Pollen morphology in the genus Veronica L. (Plantaginaceae) and its systematic significance

Pollen grains of 30, mainly annual, species from 134 populations of the genus Veronica (Plantaginaceae, formerly Scrophulariaceae) from the Mediterranean Region, have been studied with light and scanning electron microscopy. Three pollen types are defined based on pollen size and exine surface sculpture. In some cases within the study group pollen size can be useful in species determination. Hypothetical relationships of pollen grain size and aperture number with ploidy level, style length and corolla diameter are discussed. The ornamentation of the exine in Veronica, although generally a conservative character within the genus, gives some additional support to the most recent infrageneric classification of Veronica.     

Mating system variation in Veronica (Plantaginaceae): inferences from pollen/ovule ratios and other reproductive traits

The pollen–ovule ratio (P/O) is commonly used to estimate the mode of sexual reproduction in flowering plants. In previous studies, a clear correspondence has been detected between this character and the degree of autogamy. We here investigate variation in this character and its expected correlates in the genus Veronica (Plantaginaceae). Pollen–ovule ratios of 45 species representing eleven percent of all the species in the genus were investigated and compared with results from crossing experiments from previous studies. In addition, multiple populations of 17 of the 45 studied species were sampled and a controlled‐environment experiment was conducted to evaluate the extent of intraspecific variation. Moreover, relationships between P/O and other primary and secondary reproductive characters of the Veronica flower were investigated in relation to a phylogenetic hypothesis in order to determine the phylogenetic constraints on reproductive characters. The differences in P/O among species correspond well to the diversity of mating systems in Veronica and correlate well with other floral characters such as corolla size. These characters together seem to allow a powerful and fast tool to infer mating systems. However, causes for intraspecific variation of P/O, such as different cytotypes, ecotypes or different growth conditions, need to be considered.     

Is genome downsizing associated with diversification in polyploid lineages of Veronica?

The study of genome size evolution in a phylogenetic context in related polyploid and diploid lineages can help us to understand the advantages and disadvantages of genome size changes and their effect on diversification. Here, we contribute 199 new DNA sequences and a nearly threefold increase in genome size estimates in polyploid and diploid Veronica (Plantaginaceae) (to 128 species, c. 30% of the genus) to provide a comprehensive baseline to explore the effect of genome size changes. We reconstructed internal transcribed spacer (ITS) and trnL‐trnL‐trnF phylogenetic trees and performed phylogenetic generalized least squares (PGLS), ancestral character state reconstruction, molecular dating and diversification analyses. Veronica 1C‐values range from 0.26 to 3.19 pg. Life history is significantly correlated with 1C‐value, whereas ploidy and chromosome number are strongly correlated with both 1C‐ and 1Cx‐values. The estimated ancestral Veronica 1Cx‐value is 0.65 pg, with significant genome downsizing in the polyploid Southern Hemisphere subgenus Pseudoveronica and two Northern Hemisphere subgenera, and significant genome upsizing in two diploid subgenera. These genomic downsizing events are accompanied by increased diversification rates, but a ‘core shift’ was only detected in the rate of subgenus Pseudoveronica. Polyploidy is important in the evolution of the genus, and a link between genome downsizing and polyploid diversification and species radiations is hypothesized. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 178 , 243–266.     

Phylogeny of Veronica in the Southern and Northern Hemispheres based on plastid,nuclear ribosomal and nuclear low-copy DNA

The cosmopolitan and ecologically diverse genus Veronica with approximately 450 species is the largest genus of the newly circumscribed Plantaginaceae. Previous analyses of Veronica DNA sequences were in stark contrast to traditional systematics. However, analyses did not allow many inferences regarding the relationship between major groups identified, hindering further analysis of diversification and evolutionary trends in the genus. To resolve the backbone relationships of Veronica, we added sequences from additional plastid DNA regions to existing data and analyzed matching data sets for 78 taxa and more than 5000 aligned characters from nuclear ribosomal DNA and plastid DNA regions. The results provide the best resolved and supported estimate of relationships among major groups in the Northern (Veronica s. str.) and Southern Hemisphere (hebes). We present new informal names for the five main species groups within the Southern Hemisphere sect. Hebe. Furthermore, in two instances we provide morphological and karyological characters supporting these relationships. Finally, we present the first evidence from nuclear low-copy CYCLOIDEA2-region to compare results from the plastid genome with the nuclear genome.     

