Usefulness and limitations of pollen characters in environmental studies based on Viola L. species (sect. Melanium Ging.)

The aim of these studies was to determine the effect of environmental pollution on pollen development. Pollen heteromorphism (the presence of pollen morphs differing in aperture number in one flower of a plant), pollen viability (stainability) and pollen grain size in European metallophytes from sect. Melanium Ging. (Viola L., Violaceae) were analyzed by SEM and histochemical staining.Plants’ tolerance to heavy metals is positively correlated with their pollen viability, which should be termed stainability as it depends on the staining method applied and is not correlated with pollen germination. Abortive pollen can be produced as an effect of heavy metals but also may result from hybridization, a very common phenomenon in pansies. Pollen stainability in hybrids can be high (even exceeding 70%) or low (barely above 20%), and stainable pollen grains can differ greatly in size (from very small to giant), indicating a cytological imbalance resulting from disturbed meiosis. The number of pollen apertures is an adaptive character in facultative metallophytes. Plants from a metallicolous population produced a wider range of aperture number (3, 4, 5) than plants from a non-metallicolous population, which developed only 4- and 5-aperturate pollen. Three-aperturate longer-lived pollen are favored in the harsh conditions of a metal-polluted environment

The evolution of the Aristolochia pallida complex (Aristolochiaceae) challenges traditional taxonomy and reflects large-scale glacial refugia in the Mediterranean

The taxonomy of the Mediterranean Aristolochia pallida complex has been under debate since several decades with the following species currently recognized: A. pallida, A. lutea, A. nardiana, A. microstoma, A. merxmuelleri, A. croatica, and A. castellana. These taxa are distributed from Iberia to Turkey. To reconstruct phylogenetic and biogeographic patterns, we employed cpDNA sequence variation using both noncoding (intron and spacer) and protein-coding regions (i.e., trnK intron, matK gene, and trnK-psbA spacer). Our results show that the morphology-based traditional taxonomy was not corroborated by our phylogenetic analyses. Aristolochia pallida, A. lutea, A. nardiana, and A. microstoma were not monophyletic. Instead, strong geographic signals were detected. Two major clades, one exclusively occurring in Greece and a second one of pan-Mediterranean distribution, were found. Several subclades distributed in Greece, NW Turkey, Italy, as well as amphi-Adriatic subclades, and a subgroup of southern France and Spain, were revealed. The distribution areas of these groups are in close vicinity to hypothesized glacial refugia areas in the Mediterranean. According to molecular clock analyses the diversification of this complex started around 3–3.3 my, before the onset of glaciation cycles, and the further evolution of and within major lineages falls into the Pleistocene. Based on these data, we conclude that the Aristolochia pallida alliance survived in different Mediterranean refugia rarely with low, but often with a high potential for range extension, and a high degree of morphological diversity.Turkish Science Foundatio

Lack of correlation between pollen aperture number and environmental factors in pansies (Viola L., sect. Melanium Ging.) : pollen heteromorphism re-examined

The development of different pollen morphs by one specimen - pollen heteromorphism - occurs in ca. one-third species of the genus Viola. Melanium section species (pansies) stand out in producing the widest range of apertures among Viola species. Aperture number decreases with elevation increase, and faster germination of fiveaperturate pollen as compared with three-aperturate has previously been postulated. We re-examined pollen heteromorphism in the context of its viability, and made correlations with elevation (>1500 m a.s.l. versus <1500 m a.s.l.), soil type (metalliferous versus non-metalliferous; MET versus NMET) and chromosome number based on selected study criteria of ca. 20% karyologically and morphologically strongly differentiated but genetically closely related pansies. A total of 79% of analysed species were heteromorphic, forming three- to six-colp or ate pollen per individual flower. Mean aperture number and pollen viability were not affected by soil type (MET versus NMET). Mean aperture number was also not influenced by elevation or species chromosome number. Positive correlations were established between aperture number and pollen viability, negative between pollen viability and elevation (increasing altitude of 100 m decreased pollen viability by 0.4%) and lack of correlation between chromosome number and pollen viability. The varied frequencies of different pollen morphs among species are not under the general pressure of ecological conditions, as previously postulated for the species of Melanium section. Rather, this trait in pansies, similar to other floral characters (e.g. long, curved nectar spur, ‘landing platform’, posterior petals with nectar guides), is adaptive but dependent on the breeding system (inbreeding versus outbreeding) of the individual species

Insight into "serpentine syndrome" of Albanian, endemic violets (Viola L., Melanium Ging. section) : looking for unique, adaptive microstructural floral, and embryological characters

