Medium for simple, convenient and quick pythium aphanidermatum isolation

A culture medium and technology of Pythium melons and fruits, applied in the field of microorganisms, can solve the problems of difficult separation, low separation rate of target bacteria, failure to achieve separation and purification, etc., and achieve the effect of simple preparation, fast speed, practicality, quickness and cost

Active Publication Date: 2012-10-31
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In actual work, in addition to being infected by Pythium melon and fruit, the samples collected from the field inevitably often contain other non-target fungi and bacteria such as Fusarium, Rhizoctonia solani and Phytophthora, and are separated according to conventional methods In the case of tissue pathogenic bacteria, due to the contamination of these non-target fungi or bacteria, the isolation rate of the target bacteria is low or it is difficult to isolate the target bacteria
Therefore, when utilizing ordinary culture medium to isolate Pythium melon and fruit from diseased tissue or soil, the purpose of separation and purification cannot be achieved.

Method used

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  • Medium for simple, convenient and quick pythium aphanidermatum isolation
  • Medium for simple, convenient and quick pythium aphanidermatum isolation
  • Medium for simple, convenient and quick pythium aphanidermatum isolation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, three kinds of fungal inhibitors, a kind of Phytophthora inhibitor and three kinds of antibiotics are to Pythium melon and fruit inhibition

[0033] 1. Inhibitory effect of three fungal inhibitors and one Phytophthora inhibitor on Pythium melon and fruit

[0034] Using dimethylformamide as the solvent of carbendazim, using acetone or methanol as the solvent of prochloraz, pentachloronitrobenzene and flumorph, prepare 10ml of four kinds of medicament mother solutions of 5000 μg / mL respectively; The mother liquors of the three fungal inhibitors, bacterazim, pentachloronitrobenzene and prochloraz, respectively prepared a series of drug-containing PDA plates with a concentration of 50 μg / mL, 20 μg / mL and 10 μg / mL; The mother solution of the inhibitor was prepared to prepare a series of drug-containing PDA plates with a concentration of 20 μg / mL, 10 μg / mL and 5 μg / mL, and the plate with solvent was used as a control; , pentachloronitrobenzene, prochloraz and f...

Embodiment 2

[0043] Embodiment 2, three kinds of antibiotics are to the inhibitory effect of bacteria in the environment

[0044] Plants to be tested: Cultivate the susceptible cucumber variety Changchun Mici in a seedling pot with a diameter of 15 cm and a height of 12 cm, and place it in the pepper test field of the Science Park of China Agricultural University. Liquid preparation.

[0045] Soil for testing: Soil samples were collected from areas with severe damping-off disease in Daxing District, Beijing in 2011, for future use.

[0046] Test method: This test is based on the results of Example 1 to detect whether the selected three concentrations of antibiotics can effectively inhibit the bacteria in the environment. For the detection of bacteria in the environment, it is mainly aimed at the bacteria from the base of the plant stem and the diseased soil. This is based on the fact that the isolated parts of Pythium melon and fruit are mainly determined by the base of the diseased stem ...

Embodiment 3

[0049] Embodiment 3, be used for separating the PDA selective culture medium of Pythium melon and fruit

[0050] 1. Separation method

[0051] During 2011 and 2012, disease samples were collected from areas with severe damping-off disease in Daxing, Pinggu, Changping, and Shunyi districts of Beijing for the isolation of Pythium melons and fruits.

[0052] The isolation of Pythium melon and fruit is carried out as follows: select different fields in the same area, and collect damping-off disease samples (rhizome and soil) for isolation.

[0053] Separation step: clean the diseased tissue with clean water and dry it fully, cut out a small piece of tissue at the junction of the rhizome with disease and health, inoculate it on the selective medium of the present invention, and culture it in the dark at 25°C until mycelium grows; Pick the vigorously growing small piece of mycelium into clean water, and culture it at 20°C for 24-48 hours. It can be observed that a large number of s...

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Abstract

The invention discloses a selective medium for simply, conveniently and quickly isolating pythium aphanidermatum, which is particularly suitable for isolation of pythium aphanidermatum, and belongs to the technical field of microbes. The medium is obtained through adding the following disinfectant (1) or (2) into a PDA (Potato Dextrose Agar) medium: (1) is a disinfectant consisting of carbendazol, pentachloronitrobenzene, flumorph and rifampicin; and (2) is a disinfectant consisting of prochloraz, pentachloronitrobenzene, flumorph and rifampicin. Experiments show that the PDA selective medium has the characteristics of feasibility, accuracy, practicability, quickness, low cost and simplicity and convenience in preparation. When the PDA selective medium is used for isolating pythium aphanidermatum and other pythium oomycetes, the problem of serious contamination of various non-target fungus and bacteria in the isolation process of pythium aphanidermatum and other plant pathogenic pythium is solved, and the effects of quickness, high efficiency and labor and time conservation are achieved.

Description

technical field [0001] The invention relates to a selective culture medium for simple and rapid separation of Pythium. It belongs to the technical field of microorganisms. Background technique [0002] Pythium is a soil fungus distributed worldwide, with strong saprophytic ability, which can infect the roots and seedlings of more than 150 kinds of vegetable and fruit crops, causing rotten seeds, damping off, standing dry, rotten roots and rotten fruits. Among them, the damping-off disease caused by Pythium aphanidermatum is a worldwide devastating disease in the seedling stage of vegetables, flowers, medicinal plants, tobacco and beans, cotton, corn, sorghum and other plants. Usually after emergence, water-soaked dark spots appear at the base of the stem near the ground, and then expand around the stem, gradually constricting into a thin line, causing the aboveground part of the seedling to collapse due to loss of support, commonly known as stuck neck disease. When the hum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N47/18A01N43/90A01N43/84A01N33/18A01N47/38A01P3/00A01P1/00C12N1/14C12N1/02C12R1/645
Inventor 贾睿哲刘鹏飞
Owner CHINA AGRI UNIV
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