The genus Cerastium includes about 200 species that are mostly found in the temperate climates of... more The genus Cerastium includes about 200 species that are mostly found in the temperate climates of the Northern Hemisphere. Here we report the complete chloroplast genomes of Cerastium alpinum, C. arcticum and C. nigrescens. The length of cp genomes ranged from 147,940 to 148,722 bp. Their quadripartite circular structure had the same gene organization and content, containing 79 proteincoding genes, 30 tRNA genes, and four rRNA genes. Repeat sequences varied from 16 to 23 per species, with palindromic repeats being the most frequent. The number of identified SSRs ranged from 20 to 23 per species and they were mainly composed of mononucleotide repeats containing A/T units. Based on Ka/Ks ratio values, most genes were subjected to purifying selection. The newly sequenced chloroplast genomes were characterized by a high frequency of RNA editing, including both C to U and U to C conversion. The phylogenetic relationships within the genus Cerastium and family Caryophyllaceae were reconstructed based on the sequences of 71 protein-coding genes. The topology of the phylogenetic tree was consistent with the systematic position of the studied species. All representatives of the genus Cerastium were gathered in a single clade with C. glomeratum sharing the least similarity with the others.
Background The development of the plant in vitro techniques has brought about the variation ident... more Background The development of the plant in vitro techniques has brought about the variation identified in regenerants known as somaclonal or tissue culture-induced variation (TCIV). S-adenosyl-L-methionine (SAM), glutathione (GSH), low methylated pectins (LMP), and Cu(II) ions may be implicated in green plant regeneration efficiency (GPRE) and TCIV, according to studies in barley (Hordeum vulgare L.) and partially in triticale (× Triticosecale spp. Wittmack ex A. Camus 1927). Using structural equation models (SEM), these metabolites have been connected to the metabolic pathways (Krebs and Yang cycles, glycolysis, transsulfuration), but not for triticale. Using metabolomic and (epi)genetic data, the study sought to develop a triticale regeneration efficiency statistical model. The culture’s induction medium was supplemented with various quantities of Cu(II) and Ag(I) ions for regeneration. The period of plant regeneration has also changed. The donor plant, anther-derived regenerants, and metAFLP were utilized to analyze TCIV concerning DNA in symmetric (CG, CHG) and asymmetric (CHH) sequence contexts. Attenuated Total Reflectance–Fourier Transfer Infrared (ATR-FTIR) spectroscopy was used to gather the metabolomic information on LMP, SAM, and GSH. To frame the data, a structural equation model was employed. Results According to metAFLP analysis, the average sequence change in the CHH context was 8.65%, and 0.58% was de novo methylation. Absorbances of FTIR spectra in regions specific for LMP, SAM, and GSH were used as variables values introduced to the SEM model. The average number of green regenerants per 100 plated anthers was 2.55. Conclusions The amounts of pectin demethylation, SAM, de novo methylation, and GSH are connected in the model to explain GPRE. By altering the concentration of Cu(II) ions in the medium, which influences the amount of pectin, triticale’s GPRE can be increased.
Scots pine is the foundation species of diverse forested ecosystems across Eurasia and displays r... more Scots pine is the foundation species of diverse forested ecosystems across Eurasia and displays remarkable ecological breadth, occurring in environments ranging from temperate rainforests to arid tundra margins. Such expansive distributions can be favored by various demographic and adaptive processes and the interactions between them. To understand the impact of neutral and selective forces on genetic structure in Scots pine, we conducted range‐wide population genetic analyses on 2321 trees from 202 populations using genotyping‐by‐sequencing, reconstructed the recent demography of the species and examined signals of genetic adaptation. We found a high and uniform genetic diversity across the entire range (global FST 0.048), no increased genetic load in expanding populations and minor impact of the last glacial maximum on historical population sizes. Genetic‐environmental associations identified only a handful of single‐nucleotide polymorphisms significantly linked to environmental gradients. The results suggest that extensive gene flow is predominantly responsible for the observed genetic patterns in Scots pine. The apparent missing signal of genetic adaptation is likely attributed to the intricate genetic architecture controlling adaptation to multi‐dimensional environments. The panmixia metapopulation of Scots pine offers a good study system for further exploration into how genetic adaptation and plasticity evolve under gene flow and changing environment.
Background
Microspore embryogenesis is a process that produces doubled haploids in tissue cultur... more Background
Microspore embryogenesis is a process that produces doubled haploids in tissue culture environments and is widely used in cereal plants. The efficient production of green regenerants requires stresses that could be sensed at the level of glycolysis, followed by the Krebs cycle and electron transfer chain. The latter can be affected by Cu(II) ion concentration in the induction media acting as cofactors of biochemical reactions, indirectly influencing the production of glutathione (GSH) and S-adenosyl-L-methionine (SAM) and thereby affecting epigenetic mechanisms involving DNA methylation (demethylation—DM, de novo methylation—DNM). The conclusions mentioned were acquired from research on triticale regenerants, but there is no similar research on barley. In this way, the study looks at how DNM, DM, Cu(II), SAM, GSH, and β-glucan affect the ability of green plant regeneration efficiency (GPRE). Results
The experiment involved spring barley regenerants obtained through anther culture. Nine variants (trials) of induction media were created by adding copper (CuSO4: 0.1; 5; 10 µM) and silver salts (AgNO3: 0; 10; 60 µM), with varying incubation times for the anthers (21, 28, and 35 days). Changes in DNA methylation were estimated using the DArTseqMet molecular marker method, which also detects cytosine methylation.
Phenotype variability in β-glucans, SAM and GSH induced by the nutrient treatments was assessed using tentative assignments based on the Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy. The effectiveness of green plant regeneration ranged from 0.1 to 2.91 plants per 100 plated anthers. The level of demethylation ranged from 7.61 to 32.29, while de novo methylation reached values ranging from 6.83 to 32.27. The paper demonstrates that the samples from specific in vitro conditions (trials) formed tight groups linked to the factors contributing to the two main components responsible for 55.05% of the variance (to the first component DNM, DM, to the second component GSH, β-glucans, Cu(II), GPRE). Conclusions
We can conclude that in vitro tissue culture conditions affect biochemical levels, DNA methylation changes, and GPRE. Increasing Cu(II) concentration in the IM impacts the metabolism and DNA methylation, elevating GPRE. Thus, changing Cu(II) concentration in the IM is fair to expect to boost GPRE.
Metal ions in the induction medium are essential ingredients allowing green plant regeneration. F... more Metal ions in the induction medium are essential ingredients allowing green plant regeneration. For instance, Cu(II) and Ag(I) ions may affect the mitochondrial electron transport chain, influencing the Yang cycle and synthesis of S-adenosyl-L-methionine, the prominent donor of the methylation group for all cellular compounds, including cytosines. If the ion concentrations are not balanced, they can interfere with the proper flow of electrons in the respiratory chain and ATP production. Under oxidative stress, methylated cytosines might be subjected to mutations impacting green plant regeneration efficiency. Varying Cu(II) and Ag(I) concentrations in the induction medium and time of anther culture, nine trials of anther culture-derived regenerants of triticale were derived. The methylation-sensitive AFLP approach quantitative characteristics of tissue culture-induced variation, including sequence variation, DNA demethylation, and DNA de novo methylation for all symmetric-CG, CHG, an...
Green plant regeneration efficiency (GPRE) via in vitro anther culture results from biochemical p... more Green plant regeneration efficiency (GPRE) via in vitro anther culture results from biochemical pathways and cycle dysfunctions that may affect DNA and histone methylation, with gene expression influencing whole cell functioning. The reprogramming from gametophytic to sporophytic fate is part of the phenomenon. While DNA methylation and sequence changes related to the GPRE have been described, little attention was paid to the biochemical aspects of the phenomenon. Furthermore, only a few theoretical models that describe the complex relationships between biochemical aspects of GPRE and the role of Cu(II) ions in the induction medium and as cofactors of enzymatic reactions have been developed. Still, none of these models are devoted directly to the biochemical level. Fourier transform infrared (FTIR) spectroscopy was used in the current study to analyze triticale regenerants derived under various in vitro tissue culture conditions, including different Cu(II) and Ag(I) ion concentratio...
