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Microbiology 146 (2000), 893-902
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Microbiology (2000), 146, 893-902.
© 2000 Society for General Microbiology

Genomics

Chromosome mapping of the sweet potato little leaf phytoplasma reveals genome heterogeneity within the phytoplasmas

Anna C. Padovan1, Giuseppe Firraoa,1, Bernd Schneiderb,1 and Karen S. Gibb1

Northern Territory University, Faculty of Science, Darwin, Northern Territory 0909, Australia1

Author for correspondence: Anna C. Padovan. Tel: +61 8 8946 6792. Fax: +61 8 8946 6847. e-mail: anna.padovan{at}ntu.edu.au

To further understand the genomic diversity and genetic architecture of phytoplasmas, a physical and genetic map of the sweet potato little leaf (SPLL) strain V4 phytoplasma chromosome was determined. PFGE was used to determine the size of the SPLL-V4 genome, which was estimated to be 622 kb. A physical map was prepared by two-dimensional reciprocal digestions using the restriction endonucleases BssHII, SmaI, EagI and I-CeuI. Sixteen cleavage sites were located on the map. Southern hybridizations of digested SPLL-V4 chromosomal DNA were done using random clones and PCR-amplified genes as probes. This confirmed fragment positions and located the two rRNA operons and the linked fus/tuf genes encoding elongation factors G and Tu, respectively, on the physical map. An inversion of one of the rRNA operons was observed from hybridization data. Sequence analysis of one of the random clones identified a gid gene encoding a glucose-inhibited division protein. Digestions of the tomato big bud (TBB) phytoplasma chromosome with the same four enzymes revealed genome heterogeneity when compared to the closely related SPLL-V4, and a preliminary chromosome size for the TBB phytoplasma of 662 kb was estimated. This mapping information has revealed that significant genome diversity exists within the phytoplasmas.

Keywords: phytoplasma, genome mapping, ribosomal genes, pulsed-field gel electrophoresis, gid gene

Abbreviations: 2-D, two-dimensional; PFG, pulsed-field gels; SPLL, sweet potato little leaf; TBB, tomato big bud; WX, Western X

The EMBL accession number for the sequence determined in this work is AJ245515.

a Present address: Dipartimento di Biologia Applicata alla difesa delle Piante, Università di Udine, 33100 Udine, Italy.

b Present address: RWTH Aachen, Institut fuer Biologie I, Worringerweg 1, 52074 Aachen, Germany.




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