Method for rapidly and synchronously detecting wheat yellow mosaic virus and Chinese wheat mosaic virus

Abstract

The invention discloses a method for rapidly and synchronously detecting wheat yellow mosaic virus and Chinese wheat mosaic virus. According to the method, three primers comprising a reverse primer CWWY-R2 of 5'-GGTTCCMGTTATCGTACT-3' and two positive primers of CW1-F2 and WY1-F2 of 5'-GGAAGGGATGCCATACAACT-3' and 5'-ACCCTACAAACAAACTCTGC-3' are designed according to similarities and differences of two virus gene sequences, the plant general RNA serves as a template, the CWWY-R2 serve as the primer, and reverse transcription is performed to synthesize c DNA. A reaction system for synchronously detecting two viruses is established through optimization, the system comprises total 20mu L of 1.6mu L of cDNA, respectively 0.4mu L of primers (10mu mol / L), 2mu L of 10*PCR Buffer (not containing Mg<2+>), 0.4mu L of dNTPs (10mmol / L each), 0.8mu L of Taq DNA polymerase (5U / mu L), 0.8mu L of MgCl2 (25mmol) and 13.2 mu L of ddH2O; the polymerase chain reaction (PCR) has the reaction conditions of initial denaturation within 5 minutes at the temperature of 94 DEG C, 50 seconds at the temperature of 94 DEG C, 50 seconds at the temperature of 50 DEG C, and 90 seconds at the temperature of 72 DEG C within 30 cycles totally, and extension within 10 minutes at the temperature of 72 DEG C; and the PCR amplification expected target fragments are respectively WYMV 508bp and CWMV 918bp. According to the method, samples in Gaoyou, Yangzhou and Dafeng in Jiangsu are detected and are subjected to WYMV, WYMV and CWMV single infection.

Description

Technical field: [0001] The invention relates to a method for rapid and synchronous detection of wheat yellow mosaic virus and Chinese wheat mosaic virus and its application in agriculture, belonging to the field of agricultural science and technology. Background technique: [0002] Wheat is one of the most important food crops in the world, and wheat virus disease is one of the main factors affecting its production. So far, wheat-like viruses transmitted by Polymyxa graminis mainly include Chinese wheat mosaicvirus (CWMV) of the genus Furovirus, Chinese wheat mosaicvirus (CWMV), soil-borne mosaic virus of wheat (Soil -bome wheat mosaic virus (SBWMV), soil-bome cereal mosaic virus (SBCMV) and wheat yellow mosaic virus (WYMV) of the genus Bymovirus and wheat Wheat spindle streak mosaic virus (WSSMV). Among them, SBWMV is mainly distributed in the United States, France, Italy and Japan, SBCMV is distributed in France, the United Kingdom and the United States, WSSMV is distri...

AI Technical Summary

Problems solved by technology

[0005] Symptoms of WYMV and CWMV infecting wheat are mainly leaf yellowing or mosaic, dwarfing of plants, further development until withering and death, both are transmitted by Polymyxa graminearum, their biological characteristics are very similar, and it is difficult to be effective from the perspective of empirical observation Differentiation, and CWMV is often co-infected with WYMV, which increases the difficulty of disease diagnosis

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