Phylogenetic Relationships, Pathogenic Traits, and Wood-Destroying Properties of Porodaedalea niemelaei M. Fischer Isolated in the Northern Forest Limit of Larix gmelinii Open Woodlands in the Permafrost Area

The phytopathogenic and wood destroying traits were studied in a basidiomycete fungus, Porodaedalea niemelaei M. Fischer, widespread in Siberian permafrost woodlands of Gmelinii larch, Larix gmelinii (Rupr.) Rupr. Numerous stands of dying out and fallen larch trees with white-rot („corrosion rot“) were found in the study area. Butt rot incidence varied from 63 to 100 % depending on the stand age and raised up to 0.5-1.5 m above root collar on average or up to 9 m maximum. Root rot was also widespread, including larch undergrowth. The biodiversity of xylotrophic fungi was low, with a pronounced dominance of P. niemelaei. The main factors of dying out of L. gmelinii were infection by P. niemelaei promoted by mechanical damage of roots by reindeers during migration and climate anomalies. The cultures isolated from the fruiting bodies were identified as Porodaedalea niemelaei M. Fischer based on the combination of morphological, culture, and molecular genetic methods. Under laboratory conditions, the strains were characterized as psychrotolerant (temperature limit from 6 to 22 °C) and preferred cultural media based mostly on natural and plant substrates. The most active biodegradation occurred on the broadleaf wood substrates causing up to 50 % of the biomass loss accompanied by active decomposing of the lignocellulosic complex and increasing the amount of water-soluble substances. The phylogenetic analysis demonstrated that P. niemelaei is clearly different from other well-studied Porodaedalea species, such as P. chrysoloma, P. pini, and P. cancriformans, and is very close to a group of unclassified fungi isolated in Norway and Finland. The phylogenetic analysis included 43 isolates and was based on four genetic markers – ITS, nLSU, rpb2, and tef1, commonly used in fungal phylogenetic

Sequence Capture of Mitochondrial Genome with PCR-Generated Baits Provides New Insights into the Biogeography of the Genus Abies Mill.

Mitochondrial DNA (mtDNA), being maternally inherited in plants of the family Pinaceae, is an important source of phylogeographic information. However, its use is hindered by a low mutation rate and frequent structure rearrangements. In the present study, we tested the method of genomic libraries enrichment with mtDNA via the sequence capture method yielding mtDNA data which were further used to reconstruct the phylogenetic tree of the genus Abies. The baits for hybrid capture were obtained by long-range PCR using primers designed on the basis of the assembly of Abies sibirica Ledeb. mitochondrial genome. Mitochondrial genomes of Picea sitchensis (Bong.) Carr., Larix sibirica Ledeb., and Keteleeria davidiana (Bertrand) Beissn. were used as an outgroup. The resulting phylogenetic tree consists of two sister branches, including the Eurasian and American species, respectively, with some exceptions. The subclade of A. sachalinensis (F. Schmidt) Mast. and A. veitchii Lindl. (Japan and Sakhalin islands) occupies a basal position in the branch of American firs, probably due to the complex history of fir migrations from North America to Eurasia. The tree has high support for majority of clades. For species represented by more than one sample an intraspecific variability was found which is suitable to design mtDNA markers for phylogeographic and population studies

Mitochondrial DNA in Siberian conifers indicates multiple post-glacial colonization centers

The geographic variation of the mitochondrial DNA was studied in Siberian fir using the newly developed markers and compared with the phylogeographic pattern of another previously studied Siberian coniferous - Siberian larch. Similar to Siberian larch the distribution of mtDNA haplotypes in Siberian fir revealed clear differentiation among distinct geographic regions of southern Siberia and the Urals, likely indicating post-glacial re-colonization from several sources. The northern part of the range of both species was genetically homogeneous, which is probably due to its recent colonization from one of the glacial refugia. This conclusion is in agreement with published pollen and macrofossil data in Siberian fir and with the reconstruction of environmental niches indicating a dramatic reduction of the range and a likely survival of fir in certain southern areas during the last glacial maximum (LGM) – 21 thousand years ago (kya). Although the modeling of Siberian larch ecological niche reconstructed a shift of the range to the south at that period, the paleontological data indicated the presence of this species in most areas of the current range during LGM, that corresponds to the results of previous historical demography study suggesting the population expansion preceding the LGM

Comparative Genomics of Seasonal Senescence in Forest Trees

In the course of evolution, both flowering plants and some gymnosperms have developed such an adaptation to winter and unfavorable living conditions as deciduousness. Of particular interest is Siberian larch (Larix sibirica Ledeb.), which is the only species in the pine family (Pinaceae) with a seasonal deciduousness. New generation sequencing technologies make it possible to study this phenomenon at the genomic level and to reveal the genetic mechanisms of leaf and needle aging in angiosperms and gymnosperms. Using a comparative analysis of the genomes of evergreen and deciduous trees, it was found that the genes that control EXORDIUM LIKE 2 (EXL2) and DORMANCY-ASSOCIATED PROTEIN 1 (DRM1) proteins are most represented in Siberian larch, while an excess of genes that control proteins acting as immune receptors were found in evergreens. Orthologs from the family of genes that control leucine-rich repeat receptor-like kinases (LRR-RLK) contributed mostly to the distinction between evergreens and deciduous plants

