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Nuclear DNA content variation among perennial taxa of the genus Cyanus (Asteraceae) in Central Europe and adjacent areas

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The genome size of 265 plants and the GC content of 126 plants from 63 populations of the Cyanus triumfetti and Cyanus montanus groups, collected across the Carpathians, Pannonia, Bohemian Massif, and Western and Dinaric Alps were determined by PI and DAPI flow cytometry. Variation of the nuclear DNA content among homoploid species, and intraspecific and interpopulation variation were confirmed in simultaneous analyses. The 2C-value at the diploid level (the C. triumfetti group) varied from 2.53 for Cyanus dominii subsp. sokolensis to 3.06 pg for C. triumfetti s.s. (1.21-fold range). At the tetraploid level (the C. montanus group), the 2C-value varied from 5.19 for Cyanus mollis to 5.84 pg for C. montanus (1.13-fold range). High intraspecific and interpopulation variation in the amount of nuclear DNA in the C. triumfetti group correlates with the extensive morphological variation found in this group. Significant between-species differences in genome size indicate that this attribute may be used as a supportive taxonomic marker for both of the groups studied. The GC content varied by 2.93 %, from 39.46 % for “Cyanus axillaris” to 40.61 % for Cyanus adscendens; this character is of no value for taxonomic purposes. Genome size of the studied populations is significantly higher in southern parts of the distribution area and at higher elevations. Plants with smaller genomes tend to occur in dry areas at low altitudes with high diurnal and annual temperature oscillations. The GC content of the populations studied is significantly correlated with longitude, increasing from east to west; and plants with GC-rich genomes are concentrated in the coldest areas with low minimum temperatures.
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... We focused on the basic diagnostic characters repeatedly appearing in plant identification keys and taxonomic works (e.g. Dostál 1948Dostál -1949Dostál , 1958Dostál , 1989 (Španiel at al. 2008;Mereďa et al. 2011Mereďa et al. , 2016Olšavská et al. 2011Olšavská et al. , 2012Koutecký et al. 2012;Budzáková et al. 2014;Hodálová et al. 2015;Skokanová et al. 2019a, b). ...
... But by combining all studied quantitative characters of stem, leaves, and corymbs, it was possible to correctly classify more than 91% of the plants (Table 4). It is quite common phenomenon, that in taxonomic groups comprising closely related taxa with high morphological variability, combination of several morphological characters has to be used in order to determine particular taxa (Španiel et al. 2008;Olšavská et al. 2011Olšavská et al. , 2012Šingliarová et al. 2011;Koutecký et al. 2012;Budzáková et al. 2014;Hodálová et al. 2015;Mereďa et al. 2011Mereďa et al. , 2016Skokanová et al. 2019a, b). To conclude, the revealed morphological differentiation together with distinct ploidy level differentiation of the T. corymbosum group in the Slovakia allowed the recognition of two groups of populations from this area as two species T. clusii and T. corymbosum. ...
Article
The paper aims to provide a comprehensive overview of karyological and morphological variability of the Tanacetum corymbosum group in Slovakia. We estimated DNA ploidy level and relative DNA content of 530 individuals (45 populations) using DAPI flow cytometry. The morphological variation was studied by univariate as well as multivariate morphometric based on 10 morphological characters of 351 individuals (39 populations). In addition, two characters were measured on 120 achenes (six populations). Flow cytometry analyses confirmed two predominant cytotypes within T. corymbosum group in Slovakia: diploids corresponding to T. clusii and tetraploids corresponding to T. corymbosum. Further, we recorded rare occurrence (1.3% of the dataset) of minority cytotypes – presumable triploids and pentaploids, probably originating from fusion of reduced and unreduced gamete in single species populations. One triploid plant found at the only one locality with common occurrence of T. clusii and T. corymbosum is most probably of hybrid origin. Morphological analyses showed that T. clusii and T. corymbosum could not be clearly distinguished by either of the studied characters but by combining all studied quantitative characters of stem, leaves, and corymbs, it was possible to correctly classify more than 91% of the plants. Number and size of stem leaves as well as capitula, and coloration of the margin of the involucral bracts were identified as the most useful for their determination. Based on revealed karyological and morphological differentiation of T. clusii and T. corymbosum, we favour to recognize them as two separate species rather than subspecies. A key for identifying native Tanacetum species in Slovakia is presented.