Veronica: Iridoids and cornoside as chemosystematic markers

The iridoid glucoside, ajugol, and the phenylethanoid glucoside, cornoside, have been isolated from species of Veronica (Plantaginaceae) for the first time. The presence of these compounds has been screened in 18 plant accessions belonging to 15 species of Veronica (Plantaginaceae), by isolation or NMR spectroscopy of crude extracts. In addition, the distribution of iridoids in the genus has been reviewed, using mainly the published data of isolated compounds. Using the recent expansion and reclassification of the genus based on DNA-sequence results as the model, we find that the genus is rather homogeneous with regard to the distribution of iridoid glucosides, aucubin and/or catalpol as well as 6-O-esters of catalpol being universally present in 10 of the 12 subgenera for which data exist. Only the two subgenera Pocilla and Chamaedrys deviate from this pattern. Pocilla is heterogeneous; in this subgenus, species in subsect. Agrestes contain the standard iridoid garniture, while species in subsect. Biloba do not contain the 6-O-esters of catalpol, but ajugol instead. Veronica intercedens (subsect. Subracemosae) differs from the remainder of the subgenus in only containing 5-hydroxylated iridoids (melittoside and globularifolin) and is so far the only species within the genus in which such compounds have been detected. These chemical differences are clearly reflected in the DNA-based phylogram of the subgenus. Subg. Chamaedrys appears homogeneous in lacking iridoids or only containing these in small amounts, but instead half of the investigated species contained the phenylethanoid glucoside cornoside. The distribution of this compound in angiosperms is reviewed; cornoside often substitutes iridoid glucosides in plants where these are expected to be present. The chemical results of Veronica fit in very well with the phylogenetic implications of the DNA-sequence results.     

Pioneering polyploids: the impact of whole-genome duplication on biome shifting in New Zealand Coprosma (Rubiaceae) and Veronica (Plantaginaceae)

The role of whole-genome duplication (WGD) in facilitating shifts into novel biomes remains unknown. Focusing on two diverse woody plant groups in New Zealand, Coprosma (Rubiaceae) and Veronica (Plantaginaceae), we investigate how biome occupancy varies with ploidy level, and test the hypothesis that WGD increases the rate of biome shifting. Ploidy levels and biome occupancy (forest, open and alpine) were determined for indigenous species in both clades. The distribution of low-ploidy (Coprosma: 2x, Veronica: 6x) versus high-ploidy (Coprosma: 4–10x, Veronica: 12–18x) species across biomes was tested statistically. Estimation of the phylogenetic history of biome occupancy and WGD was performed using time-calibrated phylogenies and the R package BioGeoBEARS. Trait-dependent dispersal models were implemented to determine support for an increased rate of biome shifting among high-ploidy lineages. We find support for a greater than random portion of high-ploidy species occupying multiple biomes. We also find strong support for high-ploidy lineages showing a three- to eightfold increase in the rate of biome shifts. These results suggest that WGD promotes ecological expansion into new biomes.     

<Emphasis Type="Italic">In vivo</Emphasis> and <Emphasis Type="Italic">in vitro</Emphasis> antioxidant effects of three Veronica species

The aim of the present study was to examine the antioxidant activity of three Veronica species (Plantaginaceae). The antioxidant potential of various extracts obtained from aerial flowering parts was evaluated by DPPH-free (1,1-diphenyl-2-picrylhydrazyl-free) radical scavenging activity and ferric-reducing antioxidant power assays. Considerable antioxidant activity was observed in the plant samples (FRAP values ranged from 0.97 to 4.85 mmol Fe²⁺/g, and DPPH IC₅₀ values from 12.58 to 66.34 μg/ml); however, these levels were lower than the activity of the control compound butylated hydroxytoluene (BHT) (FRAP: 10.58 mmol Fe²⁺/g; DPPH IC₅₀: 9.57 μg/ml). Also, the in vivo antioxidant effects were evaluated in several hepatic antioxidant systems in rats (activities of glutathione peroxidase, glutathione reductase, peroxidase, catalase, xanthine oxidase, glutathione content and level of thiobarbituric acid reactive substances) after treatment with different Veronica extracts, or in combination with carbon tetrachloride (CCl₄). Pretreatment with 100 mg/kg b.w. of Veronica extracts inhibited CCl₄-induced liver injury by decreasing TBA-RS level, increasing GSH content, and bringing the activities of CAT and Px to control levels. The present study suggests that the extracts analyzed could protect the liver cells from CCl₄-induced liver damage by their antioxidative effect on hepatocytes.     