Babia Góra massif is the only site of occurrence of the Cerastium alpinum L. in Poland, an arctic... more Babia Góra massif is the only site of occurrence of the Cerastium alpinum L. in Poland, an arctic-alpine perennial plant with a wide distribution in North America, northwestern Asia, and Europe. To determine whether the isolated Polish populations are genetically distinct, we have performed an evaluation of C. alpinum from Babia Góra with the use of iPBS markers. A total number of 133 individuals of C. alpinum from seven populations representing four localizations of the species were analyzed, i.e., from Babia Góra (Poland), Alps (Switzerland), Nuolja massif (Sweden), and Kaffiøyra (Svalbard, Norway). Genetic analysis of all C. alpinum samples using eight PBS primers identified 262 bands, 79.4% of which were polymorphic. iPBS markers revealed low genetic diversity (average H e = 0.085) and high population differentiation (F ST = 0.617). AMOVA results confirmed that the majority of the genetic variation (62%) was recorded among populations. The grouping revealed by PCoA showed that C. alpinum from Svalbard is the most diverged population, C. alpinum from Switzerland and Sweden form a pair of similar populations, whereas C. alpinum from the Babia Góra form a heterogeneous group of four populations. Results of isolation by distance analysis suggested that the spatial distance is the most probable cause of the observed differentiation among populations. Although significant traces of a bottleneck effect were noted for all populations of C. alpinum from Babia Góra, the populations still maintain a low but significant level of genetic polymorphism. These results are of great importance for developing conservation strategies for this species in Poland.
The biological improvement of triticale, a cereal of increasing importance in agriculture, may be... more The biological improvement of triticale, a cereal of increasing importance in agriculture, may be accelerated via the production of doubled haploid lines using in vitro culture. Among the relevant factors affecting the culture efficiency are Cu(II) or Ag(I) acting, e.g., as cofactors of enzymes. The copper ions are known to positively affect green plant regeneration efficiency. However, the biochemical basis, mainly its role in the generation of in vitro-induced genetic and epigenetic variation and green plant regeneration efficiency, is not well understood. Here, we employed structural equation modeling to evaluate the relationship between de novo DNA methylation affecting the asymmetric context of CHH sequences, the methylation-sensitive Amplified Fragment Length Polymorphism related sequence variation, and the concentration of Cu(II) and Ag(I) ions in induction media, as well as their effect on S-adenosyl-L-methionine perturbations, observed using FTIR spectroscopy, and the green plant regeneration efficiency. Our results allowed the construction of a theory-based model reflecting the biological phenomena associated with green plant regeneration efficiency. Furthermore, it is shown that Cu(II) ions in induction media affect plant regeneration, and by manipulating their concentration, the regeneration efficiency can be altered. Additionally, S-adenosyl-L-methionine is involved in the efficiency of green plant regeneration through methylation of the asymmetric CHH sequence related to de novo methylation. This shows that the Yang cycle may impact the production of green regenerants.
Mitochondrial genomes have become an interesting object of evolutionary and systematic study both... more Mitochondrial genomes have become an interesting object of evolutionary and systematic study both for animals and plants, including angiosperms. Although the framework of the angiosperm phylogeny was built on the information derived from chloroplast and nuclear genes, mitochondrial sequences also revealed their usefulness in solving the phylogenetic issues at different levels of plant systematics. Here, we report for the first time the complete sequences of 26 protein-coding genes of eight Colobanthus species (Caryophyllaceae). Of these, 23 of them represented core mitochondrial genes, which are directly associated with the primary function of that organelle, and the remaining three genes represented a facultative set of mitochondrial genes. Comparative analysis of the identified genes revealed a generally high degree of sequence conservation. The Ka/Ks ratio was <1 for most of the genes, which indicated purifying selection. Only for rps12 was Ka/Ks > 1 in all studied species,...
Ilyonectria destructans is a pathogenic fungus causing root rot and other symptoms on trees and m... more Ilyonectria destructans is a pathogenic fungus causing root rot and other symptoms on trees and many crops. This paper analyses the mitochondrial genome of I. destructans and compares it with other published Nectriaceae mitogenomes. The I. destructans mitogenome appears as a circular DNA molecule of 42,895 bp and an overall GC content of 28.23%. It contains 28 protein-coding genes (15 core protein genes and 13 free-standing ORFs), two rRNAs and 27 tRNAs. The gene content and order were found to be conserved in the mitogenome of I. destructans and other Nectriaceae, although the genome size varies because of the variation in the number and length of intergenic regions and introns. For most core protein-coding genes in Nectriaceae species, Ka/Ks < 1 indicates purifying selection. Among some Nectriaceae representatives, only the rps3 gene was found under positive selection. Phylogenetic analyses based on nucleotide sequences of 15 protein-coding genes divided 45 Hypocreales species into six major clades matching the families Bionectriaceae, Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae, Hypocreaceae and Nectriaceae. I. destructans appeared as a sister species to unidentified Ilyonectia sp., closely related to C. ilicicola, N. cinnabarina and a clad of ten Fusarium species and G. moniliformis. The complete mitogenome of I. destructans reported in the current paper will facilitate the study of epidemiology, biology, genetic diversity of the species and the evolution of family Nectriace and the Hypocreales order.
Metal ions in the induction medium are essential ingredients allowing green plant regeneration. F... more Metal ions in the induction medium are essential ingredients allowing green plant regeneration. For instance, Cu(II) and Ag(I) ions may affect the mitochondrial electron transport chain, influencing the Yang cycle and synthesis of S-adenosyl-L-methionine, the prominent donor of the methylation group for all cellular compounds, including cytosines. If the ion concentrations are not balanced, they can interfere with the proper flow of electrons in the respiratory chain and ATP production. Under oxidative stress, methylated cytosines might be subjected to mutations impacting green plant regeneration efficiency. Varying Cu(II) and Ag(I) concentrations in the induction medium and time of anther culture, nine trials of anther culture-derived regenerants of triticale were derived. The methylation-sensitive AFLP approach quantitative characteristics of tissue culture-induced variation, including sequence variation, DNA demethylation, and DNA de novo methylation for all symmetric-CG, CHG, and asymmetric-CHH sequence contexts, were evaluated for all trials. In addition, the implementation of mediation analysis allowed evaluating relationships between factors influencing green plant regeneration efficiency. It was demonstrated that Cu(II) ions mediated relationships between: (1) de novo methylation in the CHH context and sequence variation in the CHH, (2) sequence variation in CHH and green plant regeneration efficiency, (3) de novo methylation in CHH sequences and green plant regeneration, (4) between sequence variation in the CHG context, and green plant regeneration efficiency. Cu(II) ions were not a mediator between de novo methylation in the CG context and green plant regeneration. The latter relationship was mediated by sequence variation in the CG context. On the other hand, we failed to identify any mediating action of Ag(I) ions or the moderating role of time. Furthermore, demethylation in any sequence context seems not to participate in any relationships leading to green plant regeneration, sequence variation, and the involvement of Cu(II) or Ag(I) as mediators.
This study investigated the details of the morphological and anatomical structure of the generati... more This study investigated the details of the morphological and anatomical structure of the generative organs of the Subantarctic flowering plant, belonging to the family Caryophyllaceae-Colobanthus apetalus (Labill.) Druce. The research material was collected in hostile natural conditions in Subantarctic regions, and also was grown in the incubators and the greenhouse of the University of Warmia and Mazury in Olsztyn (Poland). C. apetalus forms tufts with soft and grassy leaves and small greenish flowers that are more obvious than in other Colobanthus species. C. apetalus forms open (chasmogamic) flowers in greenhouse cultivation. The flowers most often form five stamens with two microsporangia. Over a dozen pollen grains are formed in each microsporangium. Studies of the plant material originated from natural conditions conducted by means of a light microscope, have shown that the ovules of the analyzed representative of the genus Colobanthus are anatropous, crassinucellar, and the monosporic embryo sac develops according to the Polygonum type (the most common type in angiosperms). C. apetalus plants underwent a full development cycle in greenhouse cultivation and produced fertile, perispermic seeds. During the C. apetalus growth in conditions at increased air humidity, the vivipary was also observed.
Deschampsia antarctica Desv. can be found in diverse Antarctic habitats which may vary considerab... more Deschampsia antarctica Desv. can be found in diverse Antarctic habitats which may vary considerably in terms of environmental conditions and soil properties. As a result , the species is characterized by wide ecotypic variation in terms of both morphological and anatomical traits. The species is a unique example of an organism that can successfully colonize inhospitable regions due to its phenomenal ability to adapt to both the local mosaic of microhabitats and to general climatic fluctuations. For this reason, D. antarctica has been widely investigated in studies analyzing morphophysi-ological and biochemical responses to various abiotic stresses (frost, drought, salinity, increased UV radiation). However, there is little evidence to indicate whether the observed polymorphism is accompanied by the corresponding genetic variation. In the present study, retrotransposon-based iPBS markers were used to trace the genetic variation of D. antarctica collected in nine sites of the Arctowski oasis on King George Island (Western Antarctic). The genotyping of 165 individuals from nine populations with seven iPBS primers revealed 125 amplification products, 15 of which (12%) were polymorphic, with an average of 5.6% polymorphic fragments per population. Only one of the polymorphic fragments, observed in population 6, was represented as a private band. The analyzed specimens were characterized by low genetic diversity (uH e = 0.021, I = 0.030) and high population differentiation (F ST = 0.4874). An analysis of Fu's F S statistics and mismatch distribution in most populations (excluding population 2, 6 and 9) revealed demographic/spatial expansion, whereas significant traces of reduction in effective population size were found in three populations (1, 3 and 5). The iPBS markers revealed genetic polymorphism of D. antarctica, which could be attributed to the mobilization of random transposable elements, unique features of reproductive biology, and/or geographic location of the examined populations.