... A great goal is also increasing the resistance to abiotic and biotic stresses, including the resistance to many types of diseases (Szabó, et al., 2010), what can be reached by higher ploidy production (Shi et al., 2015). New species and cultivars and their taxonomic relationships can be assessed upon the ploidy and relative genome size utilization (Šmarda et al., 2006;Olšavská et al., 2012). Previous studies described differences in genome size of cultivars in relation to the climatic conditions, also the correlation of the genome size and altitude, geographical situation, temperature etc. were found (Kolano et al., 2012). ...
Article
The information about genome size is implied in the studies of genetic diversity, the genome size evolution and can be used in breeding of new interesting cultivars. A total of 52 apricot cultivars, including 17 new perspective apricot hybrids planted in South Moravia region were analysed by flow cytometry. In addition, nuclear DNA content and ploidy level of 10 representative cultivars P. salicina, P. domestica, P. cerasifera and 22 interspecific hybrids of stone fruit were determined. Only the CV values lower than 5% were considered in analysis. All analysed apricot cultivars were diploid with average value of relative genome size 0.617 ± 0.014 pg/2C. The ploidy and DNA content of plums differ in botanical species. The obtained results were compared according to the geographical origin of cultivars and the theory about genome size evolution was considered. The values of relative genome size of original botanical species related to P. armeniaca (P. mume, P. sibirica, P. ansu) reached the lowest value in average compared with other cultivars in this study (0.603 pg/2C in average). Two pentaploids and one tetraploid were found in interspecific hybrids, which could be a part of interest of future studies.
... In some groups of plants, genome size is correlated with environmental conditions and/or geographical distribution (Pecinka et al. 2006, Kolář et al. 2009, Dušková et al. 2010, Olšavská et al. 2012). However, we recorded little variation in relative DNA content of S. virgaurea samples (5.8%) collected over a substantial part of its distribution area (Fig. 1) and over a wide range of altitudes (190-2130 m a.s.l.; Tables 1, 2). ...
Article
Hybridization between native and alien congeners may pose a serious threat to biodiversity and negatively affect native flora. Here we study Solidago ×niederederi, which originated and became established in Europe as a result of a cross between the alien S. canadensis and native S. virgaurea. The recent increase in the number of records of S. ×niederederi in Europe has highlighted the need to monitor its occurrence, spread and behaviour. In the present study, we tested the effectiveness of flow cytometry for detecting hybrid plants of S. ×niederederi. Sequences of the ITS region of nrDNA and the rpS15-ycf1 spacer of cpDNA were used to confirm the hybrid origin of analysed plants and to identify the maternal species. Our study included 60 single-species populations of S. canadensis, S. gigantea and S. virgaurea, and 16 mixed populations with the presence of hybrid S. ×niederederi sampled from six countries in central Europe and adjacent areas. All individuals of S. canadensis, S. ×niederederi and S. virgaurea investigated were diploid (2n~2x~18) but differed in their relative DNA content values. The DNA content of S. ×niederederi was intermediate between S. canadensis and S. virgaurea with no overlaps, with the differences between the species being statistically significant. Therefore, we conclude that flow cytometry is a reliable and efficient method for detailed screening for hybrids within mixed Solidago populations and for identifying non-flowering or morphologically ambiguous Solidago plants. Since both parental species varied only negligibly in their DNA content, it may also be applicable across a broader geographic scale. Genetic, flow cytometric and distributional data suggest that the hybrids are to a large extent early generation (likely F1) hybrids as very few cases of supposed introgressants were also inferred. The results from chloroplast rpS15-ycf1 spacer showed that hybridization has occurred in both directions.
... Nevertheless, a significant correlation between GC content and longitude was reported in Cyanus, another Asteraceae genus, where plants growing in west areas showed higher GC contents than those in the east. Moreover, plants growing in the coldest areas with low minimum temperatures tend to have higher GC contents in their genomes (Olsavska et al. 2012). ...
