Reductions in complexity of mitochondrial genomes in lichen‐forming fungi shed light on genome architecture of obligate symbioses

Symbioses among co‐evolving taxa are often marked by genome reductions such as a loss of protein‐coding genes in at least one of the partners as a means of reducing redundancy or intergenomic conflict. To explore this phenomenon in an iconic yet under‐studied group of obligate symbiotic organisms, mitochondrial genomes of 22 newly sequenced and annotated species of lichenized fungi were compared to 167 mitochondrial genomes of nonlichenized fungi. Our results demonstrate the first broad‐scale loss of atp9 from mitochondria of lichenized fungi. Despite key functions in mitochondrial energy production, we show that atp9 has been independently lost in three different lineages spanning 10 of the 22 studied species. A search for predicted, functional copies of atp9 among genomes of other symbionts involved in each lichen revealed the full‐length, presumably functional copies of atp9 in either the photosynthetic algal partner or in other symbiotic fungi in all 10 instances. Together, these data yield evidence of an obligate symbiotic relationship in which core genomic processes have been streamlined, likely due to co‐evolution.     

High selectivity in symbiotic associations of lichenized ascomycetes and cyanobacteria

The selectivity of mycobionts and cyanobionts in lichen symbioses were examined. We analyzed symbiotic cyanobionts, collected from different sample sites, and compared them to free‐living cyanobacteria Nostoc. Cyanobionts were obtained from lichens assigned to the genera Pseudocyphellaria and Sticta, in particular. Multiple gene loci were screened and direct optimization was used in the phylogenetic analyses. We show that many lichen fungi are strongly selective towards their cyanobionts. Lichenized ascomycetes seem to be able to identify and choose a specific strain, species or a species group of Nostoc with which to associate. The present analyses also suggest that some of the Nostoc taxa may be specialized in symbiotic life with only lichenized ascomycetes. Despite the selectivity observed in fungi, there appears to be no coevolution between the partners. We have also discussed the problems of using the tRNA^Leu intron as a marker in phylogenetic analyses. © The Willi Hennig Society 2006.     

The Mitochondrial Genome of Xiphinema americanum sensu stricto (Nematoda: Enoplea): Considerable Economization in the Length and Structural Features of Encoded Genes

The complete sequence of the mitochondrial genome of the plant parasitic nematode Xiphinema americanum sensu stricto has been determined. At 12626bp it is the smallest metazoan mitochondrial genome reported to date. Genes are transcribed from both strands. Genes coding for 12 proteins, 2 rRNAs and 17 putative tRNAs (with the tRNA-C, I, N, S1, S2 missing) are predicted from the sequence. The arrangement of genes within the X. americanum mitochondrial genome is unique and includes gene overlaps. Comparisons with the mtDNA of other nematodes show that the small size of the X. americanum mtDNA is due to a combination of factors. The two mitochondrial rRNA genes are considerably smaller than those of other nematodes, with most of the protein encoding and tRNA genes also slightly smaller. In addition, five tRNAs genes are absent, lengthy noncoding regions are not present in the mtDNA, and several gene overlaps are present. [Reviewing Editor: Dr. Yues van de Peer] F. Lamberti: Deceased, 2004     

Fungal specific primers for PCR‐amplification of mitochondrial LSU in lichens

Fungal specific primer sequences for the amplification of the large subunit of the mitochondrial ribosomal DNA (mtLSU) are presented in this paper. Fungal specific primers make the separation of fungal and algal cells prior to DNA‐extraction from lichens unnecessary. This is especially useful in crustose and small foliose and fruticose lichens. An example from a complex of closely related species of the crustose lichen genus Biatora shows the usefulness of mtLSU‐sequences for studies of infraspecific variability and lower level systematics of lichenized ascomycetes.     

Construction and characterization of a full-length cDNA library from mycobiont of <Emphasis Type="Italic">Endocarpon pusillum</Emphasis> (lichen-forming Ascomycota)

The first full-length cDNA library for lichenized fungi was constructed from cultured mycobiont of the arid desert lichen Endocarpon pusillum. Based on small-scale sequencing results, 111 genes of the lichenized fungi were identified for the first time, among which 11 genes shared no homology with any known fungal genes. Real-time PCR showed that the size of the mycobiont genome is 39.13 Mb and the copy number of ribosomal RNA gene repeat units is 43. The results of this study will be valuable for the ongoing lichen genome-sequencing project and the large-scale identification of functional genes from lichenized fungi.     

