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Method for identifying disease resistance of tobacco leaves with brown spot disease

A technology for identification of tobacco brown spot disease and disease resistance, which is applied in the directions of microorganism-based methods, microorganism determination/inspection, biochemical equipment and methods, etc. Stability and other issues, to achieve the effect of improving sporulation synchronization and spore quality stability, reducing identification costs, and shortening sporulation cycle

Active Publication Date: 2015-04-22
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among the existing identification methods, the field artificial disease nursery method has a long identification cycle and is easily affected by the field growth environment, making the inoculation effect unstable, and there are certain problems in the quantitative analysis of the pathogenic results
Although the identification method of the toxin of the red spot disease has the characteristics of fast processing and diversified applicable objects, it is not exactly the same as the actual pathogenesis of the tobacco red spot disease, which affects the reliability of the identification results. In addition, the determination of the toxin concentration in the actual operation It also requires more human and material resources

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] ——Resistance identification of NC89 variety to tobacco red spot disease

[0043] A. Tobacco leaf solution preparation: After picking the healthy and disease-free tobacco leaves at the first ripening stage, first soak them in 5% sodium hypochlorite solution for 1 min, rinse them with clean water, then place them in 75% ethanol solution, and soak them for 30 seconds. After drying, grind it finely in a sterilized mortar, break up the tissue, and make a 5% tobacco leaf solution with sterile water;

[0044] B. Bacteria spore culture: Cut the mycelium with Alternaria Alternaria in the petri dish into small pieces with a sterilized blade, then transfer to the Czapek medium plate, and inoculate 6 spots on each plate. Place in an incubator at 25°C for 15 days, wash the spores with tobacco leaf solution after the spores are produced, continue to stand for 8 hours to promote spore germination, and obtain a suspension of Alternaria spores with a spore concentration of 2.3×10 4 ind...

Embodiment 2

[0049] ——Resistance identification of PVH01 and PVH06 varieties to tobacco red spot disease

[0050] A. Tobacco leaf solution preparation: pick healthy and disease-free tobacco leaves at the first ripening stage, soak them in 6% sodium hypochlorite solution for 0.8 minutes, rinse them with clean water, then place them in 75% ethanol solution, and soak them for 20 seconds. After drying, grind it in a sterilized mortar, break up the tissue, and make a 4% tobacco leaf solution with sterile water;

[0051] B. Bacteria spore culture: Cut the mycelium with Alternaria Alternaria in the petri dish into small pieces with a sterilized blade, then transfer to the Czapek medium plate, and inoculate 7 spots on each plate. Place it in an incubator at 24°C for 10 days. After the spores are produced, wash the spores with tobacco leaf solution and continue to stand for 7 hours to promote the germination of the spores to obtain a suspension of Alternaria spores with a spore concentration of 1.8...

Embodiment 3

[0056] ——Identification of the resistance of new lines 9147, 9102, and 9111-21 to tobacco red spot disease

[0057] A. Tobacco leaf solution preparation: pick healthy and disease-free tobacco leaves at the first ripening stage, soak them in 4% sodium hypochlorite solution for 1.2 minutes, rinse them with clean water, then place them in 75% ethanol solution, and soak them for 40 seconds. After drying, grind it in a sterilized mortar, break up the tissue, and make a 6% tobacco leaf solution with sterile water;

[0058] B. Bacteria spore culture: Cut the mycelium with Alternaria Alternaria in the petri dish into small pieces with a sterilized blade, then transfer to the PDA medium plate, and inoculate 5 spots on each plate. Place it in an incubator at 26°C for 20 days, wash the spores with tobacco leaf solution after the spores are produced, and continue to stand for 9 hours to promote the germination of the spores to obtain a suspension of Alternaria spores with a spore concentr...

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Abstract

The invention discloses a method for identifying the disease resistance of tobacco red spot disease leaves, and belongs to the technical field of tobacco disease resistance identification. Sterilize and crush the healthy tobacco leaves at the mature stage, and make a 4-6% solution with sterile water; inoculate the spore-producing hyphae of Alternaria Alternaria on the medium plate, and cultivate in the incubator at 24-26°C for 10-20 days After spore production, wash the spores with tobacco leaf solution, let them stand for 7-9 hours to promote their germination, and obtain a bacterial suspension; when the tobacco leaves to be identified enter the mature stage, take samples, disinfect, dry and moisturize. Take 10-15 μl of the bacterial suspension and drop it on the tobacco leaves, 10-16 drops per tobacco leaf, and place the tobacco leaves in a moisture-retaining frame or an artificial climate box with a relative humidity ≥ 90% and a temperature of 24-26°C for 3-5 days. Days, after the appearance of yellow spots on the tobacco leaves, measure the diameter of the spots to determine their disease resistance. The identification method has high success rate, good reproducibility, rapidity and accuracy, convenient inspection and monitoring, no field pollution caused by artificial inoculation, and high value of popularization and application.

Description

technical field [0001] The invention belongs to the technical field of tobacco disease resistance identification, and in particular relates to a method for identifying disease resistance of tobacco red spot disease leaves with high success rate, good reproducibility, fast accuracy, convenient inspection and monitoring, and no field pollution. Background technique [0002] Tobacco red spot disease is a serious disease that endangers tobacco production. It often brings huge economic losses to tobacco leaf production in various tobacco areas in my country. It is particularly important to breed tobacco varieties resistant to red spot disease, and it is fast, accurate, reliable and simple. Easy identification methods for disease resistance can effectively shorten the breeding process. Among the existing identification methods, the field artificial disease nursery method has a long identification cycle and is easily affected by the field growth environment, making the inoculation e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/18C12R1/645
Inventor 段玉琪
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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