Primer, probe and real-time fluorescent PCR reagent case for detecting tobacco black shank bacterium

Abstract

The invention discloses a primer, probe and real-time fluorescence PCR agent box to check black shank bacterium of tobacco, which is characterized by the following: making the primer with forward primer Pn1 and backward primer Pn2; setting the base sequence as Pn1:5'- GAA CAA TGC AAC TTA TTG GAC GTT-3', Pn2:5'-AAC CGA AGC TGC CAC CCT AC -3'; setting the base sequence of probe as a random nucleic acid sequence in the zone of upper river shift two bases or lower river shift three bases on two ends of 5'-TTC ACC AGT CCA TCA CGC CAC AGC-3'; signing fluorescence report group on 5' end; signing fluorescence quenched group on 3' end; forming fluorescence labeling probe; preparing agent box with the primer and probe. This invention can quickly test black shank bacterium of tobacco in soil, which possesses important meaning.

Description

(1) Technical field [0001] The present invention relates to a bioengineering technology suitable for agricultural production, plant protection and other departments, in particular to a bioengineering technology for detecting tobacco black shank disease pathogenic fungus Phytophthora nicotianae, which is a major destructive disease in tobacco production Primers, probes and real-time fluorescent PCR kits. (2) Technical background [0002] Tobacco black shank is a major and devastating disease that affects the development of the global tobacco industry, and it is common in various parts of my country. The disease can harm all cultivated tobaccos including flue-cured tobacco, air-cured tobacco, sun-cured tobacco, burley tobacco and oriental tobacco, and can infect and harm tobacco at any growth stage, but it mainly harms tobacco plants in the field stage, and the onset of seedlings often causes tobacco. The seedlings die in pieces and become damped-off, and the onset in the fie...

AI Technical Summary

Problems solved by technology

At present, in the real-time fluorescent PCR detection of tobacco black shank, 5'-GAA CAA TGCAAC TTA TTG GAC GTT-3' and 5'-AAC CGA AGC TGC CAC CCT AC-3' are used as primers, and 5'- TTC ACC AGT CCA TCA CGC CAC AGC- Any nucleotide sequence in the region shifted 2 bases upstream or 3 bases downstream at both ends of the 3' is prepared as a probe into a corresponding real-time fluorescent PCR kit , there is no related report