Anatomical studies on selected species of Viola (Violaceae)

Cross‐sections of roots, stems, petioles and peduncles were investigated in 12 species of Viola distributed mainly in northern Iran: V. kitaibeliana, V. arvensis, V. occulta, V. tricolor (sect. Melanium), V. somchetica (sect. Plagiostigma), V. spathulata (sect. Spathulidium ined.), V. alba, V. odorata, V. sintenisii (sect. Viola subsect. Viola), V. caspia, V. reichenbachiana and V. rupestris (sect. Viola subsect. Rostratae). General anatomical features of Viola species are discussed. The following characters were found to be taxonomically informative: cross‐section shapes of the aerial stem, petiole and peduncle as well as the number of vascular bundles in the aerial stem. In taxa of sect. Melanium, stem cross‐sections are elliptic with two wings, but they are semi‐circular with two wings in taxa of sect. Viola subsect. Rostratae. Within the latter subsection, the number of vascular bundles in the stem can be used to delimit the species V. caspia, V. reichenbachiana and V. rupestris. Calcium oxalate crystals were observed in all vegetative organs of taxa belonging to sect. Viola. All examined taxa had a secondary structure with the exception of V. somchetica. Our results show that anatomical characters are useful for delimiting species, subsections, and sections within Viola.     

Morphological Characters Useful for the Delimitation of Taxa Within Viola Subsect. Viola (Violaceae): A Morphometric Study from the West Carpathians

Forty-nine morphological characters were scored or measured on 44 populations (376 individuals) of Viola subsect. Viola from the West Carpathians and adjacent areas (Slovakia, Czech Republic, Austria and Hungary). The presence of six species, namely V. alba (represented by subsp. alba), V. ambigua, V. collina, V. hirta, V. odorata and V. suavis s.l. was revealed based on pollen fertility, cytological and morphometric analyses. The morphological characters traditionally used to delimit taxa within the subsection and those revealed by our study as most reliable are widely discussed. A key for identifying the taxa and most common hybrids of subsection Viola occurring in the West Carpathians is presented. Chromosome counting and flow cytometry were used to determine the ploidy levels of the populations studied. All individuals of V. alba subsp. alba, V. collina, V. hirta and V. odorata were tetraploid, while those of V. ambigua and V. suavis s.l. were octoploid.     

Reproductive ecology of Viola mirabilis var. subglabra representing intermediate flowering characteristics between stemless and stemmed Viola

Viola (Violaceae) is one of the largest genera in angiosperms. This genus is essentially classified into stemless and stemmed groups based on growth morphology. However, Viola mirabilis var. subglabra is an exception in having intermediate flowering characteristics; cleistogamous (CL) flowers are formed in the axils of stem leaves, whereas chasmogamous (CH) flowers arise from basal rosettes (radical CH (CH(r)) flowers) and also in the axils of the stem (axially CH (CH(a)) flowers). To understand why the pattern of flower production varies in this Viola species, flower production was investigated in 10 Japanese populations from Hokkaido to the western part of Honshu in 2014 and 2015. Furthermore, flower characteristics were also compared between CH(r) and CH(a) flowers in Hokkaido. In this species, the production of CH flowers varied among individuals, and they were categorized into three groups, individuals that produced (i) only CH(r) flowers, (ii) only CH(a) flowers and (iii) both CH flowers. The frequency of these groups differed among populations, but some individuals changed the category between 2014 and 2015. Thus, the production of CH(r) and CH(a) flowers plastically changes depending on individual conditions and/or environmental factors. On the other hand, CH(r) and CH(a) flowers differed in flower size and flowering phenology. These results suggest that two types of CH flowers may play different roles in reproduction in each population, but fruit sets and seed sets did not differ between two types of CH flowers.     

