Infrageneric relationships of Korean Viola based on eight chloroplast markers

Phylogeny and infrageneric relationships were reconstructed for Korean Viola(Violaceae)using eight chloroplast sequences: the atpB-rbcL intergenic spacer,the atpF-H intergenic spacer,the matK gene,the psbA-trnH intergenic spacer,the psbK-I intergenic spacer,the rpl16 intron,the rpoC1 exon 2,and the trnL-trnF regions.The combined analyses of the eight chloroplast regions suggest that sections Dischidium and Chamaemelanium are monophyletic,whereas section Nomimium is paraphyletic; sections Dischidium and Chamaemelanium form a clade with subsections Hypocarpae and Trigonocarpae of section Nomimium,but unresolved within the clade; two subsections(subsects.Bilobatae and Vaginatae)of section Nomimium form a clade and are placed between the stem and stemless species group; and subsection Patellares of section Nomimium is monophyletic,and Viola dissecta is diverged first in the clade with high support(BS = 100,PP = 100).Our present results support the subsectional grouping based on morphological characteristics,although discordance remained at the series level.     

国产堇菜属的分子系统学研究

利用基于ITS序列的分子系统学对国产堇菜属(Viola L.)进行了系统发育重建,选择Hybanthus concolor和鳞隔堇(Scyphellandra pierrei)作为外类群,对国产堇菜属植物54种1亚种共66个序列进行分析。分子系统树中获得强烈支持的中国堇菜属属下组级分类群有Sect.Viola、Sect.Diffusae、Sect.Bilobatae、Sect.Vaginatae。Subgen.Melanium与Sect.Viola和Sect.Trigonocarpae聚在一起,说明它们的亲缘关系较近,但Sect.Trigonocarpae支获得的支持率较低。分子系统树证实了Subgen.Dischidium与Subgen.Chamaemelanium的近缘关系,但不支持根据距的长短将Subgen.Dischidium划分为Sect.Longicalcaratae和Sect.Brevicalcaratae的处理。而Pinnatae类群应该作为Sect.Adnatae下的一个亚组处理。根据分子系统发育和形态学特征对V.lucens、V.magnifica、V.dissecta、V.yunnanensis等的分类地位进行了重新评价。     

Viola suavis, a new species in the Nordic flora, with analyses of the relation to other species in the subsection Viola (Violaceae)

Evidence from cytology, enzyme markers and morphometry suggest that a species of Viola (Violaceae) collected in southeastern Norway should be referred to V. suavis , although differing in some phenotypic traits. V. suavis is not previously known from the Nordic countries, but must now be regarded as an established member of the Norwegian flora. It was probably introduced more than a century ago, providing an interesting analogy to the situation of V. alba in Öland (Sweden). Thus, it seems like human activity have played a role for expanding the distribution areas of several of the species within Viola subsection Viola (V. alba, V. hirta, V. odorata and V. suavis ). Electrophoretic evidence showed that the basal chromosome number of the subsection is x=5. V. suavis (2n=40) is allo-octoploid, and closely related tetraploid taxa were easily recognized. However, it is difficult to trace its origin to lower-ploid ancestors, and it seems likely that the complex may have contained extinct taxa with other allelic combinations that better explain the origin of V. suavis .     

Inferring species networks from gene trees in high-polyploid North American and Hawaiian violets (Viola, Violaceae)

The phylogenies of allopolyploids take the shape of networks and cannot be adequately represented as bifurcating trees. Especially for high polyploids (i.e., organisms with more than six sets of nuclear chromosomes), the signatures of gene homoeolog loss, deep coalescence, and polyploidy may become confounded, with the result that gene trees may be congruent with more than one species network. Herein, we obtained the most parsimonious species network by objective comparison of competing scenarios involving polyploidization and homoeolog loss in a high-polyploid lineage of violets (Viola, Violaceae) mostly or entirely restricted to North America, Central America, or Hawaii. We amplified homoeologs of the low-copy nuclear gene, glucose-6-phosphate isomerase (GPI), by single-molecule polymerase chain reaction (PCR) and the chloroplast trnL-F region by conventional PCR for 51 species and subspecies. Topological incongruence among GPI homoeolog subclades, owing to deep coalescence and two instances of putative loss (or lack of detection) of homoeologs, were reconciled by applying the maximum tree topology for each subclade. The most parsimonious species network and the fossil-based calibration of the homoeolog tree favored monophyly of the high polyploids, which has resulted from allodecaploidization 9-14 Ma, involving sympatric ancestors from the extant Viola sections Chamaemelanium (diploid), Plagiostigma (paleotetraploid), and Viola (paleotetraploid). Although two of the high-polyploid lineages (Boreali-Americanae, Pedatae) remained decaploid, recurrent polyploidization with tetraploids of section Plagiostigma within the last 5 Ma has resulted in two 14-ploid lineages (Mexicanae, Nosphinium) and one 18-ploid lineage (Langsdorffianae). This implies a more complex phylogenetic and biogeographic origin of the Hawaiian violets (Nosphinium) than that previously inferred from rDNA data and illustrates the necessity of considering polyploidy in phylogenetic and biogeographic reconstruction.     

