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Method for preparing culture medium appropriate for phytophthotacactorum to produce sporangiums by taking radish as raw material

A technology of Phytophthora and culture medium is applied in the field of culture medium of plant pathogen Phytophthora sporangia, which can solve problems such as contamination of sporangia, achieve simple and easy-to-learn procedures, high practical application value, and reduce the probability of contamination Effect

Inactive Publication Date: 2012-08-22
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention overcomes the shortcoming that the sporangia induced by the hydroponic method of Phytophthora viridans hyphae are easy to be polluted by miscellaneous bacteria, and provides a method for directly cultivating sporangia on the culture medium

Method used

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  • Method for preparing culture medium appropriate for phytophthotacactorum to produce sporangiums by taking radish as raw material
  • Method for preparing culture medium appropriate for phytophthotacactorum to produce sporangiums by taking radish as raw material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0008] Example 1: Comparison of the sporangia production ability difference between radish agar medium and other medium

[0009] 1. Culture medium production:

[0010] (1) Radish culture medium (100g / L): Take 100g radish, chop it, boil it in 0.5L water for 30 minutes and filter it, add 15g agar powder to the filtrate, heat to melt the agar, and then dilute to 1 liter.

[0011] (2) Radish culture medium (150g / L): Take 150g radish, chop it, boil it in 0.5L water for 30 minutes and filter it, add 15g agar powder to the filtrate, heat it to melt the agar, and then dissolve it to 1 liter.

[0012] (3) Radish culture medium (200g / L): Take 200g radish, chop it, boil it in 0.5L water for 30 minutes and filter it, add 15g agar powder to the filtrate, heat to melt the agar and then dissolve it to 1 liter.

[0013] (4) Baiyun bean culture medium: 60 g of Baiyun bean powder, boiled in 0.5 L of water for 30 minutes and filtered, added 15 g of agar powder to the filtrate, heated to melt th...

Embodiment 2

[0024] Example 2: Preparation of Zoospore Suspension in Measuring the Pathogenicity of Phytophthora victoria to Panax notoginseng Leaves

[0025] 1. Preparation of culture medium: Take 200 grams of radish, chop it, boil it in 0.5L water for 30 minutes and filter it, add 15g agar to the filtrate, heat it to melt the agar and dissolve it to 1 liter, divide it into 0.103MPa, 121℃, Autoclave for 20 minutes. Cool to about 50°C to make medium plates.

[0026] 2. Preparation of spore suspension: Use a puncher with a diameter of 5mm to take out the bacterial mass from the edge of the colony that has been cultivated for 3 days, place it on the medium plate and cultivate it at 25°C. Add 10mL of sterile water at 4°C to stimulate the release of sporangia, and zoospores will be released after about 30-60 minutes. Use a pipette to transfer the spore suspension to a beaker, measure its concentration with a hemocytometer, and adjust the final concentration to contain approximately 10 5 A s...

Embodiment 3

[0028] Embodiment three: the identification of the sporangia morphology of Phytophthora victoria

[0029] 1. Preparation of culture medium: Take 100 grams of radish, cut into small pieces, boil in 0.5L water for 30 minutes, and then filter. ℃, high pressure steam sterilization for 20min. Cool to around 50 °C onto the medium plate.

[0030] 2. Morphological identification of colonies and sporangia: use a puncher with a diameter of 5 mm to punch out the bacterial mass at the edge of the colony that has been cultivated for 3 days, and place it on a different medium plate for cultivation at 25°C. Observe the shape and color of the colony on the 7th day. The shape, size, aspect ratio of the cyst, the top structure (with or without papillae and the thickness), the shedding character of the sporangia and the length of the sporangium peduncle after shedding. The results showed that, on the radish culture medium, the primary hyphae of Phytophthora victoria were colorless, and the myc...

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Abstract

The invention discloses a method for preparing a culture medium appropriate for phytophthotacactorum to produce sporangiums by taking radish as a raw material. The concrete preparation method comprises the following steps of: taking 100-200g of radish; chopping and boiling in 0.5 L of water for 30 minutes; filtering; adding 15g of agar in filtrate, heating to melt the agar with the constant volume to be 1 L; split charging; carrying out steam sterilization at high pressure of 0.103MPa and 121 DEG C for 20 minutes; putting the sterilized culture medium into a sterilized culture dish with the diameter of 9cm so as to form a surface plate; after cooling, punching the edge of a bacterial colony cultured for 3 days through a puncher with the diameter being 5mm to obtain bacterial blocks; inoculating the bacterial blocks on the culture medium; and culturing the inoculated culture dish in darkness for 10 days at 25 DEG C so as to produce a large number of sporangiums. The probability of pollution in an operation process that a water culture method is utilized for inducing to produce the sporangiums is reduced. The raw material of the culture medium is low in cost and is easy to obtain, and the processes of preparing the culture medium and producing the sporangiums are simple and easy to learn.

Description

technical field [0001] The invention relates to a culture medium for producing sporangia of plant pathogen Phytophthora cactorum in the field of plant protection. Background technique [0002] Phytophthora cactorum Schroet, belonging to the genus Phytophthora oomycetes, is an important plant pathogen widely distributed in the world, and can infect about 200 species of plants in about 60 families and 50 genera. Phytophthora usually overwinters in diseased debris and soil as mycelium and oospores. When the conditions are suitable in the following year, the mycelium directly infects and lodges, or forms a large number of zoosporangia and spreads to the aboveground part to infect the stems and leaves. Sporangia play a very important role in the transmission and spread of the disease. Usually, the preparation of spore suspensions is required for pathogenicity assays or other related biological characteristics studies in the laboratory. At present, the method for inducing Phyto...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N3/00C12N1/14C12R1/645
Inventor 朱书生杨敏何霞红梅馨月廖静静张红骥祁蕾宋冬冬李成云朱有勇
Owner YUNNAN AGRICULTURAL UNIVERSITY
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