Method for compositely detecting tobacco common mosaic virus and potato virus Y in one step and primers thereof

Abstract

The invention relates to a method for compositely detecting tobacco common mosaic virus and potato virus Y in one step, which is characterized by comprising the following steps of: screening and comparing conservative domains in virus genome, and artificially screening and synthesizing two composite RT(Reverse Transcription)-PCR (Polymerase Chain Reaction) specific primers suitable for viral diseases; analyzing to obtain TMV (Tobacco Mosaic Virus) and PVY (potato virus Y) composite tobacco-infected samples, and integrating enzymes and buffer solution required for reverse transcription and PCR; and then establishing the method for compositely detecting the TMV and the PVY in one step by reaction condition optimization. The method has the advantages that 1) the pathogens of the TMV and the PVY are more convenient to detect, test pollution and influencing factors in the reaction are reduced, and the detection time is greatly shortened; 2) the provided TMY and PVY specific primers have high viral pathogen specificity and can effectively and independently or compositely identify the TMV and the PVY, and ensure the accuracy of the detection result; and 3) when in use, the method saves time and labor, needs low requirement on sample materials, and has less limitation on experimental conditions.

Description

technical field [0001] The invention belongs to the field of plant disease detection and prevention, and is a one-step compound detection method for Tobacco mosaic virus (TMV) and tobacco potato Y virus (Potato Y virus, PVY). Synthesis of specific nucleotide primer sequences and effective establishment of the detection method. Background technique [0002] Tobacco virus disease is the main disease in tobacco agricultural production, and the occurrence of damage will cause a great loss of tobacco yield and lead to a serious decline in the quality of tobacco leaves. Viral diseases are common and serious hazards in tobacco. According to statistics, in 2008, the loss of tobacco leaf output value caused by viral diseases in my country accounted for 23.4% of the total loss of pests and diseases. Viral diseases have become the most serious tobacco diseases (Liu Guoshun, Liu Jianli. China Tobacco Production Practical Technical Guide [M]. Beijing: Agricultural Press, 2007). Tobacco ...

AI Technical Summary

Problems solved by technology

Routine biological detection is a basic link in the research of plant viruses, mainly including determination of host range, identification of host response, temperature of virus sap, dilution limit, in vitro survival period, determination of in vitro traits, and determination of transmission routes, etc., but it is time-consuming , Fei Li (Liang Xunsheng, Xie Lianhui. Plant Virology [M]. Beijing: China Agricultural Press, 1994)

The newly developed detection methods include serological methods, immunoelectron microscopy (ISEM), and nucleic acid molecular hybridization technology (probe detection technology), which have shown great effects in the identification and detection of plant viruses, but there are some common problems. It is the high requirements for the preparation of virus sample materials, the limitation of experimental conditions and the high cost, which limits the wide application of these techniques

Images