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Susanto AH, Dwiati M, Pratiwi S. 2020. Molecular characteristics of two phenotypically identical species of Asteraceae based on the intergenic spacer trnT(UGU)-trnL(UAA). Biodiversitas 21: 5164-5169. Ogiera (Eleutheranthera ruderalis) and nodeweed (Synedrella nodiflora) are two different weed species belonging to the family Asteraceae commonly found in tropical regions. At a glance, both species show highly identical morphology, thus leading to difficulty in distinguishing between them. Therefore, molecular data based on particular markers are required. Here, we use an intergenic spacer (IGS) in the cpDNA, i.e., trnT(UGU)-trnL(UAA), as the molecular marker to reveal the difference between the two species. A pair of PCR universal primers, i.e., B48557 as the forward primer and A49291 as the reverse primer, were employed to amplify the marker. Sequence alignment was performed by the use of ClustalW implemented in Bioedit version 7.0.4.1. The results revealed that some differences with respect to both indel and base substitution were observed. Overall, this led to longer IGS trnT(UGU)-trnL(UAA) sequences of E. ruderalis than those of S. nodiflora. Although no direct relationship between the genetic and phenotypic dissimilarities was proven, coincidence seemed likely to exist. This provides molecular evidence that the two phenotypically similar species are genetically different from each other.
... In some cases, already considered in Kliment et al. (2016), thorough biosystematic studies have revealed that taxa previously thought to be endemics were in fact only poorly delimited lineages Page 5 of 15 59 of geographically more widespread species, and should not, therefore, be treated as separate entities (Kolarčik et al. 2010;Španiel et al. 2011;Slovák et al. 2012a). Conversely, an increasing number of comprehensive taxonomic investigations have provided evidence for the correct taxonomic status and thus endemism in several taxa (Goliašová 1985;Hodálová and Marhold 1998;Hodálová 1999;Mráz 2001Mráz , 2003Zhang et al. 2001;Dvořáková 2003;Zhang and Kadereit 2004;Kochjarová et al. 2006;Chrtek et al. 2007;Ronikier and Szeląg 2008;Letz 2009;Kolarčik et al. 2010;Olšavská et al. 2011Olšavská et al. , 2012Šingliarová and Mráz 2009;Šingliarová et al. 2011;Štěpánek et al. 2011;Kučera et al. 2013;Kuzmanović et al. 2013;Şuteu et al. 2013;Ferreira et al. 2015;Bureš et al. 2018;Tkach et al. 2019;Mráz et al. 2019). ...
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Effective conservation of biodiversity should build on a strong basis of taxonomic and spatial distribution knowledge. The Carpathian Mountains, an iconic centre of biodiversity in temperate Europe, harbour a remarkable number of endemic vas-cular plants. Current knowledge on their taxonomic status, spatial distribution and genetic diversity is, however, incomplete. Research and conservation efforts have mostly been country specific, resulting in contrasting chorological knowledge and taxonomic acceptance between neighbouring countries, and differing conservation policies. Urgent synchronisation of cross-border conservation measures is required. Here, we present an overview of a new international project that will address these issues, outlining the current state of knowledge and theoretical background concerning Carpathian subendemics and endemics in order to improve their conservation status. The project involves partners from eight countries, working collaboratively in conservation, research and sharing of standardised data for the Carpathian flora. Long-term seed conservation of 500 species, with a focus on endemics, regionally endangered species and range-margin populations from the Carpathians, will be used as an effective measure for the conservation of Carpathian endemics and endangered species. Research focused on the biosystematics of selected taxa will increase our knowledge of the evolutionary processes involved in the origin of the Carpathian flora. Finally, the establishment of the Carpathian Research Network (CRN) provides an official framework for pooling, sharing and standardising scientific data on Carpathian endemics from different countries. The CRN is developing an online database of the distribution of selected species (The Carpathian Endemics Distribution Database, CEDD) including their intrinsic and extrinsic traits and current opinion on their taxonomic status. This will be used to produce a monograph on the Carpathians' endemic flora.
... In some cases, already considered in Kliment et al. (2016), thorough biosystematic studies have revealed that taxa previously thought to be endemics were in fact only poorly delimited lineages of geographically more widespread species, and should not, therefore, be treated as separate entities (Kolarčik et al. 2010;Španiel et al. 2011;Slovák et al. 2012a). Conversely, an increasing number of comprehensive taxonomic investigations have provided evidence for the correct taxonomic status and thus endemism in several taxa (Goliašová 1985;Hodálová and Marhold 1998;Hodálová 1999;Mráz 2001Mráz , 2003Zhang et al. 2001;Dvořáková 2003;Zhang and Kadereit 2004;Kochjarová et al. 2006;Chrtek et al. 2007;Ronikier and Szeląg 2008;Letz 2009;Kolarčik et al. 2010;Olšavská et al. 2011Olšavská et al. , 2012Šingliarová and Mráz 2009;Šingliarová et al. 2011;Štěpánek et al. 2011;Kučera et al. 2013;Kuzmanović et al. 2013;Şuteu et al. 2013;Ferreira et al. 2015;Bureš et al. 2018;Tkach et al. 2019;Mráz et al. 2019). ...
