Cauliflower variety early screening kit for resistance to black rot

A technology for cauliflower and black rot, which is applied in the field of early screening kits for cauliflower varieties resistant to black rot, can solve the problem of less black rot disease of cauliflower, and achieve the effect of shortening the time for seed selection and saving land.

Inactive Publication Date: 2009-05-06
NANKAI UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0005] There are few reports at home and abroad on the research on cauliflower black rot, and there is no report on the use of molecular markers to assist cauliflower in breeding for disease resistance

Method used

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  • Cauliflower variety early screening kit for resistance to black rot
  • Cauliflower variety early screening kit for resistance to black rot

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Embodiment 1

[0015] Utilize known anti-black rot and black rot-sensitivity cauliflower plants to carry out detection method, comprising the following steps: (1) Extracting resistance and sensitivity to black rot cauliflower genome DNA: take 100 mg of young cauliflower leaves, cut it into pieces with scissors Add 100 μL extraction buffer solution (1% SDS, 10 mmol / L Tris, 0.3mol / LEDTA, 1% PVPP, pH 8.0) to a tissue grinder, grind thoroughly to a homogenate, then add 400 μL extraction buffer solution, mix well Afterwards, incubate in a water bath at 90°C for 20 minutes. During this period, shake it upside down from time to time, take it out and place it in an ice bath for 5 minutes, and store it at 4°C for later use. (2) Determination of DNA concentration: Two methods can be used, one is to measure by ultraviolet spectrophotometer, and the other is to measure by using a self-made known standard gel plate. The general concentration is required to be above 200ng / μL. (3) PCR amplification: add to...

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Abstract

The invention discloses a test kit of carrying out early screening on new variety for resisting cauliflower black rot and a testing method by the test kit. The test kit consists of dNTP, Taq polymerase, PCR buffer solution, DNA template, sterile deionized water, 6Xloading buffer solution and standard positive Marker. The testing method comprises the following steps: (1) one-step method extraction of cauliflower genetic group DNA; (2) electrophoretic test to sepharose gel of extraction result of the genetic group DNA; (3) PCR amplification; (4) electrophoretic test to PCR amplified product gel; and (5) result identification, namely determining situation of unknown cauliflower resisting and affecting black rot according to displace position of the standard positive Marker on gel and displace position of the PCR product. The matching rate of the result of the new variety or strain of the cauliflower resisting black rot screened and identified by the test kit with field disease resistance reaches over 96 percent. The test kit has the advantages of simple, quick and accurate test without being affected by growing season, environmental conditions and other factors, and can complete screening and identification in seedling stage, thereby saving great soil, manpower and physical resources, and greatly shortening time of seed selection. The invention has important application value in resisting cauliflower black rot breeding.

Description

technical field [0001] The invention relates to a molecular marker-assisted early screening kit for new varieties of black rot-resistant cauliflower in biotechnology and a detection method using the kit. Background technique [0002] Cauliflower black rot is caused by the invasion of Xanthomonas campestris pv. campestris (Pam.) Dowson, which is an important bacterial disease worldwide. Cauliflower black rot mainly affects the leaves or curds. When the seedlings are injured, the cotyledons are water-soaked and gradually turn brown and die; when the true leaves are infected, the germs invade through the water holes and cause the disease on the leaf edge, showing V-shaped lesions. The onset occurs at the adult plant stage, and the pathogen invades from the water holes and wounds on the leaf edge. In addition to forming a V-shaped dead spot on the leaf edge, the pathogen expands downward along the leaf veins, forming a large necrotic area or irregular yellow-brown spots. Leaf ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 宋文芹王春国郝擘陈成彬古瑜孙德岭
Owner NANKAI UNIV
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