Detection of phytoplasma infecting greengram by nested PCR

Abstract

Green gram variety LGG-460 was used to detect the phytoplasma by PCR at different stages of crop growth. Phytoplasma infected three green gram plants were collected for DNA isolation at 30, 45, 60 and 75 DAS from the same field and performed PCR with R16F2n/R16R2 primers. Samples collected at 30 and 45 DAS gave good amplification of expected product size in PCR with R16F2n/R16R2 primers, but weak bands were observed in samples collected from 60 and 75 DAS, thus indicating that the optimum age for the detection of phytoplasma in green gram is 30-45 DAS. The present result shows that the PCR techniques described here allows rapid, sensitive and accurate detection of phytoplasma in plants that are showing typical phytoplasma disease symptoms.