Genetic diversity of the endangered coastal violet <Emphasis Type="Italic">Viola grayi</Emphasis> Franchet et Savatier (Violaceae) and its genetic relationship to the species in subsection <Emphasis Type="Italic">Rostratae</Emphasis>

Viola grayi, a coastal violet, is found on sandy seashores in limited regions in Japan, but not on rocky sea cliffs or in inland areas. This stemmed violet is classified in the subsection Rostratae. Because of recent human activity on sandy seashores, this species is now recorded as endangered. The present status of this species was surveyed and its genetic diversity was analyzed using 17 newly developed simple sequence repeat markers. The results are compared with those for two commonly found inland violet species in the same subsection, V. grypoceras and V. kusanoana. The coastal violet populations showed markedly lower genetic diversity than those of the common species, V. grypoceras, whereas V. kusanoana showed intermediate genetic diversity. About a half of the total genetic variation of V. grayi is found among populations in analysis of molecular variance. In contrast, only less than 25% of the total variation is found among populations for the two common violets. These three species showed a remarkably high inbreeding coefficient, which indicates high inbreeding under natural mating conditions, although these species have both chasmogamous and cleistogamous flowers. In a population-based dendrogram, V. grayi was clustered in a single and concise group, whereas V. grypoceras and V. kusanoana were found in somewhat discrete positions. In the Bayesian analysis of genetic structure of the coastal violet, delta K was the highest at K = 7, and the seven clusters almost correspond to seven populations studied here. Implications of the conservation of this endangered coastal violet are presented.     

SPATIAL AND TEMPORAL VARIATION IN CHASMOGAMY AND CLEISTOGAMY IN OXALIS MONTANA (OXALIDACEAE)

Chasmogamous (CH) and cleistogamous (CL) flower production was investigated in natural populations of the perennial herb Oxalis montana in southern Quebec, Canada. Every 10–12 days during two flowering seasons, we recorded the reproductive output of approximately 2,000 plants distributed among five forest sites. The percentage of plants flowering, proportion of flowering plants producing CH and CL flowers, CH and CL flower number per flowering plant, and the proportion of flowers that are CL differed significantly among sites and corresponded to site differences in forest type and habitat quality. Unlike patterns in most CL species, however, reproduction by cleistogamy increased in sites with habitat conditions favorable to plant growth and sexual reproduction, and decreased in less productive sites. Chasmogamous and CL flower production increased with increasing plant size but plant size explained a significantly greater proportion of the variation in CL flower numbers. The percentage of flowering plants producing CH flowers decreased between years while the proportion of CL flowers increased at all sites during the second flowering season. The somewhat unusual aspects of reproduction in Oxalis may stem from its perennial habit which allows use of stored resources in adjusting the balance of cleistogamy versus chasmogamy in different environmental regimes.     

Phylogeny of Saxifraga section Saxifraga subsection Arachnoideae (Saxifragaceae) and the origin of low elevation shade-dwelling species

Saxifraga section Saxifraga subsection Arachnoideae is a lineage of 12 species distributed mainly in the European Alps. It is unusual in terms of ecological diversification by containing both high elevation species from exposed alpine habitats and low elevation species from shady habitats such as overhanging rocks and cave entrances. Our aims are to explore which of these habitat types is ancestral, and to identify the possible drivers of this remarkable ecological diversification. Using a Hybseq DNA-sequencing approach and a complete species sample we reconstructed and dated the phylogeny of subsection Arachnoideae. Using Landolt indicator values, this phylogenetic tree was used for the reconstruction of the evolution of temperature, light and soil pH requirements in this lineage. Diversification of subsection Arachnoideae started in the late Pliocene and continued through the Pleistocene. Both diversification among and within clades was largely allopatric, and species from shady habitats with low light requirements are distributed in well-known refugia. We hypothesize that low light requirements evolved when species persisting in cold-stage refugia were forced into marginal habitats by more competitive warm-stage vegetation. While we do not claim that such competition resulted in speciation, it very likely resulted in adaptive evolution.     