A new species of Viola L. (Violaceae) from Sichuan, China

Viola muliensis Y.S. Chen & Q.E. Yang, a new species of Viola L. (Violaceae) from Sichuan, China, is described and illustrated. Viola muliensis is endemic to Muli County, south-western Sichuan. This new species has yellow flowers with very short spurs and belongs to subsection Brevicalcaratae W. Beck., section Dischidium Ging. It is distinguished from other related species by having pedatipartite leaves.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 149 , 365–368.     

中国葡萄属植物形态学聚类分组研究

对《中国葡萄志》描述的38个中国葡萄属野生种和1个栽培种,按照《葡萄种质资源描述规范和数据标准》的要求,对18个描述符用代码数量化并进行聚类分析,根据形态特征的相似程度,将我国葡萄属野生种和欧亚种划分为8个组和5个亚组。第1组包含毛葡萄、腺枝葡萄、龙泉葡萄、美丽葡萄和庐山葡萄,第2组包含麦黄葡萄和小叶葡萄,第3组包含5个亚组21个种类,其中亚组1包含山葡萄、陕西葡萄、浙江蘡薁、蘡薁和湖北葡萄,亚组2包含绵毛葡萄和勐海葡萄,亚组3包含小果葡萄、云南葡萄和葛藟葡萄,亚组4包含毛脉葡萄、华东葡萄、武汉葡萄和井冈葡萄,亚组5包含刺葡萄、秋葡萄、桦叶葡萄、网脉葡萄、蒙自葡萄、凤庆葡萄和河口葡萄,第4组仅包含欧洲葡萄,第5组包含温州葡萄、红叶葡萄、复叶葡萄、乳源葡萄和狭叶葡萄,第6组包含东南葡萄、罗城葡萄和闽赣葡萄,第7组仅包含菱叶葡萄,第8组仅包含鸡足葡萄一个种。     

Viola hirta (Violaceae) and its relatives in Norway

Viola hirta is widespread in most of Europe but rare in Norway, occurring in four isolated enclaves around the Oslo Fjord. Allozymic and morphological patterns of variation were studied in 320 individuals of Viola hirta (2n = 20) and its relatives V. collina (2n = 20), V. odorata (2n = 20), and V. suavis (2n = 40) from 22 Norwegian populations. Ongoing introgression from both V. collina and V. odorata , as hypothesised by several authors, was not supported by our data. All hybrids proved to be primary ones and, apparently, introgression is rare. However, allozyme markers suggested that V. hirta may have been affected by ancient introgression from V. collina and possibly also from the East European V. ambigua (2n = 40) during its evolutionary history. Geographical differentiation, presumably relatively recent and post-glacial, was evident in V. hirta. Variation occurred also between and within V. odorata populations, most likely a consequence of independent introductions by man. No variation was observed in V. collina or in V suavis. A key to the Norwegian species of subsection Viola and their primary hybrids based on morphological characters is provided in Flora Nordica note No. 26.     

Chalcone synthase gene lineage diversification confirms allopolyploid evolutionary relationships of European rostrate violets

Phylogenetic relationships among and within the subsections of the genus Viola are still far from resolved. We present the first organismal phylogeny of predominantly western European species of subsection Rostratae based on the plastid trnS-trnG intron and intergenic spacer and the nuclear low-copy gene chalcone synthase (CHS) sequences. CHS is a key enzyme in the synthesis of flavonoids, which are important for flower pigmentation. Genes encoding for CHS are members of a multigene family. In Viola, 3 different CHS copies are present. CHS gene lineages obtained confirmed earlier hypotheses about reticulate relationships between species of Viola subsection Rostratae based on karyotype data. Comparison of the CHS gene lineage tree and the plastid species phylogeny of Viola reconstructed in this study indicates that the different CHS copies present in Viola are the products of both recent and more ancient duplications.     

