Efficient stable indoor inoculation method for Ustilaginoidea virens and special strain
A technology of rice smut fungus and indoor inoculation, which can be used in microorganism-based methods, botanical equipment and methods, biochemical equipment and methods, etc. Effects of vaccination methods
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Embodiment 1
[0049] The isolation, screening and identification of embodiment 1 Aspergillus oryzae bacterial strain HWD-2
[0050] 1. Isolation of Aspergillus oryzae strains
[0051] The disease-like material to be separated—the rice ball was collected by the inventor of the applicant from the paddy field of Donghu Village, Wulijie Town, Jiangxia District, Wuhan City, Hubei Province in October 2009, and was separated by conventional tissue separation method (reference: Fang Zhongda, Phytopathology, Beijing: China Agricultural Publishing House, 1998), rice balls were sterilized in 75% ethanol for 30 seconds, then surface-sterilized with 0.1% mercury chloride for 5 minutes, and rinsed with sterile water for 3 times. Then use aseptic absorbent paper to absorb the water on the surface of the rice curve ball, cut out its inner layer tissue, and place it in PSA (PSA medium composition: by g / L, potato 200g, sucrose 20g, agar powder 12g; add distilled water to 1000ml ) plate (containing 50ug / ml s...
Embodiment 2
[0074] The preparation of embodiment 2 rice smut inoculum
[0075] After the pathogenic bacteria have been cultured for 10 days, take a mycelium block with a diameter of 6mm and put it into 200ml PSB medium (PSB medium composition: in g / L: 200g potato; 20g sucrose; add distilled water to 1000ml), place at 28°C, and rotate at 180rpm Vibrating culture on a shaker for 7-10 days to obtain a mixture of mycelia and spores.
[0076] Preparation of aspergillus oryzae mycelium fragments + spore mixture (MHC solution): crush the mycelium spore mixture obtained above with a juicer (CDE-300E, Foshan Shunde Ou Ke Electric Co., Ltd.), 3000rpm Centrifuge for 5min, discard the supernatant, suspend the pellet with fresh PSB medium and dilute to spore concentration of 10 6 unit / ml, the MHC solution is obtained.
[0077] Preparation of Aspergillus oryzae parenchyma liquid: filter the mycelium spore mixture obtained above through four layers of gauze, take the filtrate, centrifuge at 3000rpm fo...
Embodiment 3
[0078] Example 3 Effects of different inoculum on the pathogenesis of rice false smut
[0079] At the booting stage of rice, inject the prepared 10 6 The parenchyma liquid 2ml of individual / ml concentration and the MHC liquid 2ml of equal spore concentration, until inoculum overflows from ear bract top. After inoculation, the rice was placed in a plant growth box (model: ZSX1500GS, Wuhan Ruihua Instrument Equipment Co., Ltd.), and kept moist for 3 days at 25°C and 95% RH, and then moved to a sprinkler irrigation room (cross-hung micro Spray system, Jiangxi Beijia Industrial Co., Ltd.), spray water once every 2 hours during the day, and spray water every 4 hours at night, keep the temperature at 25-32 ° C, and the humidity at 90%-98%. 3 weeks after inoculation, the ear rate % and disease index of rice false smut were investigated.
[0080] The results showed that injecting and inoculating the conidia liquid of Aspergillus oryzae parenchyma at the booting stage of rice can mak...
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