Real-time fluorescence quantification PCR (polymerase chain reaction) method for detecting brown rust of sugarcane

Abstract

The invention relates to a real-time fluorescence quantification PCR (polymerase chain reaction) method for detecting brown rust of sugarcane. The method comprises the steps of extraction of DNA (deoxyribonucleic acid) of the genome, establishment of a real-time fluorescence quantification PCR detection system, effectiveness judgment of experiments, and real-time fluorescence quantification PCR detection. The method has the characteristics that the sensitivity is high, the specificity is good, and the use amount of DNA is little. Compared with the technologies of conventional identification of morphological characteristics of a spore cultivated according to germs, identification of phenotype symptom of disease according to filed planting, and conventional PCR detection, the method has the advantages that the sensitivity is high, the specificity is good, fewer samples are consumed, and the PCR amplification process can be monitored intuitively at real time, and the like. A fast detection system with high sensitivity and high specificity is provided for the identification of brown rust of the sugarcane, and the method can be used for early diagnosis and detection of brown rust of the sugarcane in the field, and has a significance for the breeding of anti brown rust sugarcane and protection of genetic resources of the anti brown rust sugarcane.

Description

technical field [0001] The invention relates to a method for detecting sugarcane pathogenic bacteria, in particular to a real-time fluorescent quantitative PCR method for detecting sugarcane brown rust, and belongs to the field of biotechnology. Background technique [0002] Sugarcane brown rust disease (Sugarcane brown rust disease) is caused by the sugarcane brown rust fungus ( Puccinia melanocephala ) caused by a fungal disease. The disease is mainly transmitted by uredia spores, which attach to the surface of sugarcane leaves by wind force and invade through stomata, thereby harming sugarcane leaves, resulting in loss of cane stem yield and reduction of sucrose content. The primary source of infection is mainly from sugarcane itself or other intermediate hosts. Planting sugarcane varieties resistant to brown rust is the most economical and effective measure to control sugarcane brown rust. [0003] The detection of sugarcane brown rust and its efficiency and accuracy ...

AI Technical Summary

Problems solved by technology

Among the above three methods, the separation and cultivation technology takes a long time, is easily contaminated by miscellaneous bacteria, and the detection efficiency is not high; the field planting identification method is affected by the interaction of sugarcane, pathogenic bacteria and environmental conditions, which not only takes a long time, but also has insufficient stability in the identification results , the problem of large fluctuations; the sensitivity of conventional PCR detection technology is not high enough, and it is impossible to monitor the PCR process in real time

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