Should we hail the Red King? Evolutionary consequences of a mutualistic lifestyle in genomes of lichenized ascomycetes

The Red Queen dynamic is often brought into play for antagonistic relationships. However, the coevolutionary effects of mutualistic interactions, which predict slower evolution for interacting organisms (Red King), have been investigated to a lesser extent. Lichens are a stable, mutualistic relationship of fungi and cyanobacteria and/or algae, which originated several times independently during the evolution of fungi. Therefore, they represent a suitable system to investigate the coevolutionary effect of mutualism on the fungal genome. We measured substitution rates and selective pressure of about 2000 protein‐coding genes (plus the rDNA region) in two different classes of Ascomycota, each consisting of closely related lineages of lichenized and non‐lichenized fungi. Our results show that independent lichenized clades are characterized by significantly slower rates for both synonymous and non‐synonymous substitutions. We hypothesize that this evolutionary pattern is connected to the lichen life cycle (longer generation time of lichenized fungi) rather than a result of different selection strengths, which is described as the main driver for the Red Kind dynamic. This first empirical evidence of slower evolution in lichens provides an important insight on how biotic cooperative interactions are able to shape the evolution of symbiotic organisms.     

A Comparative Characterization of the Mitochondrial Genomes of Paramoeba aparasomata and Neoparamoeba pemaquidensis (Amoebozoa,Paramoebidae)

Marine amebae of the genus Paramoeba (Amoebozoa, Dactylopodida) normally contain a eukaryotic endosymbiont known as Perkinsela‐like organism (PLO). This is one of the characters to distinguish the genera Neoparamoeba and Paramoeba from other Dactylopodida. It is known that the PLO may be lost, but PLO‐free strains of paramoebians were never available for molecular studies. Recently, we have described the first species of the genus Paramoeba which has no parasome—Paramoeba aparasomata. In this study, we present a mitochondrial genome of this species, compare it with that of Neoparamoeba pemaquidensis, and analyze the evolutionary dynamics of gene sequences and gene order rearrangements between these species. The mitochondrial genome of P. aparasomata is 46,254 bp long and contains a set of 31 protein‐coding genes, 19 tRNAs, two rRNA genes, and 7 open reading frames. Our results suggest that these two mitochondrial genomes within the genus Paramoeba have rather similar organization and gene order, base composition, codon usage, the composition and structure of noncoding, and overlapping regions.     

EVOLUTION AND MITOCHONDRIAL DNA IN NEUROSPORA CRASSA

We studied mitochondrial DNA variability in 19 natural Neurospora crassa isolates and one wild-type isolate to examine evolution of these fungi and their mitochondrial DNA (mtDNA). We combined restriction endonuclease analysis of natural isolate mtDNA with DNA-DNA hybridization to cloned EcoR I fragments of a wild-type genome to discriminate between length mutations and site changes due to nucleotide substitution. Most variability was due to length mutations (insertions and deletions); genome size could vary 25% between pairs of isolates. Length-mutation distribution was not random, nor simply explained by the presence of coding versus noncoding regions. Restriction-site changes were few; the estimated amount of nucleotide substitution per nucleotide between the most divergent pair of isolates was 0.78%. Evolutionary relationships among isolates based on both types of mutations were compatible, and suggest that geographically distinct populations of mitochondrial DNA exist in the biological species, N. crassa. In contrast, no such correlation was shown by the previously determined distribution of nuclear heterokaryon incompatibility genes in the same isolates (Mylyk, 1975, 1976).     

Lichens of Barla Mountain in Isparta,Turkey: Diversity study and ecological assessment of the area

This study reports on 230 infrageneric lichenized and lichenicolous taxa from Barla Mountain, Isparta, Turkey and assesses the ecological features of the area using the distribution of the lichens in the region and their poleophoby and solar irradiation ecological indicator values. One lichenized fungus, Protoblastenia terricola, and one lichenicolous fungus, Zwackhiomyces dispersus, are recorded as new in Turkey and 194 taxa are reported for the first time from Barla Mountain. After this research, number of the lichenized and lichenicolous fungi taxa of Barla Mountain rose to 241. Based on assessments using the ecological indicator values, the area is dominated by natural or semi-natural and well-preserved habitats.     

