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Molecular Determination and Characterization of Phytoplasma 16S rRNA Gene in Selected Wild Grasses from Western Kenya

Adam OJ, Midega CAO, Runo S and Khan ZR

Napier grass (Pennisetum purpuruem) production for zero grazing systems has been reduced to rates of up to 90% in many smallholder fields by the Napier stunt (Ns) disease caused by phytoplasma sub-group 16SrXI in western Kenya. It is hypothesized that several other wild grasses in Kenya could be infected by phytoplasmas that would otherwise pose a significant threat to Napier, other important feeds and food crops. This study therefore sought to detect and identify phytoplasma strains infecting wild grasses in western Kenya using 16S ribosomal RNA (ribonucleic acid) gene as well as identify wild grass species hosting phytoplasmas in 646 wild grass samples that were collected in October 2011 and January 2012 during a random crossectional survey conducted in Bungoma and Busia counties of western Kenya. DNA was extracted and nested polymerase reaction (nPCR) used to detect phytoplasmas. Two sub-groups of phytoplasmas were detected in eight grass species observed to grow near infected Napier fields. Only one of the two phytoplasmas reported was related to the Ns phytoplasma. There was a strong association between proportions of phytoplasma infection and the grass species collected (p = 0.001). C. dactylon, D. scalarum, B. brizantha, poverty grass and P. maximum had high proportions of infection and were abundantly distributed in western Kenya hence considered wild phytoplasma hosts. E. indica and C. ciliaris were scarcely distributed and had low infection rates. There was statistically significant difference in proportions of infection per location of survey (p = 0.001). Phytoplasma subgroups 16SrXI and 16SrXIV were the only phytoplasma genotypes distributed among wild grasses in western Kenya. Phytoplasma subgroup 16SrXIV predominantly infects only C. dactylon and B. brizantha grasses while phytoplasma subgroup 16SrXI is broad spectrum and infects a large number of wild grasses. In general, there is a diversity of wild grasses hosting phytoplasmas in western Kenya. These host grasses may be the reason for the high rates observed in the spread of Ns disease in western Kenya by acting as reservoirs for Ns phytoplasma.