PCR-Based Sequence Analysis on Multiple Genes Other than 16S rRNA Gene for Differentiation of Phytoplasmas.
Differentiation and classification of phytoplasmas have been primarily based on the highly conserved 16S rRNA gene, for which "universal" primers are available. To date, 36 ribosomal (16Sr) groups and more than 150 subgroups have been delineated by RFLP analysis of 16S rRNA gene sequences. However, in recent years, the use of moderately conserved genes as additional genetic markers has enhanced the resolving power in delineating distinct phytoplasma strains among members of some 16Sr subgroups.This chapter describes the methodology of amplification, differentiation, and classification of phytoplasma based on less-conserved non-ribosomal genes, named rp and secY. Actual and virtual RFLP analyses of amplicons obtained by semi-universal or group-specific rp and secY gene-based primers are used for finer differentiation of phytoplasma strains within a given group. The rp and secY gene-based classification not only readily resolves 16Sr subgroups within a given 16Sr group, but also provides finer differentiation of closely related phytoplasma strains within a given 16Sr subgroup.
['Martini|Marta|M|', 'Bottner-Parker|Kristi D|KD|', 'Lee|Ing-Ming|IM|']
D001426:Bacterial Proteins D015373:Bacterial Typing Techniques D001483:Base Sequence D017124:Conserved Sequence D058885:Multilocus Sequence Typing D010802:Phylogeny D045582:Phytoplasma D010944:Plants D016133:Polymerase Chain Reaction D012336:RNA, Ribosomal, 16S
Virtual RFLP secY 16Sr group-specific primers Phytoplasma strain differentiation Restriction enzymes Semi-universal primers rplV (rpl22) rpsC (rps3)