Long-term genome diploidization in allopolyploid Nicotiana section Repandae (Solanaceae)

Here, we analyze long-term evolution in Nicotiana allopolyploid section Repandae (the closest living diploids are N. sylvestris, the maternal parent, and N. obtusifolia, the paternal parent). We compare data with other more recently formed Nicotiana allopolyploids. We investigated 35S and 5S nuclear ribosomal DNA (rDNA) chromosomal location and unit divergence. A molecular clock was applied to the Nicotiana phylogenetic tree to determine allopolyploid ages. N. tabacum and species of Repandae were c. 0.2 and 4.5 Myr old, respectively. In all Repandae species, the numbers of both 35S and 5S rDNA loci were less than the sum of those of the diploid progenitors. Trees based on 5S rDNA spacer sequences indicated units of only the paternal parent. In recent Nicotiana allopolyploids, the numbers of rDNA loci equal the sum of those of their progenitors. In the Repandae genomes, diploidization is associated with locus loss. Sequence analysis indicates that 35S and 5S units most closely resemble maternal and paternal progenitors, respectively. In Nicotiana, 4.5 Myr of allopolyploid evolution renders genomic in situ hybridization (GISH) unsuitable for the complete resolution of parental genomes.     

Cleistogamy and phylogenetic position of Viola uliginosa (Violaceae) re‐examined

Viola uliginosa is an endangered species throughout its range in Central Europe. Based on an unusual combination of morphological characters, including a reported lack of cleistogamous (CL) flowers, it was previously placed in the monotypic subsection Repentes, section Viola. Available individual phylogenetic analyses identify V. uliginosa as a lineage deeply embedded in section Viola, but hitherto its exact position was unknown. Here we confirm the occurrence of cleistogamy in V. uliginosa. Unlike other species of section Viola, V. uliginosa produced both chasmogamous and CL flowers at the same time, not at different times of the year (seasonal cleistogamy). The two flower types differed conspicuously in morphology and microstructure, especially in size and shape of the pistil and stamens as confirmed by scanning electron microscopy. Secondly, we resolved the phylogenetic position of V. uliginosa as a lineage deeply embedded in subsection Rostratae by means of a multigene multispecies coalescent analysis using *BEAST. In this analysis, V. mirabilis and V. uliginosa were successive sisters to the rest of subsection Rostratae and, because allopolyploids exist between V. mirabilis and the last one, subsection Rostratae needs to comprise all these taxa to be monophyletic. The evolution of key morphological traits is briefly discussed.     

Evolution and biogeography of the woody Hawaiian violets (Viola, Violaceae): arctic origins, herbaceous ancestry and bird dispersal

Specialists studying the genus Viola have consistently allied the Hawaiian violets comprising section Nosphinium--most of which are subshrubs or treelets--with putatively primitive subshrubs in certain South American violet groups. Hawaiian violets also possess inflorescences, a floral disposition otherwise found only in other genera of the Violaceae, thus strengthening the hypothesis of a very ancient origin for the Hawaiian species. A survey of phylogenetic relationships among infrageneric groups of Viola worldwide using nuclear rDNA internal transcribed spacer (ITS) sequences revealed a dramatically different biogeographic origin for the Hawaiian violets: A monophyletic Hawaiian clade was placed in a close sister relationship with the amphi-Beringian tundra violet, V. langsdorffii s. 1., in a highly derived position. This remarkable and unforeseen relationship received strong clade support values across analyses, and monophyly of the Hawaiian lineage was further indicated by a unique 26-base-pair deletion in section Nosphinium. The high polyploid base chromosome number (n approximately equal to 40) in the Hawaiian violets relates them to Alaskan and eastern Siberian populations in the polyploid V. langsdorffii complex. More than 50 species of the 260 allochthonous birds wintering in the Hawaiian Islands are found to breed in the Arctic, occupying habitats in which individual birds might have encountered ancestral V. langsdorffii populations and served as dispersers to the central Pacific region. Acquisition of derived morphological traits (e.g., arborescence and inflorescences), significance of a confirmed Arctic origin for a component of the Hawaiian flora, and the likelihood of other "cryptic" Arctic elements in the Hawaiian flora deserving independent molecular phylogenetic corroboration are discussed.     