The complete plastome sequences of six species were sequenced to better understand the evolutiona... more The complete plastome sequences of six species were sequenced to better understand the evolutionary relationships and mutation patterns in the chloroplast genome of the genus Colobanthus. the length of the chloroplast genome sequences of C. acicularis, C. affinis, C. lycopodioides, C. nivicola, C. pulvinatus and C. subulatus ranged from 151,050 to 151,462 bp. The quadripartite circular structure of these genome sequences has the same overall organization and gene content with 73 protein-coding genes, 30 tRNA genes, four rRNA genes and five conserved chloroplast open reading frames. A total of 153 repeat sequences were revealed. Forward repeats were dominant, whereas complementary repeats were found only in C. pulvinatus. the mononucleotide SSRs composed of A/t units were most common, and hexanucleotide SSRs were detected least often. eleven highly variable regions which could be utilized as potential markers for phylogeny reconstruction, species identification or phylogeography were identified within Colobanthus chloroplast genomes. Seventy-three protein-coding genes were used in phylogenetic analyses. Reconstructed phylogeny was consistent with the systematic position of the studied species, and the representatives of the same genus were grouped in one clade. All studied Colobanthus species formed a single group and C. lycopodioides was least similar to the remaining species.
Colletotrichum species form one of the most economically significant groups of pathogenic fungi a... more Colletotrichum species form one of the most economically significant groups of pathogenic fungi and lead to significant losses in the production of major crops-in particular, fruits, vegetables, ornamental plants, shrubs, and trees. Members of the genus Colletotrichum cause anthracnose disease in many plants. Due to their considerable variation, these fungi have been widely investigated in genetic studies as model organisms. Here, we report the complete mitochondrial genome sequences of four Colletotrichum species (C. fioriniae, C. lupini, C. salicis, and C. tamarilloi). The reported circular mitogenomes range from 30,020 (C. fioriniae) to 36,554 bp (C. lupini) in size and have identical sets of genes, including 15 protein-coding genes, two ribosomal RNA genes, and 29 tRNA genes. All four mitogenomes are characterized by a rather poor repetitive sequence content with only forward repeat representatives and a low number of microsatellites. The topology of the phylogenetic tree reflects the systematic positions of the studied species, with representatives of each Colletotrichum species complex gathered in one clade. A comparative analysis reveals consistency in the gene composition and order of Colletotrichum mitogenomes, although some highly divergent regions are also identified, like the rps3 gene which appears as a source of potential diagnostic markers for all studied Colletotrichum species.
Provenance trials were designed to analyse the quantitative responses of tree species to envi-ron... more Provenance trials were designed to analyse the quantitative responses of tree species to envi-ronmental variables found in different experiment location. However, we have still limited knowledge how natural and artificial selection affects genetic variation of the species populations gather in such experi-mental sites. We have used bulked DNA-based RAPD and ISJ analysis to investigate genetic diversity and differentiation of Scots pine populations from two Polish locations of IUFRO 1982 provenance trial placed in Kórnik and in Supraśl. Applied categories of DNA markers differed in terms of revealing genetic diversity of the species. Ten RAPD primers applied in the study yielded a total of 75 bands, of which 21 (28%) and 15 (20%) were polymorphic in Kórnik and in Supraśl, respectively. Six ISJ primers revealed 42 bands of which 4 (9.52%) and 14 (33.3%) were polymorphic in Kórnik and in Supraśl, respectively. The genetic diversity and differentiation was low, as expressed by He=0.071 and He=0.085, and by genetic distance values which ranged from 0.0 to 0.240 (on average 0.081) and from 0.017 to 0.188 (on average 0.094) for Kórnik and Su-praśl, respectively. Location of provenance trial appeared to have a significant influence on revealed level of genetic polymorphism and pattern of interpopulation differentiation. However, genetic structure found for analysed Scots pine provenances from IUFRO 1982 in Kórnik was also confirmed for Supraśl experimental site. In the light of available data we also discussed the influence of historical migration routes and gene flow on observed genetic variation of the species.
We studied an invasion of Poa annua on King George Island (Maritime Antarctic). The remoteness of... more We studied an invasion of Poa annua on King George Island (Maritime Antarctic). The remoteness of this location, its geographic isolation, and its limited human traffic provided an opportunity to trace the history of an invasion of the species. Poa annua was recorded for the first time at H. Arctowski Polish Antarctic Station in the austral summer of 1985/6. In 2008/9, the species was observed in a new locality at the Ecology Glacier Forefield (1.5 km from "Arctowski"). We used AFLP to analyze the genetic differences among three populations of P. annua: the two mentioned above (Station and Forefield) and the putative origin of the introduction, Warsaw (Poland). There was 38% genetic variance among the populations. Pairwise ФPT was 0.498 between the Forefield and Warsaw populations and 0.283 between Warsaw and Station. There were 15 unique bands in the Warsaw population (frequency from 6% to 100%) and one in the Station/Forefield populations (which appears in all analyzed i...
Antarctic pearlwort (Colobanthus quitensis) is one of the flowering plant species considered nati... more Antarctic pearlwort (Colobanthus quitensis) is one of the flowering plant species considered native to maritime Antarctica. Although the species was intensively analyzed towards its morphological, anatomical and physiological adaptation to local environment, its genetic variability is still poorly studied. In the presented study, a recently developed retrotransposon-based DNA marker system (inter Primer Binding Site – iPBS) was applied to assess the genetic diversity and differentiation of C. quitensis populations from King George Island (South Shetland Islands, West Antarctic). A total of 143 scoreable bands were detected using 7 iPBS primers among 122 plant specimens representing 8 populations. 55 (38.5%) bands were found polymorphic, with an average of 14.3% polymorphic fragments per primer. Nine of all observed fragments were represented as a private bands deployed unevenly among populations. Low genetic diversity (on average H
DNA markers have become effective tools in genetic diversity studies of forest trees. However, mo... more DNA markers have become effective tools in genetic diversity studies of forest trees. However, molecular marker analyses are associated with laborious and costly effort. One of the possibilities to overcome these constraints is to analyze bulked samples per population, rather than individual plants. We have used bulked DNA-based AFLP analysis to investigate genetic variation in Pinus sylvestris L. (Scots pine) from the IUFRO 1982 provenance trial in Kórnik (western Poland). Four AFLP primer combinations yielded a total of 309 bands, of which 208 (67.31%) were polymorphic. Thirty-six (11.65%) unique alleles were deployed randomly among the populations. Estimated genetic diversity and differentiation was high, as expressed by He = 0.238 and I = 0.356, and by genetic distance values which ranged from 0.154 to 0.363. A geographic pattern of interpopulation differentiation was observed, pointing at the individual character of populations from northeastern Europe. In the light of available data, we discuss the influence of historical migration routes, gene flow and human activity on observed genetic diversity and differentiation of Scots pine in Europe. Our results indicate that the AFLP method applied to DNA templates extracted from bulked leaf samples provides an efficient approach to elucidate genetic diversity and relationships among Scots pine populations.
The aim of the studies was to evaluate the efficiency of the KatG gene based B-SAP markers as a t... more The aim of the studies was to evaluate the efficiency of the KatG gene based B-SAP markers as a tool to distinguish morphologically diversified and geographically distant Scots pine populations and to track the routes of migrations. The 19 populations growing in the IUFRO 1982 provenance experiment and representing the natural distribution of the species in Europe were scored using 103 B-SAP loci. Among them 26% loci were polymorphic. The level of polymorphism was associated with the location of primers on the KatG template. The diversity was low, He = 0.086, and deposited mostly among populations. Seven unique markers were found that identified populations and likely they were associated with morphology. The overall genetic identity was relatively low, I = 0.933 (D = 0.069). The block of six B-SAP markers discriminated populations into two groups in agreement with their geographic origin and thereby further described as the North and the South. The North group was uniform with genetic diversity, He = 0.026 and the overall genetic distance D = 0.022. Presumably, it migrated from refugia in the Alps via France, northern Germany and Denmark, to Scandinavia and Russia. The South group was heterogeneous with He = 0.063 and D = 0.047. This group migrated from the Carpathians via Slovakia to Germany and Poland. The Balkans and Asian refugia did not take part in recolonization of Europe. The block of six B-SAP/KatG markers can be recommended for tracking postglacial history of Scots pine.
