Electron Microscopy and Molecular Characterization of Phytoplasmas Associated with Little Leaf Disease of Brinjal (Solanum melongena L.) and Periwinkle (Catharanthus roseus) in Bangladesh

In Bangladesh little leaf disease was observed in brinjal (Solanum melongena L.) and in periwinkle (Catharan- 2thus roseus). Phloem-inhabiting phytoplasmas were consistently detected in both species of diseased plants using transmission electron microscopy (TEM) and polymerase chain reaction (PCR) techniques. The shape, size and within-tissue distribution of phytoplas- mas appears to be similar in both hosts. Furthermore, the molecular characterization and identi®cations of observed phytoplasmas were carried out based on restriction fragment length polymorphism (RFLP) patterns of PCR-ampli®ed products (1200 bp) using phytoplasma-speci®c universal primers and sequencing analysis of both 16S ribosomal DNA (rDNA) and intergenic spacer region (ISR) of 16S-23S rDNA phytoplasma genes. The patterns of RFLP analysis with seven restriction enzymes exhibited a similar pattern for both phytoplasma strains. The sequence homology between these two strains showed 100% similarity based on 16S rDNA and 16S-23S ISR. Therefore, in Bangladesh the causal agents of brinjal little leaf (BLL-Bd) and periwinkle little leaf (PLL-Bd) are probably the same or closely related phytoplasma strains. These strains, are very close or identical to the strain of brinjal little leaf phytoplasma in India (BLL- In), belonging to the clover proliferation group (Lee et al., Int. J. Syst. Bacteriol. 48, 1153±1169, 1998; Seemuller et al., J. Plant Pathol. 80, 3±26, 1998).