Method and culture medium for detecting sclerotinia sclerotiorum

A technology for Sclerotinia sclerotiorum and solid culture medium, which is applied in microorganism-based methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems of low specificity, low sensitivity, strong experience type, etc. High performance and high sensitivity

Inactive Publication Date: 2014-02-12
EAST CHINA UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The purpose of the present invention is to solve the problem that the detection and identification of sclerotia in the prior art is mainly based on morphological characteristics, time-consuming, low sensitivity, low specificity and strong experience, and it is difficult to monitor and identify the occurrence of sclerotia in time. To control the spread and prevalence of pathogenic bacteria, provide a reagent and detection method for rapid detection of Sclerotinia sclerotiorum, with strong specificity, high sensitivity and high accuracy

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  • Method and culture medium for detecting sclerotinia sclerotiorum
  • Method and culture medium for detecting sclerotinia sclerotiorum
  • Method and culture medium for detecting sclerotinia sclerotiorum

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Effect test

Embodiment 1

[0045] Five kinds of Sclerotinia sclerotiorum bacteria with different physiological characteristics were cultured on PDA medium to the logarithmic growth phase, and a 6 mm diameter plate was taken from the edge of the colony of the pathogenic bacteria to be tested, and inoculated in a medium containing no or containing 50 μg / mL detection Put the reagent on a PDA medium plate and culture it at a constant temperature of 25°C for 2 days. Observe the color of the colonies with the naked eye and the color of mycelia under an optical microscope. The results are as follows: figure 1 (1)~ figure 1 (10).

[0046] figure 1 (1)~ figure 1 (5) 5 species of Sclerotinia sclerotiorum Ss01, Hm25, Hm10 tested respectively I 、Hm10 T , and Hm10 IT When grown on a plate without detection reagents, the color of the colony is white, and the hyphae are colorless.

[0047] figure 1 (6-)~ figure 1 (10) 5 species of Sclerotinia sclerotiorum Ss01, Hm25, Hm10 tested respectively I 、Hm10 T , and ...

Embodiment 2

[0050] 17 kinds of different plant pathogenic bacteria such as Rhizoctonia solani, Sclerotinia watermelon, Botrytis cinerea, etc. were cultivated on PDA medium to logarithmic growth phase, and the diameter of 6mm bacterial discs were taken at the edge of the colony of the tested pathogens. Inoculate it on a PDA medium plate that does not contain or contain 50 μg / mL detection reagent, and culture at a constant temperature of 25°C for 2 days, observe the color of the colony with the naked eye, and observe the color of mycelia under an optical microscope. The results of culture with the detection reagent are as follows: figure 1 (11)~ figure 1 (27).

[0051] figure 1 (11)~ figure 1 (27) are the colony morphology and color of the 17 plant pathogenic bacteria tested. There was no difference in the color of the colonies of these pathogens grown on plates with and without the test reagent. Among them: Rhizoctonia solani, watermelon sclerotinia, mulberry sclerotinia, cabbage anthr...

Embodiment 3

[0053] Cultivate Sclerotinia sclerotiorum on PDA medium to the logarithmic growth phase, take a 6 mm diameter plate from the edge of the colony, inoculate it in a PSB shaker flask containing 50 μg / mL detection reagent, and place it at 25 ° C, 180 rpm Cultured at constant temperature for 3 days. Observe the color of mycelium and hyphae under a light microscope. The result is as figure 2 shown.

[0054] figure 2 (a) and (c) are the mycelium of Sclerotinia sclerotiorum cultured in the PSB shake flask without detection reagent, the mycelium is white, and the mycelium is colorless; figure 2 (b) and (d) are the mycelium of Sclerotinia sclerotiorum cultured in PSB shake flasks containing 50 μg / mL detection reagent, the mycelium is green, and the mycelium is green.

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Abstract

The invention provides a method and culture medium for specifically detecting sclerotinia sclerotiorum through 3-(2-pyridyl) methyl-2-(4-chlorphenyl) imino group thiazolidine or a derivative thereof with an imido-thiazolidine structure. The method provided by the invention can be used for specifically inducing the mycelia of the sclerotinia sclerotiorum to generate outstanding green pigments in a culture medium shake flask or a flat plate which contains 0.1-100.0 micrograms/milliliter of a detection reagent and has the characteristics of high specificity and high sensitivity and stability. The method comprises the following steps: ultrasonically leaching the green mycelia by using 90% ethanol; and then regulating pH to 3.0 by using 0.1 M of HCl to obtain the green pigments, wherein the green pigments have characteristic absorption peaks in a region with the wavelength of 610-630 nanometers. The detection reagent has certain prevention and control effects on the sclerotinia rot of colza.

Description

technical field [0001] This application relates to the fields of plant protection, biotechnology, pesticide science, etc., especially the field of plant protection. Specifically, the present application relates to detection reagents and detection methods for agricultural and forestry plant Sclerotinia sclerotiorum. Background technique [0002] Sclerotinia sclerotiorum is an important agricultural disease that harms 400 species of plants in 72 families, including Brassicaceae, Asteraceae, Leguminosae, Solanaceae, Ambelliaceae, and Malvaceae, including rapeseed. In the middle and lower reaches of the Yangtze River and the main rapeseed producing areas in the southeast coast, including Jiangsu, Anhui, Shanghai, Zhejiang, Hunan, Hubei, Jiangxi and other places, the occurrence is particularly prominent, and it has become one of the main diseases of rapeseed, seriously affecting the growth of oilseed crops in my country. Production. [0003] The fungus mainly survives summer and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C09B61/00C12R1/645
Inventor 黄青春丰俊李刚月钱旭红陶黎明徐文平
Owner EAST CHINA UNIV OF SCI & TECH
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