The genus Cerastium includes about 200 species that are mostly found in the temperate climates of... more The genus Cerastium includes about 200 species that are mostly found in the temperate climates of the Northern Hemisphere. Here we report the complete chloroplast genomes of Cerastium alpinum, C. arcticum and C. nigrescens. The length of cp genomes ranged from 147,940 to 148,722 bp. Their quadripartite circular structure had the same gene organization and content, containing 79 proteincoding genes, 30 tRNA genes, and four rRNA genes. Repeat sequences varied from 16 to 23 per species, with palindromic repeats being the most frequent. The number of identified SSRs ranged from 20 to 23 per species and they were mainly composed of mononucleotide repeats containing A/T units. Based on Ka/Ks ratio values, most genes were subjected to purifying selection. The newly sequenced chloroplast genomes were characterized by a high frequency of RNA editing, including both C to U and U to C conversion. The phylogenetic relationships within the genus Cerastium and family Caryophyllaceae were reconstructed based on the sequences of 71 protein-coding genes. The topology of the phylogenetic tree was consistent with the systematic position of the studied species. All representatives of the genus Cerastium were gathered in a single clade with C. glomeratum sharing the least similarity with the others.
Background The development of the plant in vitro techniques has brought about the variation ident... more Background The development of the plant in vitro techniques has brought about the variation identified in regenerants known as somaclonal or tissue culture-induced variation (TCIV). S-adenosyl-L-methionine (SAM), glutathione (GSH), low methylated pectins (LMP), and Cu(II) ions may be implicated in green plant regeneration efficiency (GPRE) and TCIV, according to studies in barley (Hordeum vulgare L.) and partially in triticale (× Triticosecale spp. Wittmack ex A. Camus 1927). Using structural equation models (SEM), these metabolites have been connected to the metabolic pathways (Krebs and Yang cycles, glycolysis, transsulfuration), but not for triticale. Using metabolomic and (epi)genetic data, the study sought to develop a triticale regeneration efficiency statistical model. The culture’s induction medium was supplemented with various quantities of Cu(II) and Ag(I) ions for regeneration. The period of plant regeneration has also changed. The donor plant, anther-derived regenerants, and metAFLP were utilized to analyze TCIV concerning DNA in symmetric (CG, CHG) and asymmetric (CHH) sequence contexts. Attenuated Total Reflectance–Fourier Transfer Infrared (ATR-FTIR) spectroscopy was used to gather the metabolomic information on LMP, SAM, and GSH. To frame the data, a structural equation model was employed. Results According to metAFLP analysis, the average sequence change in the CHH context was 8.65%, and 0.58% was de novo methylation. Absorbances of FTIR spectra in regions specific for LMP, SAM, and GSH were used as variables values introduced to the SEM model. The average number of green regenerants per 100 plated anthers was 2.55. Conclusions The amounts of pectin demethylation, SAM, de novo methylation, and GSH are connected in the model to explain GPRE. By altering the concentration of Cu(II) ions in the medium, which influences the amount of pectin, triticale’s GPRE can be increased.
Scots pine is the foundation species of diverse forested ecosystems across Eurasia and displays r... more Scots pine is the foundation species of diverse forested ecosystems across Eurasia and displays remarkable ecological breadth, occurring in environments ranging from temperate rainforests to arid tundra margins. Such expansive distributions can be favored by various demographic and adaptive processes and the interactions between them. To understand the impact of neutral and selective forces on genetic structure in Scots pine, we conducted range‐wide population genetic analyses on 2321 trees from 202 populations using genotyping‐by‐sequencing, reconstructed the recent demography of the species and examined signals of genetic adaptation. We found a high and uniform genetic diversity across the entire range (global FST 0.048), no increased genetic load in expanding populations and minor impact of the last glacial maximum on historical population sizes. Genetic‐environmental associations identified only a handful of single‐nucleotide polymorphisms significantly linked to environmental gradients. The results suggest that extensive gene flow is predominantly responsible for the observed genetic patterns in Scots pine. The apparent missing signal of genetic adaptation is likely attributed to the intricate genetic architecture controlling adaptation to multi‐dimensional environments. The panmixia metapopulation of Scots pine offers a good study system for further exploration into how genetic adaptation and plasticity evolve under gene flow and changing environment.
Background
Microspore embryogenesis is a process that produces doubled haploids in tissue cultur... more Background
Microspore embryogenesis is a process that produces doubled haploids in tissue culture environments and is widely used in cereal plants. The efficient production of green regenerants requires stresses that could be sensed at the level of glycolysis, followed by the Krebs cycle and electron transfer chain. The latter can be affected by Cu(II) ion concentration in the induction media acting as cofactors of biochemical reactions, indirectly influencing the production of glutathione (GSH) and S-adenosyl-L-methionine (SAM) and thereby affecting epigenetic mechanisms involving DNA methylation (demethylation—DM, de novo methylation—DNM). The conclusions mentioned were acquired from research on triticale regenerants, but there is no similar research on barley. In this way, the study looks at how DNM, DM, Cu(II), SAM, GSH, and β-glucan affect the ability of green plant regeneration efficiency (GPRE). Results
The experiment involved spring barley regenerants obtained through anther culture. Nine variants (trials) of induction media were created by adding copper (CuSO4: 0.1; 5; 10 µM) and silver salts (AgNO3: 0; 10; 60 µM), with varying incubation times for the anthers (21, 28, and 35 days). Changes in DNA methylation were estimated using the DArTseqMet molecular marker method, which also detects cytosine methylation.
Phenotype variability in β-glucans, SAM and GSH induced by the nutrient treatments was assessed using tentative assignments based on the Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy. The effectiveness of green plant regeneration ranged from 0.1 to 2.91 plants per 100 plated anthers. The level of demethylation ranged from 7.61 to 32.29, while de novo methylation reached values ranging from 6.83 to 32.27. The paper demonstrates that the samples from specific in vitro conditions (trials) formed tight groups linked to the factors contributing to the two main components responsible for 55.05% of the variance (to the first component DNM, DM, to the second component GSH, β-glucans, Cu(II), GPRE). Conclusions
We can conclude that in vitro tissue culture conditions affect biochemical levels, DNA methylation changes, and GPRE. Increasing Cu(II) concentration in the IM impacts the metabolism and DNA methylation, elevating GPRE. Thus, changing Cu(II) concentration in the IM is fair to expect to boost GPRE.
Metal ions in the induction medium are essential ingredients allowing green plant regeneration. F... more Metal ions in the induction medium are essential ingredients allowing green plant regeneration. For instance, Cu(II) and Ag(I) ions may affect the mitochondrial electron transport chain, influencing the Yang cycle and synthesis of S-adenosyl-L-methionine, the prominent donor of the methylation group for all cellular compounds, including cytosines. If the ion concentrations are not balanced, they can interfere with the proper flow of electrons in the respiratory chain and ATP production. Under oxidative stress, methylated cytosines might be subjected to mutations impacting green plant regeneration efficiency. Varying Cu(II) and Ag(I) concentrations in the induction medium and time of anther culture, nine trials of anther culture-derived regenerants of triticale were derived. The methylation-sensitive AFLP approach quantitative characteristics of tissue culture-induced variation, including sequence variation, DNA demethylation, and DNA de novo methylation for all symmetric-CG, CHG, an...
Green plant regeneration efficiency (GPRE) via in vitro anther culture results from biochemical p... more Green plant regeneration efficiency (GPRE) via in vitro anther culture results from biochemical pathways and cycle dysfunctions that may affect DNA and histone methylation, with gene expression influencing whole cell functioning. The reprogramming from gametophytic to sporophytic fate is part of the phenomenon. While DNA methylation and sequence changes related to the GPRE have been described, little attention was paid to the biochemical aspects of the phenomenon. Furthermore, only a few theoretical models that describe the complex relationships between biochemical aspects of GPRE and the role of Cu(II) ions in the induction medium and as cofactors of enzymatic reactions have been developed. Still, none of these models are devoted directly to the biochemical level. Fourier transform infrared (FTIR) spectroscopy was used in the current study to analyze triticale regenerants derived under various in vitro tissue culture conditions, including different Cu(II) and Ag(I) ion concentratio...
Babia Góra massif is the only site of occurrence of the Cerastium alpinum L. in Poland, an arctic... more Babia Góra massif is the only site of occurrence of the Cerastium alpinum L. in Poland, an arctic-alpine perennial plant with a wide distribution in North America, northwestern Asia, and Europe. To determine whether the isolated Polish populations are genetically distinct, we have performed an evaluation of C. alpinum from Babia Góra with the use of iPBS markers. A total number of 133 individuals of C. alpinum from seven populations representing four localizations of the species were analyzed, i.e., from Babia Góra (Poland), Alps (Switzerland), Nuolja massif (Sweden), and Kaffiøyra (Svalbard, Norway). Genetic analysis of all C. alpinum samples using eight PBS primers identified 262 bands, 79.4% of which were polymorphic. iPBS markers revealed low genetic diversity (average H e = 0.085) and high population differentiation (F ST = 0.617). AMOVA results confirmed that the majority of the genetic variation (62%) was recorded among populations. The grouping revealed by PCoA showed that C. alpinum from Svalbard is the most diverged population, C. alpinum from Switzerland and Sweden form a pair of similar populations, whereas C. alpinum from the Babia Góra form a heterogeneous group of four populations. Results of isolation by distance analysis suggested that the spatial distance is the most probable cause of the observed differentiation among populations. Although significant traces of a bottleneck effect were noted for all populations of C. alpinum from Babia Góra, the populations still maintain a low but significant level of genetic polymorphism. These results are of great importance for developing conservation strategies for this species in Poland.
