Concerted evolution in the ribosomal RNA cistron

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Authors

Naidoo, Kershney
Steenkamp, Emma Theodora
Coetzee, Martin Petrus Albertus
Wingfield, Michael J.
Wingfield, Brenda D.

Journal Title

Journal ISSN

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Publisher

Public Library of Science

Abstract

BACKGROUND: Gene conversion is the mechanism proposed to be responsible for the homogenization of multigene families such as the nuclear ribosomal gene clusters. This concerted evolutionary process prevents individual genes in gene clusters from accumulating mutations. The mechanism responsible for concerted evolution is not well understood but recombination during meiosis has been hypothesized to play a significant role in this homogenization. In this study we tested the hypothesis of unequal crossing over playing a significant role in gene conversion events within the ribosomal RNA cistron during meiosis, mitosis or both life stages in the fungal tree pathogen Ceratocystis manginecans. METHODS: Ceratocystis manginecans, a haploid ascomycete, reproduces homothallically and was found to have two distinct sequences within the internally transcribed spacer (ITS) region of the ribosomal RNA cistron. The different ITS types were scored using PCR-RFLP assays and chi-square analyses to determine the level of significance of the changes in the ratios of the ITS types. RESULTS: The relative ratios of the two ITS sequence types changed when the fungal isolates were cultured vegetatively or allowed to produced sexual structures and spores. These active changes were shown to occur more frequently during meiosis than mitosis. CONCLUSION: The evidence presented provides concrete support for homogenization in the rRNA gene clusters found in this fungus and that the most reasonable explanation for this process is unequal crossing over.

Description

Figure S1 Aligned DNA sequence. Screen print showing the aligned DNA sequences of only ITS sequence type Y in Ceratocystis manginecans isolates CMW 13581, 13584, 23641 and 23643.
Figure S2 Aligned DNA sequence. Screen print of aligned DNA sequences showing the differences in the sequences of the two ITS sequence types (Z and Y) in Ceratocystis manginecans CMW 17568.
Figure S3 Hypothetical rRNA cistron showing meiotic and mitotic divisions. Illustration of a hypothetical situation for Ceratocystis manginecans where we assume that its rRNA cistron has 600 copies undergoing meiotic and mitotic divisions. In this example, under meiotic conditions each division represents either an increase or a decrease in the ITS sequence types. Figure S3A depicts a restricted cistron size scenario whilst Figure S3B has no size restriction.
Table S1 Summary of the observed ITS sequence types from 60 cloned amplicons across all test replicas for both meiosis and mitosis based on five sequential rounds of either mitotic transfers or sexual crosses for each isolateb.
Conceived and designed the experiments: KN BDW. Performed the experiments: KN. Analyzed the data: KN BDW MPAC ETS MJW. Contributed reagents/materials/analysis tools: BDW KN. Wrote the paper: KN BDW MPAC ETS MJW.

Keywords

Gene conversion, Nuclear ribosomal gene clusters, Ceratocystis manginecans

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Citation

Naidoo K, Steenkamp ET, Coetzee MPA, Wingfield MJ, Wingfield BD (2013) Concerted Evolution in the Ribosomal RNA Cistron. PLoS ONE 8(3): e59355. DOI: 10.1371/journal.pone.0059355