Method for high-flux identification of ralstonia solanacearum resistance

A high-throughput technology for tobacco bacterial wilt, which is applied in the field of identification of tobacco bacterial wilt resistance, can solve the differences in resistance performance between years and locations, the difficulty of controlling the temperature of the identification environment, and the demand for bacterial suspension of R. solanacearum To achieve the effects of stable incidence and disease index, completely consistent disease conditions, and stable incidence and disease index results

Inactive Publication Date: 2014-10-08
GUIZHOU TOBACCO SCI RES INST
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Problems solved by technology

This method can better reflect the resistance of varieties; but this method has a long identification period, can only be identified once a year, requires a large area of ​​disease nursery, limited tobacco materials for identification, and has a large demand for R. solanacearum suspension. The incidence is uneven, and there are problems...

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Embodiment 1-8

[0022] A method for high-throughput identification of tobacco bacterial wilt resistance, comprising the following steps:

[0023] (1) Cultivate sterile tobacco seedlings

[0024] Sterilize the quartz sand with a particle size of 2-4mm at 121°C for 30 minutes, and sow it into a sterile 12-well cell culture plate after cooling, add 10g of quartz sand to each well, and add 4ml of sterile water to each well. The flue-cured tobacco seeds to be tested (8 varieties were all provided by the Breeding Engineering Technology Center of Guizhou Tobacco Research Institute) were sown on the quartz sand of the 12-well cell culture plate. After sowing, the 12-well cell culture plate was placed at 25°C and RH>80 %, 16h light and 8h dark alternating conditions.

[0025] (2) Daily management of tobacco seedlings

[0026] After the tobacco seeds germinate, add Hoglangd nutrient solution to the 12-well microwell plate for cultivating tobacco seedlings under aseptic conditions, 2ml per well, and c...

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Abstract

The invention discloses a method for high-flux identification of ralstonia solanacearum resistance. The method comprises the following steps: sterilizing quartz sand with particle size ranging from 2 mm to 4 mm at high temperature of 121 DEG C for 30 minutes, cooling, and spreading into an aseptic 12-pore cell culture plate, wherein each pore is filled with 10 g of the quartz sand and 4 ml of aseptic water; sowing flue-cured tobacco seeds to be tested onto the quartz sand of the 12-pore cell culture plate, after sowing, and culturing by the 12-pore cell culture plate under the conditions of 25 DEG C, RH of more than 80% and alternative 16-hour lighting and 8-hour shading; after the flue-cured tobacco seeds germinate, adding a Hoagland's nutrient solution into the 12-pore cell culture plate for culturing the flue-cured tobacco seeds under the aseptic condition, continuously culturing under the conditions of 25 DEG C, RH of more than 80% and alternative 16-hour lighting and 8-hour shading, and identifying the disease resistance of the flue-cured tobacco seeds each with 5-6 leaves, wherein each pore is filled with 2 ml of the Hoagland's nutrient solution; preparing inoculating liquid and inoculating; investigating and grading the disease states. According to the method, the ralstonia solanacearum resistance can be identified with high flux and high efficiency and a large amount of flue-cured tobacco varieties can be identified in one time.

Description

Technical field [0001] The present invention is the field of crop resistance appraisal technology, which involves a method of appraisal method of tobacco blue blight resistance. [0002] Background technique [0003] Bacteria ( Ralstonia solandacearum (Smith) Yabuuucki et al) Tobacco -blue blight caused by a global violin bacterial disease, which is extremely destructive and has become the main disease that restricts tobacco production.Tobacco bliss is generally occurred in provinces and regions of major smokers.The disease is a typical vascular beam disease. All parts of the tobacco roots, stems, and leaves can be infected, but the main symptoms of the roots and stems are mainly harmful to the roots and stems of plants.At present, the cultivation and application of disease resistance is the most effective way to prevent and treat blue blight.However, in the study of disease -resistant breeding, it is necessary to quickly and accurately evaluate the resistance of a large amount o...

Claims

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Application Information

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IPC IPC(8): A01G7/00A01G31/00
Inventor 汪汉成任学良王轶蔡刘体王茂胜陆宁王仁刚
Owner GUIZHOU TOBACCO SCI RES INST
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