The biological improvement of triticale, a cereal of increasing importance in agriculture, may be... more The biological improvement of triticale, a cereal of increasing importance in agriculture, may be accelerated via the production of doubled haploid lines using in vitro culture. Among the relevant factors affecting the culture efficiency are Cu(II) or Ag(I) acting, e.g., as cofactors of enzymes. The copper ions are known to positively affect green plant regeneration efficiency. However, the biochemical basis, mainly its role in the generation of in vitro-induced genetic and epigenetic variation and green plant regeneration efficiency, is not well understood. Here, we employed structural equation modeling to evaluate the relationship between de novo DNA methylation affecting the asymmetric context of CHH sequences, the methylation-sensitive Amplified Fragment Length Polymorphism related sequence variation, and the concentration of Cu(II) and Ag(I) ions in induction media, as well as their effect on S-adenosyl-L-methionine perturbations, observed using FTIR spectroscopy, and the green plant regeneration efficiency. Our results allowed the construction of a theory-based model reflecting the biological phenomena associated with green plant regeneration efficiency. Furthermore, it is shown that Cu(II) ions in induction media affect plant regeneration, and by manipulating their concentration, the regeneration efficiency can be altered. Additionally, S-adenosyl-L-methionine is involved in the efficiency of green plant regeneration through methylation of the asymmetric CHH sequence related to de novo methylation. This shows that the Yang cycle may impact the production of green regenerants.
Mitochondrial genomes have become an interesting object of evolutionary and systematic study both... more Mitochondrial genomes have become an interesting object of evolutionary and systematic study both for animals and plants, including angiosperms. Although the framework of the angiosperm phylogeny was built on the information derived from chloroplast and nuclear genes, mitochondrial sequences also revealed their usefulness in solving the phylogenetic issues at different levels of plant systematics. Here, we report for the first time the complete sequences of 26 protein-coding genes of eight Colobanthus species (Caryophyllaceae). Of these, 23 of them represented core mitochondrial genes, which are directly associated with the primary function of that organelle, and the remaining three genes represented a facultative set of mitochondrial genes. Comparative analysis of the identified genes revealed a generally high degree of sequence conservation. The Ka/Ks ratio was <1 for most of the genes, which indicated purifying selection. Only for rps12 was Ka/Ks > 1 in all studied species,...
Ilyonectria destructans is a pathogenic fungus causing root rot and other symptoms on trees and m... more Ilyonectria destructans is a pathogenic fungus causing root rot and other symptoms on trees and many crops. This paper analyses the mitochondrial genome of I. destructans and compares it with other published Nectriaceae mitogenomes. The I. destructans mitogenome appears as a circular DNA molecule of 42,895 bp and an overall GC content of 28.23%. It contains 28 protein-coding genes (15 core protein genes and 13 free-standing ORFs), two rRNAs and 27 tRNAs. The gene content and order were found to be conserved in the mitogenome of I. destructans and other Nectriaceae, although the genome size varies because of the variation in the number and length of intergenic regions and introns. For most core protein-coding genes in Nectriaceae species, Ka/Ks < 1 indicates purifying selection. Among some Nectriaceae representatives, only the rps3 gene was found under positive selection. Phylogenetic analyses based on nucleotide sequences of 15 protein-coding genes divided 45 Hypocreales species into six major clades matching the families Bionectriaceae, Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae, Hypocreaceae and Nectriaceae. I. destructans appeared as a sister species to unidentified Ilyonectia sp., closely related to C. ilicicola, N. cinnabarina and a clad of ten Fusarium species and G. moniliformis. The complete mitogenome of I. destructans reported in the current paper will facilitate the study of epidemiology, biology, genetic diversity of the species and the evolution of family Nectriace and the Hypocreales order.
Metal ions in the induction medium are essential ingredients allowing green plant regeneration. F... more Metal ions in the induction medium are essential ingredients allowing green plant regeneration. For instance, Cu(II) and Ag(I) ions may affect the mitochondrial electron transport chain, influencing the Yang cycle and synthesis of S-adenosyl-L-methionine, the prominent donor of the methylation group for all cellular compounds, including cytosines. If the ion concentrations are not balanced, they can interfere with the proper flow of electrons in the respiratory chain and ATP production. Under oxidative stress, methylated cytosines might be subjected to mutations impacting green plant regeneration efficiency. Varying Cu(II) and Ag(I) concentrations in the induction medium and time of anther culture, nine trials of anther culture-derived regenerants of triticale were derived. The methylation-sensitive AFLP approach quantitative characteristics of tissue culture-induced variation, including sequence variation, DNA demethylation, and DNA de novo methylation for all symmetric-CG, CHG, and asymmetric-CHH sequence contexts, were evaluated for all trials. In addition, the implementation of mediation analysis allowed evaluating relationships between factors influencing green plant regeneration efficiency. It was demonstrated that Cu(II) ions mediated relationships between: (1) de novo methylation in the CHH context and sequence variation in the CHH, (2) sequence variation in CHH and green plant regeneration efficiency, (3) de novo methylation in CHH sequences and green plant regeneration, (4) between sequence variation in the CHG context, and green plant regeneration efficiency. Cu(II) ions were not a mediator between de novo methylation in the CG context and green plant regeneration. The latter relationship was mediated by sequence variation in the CG context. On the other hand, we failed to identify any mediating action of Ag(I) ions or the moderating role of time. Furthermore, demethylation in any sequence context seems not to participate in any relationships leading to green plant regeneration, sequence variation, and the involvement of Cu(II) or Ag(I) as mediators.
This study investigated the details of the morphological and anatomical structure of the generati... more This study investigated the details of the morphological and anatomical structure of the generative organs of the Subantarctic flowering plant, belonging to the family Caryophyllaceae-Colobanthus apetalus (Labill.) Druce. The research material was collected in hostile natural conditions in Subantarctic regions, and also was grown in the incubators and the greenhouse of the University of Warmia and Mazury in Olsztyn (Poland). C. apetalus forms tufts with soft and grassy leaves and small greenish flowers that are more obvious than in other Colobanthus species. C. apetalus forms open (chasmogamic) flowers in greenhouse cultivation. The flowers most often form five stamens with two microsporangia. Over a dozen pollen grains are formed in each microsporangium. Studies of the plant material originated from natural conditions conducted by means of a light microscope, have shown that the ovules of the analyzed representative of the genus Colobanthus are anatropous, crassinucellar, and the monosporic embryo sac develops according to the Polygonum type (the most common type in angiosperms). C. apetalus plants underwent a full development cycle in greenhouse cultivation and produced fertile, perispermic seeds. During the C. apetalus growth in conditions at increased air humidity, the vivipary was also observed.
Deschampsia antarctica Desv. can be found in diverse Antarctic habitats which may vary considerab... more Deschampsia antarctica Desv. can be found in diverse Antarctic habitats which may vary considerably in terms of environmental conditions and soil properties. As a result , the species is characterized by wide ecotypic variation in terms of both morphological and anatomical traits. The species is a unique example of an organism that can successfully colonize inhospitable regions due to its phenomenal ability to adapt to both the local mosaic of microhabitats and to general climatic fluctuations. For this reason, D. antarctica has been widely investigated in studies analyzing morphophysi-ological and biochemical responses to various abiotic stresses (frost, drought, salinity, increased UV radiation). However, there is little evidence to indicate whether the observed polymorphism is accompanied by the corresponding genetic variation. In the present study, retrotransposon-based iPBS markers were used to trace the genetic variation of D. antarctica collected in nine sites of the Arctowski oasis on King George Island (Western Antarctic). The genotyping of 165 individuals from nine populations with seven iPBS primers revealed 125 amplification products, 15 of which (12%) were polymorphic, with an average of 5.6% polymorphic fragments per population. Only one of the polymorphic fragments, observed in population 6, was represented as a private band. The analyzed specimens were characterized by low genetic diversity (uH e = 0.021, I = 0.030) and high population differentiation (F ST = 0.4874). An analysis of Fu's F S statistics and mismatch distribution in most populations (excluding population 2, 6 and 9) revealed demographic/spatial expansion, whereas significant traces of reduction in effective population size were found in three populations (1, 3 and 5). The iPBS markers revealed genetic polymorphism of D. antarctica, which could be attributed to the mobilization of random transposable elements, unique features of reproductive biology, and/or geographic location of the examined populations.