Siberian larch (Larix sibirica Ledeb.) chloroplast genome and development of polymorphic chloroplast markers

Abstract Background The main objectives of this study were sequencing, assembling, and annotation of chloroplast genome of one of the main Siberian boreal forest tree conifer species Siberian larch (Larix sibirica Ledeb.) and detection of polymorphic genetic markers – microsatellite loci or simple sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs). Results We used the data of the whole genome sequencing of three Siberian larch trees from different regions - the Urals, Krasnoyarsk, and Khakassia, respectively. Sequence reads were obtained using the Illumina HiSeq2000 in the Laboratory of Forest Genomics at the Genome Research and Education Center of the Siberian Federal University. The assembling was done using the Bowtie2 mapping program and the SPAdes genomic assembler. The genome annotation was performed using the RAST service. We used the GMATo program for the SSRs search, and the Bowtie2 and UGENE programs for the SNPs detection. Length of the assembled chloroplast genome was 122,561 bp, which is similar to 122,474 bp in the closely related European larch (Larix decidua Mill.). As a result of annotation and comparison of the data with the existing data available only for three larch species - L. decidua, L. potaninii var. chinensis (complete genome 122,492 bp), and L. occidentalis (partial genome of 119,680 bp), we identified 110 genes, 34 of which represented tRNA, 4 rRNA, and 72 protein-coding genes. In total, 13 SNPs were detected; two of them were in the tRNA-Arg and Cell division protein FtsH genes, respectively. In addition, 23 SSR loci were identified. Conclusions The complete chloroplast genome sequence was obtained for Siberian larch for the first time. The reference complete chloroplast genomes, such as one described here, would greatly help in the chloroplast resequencing and search for additional genetic markers using population samples. The results of this research will be useful for further phylogenetic and gene flow studies in conifers

Comparative Genomics of Seasonal Senescence in Forest Trees

In the course of evolution, both flowering plants and some gymnosperms have developed such an adaptation to winter and unfavorable living conditions as deciduousness. Of particular interest is Siberian larch (Larix sibirica Ledeb.), which is the only species in the pine family (Pinaceae) with a seasonal deciduousness. New generation sequencing technologies make it possible to study this phenomenon at the genomic level and to reveal the genetic mechanisms of leaf and needle aging in angiosperms and gymnosperms. Using a comparative analysis of the genomes of evergreen and deciduous trees, it was found that the genes that control EXORDIUM LIKE 2 (EXL2) and DORMANCY-ASSOCIATED PROTEIN 1 (DRM1) proteins are most represented in Siberian larch, while an excess of genes that control proteins acting as immune receptors were found in evergreens. Orthologs from the family of genes that control leucine-rich repeat receptor-like kinases (LRR-RLK) contributed mostly to the distinction between evergreens and deciduous plants

Симметрия транскриптома сибирской лиственницы

The paper presents a novel approach to infer a structuredness in a set of symbol sequences such as transcriptome nucleotide sequences. A distribution pattern of triplet frequencies in the Siberian larch (Larix sibirica Ledeb.) transcriptome sequences was investigated in the presented study. It was found that the larch transcriptome demonstrates a number of unexpected symmetries in the statistical and combinatorial propertiesПроанализированы структуры, выделяемые в транскриптоме лиственницы. Показано, что данный набор последовательностей обладает необычной симметрией своих статистических и комбинаторных свойст

Stepwise large genome assembly approach: a case of Siberian larch (Larix sibirica Ledeb)

Abstract Background De novo assembling of large genomes, such as in conifers (~ 12–30 Gbp), which also consist of ~ 80% of repetitive DNA, is a very complex and computationally intense endeavor. One of the main problems in assembling such genomes lays in computing limitations of nucleotide sequence assembly programs (DNA assemblers). As a rule, modern assemblers are usually designed to assemble genomes with a length not exceeding the length of the human genome (3.24 Gbp). Most assemblers cannot handle the amount of input sequence data required to provide sufficient coverage needed for a high-quality assembly. Results An original stepwise method of de novo assembly by parts (sets), which allows to bypass the limitations of modern assemblers associated with a huge amount of data being processed, is presented in this paper. The results of numerical assembling experiments conducted using the model plant Arabidopsis thaliana, Prunus persica (peach) and four most popular assemblers, ABySS, SOAPdenovo, SPAdes, and CLC Assembly Cell, showed the validity and effectiveness of the proposed stepwise assembling method. Conclusion Using the new stepwise de novo assembling method presented in the paper, the genome of Siberian larch, Larix sibirica Ledeb. (12.34 Gbp) was completely assembled de novo by the CLC Assembly Cell assembler. It is the first genome assembly for larch species in addition to only five other conifer genomes sequenced and assembled for Picea abies, Picea glauca, Pinus taeda, Pinus lambertiana, and Pseudotsuga menziesii var. menziesii