COMPARISON OF DIFFERENT DETECTION METHODS OF Rosellinia necatrix IN SOIL

To achieve a simple and sensitive Rosellinia necatrix detection method in soil, three detection methods including soil direct culture method, infected soil in culture media e. g. MEAR, MEAR + penconazole and MEAS, Nested PCR method and bait twig method were compared. Direct DNA Extraction from soil was done. Two specific primer pairs R2، R8 and R7، R10 in Nested PCR were used and the extracted DNA was purified using PVPP column. Among used methods, the direct culture method from soil was not suitable and wasn’t sensitive for detection. However, bait twig method was capable to detect, the pathogen in soil and root but it is time consuming and laborious method. In conventional PCR, using primer pairs R2-R8, 10 pg μl−1 of fungal DNA was detected; however in Nested PCR using R7-R10 primer pairs, 1fg μl−1 of fungal DNA was detected. Among used methods, the Nested PCR was better in detection of the fungus. Nested PCR method of detection in soil and root is more sensitive and reliable.