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Method for screening alternaria alternata resistant germplasm material from petri dish

A technology of tobacco red spot bacteria and red spot disease, applied in the field of agricultural biology, can solve problems such as difficult to eliminate the impact, and achieve the effect of saving manpower

Inactive Publication Date: 2013-01-16
TOBACCO RES INST CHIN AGRI SCI ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For another example, the root soaking method of seedlings is to use the toxin produced by Alternaria Alternaria Alternaria to soak the roots of tobacco plants. It is difficult to rule out the influence of bacteria in the toxin solution on the growth of young roots of plants.
Therefore, these identification methods have major defects.

Method used

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  • Method for screening alternaria alternata resistant germplasm material from petri dish
  • Method for screening alternaria alternata resistant germplasm material from petri dish
  • Method for screening alternaria alternata resistant germplasm material from petri dish

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment one, screening method is as follows

[0030] a. Cultivate Alternaria tabacum on the potato agar medium; b. After the Alternaria overgrows the culture medium, take Alternaria bacterium cake with a diameter of 5mm and inoculate it in 250ml Chapi culture medium, and culture it with shaking at 28°C for 28-30d; C, gauze filter and remove mycelia, then through 3-5 times of centrifugation, 8000r / min, microscopic examination without cells, obtain the crude extract of tobacco Alternaria toxin; d, determine the activity of toxin with the method of bioassay; e, use water Dilute the toxin crude extract by 2-16 (V / V) times, add 1.8-2.0% (W / V) agar to form a toxin-containing medium; -30min; g, sterilize the tobacco seeds; h, use tweezers to clamp the sterilized tobacco seeds onto the toxin-containing medium, seal it with a parafilm, and cultivate it for more than 14 days at 25-28°C and 12h light / d. ; i, according to whether the root of the tobacco seedling grows into the ...

Embodiment 2

[0036] Embodiment 2, screening method is as follows

[0037]a. Cultivate Tobacco Alternaria Alternaria on oatmeal agar medium; b. After Alternaria Alternaria overgrows the culture medium, take a cake of Alternaria Alternaria with a diameter of 5mm and inoculate it in 250ml Chapi culture medium, and culture it with shaking at 28°C for 28-30d C, gauze filter to remove mycelia, then through 3-5 times of centrifugation, 8000r / min, microscopic examination without cells, obtain the crude extract of tobacco Alternaria toxin; d, determine the activity of toxin with the method of bioassay; e, Dilute the toxin crude extract with water for 2-16 (V / V) times, add 1.8-2.0% (W / V) agar to form a toxin-containing medium; f, damp heat sterilization at 121°C 15-30min; g, sterilize the tobacco seeds; h, clamp the sterilized tobacco seeds onto the toxin-containing medium with tweezers, seal with a parafilm, and cultivate for 14 days at 25-28°C, 12h light / d culture conditions Above; i, identify to...

Embodiment 3

[0043] Embodiment 3, screening method is as follows

[0044] a. Cultivate Tobacco Alternaria Alternaria on corn flour agar medium; b. After Alternaria Alternaria overgrows the culture medium, take a 5mm diameter cake of Alternaria Alternaria and inoculate it in 250ml Chapi culture medium, and culture it with shaking at 28°C for 28-30d C, gauze filter to remove mycelia, then through 3-5 times of centrifugation, 8000r / min, microscopic examination without cells, obtain the crude extract of tobacco Alternaria toxin; d, determine the activity of toxin with the method of bioassay; e, Dilute the toxin crude extract with water for 2-16 (V / V) times, add 1.8-2.0% (W / V) agar to form a toxin-containing medium; f, damp heat sterilization at 121°C 15-30min; g, sterilized tobacco seeds; h, Zhou clamped the sterilized tobacco seeds onto the toxin-containing medium, sealed with a parafilm, and cultured for 14 days at 25-28°C, 12h light / d culture conditions Above; i, identify tobacco germplasm...

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Abstract

The invention discloses a method for screening an alternaria alternata resistant germplasm material from a petri dish. The method comprises the following steps of: a, culturing alternaria alternata on an agar culture medium; b, carrying out shaking culture of the alternaria alternata in a Czapek culture solution; c, coarsely extracting alternaria alternata toxin; d, determining activity of the toxin by using a biological assay method; e, adding water for diluting a toxin solution and preparing a culture medium; f, carrying out 121-DEG C moist heat sterilization on a toxin-containing culture medium for 15-30min; g, disinfecting tobacco seeds; h, culturing the tobacco seeds on the toxin-containing culture medium; and I, identifying the alternaria alternata resistant tobacco germplasm material according to the condition whether a tobacco shoot root grows in the culture medium. According to the invention, the toxin-containing culture medium is prepared by using the toxin generated by the alternaria alternata, the tobacco seeds are cultured on the culture medium, and the alternaria alternata resistant germplasm material is directly screened from the tobacco seeds rapidly, economically, conveniently and reliably. The method can replace field test screening and can be used for screening the alternaria alternata resistant germplasm material in the petri dish.

Description

Technical field: [0001] The invention relates to the field of agricultural biotechnology, in particular to a method for screening tobacco germplasm materials resistant to red spot disease in a petri dish. Background technique: [0002] Alternaria alternata (Alternaria alternata) is an important tobacco leaf spot disease. The pathogen of tobacco red spot disease is Alternaria alternata (Fries) Keissler, which belongs to Deuteromyccotina, Hyphomycetes ), Hyphomycetoles, Alternaria. The hyphae are colorless and transparent, with septa. Conidiophores grow solitary or in clusters on conidiophores, conidiophores gather in piles, with curved tops, irregular, spores are in the shape of inverted rods and hammers, separated vertically and horizontally, with 1-3 vertical cells, 3-7 horizontal cells; conidia at the end of the spore chain are smaller, elliptic, with only one septum. The size and shape of conidia vary greatly due to the age of the bacteria and the length of time to pro...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
Inventor 时焦孙丽萍孟坤张峻铨雒振宁王凤龙
Owner TOBACCO RES INST CHIN AGRI SCI ACAD
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