Hidden Compositional Heterogeneity of Fish Chromosomes in the Era of Polished Genome Assemblies

Fish chromosomes are considered homogeneous in their AT/GC nucleotide composition, and banding patterns enabling identification of homologs are largely missing. While cytogenomic approaches try to compensate for this issue by virtual karyotyping, they rely on the quality of genome assemblies available. Recently, soft-masked genome assemblies combining costly and arduous long- and short-read sequencing and new generation assemblers became available for two teleost fish species, climbing perch (Anabas testudineus) and channel bull blenny (Cottoperca gobio). Soft-masking turns repetitive sequences in a genome assembly into lower case letters, leaving unique sequences in upper case. This enables investigators to assess the proportion of guanine and cytosine nucleotides (GC%) of transposable elements as an indicator of AT/GC homogenisation in fish. We have developed a new version of our Python tool Evan, which utilises chromosome-level genome assemblies and combines the profiles of GC% and the proportion of repeats (rep%) along chromosomes. Our profiles of both of those fishes showed clear and abrupt but small-scale fluctuations in GC% along otherwise compositionally homogenised sequences. Our study also highlights the key role of the sliding window size in determining the resolution of GC% profiling. While the quality of the genome assemblies appeared to be sufficient for GC%/rep% profiling, more effective repeat masking is necessary to better distinguish to what extent repeats compositionally homogenize fish genomes.journal articl

Ecological Divergence Within the Enterobacterial Genus Sodalis: From Insect Symbionts to Inhabitants of Decomposing Deadwood

The bacterial genus Sodalis is represented by insect endosymbionts as well as free-living species. While the former have been studied frequently, the distribution of the latter is not yet clear. Here, we present a description of a free-living strain, Sodalis ligni sp. nov., originating from decomposing deadwood. The favored occurrence of S. ligni in deadwood is confirmed by both 16S rRNA gene distribution and metagenome data. Pangenome analysis of available Sodalis genomes shows at least three groups within the Sodalis genus: deadwood-associated strains, tsetse fly endosymbionts and endosymbionts of other insects. This differentiation is consistent in terms of the gene frequency level, genome similarity and carbohydrate-active enzyme composition of the genomes. Deadwood-associated strains contain genes for active decomposition of biopolymers of plant and fungal origin and can utilize more diverse carbon sources than their symbiotic relatives. Deadwood-associated strains, but not other Sodalis strains, have the genetic potential to fix N2, and the corresponding genes are expressed in deadwood. Nitrogenase genes are located within the genomes of Sodalis, including S. ligni, at multiple loci represented by more gene variants. We show decomposing wood to be a previously undescribed habitat of the genus Sodalis that appears to show striking ecological divergence

Large-scale genome sequencing of mycorrhizal fungi provides insights into the early evolution of symbiotic traits

Mycorrhizal fungi are mutualists that play crucial roles in nutrient acquisition in terrestrial ecosystems. Mycorrhizal symbioses arose repeatedly across multiple lineages of Mucoromycotina, Ascomycota, and Basidiomycota. Considerable variation exists in the capacity of mycorrhizal fungi to acquire carbon from soil organic matter. Here, we present a combined analysis of 135 fungal genomes from 73 saprotrophic, endophytic and pathogenic species, and 62 mycorrhizal species, including 29 new mycorrhizal genomes. This study samples ecologically dominant fungal guilds for which there were previously no symbiotic genomes available, including ectomycorrhizal Russulales, Thelephorales and Cantharellales. Our analyses show that transitions from saprotrophy to symbiosis involve (1) widespread losses of degrading enzymes acting on lignin and cellulose, (2) co-option of genes present in saprotrophic ancestors to fulfill new symbiotic functions, (3) diversification of novel, lineage-specific symbiosis-induced genes, (4) proliferation of transposable elements and (5) divergent genetic innovations underlying the convergent origins of the ectomycorrhizal guild. Mycorrhizal symbioses have evolved repeatedly in diverse fungal lineages. A large phylogenomic analysis sheds light on genomic changes associated with transitions from saprotrophy to symbiosis, including divergent genetic innovations underlying the convergent origins of the ectomycorrhizal guild.Peer reviewe

Molecular biology and ecology of microbial decomposition of plant-derived biopolymers in forest ecosystems

The abilities of fungi and bacteria to degrade simple and complex carbon compounds derived from different sources, such as root exudates, litter, soil organic matter or fungal mycelium were studied in this dissertation. Knowledge of functional traits, especially degradation abilities of fungi and bacteria, are important for deciphering the black box of microbial functioning in topsoil and thus aiding in modeling and predicting future directions of microbial communities development in face of global changes. Among fungal cultures form culture collection representing strains with different taxonomy and ecophysiology, the ecophysiology of fungi was more important in manifestation of functional traits than taxonomy. Among bacterial isolates from the litter and soil of spruce forest, Acidobacteria were confirmed to express multiple decomposition enzymes in high rates in vitro and were also abundant and active degraders in acidic spruce forest soil. The expression of degradation capacities of both bacteria and fungi were further studied in situ in spruce forest topsoil, that represents an important environment due to the ubiquity of coniferous forests on the Northern hemisphere. There is an obvious gap of knowledge, when comes to our understanding of seasonal effect on microbial functioning, and this is..