Application of watershed analyses and ecosystem modeling to investigate land–water nutrient coupling processes in the Guadalupe Estuary, Texas

Estuarine nutrient enrichment is thought to be controlled by land use patterns in coastal watersheds. Hence, the objective of this work was to conduct a watershed analysis in two adjacent river basins with different land use characteristics to determine their influence on estuarine ecosystem response in the Guadalupe Estuary, Texas, U.S.A. All data sources for this study were available electronically on the Internet; the data were mined, managed, analyzed and transformed to simulate the estuarine ecosystem response to watershed-derived nutrient loads. Between 1992 and 2001, developed land use/land cover increased the most while forest cover decreased the most in both basins. Two hydrologic units nearest the coast were responsible for the greatest change in land cover. Nutrient concentrations and loads were significantly higher in the San Antonio River Basin than in the Guadalupe River Basin. Both river basins exhibited the highest flows ever recorded in 1992, however the magnitude of difference in loads between the two coastal hydrologic units for a wet and dry year was much greater in the Guadalupe River Basin (GRB) than in the San Antonio River Basin (SARB); this difference supports the concept that the GRB is a nonpoint source dominated system and SARB is a point source dominated system. There was a strong correlation between developed land use and nutrient concentrations in river water; the GRB had less developed land use and lower nutrient concentrations while the SARB had more developed land use and higher nutrient concentrations. Estuarine ecosystem response differed in the timing, duration and magnitude of DIN, phytoplankton and zooplankton when nitrogen loads from the Lower Guadalupe River were used as opposed to the Lower San Antonio. The two basins studied differ in their fundamental characteristics, i.e. precipitation, flow, human population density, etc., resulting in different drivers of nitrogen loading, point sources in the San Antonio River Basin and nonpoint sources in the Guadalupe River Basin, therefore, differing estuarine ecosystem responses.     

A new species of Viola (Violaceae) from Turkey

A new species of Viola , V. kizildaghensis M. Dinç & S¸. Ẏlḋṙml̇, from Turkey is described and illustrated. It is found on the rocky slopes of K̇żldǎ, K̇żldǎ National Park, in the vilayet of Isparta, south-west Turkey, at an elevation of 1350–1600 m. It belongs to Viola , subsection Viola , and is similar to the Lebanese endemic V. libanotica Boiss., distinguished by its lanceolate, cuneate based leaves, pinkish-purple flowers with white throats, glabrous lateral petals, and peduncles bearing the mature fruit procumbent and pubescent. It is somewhat similar to another species from south-west Turkey, V. sandrasea Melchior, that is endemic to Sandras Dǎ in the vilayet Mǔla; it is distinguished from this by the characters listed above, its pubescent leaves and peduncles, and its linear-lanceolate to lanceolate stipules which have short, gland-tipped fimbriae. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 138 , 483–487.     

Polyploid and hybrid evolution in roses east of the Rocky Mountains

This study investigates the impact of hybridization and polyploidy in the evolution of eastern North American roses. We explore these processes in the Rosa carolina complex (section Cinnamomeae), which consists of five diploid and three tetraploid species. To clarify the status and origins of polyploids, a haplotype network (statistical parsimony) of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) nuclear gene was estimated for polyploids of the complex and for diploids of section Cinnamomeae in North America. A genealogical approach helped to decipher the evolutionary history of polyploids from noise created by hybridization, incomplete lineage sorting, and allelic segregation. At the diploid level, species west of the Rocky Mountains are distinct from eastern species. In the east, two groups of diploids were found: one consists of R. blanda and R. woodsii and the other of R. foliolosa, R. nitida, and R. palustris. Only eastern diploids are involved in the origins of the polyploids. Rosa arkansana is derived from the blanda-woodsii group, R. virginiana originated from the foliolosa-nitida-palustris group, and R. carolina is derived from a hybrid between the two diploid groups. The distinct origins of these polyploid taxa support the hypothesis that the three polyploids are separate species.     