The evolution of pollen heteromorphism inViola: A phylogenetic approach

Pollen heteromorphism, defined here as the production within all flowers of a plant of several pollen morphs differing in aperture number, is common in angiosperms. We have focused on the evolution of pollen heteromorphism in the genusViola, in which about 1/3 of the species are pollen-heteromorphic. We have studied the distribution of pollen heteromorphism in the genusViola using a molecular phylogeny based on ITS sequence data. We show that pollen heteromorphism has evolved independently at least six times inViola. A comparative analysis shows that the occurrence of pollen heteromorphism is correlated with sporophytic polyploidy in all sections of the genus apart from sectionMelanium. This section differs from all other sections on several aspects such as flower morphology, absence of cleistogamous flowers, and a high proportion of heteromorphic species. We discuss the possible adaptiveness of pollen heteromorphism in this section.     

No evidence of contemporary interploidy gene flow between the closely related European woodland violets Viola reichenbachiana and V. riviniana (sect. Viola,Violaceae)

• Viola reichenbachiana (2n = 4x = 20) and V. riviniana (2n = 8x = 40) are closely related species widely distributed in Europe, often sharing the same habitat throughout their overlapping ranges. It has been suggested in numerous studies that their high intraspecific morphological variability and plasticity might have been further increased by interspecific hybridisation in contact zones, given the sympatry of the species and the incomplete sterility of their hybrid. The aims of this study were to: (i) confirm that V. reichenbachiana and V. riviniana have one 4x genome in common, and (ii) determine the impact of hybridisation and introgression on genetic variation of these two species in selected European populations.
• For our study, we used 31 Viola populations from four European countries, which were analysed using AFLP and sequencing of a variable plastid intergenic spacer, trnH‐psbA.
• Our analysis revealed that V. reichenbachiana exhibited larger haplotype diversity, having three species‐specific haplotypes versus one in V. riviniana. The relationships among haplotypes suggest transfer of common haplotypes into V. riviniana from both V. reichenbachiana and hypothetically the other, now extinct, parental species. AFLP analysis showed low overall genetic diversity of both species, with V. riviniana showing higher among‐population diversity. None of the morphologically designated hybrid populations had additive AFLP polymorphisms that would have indicated recent hybridisation. Also, kinship coefficients between both species did not indicate gene flow. V. riviniana showed significant population subdivision and significant isolation by distance, in contrast to V. reichenbachiana.
• The results indicate lack of gene flow between species, high influence of selfing on genetic variability, as well as probably only localised introgression toward V. riviniana.

     

A novel case of autogamy and cleistogamy in Dendrobium wangliangii: A rare orchid distributed in the dry‐hot valley

Dendrobium wangliangii is an epiphytic orchid distributed in the Jinshajiang dry‐hot valley in Luquan County, Yunnan Province, China. Most Dendrobium spp. typically have a low fruit set, but this orchid shows a higher fruit set under natural conditions despite the lack of effective pollinators. The pollination biology of the critically endangered D. wangliangii was investigated in this study. A fruit set rate of 33.33 ± 4.71% was observed after bagging treatment in 2017 and a high fruit set rate (65.72 ± 4.44% in 2011; 50.79 ± 5.44% in 2017) was observed under natural conditions, indicating that D. wangliangii is characterized by spontaneous self‐pollination. The anther cap blocked the growing pollinium; thus, the pollinium slid down and reached the stigmatic cavity, leading to autogamous self‐pollination. Specifically, 51.50% of 162 unopened flowers (total 257 flowers) of this Dendrobium species under extreme water‐deficit conditions developed into fruits, suggesting the presence of cleistogamy in D. wangliangii. Here, cleistogamy may represent the primary mode of pollination for this orchid. Spontaneous self‐pollination and specific cleistogamous autogamy could represent major adaptions to the drought and pollinator‐scarce habitat in the Jinshajiang dry‐hot valley.     