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Effective conservation of biodiversity should build on a strong basis of taxonomic and spatial distribution knowledge. The Carpathian Mountains, an iconic centre of biodiversity in temperate Europe, harbour a remarkable number of endemic vas-cular plants. Current knowledge on their taxonomic status, spatial distribution and genetic diversity is, however, incomplete. Research and conservation efforts have mostly been country specific, resulting in contrasting chorological knowledge and taxonomic acceptance between neighbouring countries, and differing conservation policies. Urgent synchronisation of cross-border conservation measures is required. Here, we present an overview of a new international project that will address these issues, outlining the current state of knowledge and theoretical background concerning Carpathian subendemics and endemics in order to improve their conservation status. The project involves partners from eight countries, working collaboratively in conservation, research and sharing of standardised data for the Carpathian flora. Long-term seed conservation of 500 species, with a focus on endemics, regionally endangered species and range-margin populations from the Carpathians, will be used as an effective measure for the conservation of Carpathian endemics and endangered species. Research focused on the biosystematics of selected taxa will increase our knowledge of the evolutionary processes involved in the origin of the Carpathian flora. Finally, the establishment of the Carpathian Research Network (CRN) provides an official framework for pooling, sharing and standardising scientific data on Carpathian endemics from different countries. The CRN is developing an online database of the distribution of selected species (The Carpathian Endemics Distribution Database, CEDD) including their intrinsic and extrinsic traits and current opinion on their taxonomic status. This will be used to produce a monograph on the Carpathians' endemic flora.
... Regarding the basic chromosome number of the genus (n = 9), the average chromosome contains ~ 0.24 pg of DNA in diploids and triploids and ~ 0.23 pg of DNA in higher polyploids. Given that the absolute DNA content estimated with intercalating PI and the relative DNA content estimated by AT-selective DAPI dye have been proved to be highly correlated (Marhold et al., 2010;Suda et al., 2010;Olšavská et al., 2012), in further analyses we considered a plant to be euploid when its relative DNA content was within the range of ±0.24 pg from the expected value for diploids and triploids and ±0.23 pg from the expected value for tetraploids. When the relative DNA content exceeded the threshold value, the plant was classified as aneuploid. ...
Article
Populations of sympatric diploid and autopolyploid cytotypes provide a unique opportunity to study early stages of polyploid evolution. Pilosella rhodopea is a diploid–autopolyploid complex forming frequent mixed-ploidy populations, most probably representing the largest primary contact zone documented so far in angiosperms. Our aims were to elucidate: (1) the origin of autopolyploids (single vs. multiple); (2) cytotype distribution patterns at various spatial scales; and (3) potential ecological differentiation of the cytotypes by measuring several habitat variables along two elevational transects. In total, five cytotypes were found across the species range. Triploids were the most frequent (50%), followed by diploids (29%) and tetraploids (15%), whereas pentaploids and hexaploids were rare. Most populations were mixed-ploidy. Both amplified fragment length polymorphism and cytotype distribution patterns suggested multiple origins of autopolyploids and frequent intercytotype gene flow. Cytotype diversity and spatial aggregation of the cytotypes were scale dependent. At a local scale (5 m × 5 m), cytotype co-occurrence was frequent, whereas micro-scale plots (1 m × 1 m) were cytotypically more homogeneous. We did not find any evidence for ecological differentiation of the cytotypes. Our results show that P. rhodopea is a rare example of a diploid–autopolyploid complex with polytopic and ongoing polyploid formations in primary contact zones. Recurrent formation of polyploids owing to a frequent intercytotype gene flow among spatially close cytotypes might explain, at least in part, the sympatric coexistence of cytotypes.
