18 Molecular detection of Candidatus Phytoplasma trifolii.cdr

Article Information Brinjal is an important vegetable crop grown worldwide. A field survey conducted during 2012 in Kerala state of South India, the little leaf disease affected brinjal plant was observed in one of the home gardens located at Nemmara (Palakkad district). The diseased and healthy samples were collected; genomic DNA was isolated and 16SrRNA gene amplified by performing nested PCR using universal primers pairs P1/P7 and R16F2n/R2 respectively. The association of Phytoplasma was detected in all the three symptomatic brinjal samples and in Phytoplasma affected Periwinkle used as positive control. An expected amplicon size of 1250bp from nested PCR was purified and sequenced. The nucleotide sequence analysis (nBLAST) of 1250bp fragment of sequence KL-50(KP027530) revealed 97% similarity with Candidatus phytoplasma trifolii (JX104336). Phylogenetic analysis and study of virtual RFLP gel pattern of the sequence also revealed the association of Candidatus phytoplasma trifolii.