Molecular phylogenetics supports a clade of red algal parasites retaining native plastids: taxonomy and terminology revised

Parasitism is a life strategy that has repeatedly evolved within the Florideophyceae. Historically, the terms adelphoparasite and alloparasite have been used to distinguish parasites based on the relative phylogenetic relationship of host and parasite. However, analyses using molecular phylogenetics indicate that nearly all red algal parasites infect within their taxonomic family, and a range of relationships exist between host and parasite. To date, all investigated adelphoparasites have lost their plastid, and instead, incorporate a host‐derived plastid when packaging spores. In contrast, a highly reduced plastid lacking photosynthesis genes was sequenced from the alloparasite Choreocolax polysiphoniae. Here we present the complete Harveyella mirabilis plastid genome, which has also lost genes involved in photosynthesis, and a partial plastid genome from Leachiella pacifica. The H. mirabilis plastid shares more synteny with free‐living red algal plastids than that of C. polysiphoniae. Phylogenetic analysis demonstrates that C. polysiphoniae, H. mirabilis, and L. pacifica form a robustly supported clade of parasites, which retain their own plastid genomes, within the Rhodomelaceae. We therefore transfer all three genera from the exclusively parasitic family, Choreocolacaceae, to the Rhodomelaceae. Additionally, we recommend applying the terms archaeplastic parasites (formerly alloparasites), and neoplastic parasites (formerly adelphoparasites) to distinguish red algal parasites using a biological framework rather than taxonomic affiliation with their hosts.     

Viola pluviae sp. nov. (Violaceae), a member of subsect. Stolonosae in the Pacific Northwest region of North America

As hirtherto circumscribed, the allo‐octoploid Viola palustris L. exists in three distinct races in North America. The race occurring in the Pacific Northwest of North America has previously been shown to have originated by polyploidy from a different set of ancestral tetraploids than the nominal, Amphi‐Atlantic race. These two races differ also in a number of morphological characters, which has been a historical source of confusion. We here propose a new name for the western plants, V. pluviae Marcussen, H.E. Ballard & Blaxland, and typify it with a specimen collected near Mt Rainier (USA, Washington) with known chromosome number. Distribution, phylogenetic history and taxonomy are discussed and an updated morphological key to the Pacific Northwestern species of Viola is presented.     

Effects of Gibberellic Acid on Floral Development in vivo and in vitro in the Cleistogamous Species, Lamium amplexicaule L.

Using established developmental markers in the corolla and androeciumof Lamium amplexicaule L., we investigated the apparent inductionof open (chasmogamous — CH) from closed (cleistogamous— CL) flowers after application of GA₃ (0.1 mM). In vivotreatment of potentially CL flower primordia caused cell expansionbut not the increased cell division in anthers and corolla necessaryto convert a CL into a CH flower. Floral primordia that appearedto be of undetermined floral type were grown in vitro on a basalmedium supplemented with kinetin (0.1 p.p.m.) and grew to maturityas CL flowers. On media additionally supplemented with GA₃,flowers underwent anthesis, but no true CH flower was produced.Gibberellins appear to be directly responsible for anthesisin the CH flower; but additional, as yet unknown growth factorsare involved in the switch from CL to CH floral form in a developinginflorescence. floral morphogenesis, Lamium amplexicaule L, henbit, cleistogamy, gibberellin, kinetin, anthesis     

Is plasticity across seasons adaptive in the annual cleistogamous plant Lamium amplexicaule?

Background and aims Many angiosperms exhibit cleistogamy, the production of both cleistogamous flowers (CL), which remain closed and obligately self-pollinated, and chasmogamous flowers (CH), which are potentially open-pollinated. The CH proportion can be plastic. Plasticity is adaptive if environmental changes can be reliably assessed and responded to with an appropriate phenotype and if plastic genotypes have higher fitness in variable environments than non-plastic ones.Methods We studied the plastic response of four natural populations from northern and southern France of an annual cleistogamous plant, Lamium amplexicaule, to predictable seasonal variation. Plants were grown in a semi-controlled environment in spring and in autumn. We assessed the variation in flower number, phenology and cleistogamy-related traits, which were all plastic with respect to season. The CH proportion was higher in spring than in autumn in all four populations.Key Results We showed significant stabilizing selection for cleistogamy traits, with higher optimal CH proportions and more pronounced stabilizing selection in spring than in autumn. Observed CH proportions were close to the predicted optimal CH proportions in each season except in autumn for southern populations, which do not experience the autumnal growing season in nature.Conclusions These results are consistent with adaptive plasticity across seasons of cleistogamy in L. amplexicaule. We propose that adaptive plasticity of cleistogamy could be driven by pollination environment variation, with CL flowers providing reproductive assurance when pollinators are scarce and CH flowers reducing the inbreeding depression in offspring when pollinators are abundant.     

