Application of scanning electron microscopy for diagnosing phytoplasmas in single and mixed (virus-phytoplasma ) infection in Papaya

Phytoplasma and some viruses, papaya ring spot (PRSV) and papaya mosaic (PapMV) have been reported in papaya, from different Mexican states. Some symptoms of yellow type diseases, such as mosaics, stunting, bunchy top and leaf chlorosis, necrosis and malformations are somewhat similar in appearance, but caused by distinct pathogens. Using a scanning electron microscopy (SEM) technique phytoplasmas were detected in the phloem tissues of field and greenhouse-indexed papaya plants from Baja California Sur (BCS). Samples from 32 local varieties, as well as cv. Maradol, showing numerous symptoms of dieback, mosaics, bunchy top, and yellow crinkle were analyzed. The pathogen was detected in stems, leafstalks, roots, axillary leaflets, leaf veins and flowers. Phytoplasma was also detected in dry and in germinated seeds within the fruit, suggesting seed transmission of the pathogen. Some ultrastructural peculiarities of phytoplasma in infected tissues were also observed. No viral infection with PRSV and PapMV was revealed neither in test-plants nor by molecular techniques. Application of SEM technique for analysis of papaya samples from Veracruz and Irapuato, both from field-grown and mechanically inoculated plants with PRSV and PapMV in various combinations also revealed phytoplasmas in the phloem of most of tested samples. In some cases, along with phytoplasmas, rod-shaped bacteria were distinguished.Keywords: Papaya, phytoplasma, papaya ringspot virus, papaya mosaic virus, scanning electron microscopy, Mexic

Phytoplasma transmission by seed

The transmission of phytoplasmas by seed was reported in several herbaceous and some woody plant species. The seedlings were usually analyzed for the presence of phytoplasmas in different stages of growth by nested-PCR/RFLP analysis and phytoplasmas belonging to different ribosomal groups according with species and geographical distribution were detected. The phytoplasma isolation from corn seedlings confirms the seed transmission of viable phytoplasma cells. The sudden epidemic events associated with the presence of phytoplasmas molecularly undistinguishable, in very distant geographic areas, on the same herbaceous species, provides further indications of transmission of these prokaryotes also by seeds. In all the analysed species, the phytoplasma detected in seedlings belong to different ribosomal groups, in some cases they are found also in mixed infection. Since phytoplasma presence is not contemplated in propagation material by the any of the worldwide plant protection quarantine protocols nor by seed producers, the movement of seeds from infected plants implies a wide dissemination of the pathogens and therefore of the associated diseases in still uncontaminated areas