Plastome comparison and evolution within the tribes of Plantaginaceae: Insights from an Asian gypsyweed

In spite of availability of several plastomes representing different tribes of Plantaginaceae, sparse attempts have been made to understand the plastome structure, evolution, and phylogenomics. In the present study, we have made an effort to understand the gene content and plastome evolution in the family Plantaginaceae using the newly generated plastome sequence of Veronica ovata subsp. kiusiana, a taxon native to SE Asia. In the first-ever attempt, plastomes of seven out of 10 tribes of Plantaginaceae have been compared to understand the evolution across the tribes of Plantaginaceae. The size of the plastome of V. ovata subsp. kiusiana is 152,249 bp, showing a typical quadripartite structure containing LSC, SSC, and two IRs with the sizes of 83,187, 17,704, and 25,679 respectively. The plastome comparison revealed the unique deletions in ycf2 and ndhF genes of members of different tribes, and also revealed high nucleotide variable hotspots. The study also revealed six highly variable genes and intergenic spacer viz. rps16, rps15-ycf1, ccsA-ndhD, ndhC-trnV, petN-psbM, and ycf1-trnN as potential DNA barcodes for the genus Veronica. The phylogenomic study revealed the sister relationship between V. ovata subsp. kiusiana and V. persica and also suggested the tentative placement of seven tribes in the family Plantaginaceae.     

Veronica: Parallel morphological evolution and phylogeography in the Mediterranean

The genus Veronica s. lat. comprises about 450 species (including about 180 species from the southern hemisphere Hebe-complex), many of which grow in the Mediterranean area. Their extreme variability in morphology, life form and habitats has led to many suggestions regarding evolution and biogeography. Difficulties arise from parallel syndromes, widespread among alpine species and lowland perennials, and particularly among annual species of the genus. We have used sequences of the plastid trnL-F region and nuclear ribosomal ITS sequences to differentiate between different clades of Veronica and reveal cases of parallel evolution. Based on this data, cases of parallel evolution have been found in biogeographical patterns among the alpine species of Veroniceae, in which species from European mountains have affinities to those in the Central Asian/Himalayan region whereas alpine species from Turkey are probably more recently derived from lowland southwestern Asian taxa. Different subspecies of Veronica bombycina gained their characteristic morphology independently and parallel in adaptation to their alpine environment. Pinnatifid leaves have been gained parallel in perennial grassland species of Veronica. Finally, parallel evolutionary trends in many characters, not only morphological but also molecular characters, are common among annual species of Veronica.We wish to thank the Studienstiftung des deutschen Volkes for a doctoral scholarship to DCA and the Spanish Ministerio de Ciencia y Tecnologia for a postdoctoral grant (programa FPI) to MMO. This work was partially supported by the Junta de Castilla y Leon and the European Union (FSE) through the research project SA117/01. We also thank Prof. F. Ehrendorfer for critical comments and valuable discussions. Further, DCA thanks Daniela Hanfland and Yoshiki Nakamoto for help in the field. Manfred A. Fischer, Lena Struwe, Bernhard Dickore, Christoph Dobes, Eberhard Fischer, Niels Köster, Gerald Schneeweiss, Tod Stuessy, Tim Utteridge and Matsugo Yokota have provided plant material for this study.     

Veronica: Chemical characters for the support of phylogenetic relationships based on nuclear ribosomal and plastid DNA sequence data

Molecular phylogenetic analyses have revealed many relationships in Veronica (Plantaginaceae) never anticipated before. However, phytochemical characters show good congruence with DNA-based analyses. We have analysed a combined data set of 49 species and subspecies derived from the nuclear ribosomal ITS-region (18 new sequences) and the plastid trnL-F region (12 new sequences) of Veronica emphasizing subgenera Chamaedrys and Pocilla and separate analyses of subgenera Pentasepalae (ITS only) and Pseudolysimachium. Results for subgenus Chamaedrys show that European and Asian perennial species are monophyletic sister groups with the annual species consecutive sisters to them. All species of Veronica that contain cornoside are found in this subgenus, although some species seem to have secondarily lost the ability to produce this compound. Subgenera Pocilla and Pentasepalae are well supported sister groups characterized by the occurrence of 8-hydroxyflavones. The traditional subsection Biloba of subgenus Pocilla is biphyletic with Veronica intercedens being clearly separate from the rest of the group. This result is mirrored by the unusual phytochemical arsenal of V. intercedens, which is the only species in the genus analysed to date to contain melittoside and globularifolin. Subgenus Pentasepalae appears to be a clade of diverse lineages from southwestern Asia and a single European clade. Species shown to have 6-hydroxyflavones do not form a monophyletic group. Subgenus Pseudolysimachium seems to have originated in Eastern Asia. 6-Hydroxyflavones acylated with phenolic acids are common in this subgenus but may have originated only later in the evolution of the group. Possible chemotaxonomic markers for other groups are discussed.     

