Detection kit and detection method for cabbage oxysporum

Abstract

The invention discloses a detection kit for cabbage oxysporum. A reagent of the kit comprises a detection primer; the detection primer comprises 1 mul of upstream primer Foc-R and 1 mul of downstream primer Foc-S both of which the concentration is 10 pmol / mul, 2.0 mul of 10*PCR (Polymerase Chain Reaction) Easy Taq buffer solution, 0.5mul of 10mM dNTPs (diethyl-nitrophenyl thiophosphates), 0.4 mul of Taq polymerase of which the active enzyme concentration is 5U / ml, 1mul of cabbage oxysporum positive control DNA (deoxyribonucleic acid), and 20 mul of ultrapure water of which the weight concentration is not smaller than 99%; the sequence of Foc-R is 5'-TCAATGATAGTGACAAGGGTTT-3', and the sequence of the Foc-S is 5'-AATTTGCTGTGATAGGTGGAT-3'. The invention further discloses a detection method which comprises the following steps of: (1) extracting the DNA of a plant tissue or soil; (2) carrying out PCR amplification on the DNA; and (3) carrying out electrophoretic separation on the amplified product, and after separation, judging the results according to the size of the amplified product under a UV (ultraviolet) lamp after the product is colored by ethidium bromide. The kit and the method for quick molecular detection of cabbage oxysporum are high in accuracy, simple and convenient to operate, strong in specificity and high sensitivity.

Description

technical field [0001] The invention belongs to the fields of detection and identification of crop diseases and plant quarantine, and in particular relates to a detection kit and a detection method for cabbage wilt. Background technique [0002] Now known cabbage wilt is exactly Fusarium oxysporum sticky group specialization type, and the cabbage wilt disease caused by Fusarium oxysporum f.sp.Conglutinans is a devastating soil-borne disease. It occurs in most of the cabbage producing areas in the world. Since the first report of the disease in my country in 2001, it has occurred in Zhangjiakou City, Hebei Province, Jinzhong City, Shanxi Province, and Datong City, Shanxi Province. The occurrence in my country is spreading. It has become an important disease affecting the quality and yield of cabbage in my country. Cabbage wilt is a soil-borne disease. The pathogenic bacteria can overwinter in the soil and diseased residues to form an infection cycle, and its infectivity is str...

AI Technical Summary

Problems solved by technology

[0005] In order to solve the technical defects of the above-mentioned detection of Fusarium wilt of cabbage, the present invention discloses a molecular detection reagent for Fusarium wilt of cabbage and its detection kit, the purpose of which is to meet the needs of the biological detection method of Fusarium wilt of cabbage in the prior art. The period is long, and there is no molecular detection method for Fusarium wilt of cabbage at present. Through the whole genome sequencing of Fusarium wilt of cabbage, and the method of comparative genomics to find out the genome fragments of Fusarium wilt of cabbage specific to other fungus, and then this is aimed at A pair of highly specific primers were designed for the specific fragment base sequence of Fusarium wilt of Brassica oleracea, which is used for high-sensitivity and rapid molecular detection of plant tissues and soils with Fusarium wilt of cabbage, and provides a high-accuracy, easy-to-operate, and specific A kit for rapid molecular detection of Fusarium wilt of cabbage with high sensitivity, which can be used for high-sensitivity rapid molecular detection of plant tissues and soils with bacteria, and is very important for early detection and diagnosis before Fusarium wilt of cabbage causes disease symptoms significance

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