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Mass culture method of pyricularia grisea conidia
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A technology of conidia and rice blast fungus, applied in the field of plant pathology, can solve the problems of less conidia, easy contamination of miscellaneous bacteria, waste of time and culture medium, etc.
Active Publication Date: 2021-06-22
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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Problems solved by technology
The first type of method is to use relevant nutrients such as oat juice, rice bran juice, rice straw juice and corn flour to make agar medium, transfer the mycelium to the surface of the medium, and scrape the mycelium after it is covered with mycelium In addition, the conidia can be obtained by continuing to cultivate, this method is not easy to pollute, but the amount of conidia obtained is less
The second type of method is to use food crop grains such as barley and sorghum to make medium, transfer the mycelium to the grains, after the grains are covered with mycelia, wash off the mycelia on the surface of the grains with water, and the grains can be cultivated again. Obtain a large amount of conidia, but this method is easy to contaminate miscellaneous bacteria during the cultivation of conidia
At present, in a culture cycle of blast fungus conidia, usually only 1-2 conidia are washed out before the medium is disposed of, which will result in only a small amount of spores being obtained in one culture cycle, wasting time and cultivation base
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Embodiment 1
[0026] A method for mass culture of conidia of rice blast fungus, comprising the following steps:
[0028] Wash and cook the fresh barley grains, drain the surface water of the barley grains, pick out the cracked grains, put the remaining grains into a triangular flask, and sterilize them for later use;
[0029] 2) Cultivate conidia:
[0030] Activate Magnaporthe grisea on YSA medium. After 7 days, pick fresh mycelium pieces and place them in the center of the surface layer of barley grains in a triangular flask, and place them in a biochemical incubator at 28°C for dark cultivation. After 8 days, shake the grains in the bottle evenly , continue culturing at 28°C for 9 days to produce conidia;
[0031] The YSA medium is made of 10g / L soluble starch, 2g / L yeast extract, and 19g / L agar;
[0033] Sterilize all equipment and test environment involved in the process of eluting conidia;
[0034] Pou...
Embodiment 2
[0038] Step 2) Cultivate conidia:
[0039] Activate Magnaporthe grisea on YSA medium. After 6 days, pick a fresh mycelium block and place it in the center of the surface layer of barley grains in a triangular flask, and place it in a biochemical incubator at 26°C for dark cultivation. After 9 days, shake the grains in the bottle evenly , continue to culture at 26°C for 10 days to produce conidia;
[0040] The YSA medium is made of 10g / L soluble starch, 2g / L yeast extract, and 18g / L agar;
[0041] Step 3) Elution of conidia: continue culturing at 26°C to produce conidia;
[0042] The spores were eluted once every 36 hours at intervals, and the method was the same as the first elution, and each bottle of grains was eluted 7 times; other steps were the same as in Example 1.
Embodiment 3
[0044] Step 2) Cultivate conidia:
[0045] Activate Magnaporthe grisea on YSA medium. After 8 days, pick fresh mycelium pieces and place them in the center of the surface layer of barley grains in a triangular flask, and place them in a biochemical incubator at 27°C for dark cultivation. After 8 days, shake the grains in the bottle evenly , continue to culture at 27°C for 10 days to produce conidia;
[0046] The YSA medium is made of 10g / L soluble starch, 2 g / L yeast extract, and 20g / L agar;
[0048] Placed at 27°C to continue culturing to produce conidia;
[0049] The spores were eluted once every 40 h at intervals, and the method was the same as the first elution, and each bottle of grains was eluted 6 times; other steps were the same as in Example 1.
[0050] The beneficial effects of the present invention are further illustrated below through corresponding test data.
[0051] 1. Materials and methods
[0052] 1. Tested strains
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Abstract
The invention discloses a mass culture method for conidia of magnaporthe oryzae, and belongs to the technical field of phytopathology. The method comprises the following steps: 1) preparing a barley grain culture medium; 2) culturing conidia; (3) eluting the conidia: sterilizing all appliances and test environments involved in the process of eluting the conidia, wherein the spores are eluted once every 36-48 hours of culture in a manner as same as that of the first elution, each bottle of wheat grains being totally eluted for 5-7 times. According to the mass culture method of the pyricularia oryzae conidia, a barley grain culture medium is used for culturing the pyricularia oryzae conidia, barley grains cultured in a triangular flask are subjected to conidia elution with sterile water, and the conidia are eluted once every 36-48 h of culture; therefore, it is fully guaranteed that the barley grains are prevented from being polluted by infectious microbes in the elution and culture process, all strains to be tested can be eluted for 5-7 times, and therefore a large number of conidia can be obtained.
Description
technical field [0001] The invention belongs to the technical field of phytopathology, and specifically relates to a mass cultivation method of conidia of rice blast fungus. Background technique [0002] Magnaporthe grisea ( Magnaporthe oryzae ) is the most destructive pathogenic bacteria on rice production, [0003] The research on the biology and genetics of the rice blast fungus, the genetic mechanism of the interaction between the rice blast fungus and rice, and the research and development of rice blast control technology and control agents all require the conidia of the rice blast fungus, and the resistance of rice to rice blast Relevant tests such as the identification, evaluation and resistance breeding of M. oryzae require a large amount of conidia of Magnaporthe grisea. [0004] At present, there are two main methods for cultivating conidia of Magnaporthe grisea, each with advantages and disadvantages. The first type of method is to use relevant nutrients such a...
Claims
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Application Information
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