A new species of <Emphasis Type="Italic">Macroptilium</Emphasis> (Benth.) Urb. (<Emphasis Type="Italic">Leguminosae</Emphasis>: <Emphasis Type="Italic">Papilionoideae</Emphasis>: <Emphasis Type="Italic">Phaseolinae</Emphasis>) from North-Eastern Brazil

Summary  A new species of Macroptilium sect. Microcochle (Benth.) J. A. Lackey is herein described from the states of Piauí and Bahia, Brazil. Macroptilium cochleatum is characterized by few-flowered inflorescences, calyx teeth longer than the tube, a tightly four-coiled keel, and linear, patent fruits. The discovery of this unique Macroptilium species, with its distally coiled keel, expands the diagnostic features of the genus. A key to the Brazilian species of sect. Microcochle is provided.     

Evolutionary Rates in Veronica L. (Plantaginaceae): Disentangling the Influence of Life History and Breeding System

The evolutionary rate at which DNA sequences evolve is known to differ between different groups of organisms. However, the reasons for these different rates are seldom known. Among plants, the generation-time hypothesis, which states that organisms that reproduce faster also have more DNA substitutions per time, has gained most popularity. We evaluate the generation-time hypothesis using 131 DNA sequences from the plastid trnLF region and the nuclear ribosomal ITS region of the genus Veronica (Plantaginaceae). We also examine the alternative hypothesis that a higher substitution rate is correlated with selfing breeding system. Selfing is associated with annual life history in many organisms and may thus often be the underlying reason for observed correlations of annual life history with other characters. We provide evidence that annual life history is more likely to be the responsible factor for higher substitution rates in Veronica than a selfing breeding system. Nevertheless, the way in which annual life history may influence substitution rate in detail remains unknown, and some possibilities are discussed.     

<Emphasis Type="Italic">Mastigostyla</Emphasis> I. M. Johnst. in Bolivia: three new species and new data on <Emphasis Type="Italic">M. cardenasii</Emphasis> R. C. Foster

Four species of Mastigostyla (Iridaceae) from Bolivia are described, illustrated and maps of their distribution provided: M. cardenasii R. C. Foster, M. chuquisacensis Huaylla & Wilkin, M. torotoroensis Huaylla & Wilkin and M. woodii Huaylla & Wilkin, the last three as new species. Mastigostyla gracilis R. C. Foster is synonymised under M. cardenasii R. C. Foster. Notes on conservation status, nomenclature and identification are included. Although some uncertainty exists over generic placement due to phylogenetic systematic research now in progress, marked infraspecific variation is documented for the first time within South American Tigridieae. This draws attention to the need for more extensive sampling if adequate species delimitation is to be achieved in Tigridieae. Photographs illustrate morphological variation in all four species. A key is provided to facilitate identification of Bolivian Mastigostyla.     

New species of Metepsilonema (Nemata,Epsilonematidae) from the Channel and the Mediterranean Sea with a polytomous key for the identification of species of the genus

Four new species of Metepsilonema are described from sublittoral stations from the Channel and are mainly characterized: M. volutum sp.nov. by the large body size for the genus up to 460 m, 118–121 well overlapping annuli and wide multispiral amphids; M. amphidoxum sp.nov. by 127–133 shortly overlapping annuli and sexual dimorphism in amphid size; M. comptum sp.nov. by 133–140 non-overlapping annuli, anteriormost annuli provided with a wide lumen and borders ornamented with ridges and M. corrugatum sp.nov. by 108–111 overlapping annuli provided with ridges). Additional information is provided for M. callosum together with a discussion on intraspecific variability. A new species of Metepsilonema spec. based on females only, is described but remain unnamed. M. comptum sp.nov. and M. corrugatum sp.nov. are also occurring in the Mediterranean Sea. A polytomous key to species level is presented.     