Cleistogamy in the rare high Andean perennial herb Cryptantha capituliflora (Boraginaceae)

 Axial and apical flowers of Cryptantha capituliflora were analyzed with regard to morphology and pollen tube growth to assess the occurrence of cleistogamy. Although intermediate floral forms do occur, cleistogamous flowers were significantly smaller than chasmogamous flowers, had fewer anthers, and showed a distinctive stigmatic surface. Chasmogamous flowers can be cross-pollinated. Nevertheless, the growth of self-pollen tubes in few chasmogamous buds jointly with flower characters suggests that these flowers can probably produce fruits through autonomous selfing. The mean seed number per fruit did not differ between fruits from chasmogamous and cleistogamous flowers. Cleistogamous flowers were only observed in axial inflorescences, which are completely covered by the leaf. Other species of section Cryptantha also show the same trend, with cleistogamous flowers located in the lower half of the stems. This pattern is discussed in relation to dissimilarities in the outcrossing opportunities between flower types within the plant. Received May 22, 2002; accepted November 14, 2002 Published online: March 20, 2003     

VARIATION IN THE DEGREE OF CLEISTOGAMY WITHIN AND AMONG SPECIES OF THE GRASS DANTHONIA

North American species of the grass Danthonia bear distinct chasmogamous and cleistogamous flowers. Four taxa occurring in North Carolina (D. compressa, D. epilis, D. sericea, and D. spicata) were examined for variation in the degree of cleistogamy. The flowering culms bear a single terminal panicle consisting of 4 to 12 spikelets of chasmogamous flowers. In each leaf axil, at the nodes of the flowering culm, is a single spikelet of cleistogamous flowers completely surrounded by the leaf sheath. The percentage of cleistogamous flowers produced on any culm depends on the number of chasmogamous and cleistogamous spikelets and the number of flowers in each type of spikelet. All four characters vary among the taxa examined. Danthonia compressa produces, on the average, 50% cleistogamous flowers; D. spicata, 25% cleistogamous flowers; D. sericea and D. epilis, 5% cleistogamous flowers. The degree of cleistogamy in D. spicata is associated with certain habitat features. Populations from mountain sites, disturbed sites, and non-woodland sites produced higher percentages of cleistogamous flowers than did populations from piedmont, undisturbed, and woodland sites, respectively. Grazing may favor increased cleistogamy because cleistogamous flowers are produced lower on the plant. In North Carolina, the most frequently grazed Danthonia taxa also produce the highest percentage of cleistogamous flowers.     

Die hypopeltaten Sepalen vonViola arvensis undViola mirabilis

Zusammenfassung Die Sepalen vonViola arvensis undViola mirabilis und möglicherweise allerViola-Arten sind, wie die Untersuchungen der äußeren Morphologie, der Ontogenie, der Histogenie und der Bündelmorphologie ergeben, hypopeltat-schildförmige bzw.-schlauchförmige Blätter, deren dorsale Querzone zu dem basalen Anhängsel auswächst. Durch diesen Spreitenbau, der beiViola nur bei den Kelchblättern auftritt, erweisen sich diese als laminale Kelchblätter.     

