Culture method for rapid and mass production of spores from pathogenic bacteria of sugarcane stripe
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A cultivation method and technology of brown streak disease, which is applied in the field of rapid and massive spore production of the sugarcane brown streak disease pathogen, can solve the problems of less spore production and long spore production time, etc.
Pending Publication Date: 2021-09-10
SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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[0005] Aiming at the deficiencies in the prior art, the present invention provides a method for rapidly mass-producing sporulation of sugarcane brown streak pathogen, which can effectively solve the problems of long sporulation time and low sporulation amount existing in existing sporulation methods
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Embodiment 1
[0035] (1) Activation of sugarcane brown streak pathogenic bacteria: inoculate sugarcane brown streak pathogenic bacteria on potato dextrose agar medium, seal the petri dish with parafilm, put it upside down in a constant temperature incubator at 28°C, and cultivate in the dark for 4 days to obtain pathogenic bacteria. spare;
[0036] (2) Preparation of sporulation medium: take 30 g of oat flakes, add ultrapure water to it, water bath at 80° C. for 120min, stir continuously during the period, filter with four layers of gauze, take the filtrate, add ultrapure water to make the volume to 1000mL, add to it Add 17g agar powder, mix well, sterilize by autoclaving at 121°C, 105KPa for 20min, take out and cool to 50°C, add ampicillin with a final concentration of 0.01mg / mL in the ultra-clean workbench (ampicillin in the sporulation medium) concentration), mix well, and distribute it in petri dishes to obtain a sporulation medium;
[0037] (3) Inoculation: take the activated sugarcan...
Embodiment 2
[0042] Traditional sporulation culture method
[0043](1) Activation of the pathogenic bacteria of brown streak disease on sugarcane: inoculate the pathogenic bacteria of brown streak on sugarcane on the potato dextrose agar medium, seal the petri dish with parafilm, put it upside down in a constant temperature incubator at 28°C, and cultivate in the dark for 4 days. Get the pathogenic bacteria for backup;
[0044] (2) Peel and chop 200g of potatoes, simmer for 30min until the potatoes are soft and rotten, filter out the residue with four layers of gauze, add 20g of agar powder and 20g of glucose to the filtrate, add ultrapure water after dissolving and dilute to 1000mL, 121°C, Autoclave at 105KPa for 20min, and distribute them in petri dishes. The activated sugarcane brown streak pathogen was inoculated into the medium, cultivated in the dark in a constant temperature incubator at 28°C, and the colony growth was observed on the 20th day (see figure 1 Left), eluted spores, f...
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Abstract
The invention discloses a culture method for rapid and mass production of spores from pathogenic bacteria of sugarcane stripe and belongs to the technical field of phytopathogen-induced spore production. The method comprises the following steps: firstly, preparing a spore production culture medium mainly comprising oatmeal, agar powder and ultrapure water, then, inoculating the spore production culture medium with activated pathogenic bacteria of the sugarcane stripe, carrying out dark culture at a constant temperature of 28 DEG C, scraping off aerial hyphae by using a sterile cotton swab when a diameter of a bacterial colony reaches 5cm, carrying out dark culture for 7 days at the constant temperature of 28 DEG C again, eluting conidia with sterile water, and measuring the number of the spores under a microscope. By adopting the method to culture the pathogenic bacteria of the sugarcane stripe, the spore production efficiency of the pathogenic bacteria of the sugarcane stripe can be greatly improved, the method has the advantages of high bacterial colony growth speed, large sporulation quantity, simplicity in operation and the like, an identification cycle for sugarcane-stripe-resistant varieties is shortened, human and material resources are saved, the method has a relatively high application value, and a scientific and effective technical support is provided for in-depth research on occurrence rules and prevention and control means of sugarcane stripe diseases.
Description
technical field [0001] The invention belongs to the technical field of inducing sporulation by plant pathogenic bacteria, and in particular relates to a culture method for rapid and mass sporulation of sugarcane brown streak pathogenic bacteria. Background technique [0002] Sugarcane brown streak disease is an important fungal disease that harms sugarcane leaves. It was first discovered in Cuba in 1924, and then in many sugarcane-growing countries such as the United States, China, India, Australia, Brazil, Fiji, Japan, Panama, South Africa, Pakistan, and Thailand. There are reports of occurrence and damage to different degrees, which bring different degrees of economic losses to the local cane sugar industry. In recent years, with the large-scale planting of susceptible varieties such as Xintaitang No. 25 and Yuetang 93-159, and the increase of rainy and high-humidity weather, sugarcane brown streak disease has broken out and endangered large areas in major sugarcane produc...
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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N3/00C12R1/645
CPCC12N1/14C12N3/00
Inventor 单红丽黄应昆高三基张慧丽王长秘李婕王晓燕张荣跃李银煳
Owner SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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