The complete plastome sequences of six species were sequenced to better understand the evolutiona... more The complete plastome sequences of six species were sequenced to better understand the evolutionary relationships and mutation patterns in the chloroplast genome of the genus Colobanthus. the length of the chloroplast genome sequences of C. acicularis, C. affinis, C. lycopodioides, C. nivicola, C. pulvinatus and C. subulatus ranged from 151,050 to 151,462 bp. The quadripartite circular structure of these genome sequences has the same overall organization and gene content with 73 protein-coding genes, 30 tRNA genes, four rRNA genes and five conserved chloroplast open reading frames. A total of 153 repeat sequences were revealed. Forward repeats were dominant, whereas complementary repeats were found only in C. pulvinatus. the mononucleotide SSRs composed of A/t units were most common, and hexanucleotide SSRs were detected least often. eleven highly variable regions which could be utilized as potential markers for phylogeny reconstruction, species identification or phylogeography were identified within Colobanthus chloroplast genomes. Seventy-three protein-coding genes were used in phylogenetic analyses. Reconstructed phylogeny was consistent with the systematic position of the studied species, and the representatives of the same genus were grouped in one clade. All studied Colobanthus species formed a single group and C. lycopodioides was least similar to the remaining species.
Colletotrichum species form one of the most economically significant groups of pathogenic fungi a... more Colletotrichum species form one of the most economically significant groups of pathogenic fungi and lead to significant losses in the production of major crops-in particular, fruits, vegetables, ornamental plants, shrubs, and trees. Members of the genus Colletotrichum cause anthracnose disease in many plants. Due to their considerable variation, these fungi have been widely investigated in genetic studies as model organisms. Here, we report the complete mitochondrial genome sequences of four Colletotrichum species (C. fioriniae, C. lupini, C. salicis, and C. tamarilloi). The reported circular mitogenomes range from 30,020 (C. fioriniae) to 36,554 bp (C. lupini) in size and have identical sets of genes, including 15 protein-coding genes, two ribosomal RNA genes, and 29 tRNA genes. All four mitogenomes are characterized by a rather poor repetitive sequence content with only forward repeat representatives and a low number of microsatellites. The topology of the phylogenetic tree reflects the systematic positions of the studied species, with representatives of each Colletotrichum species complex gathered in one clade. A comparative analysis reveals consistency in the gene composition and order of Colletotrichum mitogenomes, although some highly divergent regions are also identified, like the rps3 gene which appears as a source of potential diagnostic markers for all studied Colletotrichum species.
Provenance trials were designed to analyse the quantitative responses of tree species to envi-ron... more Provenance trials were designed to analyse the quantitative responses of tree species to envi-ronmental variables found in different experiment location. However, we have still limited knowledge how natural and artificial selection affects genetic variation of the species populations gather in such experi-mental sites. We have used bulked DNA-based RAPD and ISJ analysis to investigate genetic diversity and differentiation of Scots pine populations from two Polish locations of IUFRO 1982 provenance trial placed in Kórnik and in Supraśl. Applied categories of DNA markers differed in terms of revealing genetic diversity of the species. Ten RAPD primers applied in the study yielded a total of 75 bands, of which 21 (28%) and 15 (20%) were polymorphic in Kórnik and in Supraśl, respectively. Six ISJ primers revealed 42 bands of which 4 (9.52%) and 14 (33.3%) were polymorphic in Kórnik and in Supraśl, respectively. The genetic diversity and differentiation was low, as expressed by He=0.071 and He=0.085, and by genetic distance values which ranged from 0.0 to 0.240 (on average 0.081) and from 0.017 to 0.188 (on average 0.094) for Kórnik and Su-praśl, respectively. Location of provenance trial appeared to have a significant influence on revealed level of genetic polymorphism and pattern of interpopulation differentiation. However, genetic structure found for analysed Scots pine provenances from IUFRO 1982 in Kórnik was also confirmed for Supraśl experimental site. In the light of available data we also discussed the influence of historical migration routes and gene flow on observed genetic variation of the species.
We studied an invasion of Poa annua on King George Island (Maritime Antarctic). The remoteness of... more We studied an invasion of Poa annua on King George Island (Maritime Antarctic). The remoteness of this location, its geographic isolation, and its limited human traffic provided an opportunity to trace the history of an invasion of the species. Poa annua was recorded for the first time at H. Arctowski Polish Antarctic Station in the austral summer of 1985/6. In 2008/9, the species was observed in a new locality at the Ecology Glacier Forefield (1.5 km from "Arctowski"). We used AFLP to analyze the genetic differences among three populations of P. annua: the two mentioned above (Station and Forefield) and the putative origin of the introduction, Warsaw (Poland). There was 38% genetic variance among the populations. Pairwise ФPT was 0.498 between the Forefield and Warsaw populations and 0.283 between Warsaw and Station. There were 15 unique bands in the Warsaw population (frequency from 6% to 100%) and one in the Station/Forefield populations (which appears in all analyzed i...
Antarctic pearlwort (Colobanthus quitensis) is one of the flowering plant species considered nati... more Antarctic pearlwort (Colobanthus quitensis) is one of the flowering plant species considered native to maritime Antarctica. Although the species was intensively analyzed towards its morphological, anatomical and physiological adaptation to local environment, its genetic variability is still poorly studied. In the presented study, a recently developed retrotransposon-based DNA marker system (inter Primer Binding Site – iPBS) was applied to assess the genetic diversity and differentiation of C. quitensis populations from King George Island (South Shetland Islands, West Antarctic). A total of 143 scoreable bands were detected using 7 iPBS primers among 122 plant specimens representing 8 populations. 55 (38.5%) bands were found polymorphic, with an average of 14.3% polymorphic fragments per primer. Nine of all observed fragments were represented as a private bands deployed unevenly among populations. Low genetic diversity (on average H
DNA markers have become effective tools in genetic diversity studies of forest trees. However, mo... more DNA markers have become effective tools in genetic diversity studies of forest trees. However, molecular marker analyses are associated with laborious and costly effort. One of the possibilities to overcome these constraints is to analyze bulked samples per population, rather than individual plants. We have used bulked DNA-based AFLP analysis to investigate genetic variation in Pinus sylvestris L. (Scots pine) from the IUFRO 1982 provenance trial in Kórnik (western Poland). Four AFLP primer combinations yielded a total of 309 bands, of which 208 (67.31%) were polymorphic. Thirty-six (11.65%) unique alleles were deployed randomly among the populations. Estimated genetic diversity and differentiation was high, as expressed by He = 0.238 and I = 0.356, and by genetic distance values which ranged from 0.154 to 0.363. A geographic pattern of interpopulation differentiation was observed, pointing at the individual character of populations from northeastern Europe. In the light of available data, we discuss the influence of historical migration routes, gene flow and human activity on observed genetic diversity and differentiation of Scots pine in Europe. Our results indicate that the AFLP method applied to DNA templates extracted from bulked leaf samples provides an efficient approach to elucidate genetic diversity and relationships among Scots pine populations.
The aim of the studies was to evaluate the efficiency of the KatG gene based B-SAP markers as a t... more The aim of the studies was to evaluate the efficiency of the KatG gene based B-SAP markers as a tool to distinguish morphologically diversified and geographically distant Scots pine populations and to track the routes of migrations. The 19 populations growing in the IUFRO 1982 provenance experiment and representing the natural distribution of the species in Europe were scored using 103 B-SAP loci. Among them 26% loci were polymorphic. The level of polymorphism was associated with the location of primers on the KatG template. The diversity was low, He = 0.086, and deposited mostly among populations. Seven unique markers were found that identified populations and likely they were associated with morphology. The overall genetic identity was relatively low, I = 0.933 (D = 0.069). The block of six B-SAP markers discriminated populations into two groups in agreement with their geographic origin and thereby further described as the North and the South. The North group was uniform with genetic diversity, He = 0.026 and the overall genetic distance D = 0.022. Presumably, it migrated from refugia in the Alps via France, northern Germany and Denmark, to Scandinavia and Russia. The South group was heterogeneous with He = 0.063 and D = 0.047. This group migrated from the Carpathians via Slovakia to Germany and Poland. The Balkans and Asian refugia did not take part in recolonization of Europe. The block of six B-SAP/KatG markers can be recommended for tracking postglacial history of Scots pine.
The crucial aim of this work was to compare four relict Scots pine populations in the Pieniny Wła... more The crucial aim of this work was to compare four relict Scots pine populations in the Pieniny Właściwe range by using morphological characteristic of their needles. The material was taken from the trees growing in the following stands: Sokolica, Czerwone Skałki, Biała Skała, Zamkowa Góra. Ten needles collected from twelve different trees found on each stand were taken for measurement in October 2003. Seven morphological traits of two-year old needles were studied: length of the needles, number of stomatal rows on the flat and convex sides of the needle, number of stomata per 2 mm of the needle length on its fl at and convex sides, number of serrations per 2 mm of the needle length on its right and left edges.
