Differential amplification of sequence heterogeneous ribosomal RNA genes and classification of the ‘Fragaria multicipita’ phytoplasma

Authors

Robert E. Davis, USDA-ARS
Rasa Jomantiene, USDA-ARS
Audrone Kalvelyte, Institute of Biochemistry
Ellen L. Dally, USDA-ARS

Date of this Version

2003

Comments

Published in Microbiol. Res. (2003) 158, 229–236.

Abstract

Ribosomal (r) RNA interoperon sequence heterogeneity in the ‘Fragaria multicipita’ phytoplasma, a member of group 16SrVI, was initially observed in RFLP patterns of rDNA amplified in the polymerase chain reaction (PCR), and was confirmed through sequence analysis of cloned rDNA. Sequences from operons rrnA and rrnB were amplified in PCR primed by primer pair P1/P7 but from only rrnA in PCR primed by primer pair R16mF2/R16mR1. Preferential amplification of DNAfrom operon rrnA was explained by base mismatches between the R16mF2/R16mR1 primers and primer annealing sites in rrnB. The results revealed potential for classification of a phytoplasma into two different subgroups within a 16S rRNA group, if the phytoplasma’s 16S rRNA gene sequences are independently characterized. It is suggested that the rRNA operon containing species-specific signature sequence(s) should be specified, and where possible sequences from both 16S rRNA genes should be included, in descriptions of new ‘Candidatus Phytoplasma species’.