16S rRNA interoperon sequence heterogeneity distinguishes strain populations of palm lethal yellowing phytoplasma in the Caribbean region

Abstract

DNA of phytoplasmas in lethal yellowing (LY)-diseased palms was detected by a nested polymerase chain reaction (PCR) assay employing rRNA primer pair P1/P7 followed by primer pair LY16Sf/LY 16-23Sr. Polymorphisms revealed by HinfI endonuclease digestion of rDNA products differentiated coconut-infecting phytoplasmas in Jamaica from those detected in palms in Florida, Honduras and Mexico. A three fragment profile was generated for rDNA from phytoplasmas infecting all 21 Jamaican palms whereas a tire fragment profile was evident for phytoplasmas infecting the majority of Florida (20 of 21), Honduran (13 of 14) and Mexican (5 of 5) palms. The RFLP profile indicative of Florida LY phytoplasma was resolved by cloning into two patterns, one of three bands and the other of four bands, that together constituted the five fragment profile. The two patterns were attributed to presence of two sequence heterogeneous rRNA operons, rrnA and rrnB, in most phytoplasmas composing Florida, Honduran and Mexican LY strain populations. Unique three and four fragment RFLP profiles indicative of LY phytoplasmas infecting Howea forsteriana and coconut palm in Florida and Honduras, respectively, were also observed. By comparison, the Jamaican LY phytoplasma population uniformly contained one or possibly two identical rRNA operons. No correlation between rRNA interoperon heterogeneity and strain variation in virulence of the LY agent was evident from this study.