Relationships among pansies (Viola section Melanium) investigated using ITS and ISSR markers

Sequences of the nuclear region ITS and the variable molecular markers ISSR were used to estimate the phylogeny of the section Melanium of the genus Viola. We confirm that the so-called pansies form a derived and monophyletic group. Two floral characters, the upturned side petals and the large size of pollen grains appear to be synapomorphies in Melanium. The Melanium species are very closely related, as shown by the reduced genetic variation compared to the other sections of Viola. Our analyses suggest x=5 or x=7 as the possible base chromosome number of the section Melanium. Polyploidy and hybridization would have played an important role in the evolutionary history of this clade resulting in a wide range of chromosome number. The low genetic differentiation and the complex cytological evolution suggest that diversification in Melanium is the result of a reticulate evolution and rapid radiation in Europe and Northern Africa.     

Cleistogamy: A tool for the study of floral morphogenesis,function and evolution

Cleistogamy—the production of open (chasmogamous—CH) and closed (cleistogamous—CL) floral forms by a species—is widespread among the angiosperms. While the CL flower is autogamous, the CH flower may provide a means for outcrossing. The term “cleistogamy” has also been used to describe other phenomena. A classification of types of cleistogamy is proposed. In this review, a restricted definition of cleistogamy is used to refer to species which show real floral dimorphisms, with divergent developmental pathways leading to CL and CH as well as intermediate floral forms. Reductions in the androecium and corolla are the most common feature of the CL flowers. The structural, developmental, and functional aspects of cleistogamy are reviewed. Evidence is presented to show that the CL flowers have modifications in their development which ensure self pollination. A proposal is made for using this phenomenon of dimorphic flower production as a system for the study of floral morphogenesis, function and evolution.     

Plant dispersion,pollination and gene flow in Viola

Summary The effect of pollinator activity on gene flow in colonies of Viola were examined by measuring pollinator flight distances, the frequency of interplant flights and percent pollination under different plant spacing patterns. Pollinator flight distances were directly proportional to spacing parameters while the frequency of interplant flights and percent pollination were inversely proportional to spacing parameters. These findings show that gene flow is reduced by pollinator activity over a wide range of spacing parameters but in populations with low spacing means highly localized gene exchange can occur within the colony. Isolation of colonies may be expected under these circumstaces and cleistogamy may be the optimal breeding system. However, chasmogamous flowers may be important both in promoting within-colony gene exchange and long distance between-colony gene exchange corresponding to the sexual functions proposed in several recent models. Viola colonies appear to be semi-isolated demes with pollinator service which can bring adaptive genes to high localized frequencies, but which maintains low frequency, long-distance gene dispersal. This pattern corresponds to the Shifting Balance view of evolution.     

滇水金凤花距发育相关基因ABP的克隆及表达分析

为探究滇水金凤(Impatiens uliginosa)ABP基因的结构和表达特征,该研究以滇水金凤为材料,采用RT-PCR 技术对滇水金凤ABP基因进行克隆,运用DNAMAN和MEGA对其所编码的蛋白序列进行同源性分析和系统进化分析,并利用qRT-PCR分析ABP基因的时空表达模式。结果表明:(1)滇水金凤ABP基因的cDNA 全长为627 bp,编码208 aa,命名为IuABP基因,其蛋白具有Cupin超家族蛋白的典型结构。(2)同源性分析表明滇水金凤ABP基因的氨基酸序列与喜马拉雅凤仙花(I. glandulifera)、月季(Rose chinensis)、木薯(Manihot esculenta)等物种的同源性均达71%; 系统进化分析表明IuABP与喜马拉雅凤仙花(Impatiens glandulifera)聚为一支,亲缘关系最近。(3)qRT-PCR分析表明IuABP基因在滇水金凤花距发育的3个时期及2个部位均有表达。随着花距的发育,IuABP基因在滇水金凤花距檐部的表达量呈先下降后上升的趋势,在盛花期时达最高,而在花距距部的表达量逐渐下降。以上结果为进一步研究滇水金凤ABP基因在花距发育中的功能及其表达调控机制提供了一定的理论参考。