Molecular Detection and Diagnosis of Phytoplasma Infection in Cassava (Manihot esculenta Crantz) in Eastern Visayas, Philippines
- DNA extraction,
- PCR detection,
Phytoplasma disease is a threat to the cassava industry but little is known about
the disease in the country. This study was conducted to develop and optimize a protocol
for the detection of phytoplasma and determine the existence of phytoplasma strains in
the different cassava growing areas in Eastern Visayas, Philippines.
PCR reaction using primer pair PA2F/R and nested primer pair NPA2F/R as
well as the use of primer pair P1/P7 followed by nested primer pair R16F2n/R2 with
slight modifications in the thermocycling conditions detected phytoplasma infecting
Ten out of the eleven areas in Eastern Visayas, a scale insect and a cassava
grown from tissue culture tested positive for phytoplasma using nested primer pairs
NPA2F/R and R16F2n/R16R2. Six band patterns were detected when NPA2F/R primer
amplicons were digested with RsaI and MspI restriction enzymes suggesting that more
than one strain of phytoplasma infect cassava.