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A technology of rice smut fungus and separation method, applied in the field of microorganisms, to achieve the effects of ensuring germination ability, avoiding killing effect, and weakening interference effect
Inactive Publication Date: 2013-05-15
INST OF PLANT PROTECTION FAAS
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A simple rapid separation method of rice koji bacteria (ZL201010301356.0) is an invention patent of the applicant. Its main technical feature is that after the rice koji ball surface is treated with a disinfectant, the rice koji ball is crushed with sterile water to make the mashing liquid Containing chlamydospores and inner tissue pieces, and then washing the surface of Wakimoto Zhe's medium for separation with a crushing liquid, the rice smut can be quickly isolated, but this method uses ethanol and mercuric chloride for disinfection during the separation process Because chlamydospores are more sensitive to disinfectants (Wang Yongqiang, 2010, comparative study on the isolation methods of Chlorosclerotinum oryzae (Oryza sativa), Acta Mycophyta Sinica 2010, 29 (1), using this method for longer storage time, The isolation of Aspergillus oryzae with low chlamydospore germination ability has obvious influence
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Embodiment 1
[0025] (1) The source of the rice curved ball standard sample: rice curved balls stored at room temperature for 3 months.
[0026] (2) Culture medium preparation
[0027] A: The medium used for separation is Wakimoto Tetsu’s medium: 300g potato, 5g peptone, 15g sucrose, 0.5g Ca(NO3)2 4H2O, 2.0g Na2HPO4 12H:0, 16g agar, add water to 1000mL . The culture medium was sterilized by high-pressure damp heat at 121°C for 20 minutes, and when the culture medium was cooled to 45°C-50°C, add chloramphenicol (50 μg / ml) to inhibit bacterial growth, mix well, and pour it into a petri dish, and the medium was solidified for later use use.
[0028] B: The culture medium used is PSA solid medium and PS liquid medium. The PSA solid medium is 200g of potatoes, 20g of agar, 20g of sucrose, and dilute to 1000mL with water. PS liquid medium is 200g of potatoes, 20g of sucrose, and dilute to 1000mL with water. All the above culture media were sterilized under high pressure and damp heat at 121°...
Embodiment 2
[0038] Example 2 Comparison of the results of several methods for the isolation of rice smut bacteria
[0039] 1, the effective isolation method of rice false smut bacteria of the present invention, its step is as follows:
[0040] 1) Isolation of rice smut bacteria
[0041] Same as Example 1
[0042] 2) The characteristics of growth and conidia germination of rice false smut and microscopic observation
[0043] Same as Example 1
[0044] 2. Chlamydospore suspension method
[0045] Referring to Zhou Yongli's 1999 method (Zhou Yongli, 1999, A Preliminary Study on the Isolation Technology of Rice Algeria, China Rice Science, 1999.13(3): l86-188), that is, the rice balls were sterilized with 75% ethanol for 30 seconds, and then sterile Wash 3 times with water, then place in an Eppendorf tube and vibrate to prepare a chlamydospore suspension, take 2 to 3 drops and add it to a petri dish, and pour it into a petri dish when the culture medium is cooled to 45°C to 50°C, gently S...
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Abstract
The invention relates to an effective Ustilaginoidea virens separation method, and belongs to the technical field of microbe. The method comprises the steps of: scraping a small amount of chlamydospore powder on surfaces of false smut balls with a pricking needle; placing the powder in a sterile petri dish; adding 0.5mL of sterile water suspension chlamydospores; then diluting the chlamydospore suspension by 2-3 gradients, with the ratio of the chlamydospore suspension to sterile water being 1:5 in a 1.8mL Eppendorf tube through a gradient dilution method; transferring 0.2mL of the diluted chlamydospore suspension to surface of a Wakimoto toceshi solid culture medium for separation (containing 50 mug / ml chloramphenicol); smearing evenly; inversely placing the petri dish into an incubator at 28 DEG C for dark culture; and effectively separating out target pathogens six days later. The method provided by the invention obviously weakens interference effect of infectious microbe in chlamydospores of rice false smut balls, has very simple operation procedure, is suitable for separation of rice false smut balls stored at normal temperature or 4 DEG C in a refrigerator within 3 months, and has the best separation effect on rice false smut ball bacteria just acquired in the field.
Description
technical field [0001] The invention relates to an effective method for isolating rice smut bacteria and belongs to the technical field of microorganisms. Background technique [0002] Rice false smut, commonly known as harvest disease, is an important disease of rice spikes, and its pathogen is Deuteromycotina genus Chlorosclerotinia Ustilaginoidea virens (Cook.) Tak. In recent years, with the vigorous promotion of high-yield and high-quality rice with dense panicles, the area and damage degree of rice false smut have gradually increased. The occurrence of rice false smut has a significant impact on rice yield and quality. In 1-10 grains, the most will reach 30-50 grains. The rice chlamydospore powder is dark green or black. After being mixed into the rice, it will seriously pollute the rice, affect the quality and appearance of the rice, and be mixed with the rice koji in the diseased rice. Balls and germs contain toxins that are toxic to humans and animals. Rice false ...
Claims
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