The invention provides RPA (
recombinase polymerase amplification) detection primers for
colletotrichum gloeosporioides of green vegetables and application of the RPA detection primers and belongs to the technical field of
agriculture. In order to solve the problems of long time consumption (2-3 hours), low sensitivity (5 ng / mu L) and dependence on precise equipment in the existing PCR detection, the invention provides RPA primers 56F1 (5 '-GAAAAGCGGCAT-3') and 56R1 (5 '-CGTTGGAAAATACA-3') of targeted
pathogenic bacteria specific genes NC030956, as well as an RPA primer 62F1 / F1 and an RPA primer 62F2 / F2, and establishes a 39 DEG
C constant-temperature 20-minute amplification
system. The method realizes three major breakthroughs: 1, the detection sensitivity reaches 0.05 ng / mu L and is improved by 100 times compared with PCR (
Polymerase Chain Reaction); 2, seven related pathogens such as
colletotrichum nicotianae are strictly distinguished, zero cross reaction is realized, and the sample accuracy rate is 100%; 3, the whole process only needs 1.5 hours, the
DNA crude extraction time of diseased leaves is 30 minutes, the RPA amplification time is 20 minutes, the
electrophoresis detection time is 35 minutes, the cost is low, and a
thermal cycler is not needed. The invention provides an on-site solution with the characteristics of rapidness, accuracy and saving for prevention and control of the cruciferous anthracnose, and solves the problem that the traditional technology cannot give consideration to both speed and sensitivity.