Über Cyanotrophie bei Flechten

In contrast to the well-known blue-green algal containing lichens several green algal containing lichens, belonging to very different genera, show regular connections to free-living or ± lichenized blue-green algae, mainlyStigonema. Most of these lichens have squamulose thalli. This lichen-algal relationship, regarded as cyanotrophy, may be either facultative or obligate. Some of the species occur only on very poor, acidic rocks onStigonema, while they occur independent ofStigonema in high nutrient biotops. Obligate species cover the blue-green algae with hyphae and some of these species cover the algae so extensively that one can call these connections paracephalodia. — Two species and one variety are new to science from the mainly Himalayan genusBryonora. They occur in high elevations in Nepal and are cyanotrophic.Bryonora selenospora has thick, halfmoon-shaped to slightly twisted ascospores. The other two new taxa areB. reducta andB. rhypariza var.cyanotropha. There are several other cyanotrophic lichen taxa besides the ones described here. They will be introduced at a later occasion.

Frau Prof. Dr.Elisabeth Tschermak-Woess zu ihrem 70. Geburtstag gewidmet.     

PHYLOGENETIC DIVERSITY OF TRENTEPOHLIALEAN ALGAE ASSOCIATED WITH LICHEN‐FORMING FUNGI1

Nearly one‐fourth of the lichen‐forming fungi associate with trentepohlialean algae, yet their genetic diversity remains unknown. Recent work focusing on free‐living trentepohlialean algae has provided a phylogenetic context within which questions regarding the lichenization of these algae can be asked. Here, we concentrated our sampling on trentepohlialean algae from lichens producing a diversity of growth forms (fruticose and crustose) over a broad geographic substratum, ecological, and phylogenetic range. We have demonstrated that there is no evidence for a single clade of strictly lichenized algae; rather, a wide range demonstrated the ability to associate with lichenized fungi. Variation was also observed among trentepohlialean algae in lichens from a single geographic area and tree, suggesting that fungi in close proximity can associate with different trentepohlialean algae, consistent with the findings of trebouxiophycean algae and cyanobacteria.     

Promiscuous mitochondrial group II intron sequences in plant nuclear genomes

Gene translocations from the organelles to the nucleus are postulated by the endosymbiont hypothesis. We here report evidence for sequence insertions in the nuclear genomes of plants that are derived from noncoding regions of the mitochondrial genome. Fragments of mitochondrial group II introns are identified in the nuclear genomes of tobacco and a bean species. The duplicated intron sequences of 75–140 bp are derived from cis- and trans-splicing introns of genes encoding subunits 1 and 5 of the NADH dehydrogenase. The mitochondrial sequences are inserted in the vicinities of a lectin gene, different glucanase genes and a gene encoding a subunit of photosystem II. Sequence similarities between the nuclear and mitochondrial copies are in the range of 80 to 97%, suggesting recent transfer events that occurred in the basic glucanase genes before and in the lectin gene after the gene duplications in the evolution of the nuclear gene families. Overlapping regions of the same introns are in two instances also involved in intramitochondrial sequence duplications. Correspondence to: V. Knoop     

MOLECULAR PHYLOGENY OF STAURASTRUM MEYEN EX RALFS AND RELATED GENERA (ZYGNEMATOPHYCEAE,STREPTOPHYTA) BASED ON CODING AND NONCODING RDNA SEQUENCE COMPARISONS1

Nuclear‐encoded small subunit rDNA, 1506 group I intron, and internal transcribed spacer sequences were obtained from 39 strains representing five core desmid genera, Staurastrum, Staurodesmus Teil., Cosmarium Corda ex Ralfs, Xanthidium Ehr. ex Ralfs, and Euastrum Ehr. ex Ralfs (Desmidiaceae, Zygnematophyceae), and used individually and concatenated to assess phylogenetic relationships between putatively allied members of the family. To identify positional homology between divergent noncoding sequences, secondary structure models were generated and their reliability assessed by screening the alignment for compensating base changes. The phylogeny based on coding and noncoding sequence comparisons confidently resolved a monophyletic core of the genus Staurastrum but also revealed the artificial nature of the traditional genus. Twenty distinct species representing a wide range of morphotypes of Staurastrum formed a strongly supported generic clade that was further split into three well‐resolved lineages. The phylogenetic relationships revealed within Staurastrum were in conflict with all previous formal or informal classifications of the genus. The genera Staurodesmus and Cosmarium were shown to be highly polyphyletic, and some morphologically similar taxa displayed high sequence divergence that exceeded generic boundaries. Apparently, the taxonomic significance of some morphological characters in Staurastrum and other desmid genera has been greatly overestimated.     

