Real-time fluorescence PCR detection probe and kit for soyabean phytophthora and the detection method thereof

Abstract

The present invention provides primers, probes and reagent kit for real-time fluorescent detection of soybean phytophthora, as one quick detection method specially for soybean phytophthora. It has detection time of only one day, high sensitivity, high specificity and high stability and reliability. The method is superior to available ELISA method and conventional PCR detection method. The method is suitable for detecting and / or identifying single oospore, single swarm sporangium and trace amount of pathogen spore in soil, soybean in both field and port.

Description

technical field [0001] The present invention relates to a kind of biotechnology, specifically, relate to a kind of simple, fast, micromolecular detection probe and test kit to soybean blight (Phytophthora sojae), be suitable for departments such as port inspection and quarantine, agricultural production, plant protection use. Background technique [0002] Soybean is the main agricultural product in my country. Soybean blight is a devastating disease on soybean. This kind of pathogen is a kind of quarantine pest that is prohibited from entering my country's imported plants. [0003] Soybean blight was first reported in the United States. In 1957, soybean blight alone caused a loss of 1.5 million U.S. dollars in Ohio. In 1978, soybean blight broke out for the second time in the United States, and the national disease area reached 8 million hectares. So far, soybean blight Has been widely distributed in the United States. At present, the disease has been widely distributed in...

AI Technical Summary

Problems solved by technology

[0005] For the detection or identification of soybean blight, there are currently two main methods at home and abroad, namely: (1) soil trapping method. The soil trapping method relies on the parasitic specialization of Phytophthora sojae. Trapping pathogenic bacteria in the soil, and then separating, cultivating and determining the pathogenicity of the trapped pathogenic bacteria; (2) ELISA method, ELISA method relies on the reaction of antigen and antibody, its disadvantage is: low sensitivity

Although this method can obtain reliable detection results, it has the following disadvantages: 1) it takes a long time, and it generally takes 15 to 20 days to complete the entire detection process from soil cultivation to isolation cultivation, and then to pathogenicity determination. test result

2) For the quality supervision, inspection and quarantine department, since there are very few soil samples obtained at entry and exit, fewer oospores can be detected in the soil, and it is difficult and time-consuming to trap soybean blight using susceptible soybean leaves. Meet the needs of animal and plant quarantine inspection departments

3) In the agricultural production sector, the detection of soybean blight in the past often used methods of isolating and cultivating diseased plants or trapping pathogenic bacteria from the soil. The former used diseased tissues to isolate soybean blight, which was difficult and took a long time. The difficulties encountered by the method of soil trapping are the same as those encountered by the quality supervision, inspection and quarantine department

Images