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Polyploidy is a major force in plant evolution. The possession of more than two chromosome complements may affect the genetic and genomic constitution and the phenotypes of polyploids, having consequences for their ecology, geography and diversification. Here, we used Turnera sidoides autopolyploid complex to assess the effect of polyploidy on genome size and on the diversification of this species. The remarkable diversity in habitat preferences, ploidy levels and multiple independent origins of polyploids, make this complex a suitable model for disentangling the effects of phylogenetic relationships and environmental conditions on the variation in genome size. We used an integrative approach comprising genome size estimations in 53 (diploid to hexaploid) populations of the different subspecies andmorphotypes of T. sidoides, a molecular phylogenetic reconstruction and a biogeographical analysis to identified closely related diploids and polyploids that remained in the same habitat aiming to answer the following questions: 1) does polyploidy per se induce a significant change of the Cx-value?; 2) is the Cx-value variation an adaptive response/consequence to different environmental gradients?; and 3) does polyploidy enable the range expansion of diversified diploid lineages within the same ecoregion?. Comparisons of Cx-values among phylogenetically related diploids–polyploids provided evidence that polyploidy is not the main factor determining the Cx variation. Instead, the Cx-values varied associated with few climatic variables along latitudinal and climatic gradients, suggesting that variation in genome size would have been an adaptive response to different habitats. The analysis of the environmental preferences of diploid and polyploids within each lineage provides evidence that autopolyploidy further promoted the range expansion of the already diversified diploids. These results together with the morphological constancy among different cytotypes within lineages suggest that autopolyploidy per se did not contribute significantly to the morphological and taxonomic diversification, but enabled further range expansion of lineages within the complex.
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Multivariate morphometrics and an assessment of genetic diversity obtained using amplified fragment length polymorphism (AFLP) were used to determine the variability of the polymorphic group Cyanus triumfetti in Central Europe. The ploidy level of the populations studied was also determined; all individuals from the C. triumfetti group were diploid (2n ̃ 2x ̃ 22) and all those of the related C. montanus group were tetraploid (2n ̃ 4x ̃ 44). A multivariate morphometric study of 71 populations revealed that three species from the C. triumfetti group occur in Central Europe, namely 'Cyanus axillaris', C. strictus and C. dominii. Three subspecies are recognized within the latter species, namely C. dominii subsp. dominii, C. dominii subsp. slovenicus and C. dominii subsp. sokolensis. Morphological characters of leaves are the best features for delimiting these taxa; a shift in characters caused by cultivation did not affect the value of key characters and differences among the taxa remained. AFLP analysis of 38 populations from the C. triumfetti group and two from the C. montanus group revealed a contrasting pattern of genetic variation that was related to the geographic distribution of the populations rather than the morphological variation in the C. triumfetti group. The AFLP data revealed the following three genetically differentiated and allopatric groups: (i) C. triumfetti s.s. and C. montanus from the Western Alps, (ii) 'C. axillaris' from Austria and the Czech Republic (except the Carpathians) and (iii) 'C. axillaris', C. strictus and C. dominii from the Western Carpathians and Pannonia. The striking genetic gap between the Austrian-Czech and theW Carpathian-Pannonia groups and the high genetic diversity and weak genetic differentiation within the latter group are discussed in the light of potential glacial refuges, postglacial migration routes and/or the probability of hybridization events occurring during the evolutionary history of this group. An identification key for the taxa of the C. triumfetti group in Central Europe is presented.
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Genome size has been suggested as one of the traits associated with invasivenessof plant species. To provide aquantitative insight into the role of this trait, we estimated nuclear DNA contentin 93 alien species naturalized in the Czech Republic, belonging to 32 families, by using flow cytometry, and compared it with the values reported for non-invading congeneric and confamilial species from the Plant DNA C-values database. Species naturalized in the Czech Republic have significantly smaller genomes than their congeners not known to be naturalized or invasive in any part of the world. This trend is supported at the family level: alien species naturalized in the Czech flora have on average a smaller genome than is the mean value for non-invading confamilials. Moreover, naturalized and non-invading species clearly differed in the frequency offive genome size categories; this difference was mainly due to very small genomes prevailing and intermediate to very large genomes underrepresented in the former group. Our results provide the first quantitative support for association of genome size with invasiveness, based on a large set of alien species across a number of plant families. However, there was no difference in the genome size of invasive species compared to naturalized but non-invasive. This suggests that small genome size provides alien plants with an advantage already at the stage of naturalization and need not be necessarily associated with the final stage of the process, i.e. invasion.