Comparative Metabolism of Free‐living Bodo saltans and Parasitic Trypanosomatids

Comparison of the genomes of free‐living Bodo saltans and those of parasitic trypanosomatids reveals that the transition from a free‐living to a parasitic life style has resulted in the loss of approximately 50% of protein‐coding genes. Despite this dramatic reduction in genome size, B. saltans and trypanosomatids still share a significant number of common metabolic traits: glycosomes; a unique set of the pyrimidine biosynthetic pathway genes; an ATP‐PFK which is homologous to the bacterial PP_i‐PFKs rather than to the canonical eukaryotic ATP‐PFKs; an alternative oxidase; three phosphoglycerate kinases and two GAPDH isoenzymes; a pyruvate kinase regulated by fructose‐2,6‐bisphosphate; trypanothione as a substitute for glutathione; synthesis of fatty acids via a unique set of elongase enzymes; and a mitochondrial acetate:succinate coenzyme A transferase. B. saltans has lost the capacity to synthesize ubiquinone. Among genes that are present in B. saltans and lost in all trypanosomatids are those involved in the degradation of mureine, tryptophan and lysine. Novel acquisitions of trypanosomatids are components of pentose sugar metabolism, pteridine reductase and bromodomain‐factor proteins. In addition, only the subfamily Leishmaniinae has acquired a gene for catalase and the capacity to convert diaminopimelic acid to lysine.     

Intron–Genome Size Relationship on a Large Evolutionary Scale

The intron–genome size relationship was studied across a wide evolutionary range (from slime mold and yeast to human and maize), as well as the relationship between genome size and the ratio of intervening/coding sequence size. The average intron size is scaled to genome size with a slope of about one-fourth for the log-transformed values; i.e., on the global scale its increase in evolution is lower than the increase in genome size by four orders of magnitude. There are exceptions to the general trend. In baker's yeast introns are extraordinarily long for its genome size. Tetrapods also have longer introns than expected for their genome sizes. In teleost fish the mean intron size does not differ significantly, notwithstanding the differences in genome size. In contrast to previous reports, avian introns were not found to be significantly shorter than introns of mammals, although avian genomes are smaller than genomes of mammals on average by about a factor of 2.5. The extra-/intragenic ratio of noncoding DNA can be higher in fungi than in animals, notwithstanding the smaller fungal genomes. In vertebrates and invertebrates taken separately, this ratio is increasing as the increase in genome size. Two hypotheses are proposed to explain the variation in the extra-/intragenic ratio of noncoding DNA in organisms with similar numbers of genes: transition (dynamic) and equilibrium (static). According to the transition model, this variation arises with the rapid shift of genome size because the bulk of extragenic DNA can be changed more rapidly than the finely interspersed intron sequences. The equilibrium model assumes that this variation is a result of selective adjustment of genome size with constraints imposed on the intron size due to its putative link to chromatin structure (and constraints of the splicing machinery). Received: 23 October 1997 / Accepted: 14 April 1999     

Stripped-down DNA repair in a highly reduced parasite

Background  

Encephalitozoon cuniculi is a member of a distinctive group of single-celled parasitic eukaryotes called microsporidia, which are closely related to fungi. Some of these organisms, including E. cuniculi, also have uniquely small genomes that are within the prokaryotic range. Thus, E. cuniculi has undergone a massive genome reduction which has resulted in a loss of genes from diverse biological pathways, including those that act in DNA repair.     

Quantifying the climatic niche of symbiont partners in a lichen symbiosis indicates mutualist‐mediated niche expansions

The large distributional areas and ecological niches of many lichenized fungi may in part be due to the plasticity in interactions between the fungus (mycobiont) and its algal or cyanobacterial partners (photobionts). On the one hand, broad‐scale phylogenetic analyses show that partner compatibility in lichens is rather constrained and shaped by reciprocal selection pressures and codiversification independent of ecological drivers. On the other hand, sub‐species‐level associations among lichen symbionts appear to be environmentally structured rather than phylogenetically constrained. In particular, switching between photobiont ecotypes with distinct environmental preferences has been hypothesized as an adaptive strategy for lichen‐forming fungi to broaden their ecological niche. The extent and direction of photobiont‐mediated range expansions in lichens, however, have not been examined comprehensively at a broad geographic scale. Here we investigate the population genetic structure of Lasallia pustulata symbionts at sub‐species‐level resolution across the mycobiont's Europe‐wide range, using fungal MCM7 and algal ITS rDNA sequence markers. We show that variance in occurrence probabilities in the geographic distribution of genetic diversity in mycobiont‐photobiont interactions is closely related to changes in climatic niches. Quantification of niche extent and overlap based on species distribution modeling and construction of Hutchinsonian climatic hypervolumes revealed that combinations of fungal–algal interactions change at the sub‐species level along latitudinal temperature gradients and in Mediterranean climate zones. Our study provides evidence for symbiont‐mediated niche expansion in lichens. We discuss our results in the light of symbiont polymorphism and partner switching as potential mechanisms of environmental adaptation and niche evolution in mutualisms.     