The genus Viola, particularly Melanium section, rich in metallophytes, is an excellent taxon for study of microevolutionary and adaptation processes. Pollen, ovule, and microstructural floral characters were investigated by LM, SEM, and CLSM in seven endemic Albanian violets, five serpentinophytes (Viola albanica, V. dukadjinica, V. albanica × V. dukadjinica, V. raunsiensis, and V. macedonica), two from chalk soil (V. aetolica and V. schariensis), and in their closest relatives (V. lutea ssp. sudetica, V. tricolor ssp. tricolor, and V. arvensis) for their taxonomic usefulness and adaptive value. Three among analyzed characters were common in all Albanian violets however not unique. Serpentinophytes, V. aetolica and V. schariensis possessed hairs deep inside the spur, developed pollen heteromorphism, both increase the chance of pollination in unpredictable conditions and had strongly developed tannin rich layer in the outer integument of the young ovules with a protective role. They also all exhibited high pollen viability (86.9 ± 10.2%), high frequency of normally developed, enlarged (fertilized) ovules in ovary (65.0 ± 24.0%), but also high frequency of degenerations in developing ovules (40.4 ± 9.8%). Several flower characters may be adaptive in the unfavorable, high altitude environment, including serpentine soils. High pollen viability and normally developed fertilized ovules are sufficient for Albanian species maintenance

Insight into “serpentine syndrome” of Albanian, endemic violets (<i>Viola</i> L., <i>Melanium</i> Ging. section) – Looking for unique, adaptive microstructural floral, and embryological characters

<p>The genus <i>Viola</i>, particularly <i>Melanium</i> section, rich in metallophytes, is an excellent taxon for study of microevolutionary and adaptation processes. Pollen, ovule, and microstructural floral characters were investigated by LM, SEM, and CLSM in seven endemic Albanian violets, five serpentinophytes (<i>Viola albanica</i>, <i>V. dukadjinica</i>, <i>V. albanica</i> × <i>V. dukadjinica</i>, <i>V. raunsiensis,</i> and <i>V. macedonica</i>), two from chalk soil (<i>V. aetolica</i> and <i>V. schariensis</i>), and in their closest relatives (<i>V. lutea</i> ssp<i>. sudetica</i>, <i>V. tricolor</i> ssp. <i>tricolor,</i> and <i>V. arvensis</i>) for their taxonomic usefulness and adaptive value. Three among analyzed characters were common in all Albanian violets however not unique. Serpentinophytes, <i>V. aetolica</i> and <i>V. schariensis</i> possessed hairs deep inside the spur, developed pollen heteromorphism, both increase the chance of pollination in unpredictable conditions and had strongly developed tannin rich layer in the outer integument of the young ovules with a protective role. They also all exhibited high pollen viability (86.9 ± 10.2%), high frequency of normally developed, enlarged (fertilized) ovules in ovary (65.0 ± 24.0%), but also high frequency of degenerations in developing ovules (40.4 ± 9.8%). Several flower characters may be adaptive in the unfavorable, high altitude environment, including serpentine soils. High pollen viability and normally developed fertilized ovules are sufficient for Albanian species maintenance.</p

Insight into “serpentine syndrome” of Albanian, endemic violets (<i>Viola</i> L., <i>Melanium</i> Ging. section) – Looking for unique, adaptive microstructural floral, and embryological characters

<p>The genus <i>Viola</i>, particularly <i>Melanium</i> section, rich in metallophytes, is an excellent taxon for study of microevolutionary and adaptation processes. Pollen, ovule, and microstructural floral characters were investigated by LM, SEM, and CLSM in seven endemic Albanian violets, five serpentinophytes (<i>Viola albanica</i>, <i>V. dukadjinica</i>, <i>V. albanica</i> × <i>V. dukadjinica</i>, <i>V. raunsiensis,</i> and <i>V. macedonica</i>), two from chalk soil (<i>V. aetolica</i> and <i>V. schariensis</i>), and in their closest relatives (<i>V. lutea</i> ssp<i>. sudetica</i>, <i>V. tricolor</i> ssp. <i>tricolor,</i> and <i>V. arvensis</i>) for their taxonomic usefulness and adaptive value. Three among analyzed characters were common in all Albanian violets however not unique. Serpentinophytes, <i>V. aetolica</i> and <i>V. schariensis</i> possessed hairs deep inside the spur, developed pollen heteromorphism, both increase the chance of pollination in unpredictable conditions and had strongly developed tannin rich layer in the outer integument of the young ovules with a protective role. They also all exhibited high pollen viability (86.9 ± 10.2%), high frequency of normally developed, enlarged (fertilized) ovules in ovary (65.0 ± 24.0%), but also high frequency of degenerations in developing ovules (40.4 ± 9.8%). Several flower characters may be adaptive in the unfavorable, high altitude environment, including serpentine soils. High pollen viability and normally developed fertilized ovules are sufficient for Albanian species maintenance.</p