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Efficient stable indoor inoculation method for Ustilaginoidea virens and special strain

A technology of rice smut fungus and indoor inoculation, which is applied in microorganism-based methods, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problems of unstable incidence of artificial inoculation, and achieve stable and efficient inoculation effect, optimization Effects of vaccination methods

Inactive Publication Date: 2012-07-25
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to overcome the defects of the prior art, to propose a method for indoor artificial inoculation of rice smut with high efficiency and stability and a bacterial strain dedicated to the method, and to solve the difficult problem that the incidence of artificial inoculation of rice false smut is unstable at present

Method used

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  • Efficient stable indoor inoculation method for Ustilaginoidea virens and special strain
  • Efficient stable indoor inoculation method for Ustilaginoidea virens and special strain
  • Efficient stable indoor inoculation method for Ustilaginoidea virens and special strain

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Effect test

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, Ou Ke Electric Co., Ltd., Shunde District, Foshan City), under 3000rmp Centrifuge for 5min, discard the supernatant, suspend the pellet with fresh PSB medium and dilute to spore concentration of 10 6 individual / 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 filtra...

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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Abstract

The invention belongs to the technical field of plant protection, and particularly relates to an efficient stable indoor inoculation method for Ustilaginoidea virens and a special strain. The method includes: subjecting a hyphal mass (6mm) cultivated for 7 days on PSA (potato dextrose agar) to shake cultivation through PSB (photosynthetic bacteria) for 7-10 days so as to prepare hypha and spore mixed liquor, and filtering the mixed liquor with four layers of filter cloth to obtain spore liquor inoculants 106 / ml; inoculating rice ear bracts by injecting at the seventh stage of rice young ear differentiation; and keeping moist for three days at 25 DEG C and under 95%RH (relative humidity), and placing in a spray irrigation net chamber for cultivation at 25-32 DEG C and under 90%-98%RH. By the method, the rate of diseased ears under artificial inoculation of Ustilaginoidea virens can reach more than 90%. By the method compared with the outdoor artificial inoculation techniques, efficientstable incidence of Ustilaginoidea virens can be obtained under controllable conditions.

Description

technical field [0001] The invention belongs to the technical field of plant protection, and in particular relates to an efficient and stable indoor inoculation method of rice false smut bacteria and a special bacterial strain applying the method. Background technique [0002] Rice false smut is a disease of rice panicle grains caused by Ustilaginoidea virens. In recent years, with the change of rice cultivation mode, the increase of nitrogen fertilizer application in paddy fields and the large-scale promotion of new high-yielding dense-panicle hybrid rice, rice false smut has become increasingly serious in various rice-producing areas around the world. Rice false smut not only affects the yield of rice, but also pollutes rice with toxins produced by pathogenic bacteria, affects rice quality, reduces commodity value, and seriously threatens food safety around the world. So far, many scholars have carried out extensive research on rice smut, and initially revealed the biolog...

Claims

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Application Information

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IPC IPC(8): A01G16/00C12N1/14C12R1/645A01G22/22
Inventor 黄俊斌罗朝喜贾切郑露
Owner HUAZHONG AGRI UNIV
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