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Close relatedness of Acanthamoeba pustulosa with Acanthamoeba palestinensis based on isoenzyme profiles and rDNA PCR-RFLP patterns
Young Ho Kim,1Mee Sun Ock,2Ho Cheol Yun,1Mee Yul Hwang,1Hak Sun Yu,1Hyun Hee Kong,1 and Dong Il Chung*1
1Department of Parasitology, Kyungpook National University School of Medicine, Taegu 700-422, Korea.
2Department of Parasitology, College of Medicine, Kosin University, Pusan 602-030, Korea.
Received September 06, 1996; Accepted November 29, 1996.
Abstract
The taxonomic validity of morphological group III Acanthamoeba spp. is uncertain. In the present study, six type strains of group III Acanthamoeba spp., Acanthamoeba culbertsoni, A. healyi, A. pustulosa, A. palestinensis, A. royreba and A. lenticulata were subjected for the evaluation of their taxonomic validity by comparison of the isoenzyme patterns by isoelectic focusing on polyacrylamide gels, mitochondrial DNA (Mt DNA) restriction fragment length polymorphism (RFLP), and small subunit ribosomal DNA (ssu rDNA) PCR-RFLP patterns. The Mt DNA RFLP patterns were heterogeneous between species. The type strains of A. palestinensis and A. pustulosa showed almost identical patterns of isoenzymes and rDNA PCR-RFLP with an estimated sequence divergence of 2.6%. The other species showed heterogeneous patterns of isoenzymes and rDNA PCR-RFLP. It is likely that A. pustulosa is closely related with A. palestinensis and that the former may be regarded as a junior synonym of the latter.
Figures
Fig. 1 Microagraphs of Acanthamoeba cysts. A, A. culbertdoni; B, A. healyi; C, A. pustulosa; D, A. palestinensis; E, A. royreba; F, A. lenticulata. Bar = 10.0 µm.
Fig. 2 Polyacrylamide gel isoelectric focusing patterns for five kinds of isoenzymes of 6 strains of Acanthamoeba. AcP, acid phosphatase; ADH, alcohol dehydrogenase; GPI, glucose phosphate isomerase; LAP, leucine aminopeptidase; MDH, malate dehydrogenase. Lanes: 1, A. culbertdoni; 2, A. healyi; 3, A. pustulosa; 4, A. palestinensis; 5, A. lenticulata; 6, A. royreba.
Fig. 3 Agarose gel electrophoretic restriction fragment patterns for mitochondrial DNA from 6 strains of Acanthamoeba. Lanes: 1, A. culbertdoni; 2, A. healyi; 3, A. pustulosa; 4, A. palestinensis; 5, A. lenticulata; 6, A. royreba; M, Hind III digested lambda DNA for molecular size standard.
Fig. 4 Agarose gel electrophoretic restriction fragment patterns for PCR amplified ssu rDNA from 6 strains of Acanthamoeba. Lanes: 1, A. culbertdoni; 2, A. healyi; 3, A. pustulosa; 4, A. palestinensis; 5, A. royreba; 6, A. lenticulata; M, DNA molecular size stanard.
Fig. 5 Phylogenetic tree of Acanthamoeba isolates based on genetic divergence estimates. The matrix of divergence estimates in Table 2 was used to construct this tree using UPGMA.
Tables
Table 1 Morphology and sources of Acanthamoeba strains analyzed in this study
Table 2 Proportions of homologous fragments and estimated average numbers of changes per nucleotide position
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