On Paphiopedilum malipoense sp. nov.—An Intermediate Form Between Paphiopedilum and Cypripedium,with a Discussion on the Origin of the genus

Paphiopedilum malipoense S. C. Chen et Tsi is a very interesting new species with its flower similar to that of Cypripedium, especially section Cypripedium. It belongs to subgenus Brachypetalum, the most primitive group of Paphiopedilum, but differs from its allied species in hgniva elliptic-lanceolate sepal with cuspidately acuminate apex, rather narrow petals and horizontal lip, which are of common occurrence in many cypripediums, but very rare in paphiopedilums. Apparently, this is an intermediate form, or a link, between Paphiopedilum and Cypripedium, but it does not seem to arise from hybridization between them, because no Cypripedium has been found wherever Paphiopedilum occurs. The new species is distributed in southeastern Yunnan of China. In this area, as well as in river valleys of western Yunnan or the Hengduan Mountains, there have been four species of the same genus reported before. As we know, the Hengduan Mountains and their adjacent areas are rich in Cypripedium. The differentiation of the genus there is remarkable. All five sections it contains occur there and three of them are quite distinctive. For example, the general appearance of the section Bracleosa is dissimilar to that of any other cypripediums, but closely resembles that of Listera. It appears that the difference between sect. Bracteosa of Cypripedium and sect. Brachypetalum of Paphiopedilum is not necessarily wider than that between sect. Bracteosa and sect. Cypripedium of the same genus. Apparently, it is reasonable to consider Paphiopedilum to be an evolutional branch of Cypripedium extending into tropical area, with its primitive group (subgenus Brachypetalum) still remaining in its northern fringe area. This primitive subgenus has eight species, distributed from western Yunnan to the Malay Peninsula. Five of them, including the intermediate and primitive form published here, are found in the hilly land of southeastern Yunnan and the river valleys of western Yunnan. All these facts suggest its area of origin: the river valleys of the Hengduan Mountains and the lower hilly land contiguous to the southof them.     

On the origin of the sweet-smelling Parma violet cultivars (Violaceae): wide intraspecific hybridization, sterility, and sexual reproduction

Parma violets are reputed for their double, fragrant flowers and have been cultivated for centuries in Europe. However, due to a rather atypical morphology their taxonomic affinity has not been clarified. Authors have proposed an origin from three possible species, Viola alba, V. odorata, or V. suavis, or a hybrid origin. Using both ITS sequence variation and allozyme variation in 14 putative loci, we showed that the Parma violet cultivars have their origin within Viola alba and that they are best included in the Mediterranean subsp. dehnhardtii. There is no trace of interspecific hybridization. However, the cultivars appear to have a single origin in a wide hybrid within V. alba, involving parental plants from the eastern and western Mediterranean region; historical literature sources seem to indicate Turkey and Italy, respectively. The Parma violet cultivars possess high levels of allozyme heterozygosity and to some extent also within-individual ITS sequence variation. Losses of heterozygosity and within-individual ITS sequence variation in some of the cultivars indicate subsequent rare events of sexual reproduction, presumably through cleistogamous seed set. We unambiguously identify the closest wild relative of this group of cultivars, allowing growers to develop new selection procedures, and show a peculiar molecular process associated with human selection.     

Unusual patterns of hybridization involving a narrow endemic Rhododendron species (Ericaceae) in Yunnan, China

? Premise of the study: One potential threat to rare species is genetic swamping caused by hybridization, but few studies have quantified this threat. Rhododendron cyanocarpum is a narrow endemic species that occurs sympatrically with potentially interfertile congeners throughout its range within Yunnan, China. We searched the entire distribution of R. cyanocarpum for hybrids and examined the patterns of hybridization to assess potential threat from hybridization. ? Methods: In a comprehensive field survey, we detected only one instance of hybridization involving R. cyanocarpum, with R. delavayi, at Huadianba near Dali. Material of both species and putative hybrids was examined using morphology, chloroplast DNA, nuclear ribosomal DNA, and Bayesian analysis of AFLP profiles. ? Key results: Of 10 putative hybrids, two were F(1)(')s and at least seven were F(2)(')s. Four backcrosses to R. delavayi were detected among material with R. delavayi-like morphology within the hybrid zone. Backcrosses to R. cyanocarpum were not detected. Therefore F(2)(')s outnumbered all other classes within the hybrid zone, a situation not previously confirmed for plants and extremely rare generally. Hybridization was asymmetrical, with R. delavayi as the maternal parent in all but one of the hybrids detected. ? Conclusions: Although natural hybridization is common in Rhododendron, it is rare in R. cyanocarpum and is apparently not accompanied by backcrossing toward R. cyanocarpum. Hence, there is no immediate risk of genetic swamping, unless habitat disturbance increases and changes the patterns of hybridization. Our study is the first to report a plant hybrid zone dominated by F(2) hybrids. This pattern might contribute to species barrier maintenance.     