Molecular Evolution of the mtDNA Encoded <Emphasis Type="Italic">rps3</Emphasis> Gene Among Filamentous Ascomycetes Fungi with an Emphasis on the Ophiostomatoid Fungi

The mitochondrial ribosomal protein S3 (rps3) gene within the fungi is extremely diverse in location and organization, some versions of this gene have been incorporated into a group I intron, others appear to have gained large insertions, microsatellite expansions, or have been invaded by homing endonucleases. Among the ascomycetes fungi the group I intron encoded version of rps3 appears to have a rather complex evolutionary history including first the acquisition of rps3 by a group I intron (mL2449), the loss of the mL2499 intron and the establishment of rps3 as a free-standing gene, and the eventual loss of the intron derived version of rps3.     

A workflow of massive identification and application of intron markers using snakes as a model

Relative to the commonly used mitochondrial and nuclear protein‐coding genes, the noncoding intron sequences are a promising source of informative markers that have the potential to resolve difficult phylogenetic nodes such as rapid radiations and recent divergences. Yet many issues exist in the use of intron markers, which prevent their extensive application as conventional markers. We used the diverse group of snakes as an example to try paving the way for massive identification and application of intron markers. We performed a series of bioinformatics screenings which identified appropriate introns between single‐copy and conserved exons from two snake genomes, adding particular constraints on sequence length variability and sequence variability. A total of 1,273 candidate intron loci were retrieved. Primers for nested polymerase chain reaction (PCR) were designed for over a hundred candidates and tested in 16 snake representatives. 96 intron markers were developed that could be amplified across a broad range of snake taxa with high PCR successful rates. The markers were then applied to 49 snake samples. The large number of amplicons was subjected to next‐generation sequencing (NGS). An analytic strategy was developed to accurately recover the amplicon sequences, and approximately, 76% of the marker sequences were recovered. The average p‐distances of the intron markers at interfamily, intergenus, interspecies, and intraspecies levels were .168, .052, .015, and .004, respectively, suggesting that they were useful to study snake relationships of different evolutionary depths. A snake phylogeny was constructed with the intron markers, which produced concordant results with robust support at both interfamily and intragenus levels. The intron markers provide a convenient way to explore the signals in the noncoding regions to address the controversies on the snake tree. Our improved strategy of genome screening is effective and can be applied to other animal groups. NGS coupled with appropriate sequence processing can greatly facilitate the extensive application of molecular markers.     

Two new decapod (Crustacea,Malacostraca) complete mitochondrial genomes: bearings on the phylogenetic relationships within the Decapoda

Recent advances in molecular phylogenetics are continuously changing our perception of decapod phylogeny. Although the two suborders Dendrobranchiata and Pleocyemata within the Decapoda are widely accepted, this taxonomic view is now challenged when using mitochondrial protein‐coding genes to investigate the decapod phylogeny, especially for the basal pleocyematan groups. Here, we enhanced taxonomic coverage by sequencing the genomes of two basal decapod taxa Alpheus distinguendus and Panulirus ornatus, representing two infraorders, Caridea and Achelata, respectively. Based on these two and other available mitochondrial genomes, we evaluated the usefulness of protein‐coding genes in resolving deep phylogenetic relationships of the Decapoda using maximum likelihood and Bayesian analyses. The mt genomic results revealed a novel gene order because of the reverse transposition of trnE (transfer, trn for Glutamate) and a pseudogene‐like trnS (AGN) [trn for Serine (S₁, AGN)] in the mitochondrial genome of A. distinguendus, and a duplicate of 89 bp sequences in the putative noncoding region of P. ornatus. Our phylogenetic inferences suggest monophyly of the Decapoda and its two suborders, and that several lineages within the Reptantia are consistently recovered with high nodal supports. Our findings suggest that the best mitochondrial genome phylogeny can be found on the premise that systematic errors should be minimized as much as possible. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 162 , 471–481.