Detection of phytoplasmas in plant internal vascular fluids: a cell-lysis-free method

In the present study, techniques for detecting phytoplasma, a destructive plant pathogen were optimized by analyzing internal vascular fluids rather than traditional tissue samples. The study compares different sap extraction techniques like root pressure and vacuum pump extraction methods, determining that the vacuum approach using only the stem yields the most efficient sap recovery. The sap fluid contains a mixture of xylem and phloem; it provides a reliable medium for detection without the need for complex cell lysis. Testing revealed that nested PCR combined with the CTAB DNA extraction method was essential for successful pathogen detection. These findings offer a faster and more accurate alternative for monitoring crop health.