Abstract
A nested polymerase chain reaction (PCR) assay was used to amplify 16S-23S intergenic spacer (IGS) region from DNA samples individually extracted from 25 Sabal palmetto (cabbage palms) showing symptoms of Texas Phoenix decline (TPD) in West Central Florida. The IGS region was also amplified from DNA from other palm species showing symptoms of TPD and lethal yellowing (LY). A subset of the aforementioned phytoplasma DNA samples (Sabal and other palm species) together with additional samples from various hosts collected from different geographical localities were further studied to compare the collected phytoplasma strains using sequence analysis of the glycoprotease (gcp) genes. Restriction fragment length polymorphisms (RFLP) analysis of the PCR-amplified 16S-23S IGS region and thegcp gene using a three restriction enzymes showed that the population of the phytoplasmas infecting S. palmetto in West Central Florida is probably homogenous. The S. palmetto phytoplasma also appeared similar to all the 16SrIV-D phytoplasmas infecting other palm species and different from all phytoplasmas belonging to the 16SrIV-A subgroup. We recommend more work using genes or genomic regions other than the 16S-23S IGS region and the gcp gene to be done. Key words: 16S-23S intergenic spacer region, glycoprotease gene, phytoplasma, Texas Phoenix decline, lethal yellowing.
Highlights
Until 2005, the only phytoplasma disease of palms known to occur in Florida was Lethal Yellowing (LY) caused by phytoplasma subgroup 16SrIV-A
Similarity analysis of the assembled nucleotide sequences derived from these polymerase chain reaction (PCR) fragments demonstrated the sequences were of phytoplasma origin, giving assurance that the PCR fragments were amplified from phytoplasmas
Based on analysis of Restriction fragment length polymorphisms (RFLP) profiles generated by digestion of each amplified PCR fragment with AseI restriction enzyme, 16SrIV-D strains from cabbage palms all had the same profiles, except for SP7 which had an additional band (Figure 2A to C)
Summary
Until 2005, the only phytoplasma disease of palms known to occur in Florida was Lethal Yellowing (LY) caused by phytoplasma subgroup 16SrIV-A. (coconut palm) is the primary palm susceptible to this subgroup, the disease has been observed in 35 other palm species (Harrison and Jones, 2004). During the 40 plus years that LY has been active in Florida, no palm species native to Florida has been affected by this disease. Even with the detection of a decline phytoplasma disease in Florida in 2005, native palm species remained unaffected (Harrison et al, 2008). It was a surprise in 2008 to determine that the lethal decline of Sabal palmetto (Walter) Lodd.
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