Screening Method of Virus-free Potato Plantlets Against Early Blight

Abstract

The invention relates to the technical field of screening methods for anti-early blight materials, and is a screening method for anti-early blight of virus-free potato test-tube vaccines; The third step is the preparation of sporangia suspension; the fourth step is inoculation. Compared with the traditional screening method, the screening method for anti-early blight of the virus-free potato test-tube vaccine of the invention has less error, shorter screening time and lower screening cost; at the same time, the screening method for the anti-early blight of the virus-free potato test-tube vaccine of the present invention makes up for the early blight resistance of potato in Xinjiang region. Due to the gaps in research and prevention, a set of screening methods for potato anti-early blight test-tube vaccines in Xinjiang has been established, which has greatly improved the timeliness, effectiveness and accuracy of early blight identification and is of great significance in the rapid identification and prevention of blight. .

Description

technical field

[0001] The invention relates to the technical field of screening methods for anti-early blight materials, and relates to a screening method for virus-free potato tube seedlings against early blight. Background technique

[0002] Potato is an annual herbaceous tuber plant of Solanaceae, and it is an important grain and vegetable crop. Potato early blight caused by Alternaria solani infection is a serious disease in potato production. It occurs in all potato production areas and tends to increase year by year. Especially in developing countries, it is considered to be the second largest potato disease after potato late blight. In terms of production, there are many evaluations on the resistance to potato late blight and viral diseases, but less research on early blight. Moreover, the existing early blight evaluation methods have large deviations, long detection time and high cost, which are not suitable for the evaluation of virus-free potato tube seedlings a...

Examples

Embodiment 1

[0015] Embodiment 1, the screening method of this detoxified potato test-tube plantlet anti-early blight, is carried out according to the following steps: the first step, the isolation of pathogenic bacteria, collects the potato early blight diseased leaf in the field of different regions, gathers the potato early blight diseased leaf After rinsing with tap water for 1 to 3 times, soak in ethanol aqueous solution for 20s to 30s after soaking, then soak in mercuric chloride aqueous solution for 10s to 20s, rinse with sterile water for 3 to 5 times, and then use sterilized Take out the leaves at the junction of disease and health with a bacterial scalpel, place the back of the leaves at the junction of disease and health on the PDA plate medium, put 3 to 4 leaves in each dish, and place them in an incubator at 25°C for 3 days to 3 days in the dark. 5d, after culturing, isolate the pathogenic bacteria on the leaves to obtain the pathogenic bacteria;

[0016] The second step is th...

Embodiment 2

[0021] Example 2, as an optimization of the above example, the PDA medium is obtained as follows: Weigh 200g of fresh potatoes, wash and peel the slices, add 1000mL of deionized water and boil for 15min, filter with gauze and add deionized water to make up to 1000mL , then add 12g glucose and 10g to 12g agar powder, boil to obtain PDA medium after the agar powder is completely melted. Boil until the agar powder is completely melted, then subpackage; the subpackaged PDA medium should be sterilized within 1 hour, using high-pressure steam for moist heat sterilization, under 0.1MPa steam pressure, the temperature reaches 121°C, and maintains for 20 minutes.

[0022] Packing method: A. Divided into Erlenmeyer flasks (conical flasks); use 250 mL Erlenmeyer flasks for dispensing, each bottle is divided into about 200 mL of culture medium, and the mouth of the bottle is covered with a ventilated and sterilized sealing film, which is tied tightly with cotton thread.

[0023] B. Test...

Embodiment 3

[0026] Embodiment 3, as the optimization of above-mentioned embodiment, the blade size of sick and healthy junction is 0.5cm 2 to 1cm 2 .

technical field

[0001] The invention relates to the technical field of screening methods for anti-early blight materials, and relates to a screening method for virus-free potato tube seedlings against early blight. Background technique

[0002] Potato is an annual herbaceous tuber plant of Solanaceae, and it is an important grain and vegetable crop. Potato early blight caused by Alternaria solani infection is a serious disease in potato production. It occurs in all potato production areas and tends to increase year by year. Especially in developing countries, it is considered to be the second largest potato disease after potato late blight. In terms of production, there are many evaluations on the resistance to potato late blight and viral diseases, but less research on early blight. Moreover, the existing early blight evaluation methods have large deviations, long detection time and high cost, which are not suitable for the evaluation of virus-free potato tube seedlings a...

Examples

Embodiment 1

[0015] Embodiment 1, the screening method of this detoxified potato test-tube plantlet anti-early blight, is carried out according to the following steps: the first step, the isolation of pathogenic bacteria, collects the potato early blight diseased leaf in the field of different regions, gathers the potato early blight diseased leaf After rinsing with tap water for 1 to 3 times, soak in ethanol aqueous solution for 20s to 30s after soaking, then soak in mercuric chloride aqueous solution for 10s to 20s, rinse with sterile water for 3 to 5 times, and then use sterilized Take out the leaves at the junction of disease and health with a bacterial scalpel, place the back of the leaves at the junction of disease and health on the PDA plate medium, put 3 to 4 leaves in each dish, and place them in an incubator at 25°C for 3 days to 3 days in the dark. 5d, after culturing, isolate the pathogenic bacteria on the leaves to obtain the pathogenic bacteria;

[0016] The second step is th...

Embodiment 2

[0021] Example 2, as an optimization of the above example, the PDA medium is obtained as follows: Weigh 200g of fresh potatoes, wash and peel the slices, add 1000mL of deionized water and boil for 15min, filter with gauze and add deionized water to make up to 1000mL , then add 12g glucose and 10g to 12g agar powder, boil to obtain PDA medium after the agar powder is completely melted. Boil until the agar powder is completely melted, then subpackage; the subpackaged PDA medium should be sterilized within 1 hour, using high-pressure steam for moist heat sterilization, under 0.1MPa steam pressure, the temperature reaches 121°C, and maintains for 20 minutes.

[0022] Packing method: A. Divided into Erlenmeyer flasks (conical flasks); use 250 mL Erlenmeyer flasks for dispensing, each bottle is divided into about 200 mL of culture medium, and the mouth of the bottle is covered with a ventilated and sterilized sealing film, which is tied tightly with cotton thread.

[0023] B. Test...

Embodiment 3

[0026] Embodiment 3, as the optimization of above-mentioned embodiment, the blade size of sick and healthy junction is 0.5cm 2 to 1cm 2 .