Charakterizace společenstva hub, podílejícího se na rozkladu opadu v jehličnatých lesích Národního parku Šumava

Understanding of carbon cycling in coniferous forests that represent a large carbon sink is crucial for our understanding of natural processes under global climate change. Recognition of fungi as fundamental decomposers can contribute to this understanding. Fungi are able to decompose numbers of substrates and possess a variety of enzymes to do so In this study I present litter decomposing fungi in mountain spruce forest from national park Šumava. The aim of my thesis was to follow succession and community changes of fungi from the early stages of decomposition of Picea abies needles until degradation of organic matter in the organic horizon of the soil. This aim was accomplished partly by recording the extracellular enzyme production of fungi in different stages of decomposition from needles attached to the twigs of a fallen tree to a litter material in later stages of decomposition on the soil surface. In addition to testing of fungi on their natural substrata - needle litter, enzyme activities were also measured in laboratory agar cultures, which allow comparison of diverse fungi with different origins. Enzyme activities were aimed at enzymes decomposing cellulose and compounds found in litter. Although ecology of endophytic and saprothrophic fungi suggest differences in enzyme production, these..

Single nucleotide polymorphism as a biomarker of colorectal cancer predispositon

Department of Cell BiologyKatedra buněčné biologieFaculty of SciencePřírodovědecká fakult

Molekulární biologie a ekologie rozkladu rostlinných biopolymerů v lesních ekosystémech mikroorganismy

V této dizertační práci byly studovány schopnosti hub a bakterií degradovat jednoduché i složité uhlíkaté sloučeniny získané z různých zdrojů, jako jsou kořenové exsudáty, opad, půdní organická hmota nebo houbové mycelium. Znalost funkčních vlastností, zejména degradačních schopností hub a bakterií, je důležitá pro dešifrování černé skříňky fungování mikrobů v půdě, a tím pomáhá při modelování a předpovídání budoucích směrů vývoje mikrobiálních společenstev tváří v tvář globálním změnám. U sbírkových kmenů hub s různou taxonomií a ekofyziologií byla ekofyziologie důležitější při projevu funkčních vlastností než taxonomie. U bakteriálních izolátů z opadu a půdy smrkového lesa, bylo potvrzeno, že Acidobakterie produkují širokou škálu enzymů ve velkém množství in vitro, byly také hojně zastoupeny a také aktivní v procesech rozkladu v kyselé smrkové půdě. Exprese degradačních enzymů u bakterií i hub byly dále studovány in situ v půdě smrkového lesa, která představuje důležité životní prostředí vzhledem k širokému rozšíření jehličnatých lesů na severní polokouli. Naše znalosti, pokud jde o pochopení sezónního vlivu na mikrobiální činnost, jsou nedostatečné. To je důvod, proč byl studovaný vliv sezónnosti na transkripci všech funkčních genů u hub a bakterií. Stejné experimentální uspořádání bylo použito...The abilities of fungi and bacteria to degrade simple and complex carbon compounds derived from different sources, such as root exudates, litter, soil organic matter or fungal mycelium were studied in this dissertation. Knowledge of functional traits, especially degradation abilities of fungi and bacteria, are important for deciphering the black box of microbial functioning in topsoil and thus aiding in modeling and predicting future directions of microbial communities development in face of global changes. Among fungal cultures form culture collection representing strains with different taxonomy and ecophysiology, the ecophysiology of fungi was more important in manifestation of functional traits than taxonomy. Among bacterial isolates from the litter and soil of spruce forest, Acidobacteria were confirmed to express multiple decomposition enzymes in high rates in vitro and were also abundant and active degraders in acidic spruce forest soil. The expression of degradation capacities of both bacteria and fungi were further studied in situ in spruce forest topsoil, that represents an important environment due to the ubiquity of coniferous forests on the Northern hemisphere. There is an obvious gap of knowledge, when comes to our understanding of seasonal effect on microbial functioning, and this is...Katedra genetiky a mikrobiologieDepartment of Genetics and MicrobiologyPřírodovědecká fakultaFaculty of Scienc

Single nucleotide polymorphism as a biomarker of colorectal cancer predispositon

Department of Cell BiologyKatedra buněčné biologieFaculty of SciencePřírodovědecká fakult

Data from: The rpb2 gene represents a viable alternative molecular marker for the analysis of environmental fungal communities

Although the commonly used internal transcribed spacer region of rDNA (ITS) is well suited for taxonomic identification of fungi, the information on the relative abundance of taxa and diversity is negatively affected by the multicopy nature of rDNA and the existence of ITS paralogues. Moreover, due to high variability, ITS sequences cannot be used for phylogenetic analyses of unrelated taxa. The part of single-copy gene encoding the second largest subunit of RNA polymerase II (rpb2) was thus compared with first spacer of ITS as an alternative marker for the analysis of fungal communities in spruce forest topsoil, and their applicability was tested on a comprehensive mock community. In soil, rpb2 exhibited broad taxonomic coverage of the entire fungal tree of life including basal fungal lineages. The gene exhibited sufficient variation for the use in phylogenetic analyses and taxonomic assignments, although it amplifies also paralogues. The fungal taxon spectra obtained with rbp2 region and ITS1 corresponded, but sequence abundance differed widely, especially in the basal lineages. The proportions of OTU counts and read counts of major fungal groups were close to the reality when rpb2 was used as a molecular marker while they were strongly biased towards the Basidiomycota when using the ITS primers ITS1/ITS4. Although the taxonomic placement of rbp2 sequences is currently more difficult than that of the ITS sequences, its discriminative power, quantitative representation of community composition and suitability for phylogenetic analyses represent significant advantages