Floral Biology and Pollination Mechanisms in Two Viola Species—From Nectar to Pollen Flowers?

The genus Viola is represented by four related species in Brazil belonging to section Leptidium, one of the most primitive sections in the genus. Floral biology and pollination by bees were studied in Viola cerasifolia and V. subdimidiata in high-altitude areas in south-eastern Brazil. Flowers are zygomorphic and spurred. The five stamens are arranged in a cuff around the ovary, and pollen is released by means of apical connective projections, which form a cone surrounding the base of the style. The connective projections of the inferior stamens are elongated and curved to form a hook-shaped structure. Nectar-secreting tissue can occur in the basal connective appendages of the inferior stamens, which project into the spur. Flowers of V. subdimidiata secreted a mean volume of 0.14 micro l nectar over a 24-h period; approx. 40 % of flowers did not secrete any nectar. The main pollinators of these Viola species are female bees belonging to the genus Anthrenoides (Andrenidae), which search mainly for pollen. These bees seem to be oligolectic and obtain large amounts of pollen from Viola by vibrating the flowers or by moving the hook repeatedly back and forth. Males of Anthrenoides patrol Viola clusters and also feed on nectar, acting as secondary pollinators. The basic floral structure in the genus Viola fits that of 'nectar flowers'. The uncommon hook-shaped projections, scanty nectar production, and behaviour of pollinators suggest that V. cerasifolia and V. subdimidiata are shifting their reward for pollinators from nectar to pollen. Based on floral morphology, this shift may be widespread in Viola sect. Leptidium.     

A new violet species (Violaceae) from the south-west Alps

Viola laricicola Marcussen sp. nov. is described from collections from four French and Italian localities in the south-west Alps. It is closely related to V. riviniana (section Trigonocarpea ) but differs in a few morphological characters and by the chromosome number 2 n  = 20. Viola laricicola is endemic to the montane and subalpine coniferous forests in the continental parts of the Alps.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 142 , 119–123.     

Species delimitation in the Ponto-Caucasian Viola sieheana complex, based on evidence from allozymes, morphology, ploidy levels, and crossing experiments

Information from allozymes, ploidy levels, morphology, cross-compatibility and biogeography suggests that the easternmost occurrences of the Ponto-Caucasian Viola sieheana should be considered to belong to a separate species, V. caspia. Morphological characters delimitating V. sieheana and V. caspia are presented and discussed. The main distribution of V. sieheana s. str. is in the eastern Balkans, Turkey, Cyprus, and The Lebanon, and of V. caspia in the Caspio-Caucasian region westwards to Crimea and north-eastern Turkey. Their distributions in the intermediate areas remain unknown. Plants of V. sieheana s. str. from Cyprus, Greece, and Turkey were all blue-flowered and had the chromosome number 2n?=?12x?=?60. Its presence in the Peloponnese (Parnon, Ta?getos, Menalon) and in the Pindhos mountains, Greece, was confirmed. Reports of plants with 2n?=?8x?=?40 from these stations seem to refer to the sterile hybrid V. reichenbachiana?×?sieheana which is locally abundant. V. caspia from Azerbaijan had 2n?=?8x?=?40 and had either whitish or blue flowers. In spite of sympatry and almost complete cross-compatibility, the two corolla colour morphs showed strong allozymic differentiation in four out of seven isozyme systems, suggesting reproductive isolation and incipient speciation. Within section Viola, V. caspia and V. sieheana seem no more closely related to each other than to either of the other investigated European species. The possibility that V. sieheana is an ancient allopolyploid involving the Greek narrow endemic V. dirphya (2n?=?40) is discussed.