For the investigation two populations of Scots pine were selected, which originated from Puszcza ... more For the investigation two populations of Scots pine were selected, which originated from Puszcza Notecka: 1 (Chlebowo), appearing on peat-based soils and 2 (Bąblin) - trees of this population grow on podzolic soils. For the analyses of morphological needle characteristics, three hundred needles were sorted out from each of the thirty trees of population in ten repetitions per each tree. Seven morphological needle characteristics were analyzed, describing: their length, number of stomatal rows on the flat and the convex side of the needle, average number of stomata per 2 mm of the needle length on both sides and the number of characteristic serrations on both edges of the needle. The use of Snedecore’s F statistics has revealed that the two populations differ significantly from each other as regards the needle length and stomatal row number on both sides of the needle. Each of the population has characteristic, individual character correlations.
We investigated the variation of five needle morphological traits of the Scots pine (Pinus sylves... more We investigated the variation of five needle morphological traits of the Scots pine (Pinus sylvestris L.) in four populations from the Tatra Mts. The greatest variation was found in needle length, number of stomatal rows, both on the adaxial and abaxial side of leaf, whereas the number of stomata per 2 mm of the needle length, for both sides of the leaf, showed only small intrapopulational variation. The results obtained by discriminant analysis show the distinct character of Scots pine populations from the Tatra National Park, particularly from the 'Filipczańskie Skałki' locality. 'Filipczańskie Skałki' is a small, isolated population differing in the number of stomata and stomatal rows.
The crucial aim of this work was to compare four relict Scots pine populations in the Pieniny Wła... more The crucial aim of this work was to compare four relict Scots pine populations in the Pieniny Właściwe range by using morphological character-istic of their needles. The material was taken from the trees growing in the following stands: Sokolica, Czerwone Skałki, Biała Skała, Zamkowa Góra. Ten needles collected from twelve different trees found on each stand were taken for measurement in October 2003. Seven morphological traits of two-year old needles were studied: length of the needles, number of stomatal rows on the fl at and convex sides of the needle, number of stomata per 2 mm of the needle length on its fl at and convex sides, number of serrations per 2 mm of the needle length on its right and left edges.
To evaluate the geographic variability of Pinus sylvestris populations seven morphological traits... more To evaluate the geographic variability of Pinus sylvestris populations seven morphological traits of needles of pines from IUFRO 1982 provenance trial have been analyzed. The studied populations originated from northern (>55°N in Russia, Sweden and Latvia), central (55-47°N in Poland, Germany, Belgium, France, Slovakia) andsouthern (<47°N in Hungary, Bosnia, Montenegro andTurkey) European ranges of Scots pine. The analyzedprovenance trial experimental areas were locatedin Kórnik (western Poland) andin Supraśl (north-eastern Poland). The greatest variation was found in needle length and number of stomata rows on the flat and convex side of a needle, whereas number of stomata per 2mmof needle length on flat and convex side of a needle was stable, with minor interpopulational variation. Biometrical analyses revealed a significant population × location interaction anda geographical pattern in interpopulational differentiation in both experimental sites, with the northern andsouthern European Scots pine groups of provenancesdiffering significantly from the group of central origin. The results obtainedare compatible with previous results of studies on provenance variability of the Scots pine from IUFRO 1982. In the light of available data, the influence of the Balkan glacial refugia of Pinus sylvestris on a present genetic diversity of this species in Europe and the reconstruction of Scots pine migration routes after the last glacial period are discussed.
Efficient use of any breeding resources requires a good understanding of the genetic value of the... more Efficient use of any breeding resources requires a good understanding of the genetic value of the founder breeding materials for predicting the gain and diversity in future generations. This study evaluates the distribution of genetic variation and level of relatedness among and within nine breeding populations of Norway spruce for Northern Sweden using nuclear microsatellite markers. A sample set of 456 individuals selected from 140 stands were genotyped with 15 SSR loci. Over all loci each individual was identified with unique multilocus genotype. High genetic diversity (average He=0.820) and low population differentiation (FST=0.0087) characterized this material. Although low in FST, the two northernmost populations were clustered as a distinct group diverged from the central populations. The population differentiation pattern corresponds well with the post glacial migration history of Norway spruce and the current gene flow and human activity in the region. The average inbreeding coefficient was 0.084 after removal loci with high frequency of null alleles. The estimated relatedness of the trees gathered in the breeding populations was very low (average kinship coefficient 0.0077) and not structured. The high genetic variation and low and not structured relatedness between individuals found in the breeding populations confirm that the Norway spruce breeding stock for northern Sweden represent valuable genetic resources for both long-term breeding and conservation programs.
The Antarctic and the Southern Ocean are a critically important part of the Earth system. The cli... more The Antarctic and the Southern Ocean are a critically important part of the Earth system. The climatic, physical, and biological properties of this region are closely linked to other parts of the global environment. 200 years of direct human impact, recent climate amelioration and changes in the main sources and circulation of biogenic compounds as well as accumulation of industrial contaminants have significantly affected the whole ecosystem. Particularly sensitive is the region of the Western Antarctic Peninsula, which is considered to be one of the hot spots of the Earth. In this paper, we review recent literature and compare it with historical data to estimate and predict the consequences of this process. The Antarctic ecosystems can no longer be regarded as pristine. Global as well as local human influence has transgressed the barriers isolating that continent from the rest of the World, causing previously observed changes to accelerate
The effect of methanesulfonic acid (MSA) on the morphophysiology and biochemistry of the subantar... more The effect of methanesulfonic acid (MSA) on the morphophysiology and biochemistry of the subantarctic species Colobanthus apetalus and the Antarctic species Colobanthus quitensis was examined. We evaluated the effects of various concentrations of MSA on the germination capacity and germination rate of seeds, seedling growth, chlorophyll fluorescence in cotyledons, and the proline content of seedlings under laboratory conditions at temperatures of 20°C (day) and 10°C (night) with a 12/12 h photoperiod. The examined C. apetalus seeds were grown in a greenhouse, and C. quitensis seeds were harvested in Antarctica and grown in a greenhouse (Olsztyn, Poland). The seeds of C. apetalus were characterized by the highest germination capacity and the highest germination rate, whereas C. quitensis seedlings were characterized by the most favorable growth and development. Only the highest concentrations of MSA decreased the intensity of chlorophyll fluorescence in the cotyledons of both Colobanthus species. The proline content of C. apetalus and C. quitensis seedlings increased significantly after MSA treatments. The results of this study clearly indicated that Colobanthus quitensis is more resistant to chemical stress induced by MSA. This is a first study to investigate the influence of MSA on the morphophysiology and biochemistry of higher plants.
There is only one species representing Magnoliopsida which is considered as native to the Antarct... more There is only one species representing Magnoliopsida which is considered as native to the Antarctic, i.e., Antarctic pearlwort (Colobanthus quitensis). Although it was intensively studied toward the morphophysiological adaptation to extreme environmental conditions of that area, there is still a lack of sufficient data on its genetic variability. Nine C. quitensis populations from Chile and the Maritime Antarctic were sampled to estimate the pattern of genetic variation in relation to the geographic distribution of analyzed populations and postglacial history of the species. The retrotransposon-based DNA marker system used in our studies appeared to be effective in revealing genetic polymorphism between individuals and genetic differentiation among populations. Although the level of polymorphism was low (9.57%), the Analysis of Molecular Variance showed that overall population differentiation was high (FST = 0.6241) and revealed significant differentiation between the Northern and Southern Group of populations as well as the population from Conguillio Park. The observed genetic subdivision of C. quitensis populations was confirmed by Bayesian clustering and results of Principal Coordinates Analysis. The Southern Group of populations was characterized by generally higher genetic diversity, which was expressed by the values of the effective number of alleles, expected heterozygosity and by the distribution of private alleles. Our results suggest that the species may have survived the Last Glacial Maximum in refugia located both on the South American continent and in geographically isolated islands of the Maritime Antarctic, i.e., they support the concept of the multiregional origin of C. quitensis in Antarctica.
