick the sweet potato black spot fungus strains, inoculate them in the center of the P+S medium, and cultivate them in the dark at 25°C, then select the sweet potato black spot fungus strains that have been cultured for 10 days and 16 days respectively, and rinse the surface of the culture medium with sterile water. Bacteria, to obtain the washing solution containing conidia, then use sterile water to adjust the concentration of conidia in the washing solution, microscopic examination, and finally prepare the concentration of conidia as 1 × 10 6 / ml of conidia suspension. Sweet potato leaf tissue fluid was added to the two conidia suspensions respectively, and the amount of tissue fluid added was 20 μl of tissue fluid per 1 ml of conidia suspension. Continue culturing the conidia suspension added to the tissue fluid for 1h, 2h, 2.5h, 3...

Embodiment 3

[0036] Embodiment 3 The relationship between the concentration of sweet potato leaf tissue fluid and the germination rate of conidia

[0037] Pick the strain of sweet potato black spot fungus, inoculate it in the center of the P+S medium, cultivate it under dark conditions at 25°C for 8 days, then wash the bacteria on the surface of the medium with sterile water to obtain a washing solution containing conidia, Then use sterile water to adjust the concentration of conidia in the flushing liquid, examine with a microscope, and finally prepare the concentration of conidia to be 1×10 6 / ml of conidia suspension. Different amounts of tissue fluid were added to the conidia suspension, and the amount of tissue fluid added was 0, 1, 3, 6, 12, 15, 20, and 30 μl of tissue fluid per 1 ml of conidia suspension. Continue culturing the conidia suspensions added with different amounts of tissue fluid for 3 h at 25°C in the dark, and perform microscopic examination on the germination of coni...

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Abstract

The invention discloses a method for promoting stable germination of Ophiostoma fimbriatum conidia and obtaining the optimum observation effect. The method comprises the following steps: inoculating Ophiostoma fimbriatum strain to a P+S medium, culturing for 7-21 d, and preparing a conidial suspension; adding a sweet potato leaf tissue fluid into the conidial suspension, continuously culturing until Ophiostoma fimbriatum conidia in the conidial suspension are germinated and the optimum observation effect is obtained, and carrying out microscopic examination. By combining characteristics of Ophiostoma fimbriatum and through repeated experimental demonstrations, a method for promoting stable germination of Ophiostoma fimbriatum conidia is obtained. by the method, germination rate of Ophiostoma fimbriatum conidia can be stabilized at 90% and above, the optimum observation time is determined, requirements of a fungicide screening test are met, and a guarantee is provided for screening a medicament for controlling Ophiostoma fimbriatum, evaluating drug resistance risk of the medicament and smoothly conducting a test by means of germination of conidia.

Description

technical field [0001] The invention relates to a method for promoting the stable germination of conidia of sweet potato black spot fungus and obtaining the best observation effect, in particular to a method for promoting the stable germination of sweet potato black spot fungus conidia under specific culture conditions and obtaining the best observation effect. The best way to observe the effect. Background technique [0002] Sweet potato black spot is the main disease of sweet potato in field cultivation period and storage period. The yield loss caused by this disease is generally 5%-10% every year, and it can reach 20%-50% or even higher when the damage is serious. At the same time, diseased potatoes will also produce toxic substances such as ipomeamarone and ipomeanine, which can cause poisoning and even death after eating by humans and livestock. The focus of disease control. [0003] The control of sweet potato black spot mainly depends on the use of fungicides, but t...

Claims

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Application Information

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IPC IPC(8): C12Q1/18C12Q1/04C12N1/14C12R1/645
CPCC12N1/14C12Q1/045C12Q1/18C12N1/145C12R2001/645
Inventor 张德胜张振臣王爽秦艳红乔奇田雨婷王永江白瑞英
Owner INST OF PLANT PROTECTION HENAN ACAD OF AGRI SCI
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