Method for phytophthora nicotianae inoculation of tobacco

A technology of Phytophthora tobacco and tobacco, applied in the field of plant disease resistance identification, can solve problems such as large occupied space, and achieve the effects of small occupied space, saving labor costs, and shortening experimental period

Inactive Publication Date: 2015-03-11
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

Bacteria and mycelium block trauma inoculation requires the tobacco varieties to be tested to be sown and cultivated until they become seedlings, and

Method used

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  • Method for phytophthora nicotianae inoculation of tobacco
  • Method for phytophthora nicotianae inoculation of tobacco

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preparation example Construction

[0015] B. Preparation of Phytophthora Nicotiana Zoospore Suspension: Inoculate Phytophthora nicotianae on OA medium plates, cultivate in the dark at 25~28℃ for 7~10 days until the hyphae overwhelm the entire plate, and cut the hyphae into small pieces. Block to obtain hyphae block, liquid culture induces zoosporangium, low temperature promotes zoosporangium release zoospores, adjust the zoospore concentration to 10 with sterile water 4~6 Pieces / ml to obtain a zoospore suspension of Phytophthora tabaci;

[0016] C. Root irrigation inoculation; take the zoospore suspension of Phytophthora nicotianae obtained in step B and slowly drip it onto the germination bed, allowing it to penetrate into the root of each cotyledon seedling;

[0017] D. Count the incidence and evaluate the results of disease resistance: After inoculation, the cotyledon seedlings will be cultured for 3 to 5 days at a relative humidity of 80~85%, a light intensity of 1000~2000Lx, and a temperature of 25~28℃. Investig...

Embodiment 1

[0026] Phytophthora Phytophthora Example 1: Experimental procedure and inoculation amount of zoospores inoculating cotyledon seedlings

[0027] In this example, Phytophthora Nicotiana race 0 (People's Republic of Tobacco Industry Standard YC / T39-1996) was used as the experimental strain, and the disease-resistant variety RBST and the susceptible variety Honghua Dajinyuan were used as the experimental tobacco varieties.

[0028] The specific operations are as follows:

[0029] (1) Preparation of tobacco cotyledon seedlings: After the surface of the disease-resistant variety RBST and the susceptible variety Honghua Dajinyuan tobacco seeds are disinfected, a 10×10 square array is spotted on the germination bed of a petri dish made of sponge and filter paper at a relative humidity of 80 ~85%, light intensity 1000~2000Lx, temperature 25~28℃ in the artificial climate box for 7~10 days after germinating to form cotyledon seedlings, standby;

[0030] (2) Preparation of Phytophthora tabaci zoo...

Embodiment 2

[0035] Example 2: Zoospores inoculate cotyledon seedlings to screen for resistance

[0036] In this example, 15 materials are used, and race 0 is used. The specific steps are as follows:

[0037] (1) Preparation of tobacco cotyledon seedlings: After the surface of the tobacco seeds are disinfected, a 10×10 square array is spotted on the germination bed of a petri dish made of sponge and filter paper. The relative humidity is 80-85%, the light intensity is 2000LX, and the temperature is 28℃. After 10 days of incubation in the climate chamber / indoor, they will germinate and form cotyledon seedlings for use;

[0038] (2) Preparation of Phytophthora nicotiana zoospore suspension: Inoculate Phytophthora nicotianae on an OA medium plate, culture at a constant temperature of 28℃ for 10 days in the dark, and cut the hyphae into 2mm×2mm after the hyphae grow over the entire plate Add the small pieces of 10% V8 juice to the liquid culture medium in the shallow dish of the 10% V8 juice culture...

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Abstract

The invention discloses a method for phytophthora nicotianae inoculation of tobacco. The method for the phytophthora nicotianae inoculation of tobacco includes steps that (1) preparing for tobacco seed plantlets; (2) preparing phytophthora nicotianae zoospore suspension; (3) irrigating the roots to inoculate; (4) counting the morbidity situation, and evaluating the disease resistant result. The zoospore inoculated to the roots of the seed plantlets can be quantified rigidly, the resistant difference between the same variety due to excessive inoculation amount is avoided, and the resistance identification is more precise and effective; in the actual operation, the inoculation can be performed in quantities, the space usage is small, the labor cost is saved, and the experiment period is shortened. The method for the phytophthora nicotianae inoculation of tobacco is suitable for popularization.

Description

technical field [0001] The invention belongs to the technical field of plant disease resistance identification, and in particular relates to a method for inoculating tobacco with Phytophthora tabacum. Background technique [0002] Tobacco black shank is caused by Phytophthora nicotianae ( Phytophthora nicotianae ) is a soil-borne disease, which is one of the main diseases affecting the yield and quality of tobacco. The safest and most effective measure to prevent and control this disease is to use disease-resistant varieties. Disease-resistant varieties need to be verified through resistance identification. In the identification of variety resistance, field identification often involves fewer varieties, and the ones suitable for large-scale resistance identification are mainly identification at the seedling stage, and the fungus grain method described in GB / T 23224 and the mycelium block are more widely used. Wound inoculation (Chinese invention patent ZL 200610048658; Hu...

Claims

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Application Information

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IPC IPC(8): A01G7/06
CPCA01G7/06
Inventor 方敦煌陈学军肖炳光童治军杨大海吴兴富
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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