The phenotypic and genetic relationships among five Pinus sylvestris populations in the western p... more The phenotypic and genetic relationships among five Pinus sylvestris populations in the western part of the Carpathian Mountains were analyzed based on seven morphological needle traits and 67 loci using 10 RAPD primers. Three of the studied populations were on the western side (Macelowa Góra, Czubatka, Łazowa Skałka) and two were located in the central part (Sokolica, Zamkowa Góra) of the Pieniny National Park. The examined populations had short needles, which is characteristic for mountain habitats of the Scots pine. The populations from the Pieniny Mts. had fewer rows of stomata on both sides of the needles than others studied stands of Eurosiberian Scots pine. The number of stomata on both flat and convex sides of the needles was similar to the values observed for populations growing on the nearby Tatra Mountains. Moreover, when these two traits are considered, the populations from Sokolica and Łazowa Skałka share the highest degree of similarity.. Nei’s genetic distances and genetic identities between the populations were calculated on the basis of RAPD loci. The results show that the lowest Nei’s distance exists between the populations from the western part of the Pieniny Mts. – Macelowa Góra and Czubatka, and its central part – Sokolica and Zamkowa Góra, with the highest values recorded between the populations of Zamkowa Góra and Czubatka, and Łazowa Skałka and Czubatka. A similar pattern of phenotypic and genetic relations that was observed between the populations in both regions of the Pieniny Mountains could be the result of planting-seed material sourced from the central Pieniny Mts. in the area of western Pieniny Mts.
The phytotoxic effects of fluoride and its derivatives on the seeds and seedlings of the Colobant... more The phytotoxic effects of fluoride and its derivatives on the seeds and seedlings of the Colobanthus apetalus and Colobanthus quitensis were studied. This is a first study to evaluate the influence of sodium fluoride (NaF) on the morphophysiological and biochemical processes on two Colobanthus species. The influence of various concentrations of NaF (9 mM, 19 mM, 29 mM) on the germination capacity and germination rate of seeds, seedlings growth and the proline content of plant tissues was analyzed under laboratory conditions (20/10°C, 12/12 h). The seeds of C. apetalus were collected from a greenhouse, whereas the seeds of C. quitensis were collected in Antarctica and in a greenhouse (Olsztyn, Poland). The tested concentrations of NaF did not suppress the germination of C. apetalus seeds, but the germination of C. quitensis seeds was inhibited. Sodium fluoride mainly inhibited root growth of C. apetalus and C. quitensis. In both analyzed species, the free proline content of seedlings increased significantly under exposure to NaF. The results of this study clearly indicate that C. apetalus and C. quitensis are highly resistant to NaF stress.
Vascular plants developed complex mechanisms to respond and adapt to recurring biotic and abiotic... more Vascular plants developed complex mechanisms to respond and adapt to recurring biotic and abiotic stresses (Bruce et al., 2007). There are a number of possible molecular mechanisms which may lead to genetically-determined phenotypic variability (Ingram & Bartels, 1996; Kreps et al., 2002; Tardif et al., 2007). In response to environmental stress, several types of morphological variants may arise, from which only the beneficial ones can be passed on to the next generation through natural selection (Piacentini et al., 2014). One of the mechanisms responsible for these evolutionary processes is transposon activation in response to severe environmental change which can disorganize the mechanism of transposon silencing (Kalendar et al., 2000; Piacentini et al., 2014). As a result, the explosion of transposon activity may be observed followed by the creation of genetic variability and associated macro-evolutionary processes (Rebollo et al., 2010; Schrader et al., 2014).
Uploads
Papers by Piotr Androsiuk
Microspore embryogenesis is a process that produces doubled haploids in tissue culture environments and is widely used in cereal plants. The efficient production of green regenerants requires stresses that could be sensed at the level of glycolysis, followed by the Krebs cycle and electron transfer chain. The latter can be affected by Cu(II) ion concentration in the induction media acting as cofactors of biochemical reactions, indirectly influencing the production of glutathione (GSH) and S-adenosyl-L-methionine (SAM) and thereby affecting epigenetic mechanisms involving DNA methylation (demethylation—DM, de novo methylation—DNM). The conclusions mentioned were acquired from research on triticale regenerants, but there is no similar research on barley. In this way, the study looks at how DNM, DM, Cu(II), SAM, GSH, and β-glucan affect the ability of green plant regeneration efficiency (GPRE).
Results
The experiment involved spring barley regenerants obtained through anther culture. Nine variants (trials) of induction media were created by adding copper (CuSO4: 0.1; 5; 10 µM) and silver salts (AgNO3: 0; 10; 60 µM), with varying incubation times for the anthers (21, 28, and 35 days). Changes in DNA methylation were estimated using the DArTseqMet molecular marker method, which also detects cytosine methylation.
Phenotype variability in β-glucans, SAM and GSH induced by the nutrient treatments was assessed using tentative assignments based on the Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy. The effectiveness of green plant regeneration ranged from 0.1 to 2.91 plants per 100 plated anthers. The level of demethylation ranged from 7.61 to 32.29, while de novo methylation reached values ranging from 6.83 to 32.27. The paper demonstrates that the samples from specific in vitro conditions (trials) formed tight groups linked to the factors contributing to the two main components responsible for 55.05% of the variance (to the first component DNM, DM, to the second component GSH, β-glucans, Cu(II), GPRE).
Conclusions
We can conclude that in vitro tissue culture conditions affect biochemical levels, DNA methylation changes, and GPRE. Increasing Cu(II) concentration in the IM impacts the metabolism and DNA methylation, elevating GPRE. Thus, changing Cu(II) concentration in the IM is fair to expect to boost GPRE.
activity on observed genetic diversity and differentiation of Scots pine in Europe. Our results indicate that the AFLP method applied to DNA templates extracted from bulked leaf samples provides an efficient approach to elucidate genetic diversity and relationships among Scots pine populations.
Microspore embryogenesis is a process that produces doubled haploids in tissue culture environments and is widely used in cereal plants. The efficient production of green regenerants requires stresses that could be sensed at the level of glycolysis, followed by the Krebs cycle and electron transfer chain. The latter can be affected by Cu(II) ion concentration in the induction media acting as cofactors of biochemical reactions, indirectly influencing the production of glutathione (GSH) and S-adenosyl-L-methionine (SAM) and thereby affecting epigenetic mechanisms involving DNA methylation (demethylation—DM, de novo methylation—DNM). The conclusions mentioned were acquired from research on triticale regenerants, but there is no similar research on barley. In this way, the study looks at how DNM, DM, Cu(II), SAM, GSH, and β-glucan affect the ability of green plant regeneration efficiency (GPRE).
Results
The experiment involved spring barley regenerants obtained through anther culture. Nine variants (trials) of induction media were created by adding copper (CuSO4: 0.1; 5; 10 µM) and silver salts (AgNO3: 0; 10; 60 µM), with varying incubation times for the anthers (21, 28, and 35 days). Changes in DNA methylation were estimated using the DArTseqMet molecular marker method, which also detects cytosine methylation.
Phenotype variability in β-glucans, SAM and GSH induced by the nutrient treatments was assessed using tentative assignments based on the Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy. The effectiveness of green plant regeneration ranged from 0.1 to 2.91 plants per 100 plated anthers. The level of demethylation ranged from 7.61 to 32.29, while de novo methylation reached values ranging from 6.83 to 32.27. The paper demonstrates that the samples from specific in vitro conditions (trials) formed tight groups linked to the factors contributing to the two main components responsible for 55.05% of the variance (to the first component DNM, DM, to the second component GSH, β-glucans, Cu(II), GPRE).
Conclusions
We can conclude that in vitro tissue culture conditions affect biochemical levels, DNA methylation changes, and GPRE. Increasing Cu(II) concentration in the IM impacts the metabolism and DNA methylation, elevating GPRE. Thus, changing Cu(II) concentration in the IM is fair to expect to boost GPRE.
activity on observed genetic diversity and differentiation of Scots pine in Europe. Our results indicate that the AFLP method applied to DNA templates extracted from bulked leaf samples provides an efficient approach to elucidate genetic diversity and relationships among Scots pine populations.
material was taken from the trees growing in the following stands: Sokolica, Czerwone Skałki, Biała Skała, Zamkowa Góra. Ten needles collected from twelve different trees found on each stand were taken for measurement in October 2003. Seven morphological traits of two-year old needles were studied: length of the needles, number of stomatal rows on the flat and convex sides of the needle, number of stomata per 2 mm of the needle length on its fl at and convex sides, number of serrations per 2 mm of the needle
length on its right and left edges.
study to evaluate the influence of sodium fluoride (NaF) on the morphophysiological and
biochemical processes on two Colobanthus species. The influence of various concentrations of NaF (9 mM, 19 mM, 29 mM) on the germination capacity and germination rate of seeds, seedlings growth and the proline content of plant tissues was analyzed under laboratory conditions (20/10°C, 12/12 h). The seeds of C. apetalus were collected from a greenhouse, whereas the seeds of C. quitensis were collected in Antarctica and in a greenhouse (Olsztyn, Poland). The tested concentrations of NaF did not suppress the germination of C. apetalus seeds, but the germination of C. quitensis seeds was inhibited. Sodium fluoride mainly inhibited root growth of C. apetalus and C. quitensis. In both analyzed species, the free proline content of seedlings increased significantly under exposure to NaF. The results of this study clearly indicate that C. apetalus and C. quitensis are highly resistant to NaF stress.