Development of a rapid method for the diagnosis of citrus greening disease using the polymerase chain reaction

The fastidious bacterium causing citrus greening disease occurs in uneven and low concentrations in the sieve tubes of host plants. A rapid and sensitive assay based on the polymerase chain reaction (PCR) has been developed using the primers derived from the sequences of the cloned DNA fragment of greening fastidious bacterium (GFB) to detect GFB infection in citrus. One set of the primer pairs (named 226-primer pair), which generates a 226 bp GFB-specific fragment from total DNA templates purified from diseased citrus plants, was tested and chosen for PCR amplification. The PCR-based assay using this 226-primer pair effectively detected GFB infection in various citrus cultivars collected from different Asian countries. This detection technique, which can be completed within 6 h, offers a rapid and efficient method for accurate diagnosis of citrus greening disease.