Two new species of <Emphasis Type="Italic">Lonchocarpus</Emphasis> (Leguminosae,Papilionoideae, Millettieae) from Brazil

Two new species of Lonchocarpus from Brazil, assigned to sect. Laxiflori Benth., are described and illustrated and their affinities, geographical distribution and conservation status are discussed. Lonchocarpus laticiferus from arboreal caatinga vegetation of Bahia State, and deciduous forest of Minas Gerais State is related to L. montanus A. M. G. Azevedo ex M. J. Silva & A. M. G. Azevedo. Lonchocarpus graciliflorus, known only from the rain forest of Paraná, is morphologically similar to L. longiunguiculatus M. J. Silva & A. M. G. Azevedo.     

Dietary specialization and the effects of plant species on potential multitrophic interactions of three species of nymphaline caterpillars

The diet breadth of insect herbivores influences their response to variation in plant quality, and these bitrophic interactions have implications for the higher‐level trophic interactions between herbivores and their natural enemies. In this comparative study, we examined the role of host plant species and plant secondary chemistry on the potential interactions between three species of nymphaline caterpillars and their natural enemies. The caterpillar species (all Lepidoptera: Nymphalidae) varied in their degree of specialization: the buckeye, Junonia coenia Hübner, is a specialist on plants that contain iridoid glycosides (IGs); the white peacock, Anartia jatrophae L., feeds on plants in five families, some of which contain IGs and some of which do not; and the painted lady, Vanessa cardui L., is a generalist, feeding on plants in at least 15 families. Each species was reared on leaves of an introduced host plant, Plantago lanceolata L. (Plantaginaceae), which produces two IGs, aucubin and catalpol, and on another plant species that is a common host plant. These alternate host plants were Plantago major L. (Plantaginaceae) for J. coenia, Bacopa monnieri (L.) Pennell (Plantaginaceae) for A. jatrophae, and Malva parviflora L. (Malvaceae) for V. cardui. We examined growth, sequestration, and immune response of these caterpillars on the different host plant species. Junonia coenia developed more rapidly and sequestered higher IG concentrations when reared on P. lanceolata, whereas both other species grew more slowly on P. lanceolata. Host plant did not influence immune response of J. coenia or A. jatrophae, whereas V. cardui immune response was weaker when reared on P. lanceolata. Junonia coenia was most efficient at IG sequestration and A. jatrophae was least efficient, when all three species were reared on P. lanceolata. These results indicate that diet breadth may play an important role in structuring tritrophic interactions, and this role should be further explored.     

Morphological,molecular and biological evidence reveal two cryptic species in Mecinus janthinus Germar (Coleoptera,Curculionidae), a successful biological control agent of Dalmatian toadflax,Linaria dalmatica (Lamiales,Plantaginaceae)

A combined morphological, molecular and biological study shows that the weevil species presently named Mecinus janthinus is actually composed of two different cryptic species: M. janthinus Germar, 1821 and M. janthiniformis To?evski & Caldara sp.n. These species are morphologically distinguishable from each other by a few very subtle morphological characters. On the contrary, they are more readily distinguishable by both molecular and biological characters. A molecular assessment based on the mitochondrial DNA cytochrome oxidase subunit II gene revealed fixed differences between the two species with p‐distances between samples of both species ranging from 1.3 to 2.4%. In addition to this, the larvae of the two species are found to develop on different species within the genus Linaria (Plantaginaceae): M. janthinus is associated with yellow toadflax (L. vulgaris) and M. janthiniformis with broomleaf toadflax (L. genistifolia) and Dalmatian toadflax (L. dalmatica). Molecular and host use records further suggest the occurrence of a third species associated with L. vulgaris within M. janthinus, sampled from north Switzerland, central Hungary and east Serbia. The significance of these new findings is of particular importance because species of the M. janthinus group are used, or are potential candidates, for the biological control of invasive toadflaxes in North America.     

<Emphasis Type="Italic">Tabaroa</Emphasis>, a new genus of Leguminosae tribe Brongniartieae from Brazil

A new genus of Leguminosae in tribe Brongniartieae is proposed based on a new species endemic to the Caatinga of Bahia state. The new genus is named as Tabaroa L. P. Queiroz, G. P. Lewis & M. F. Wojc. and the new species as Tabaroa caatingicola L. P. Queiroz, G. P. Lewis & M. F. Wojc. A phylogenetic study of the Brongniartieae based on nuclear rDNA ITS and plastid matK sequences supports a closer relationship between Tabaroa and Harpalyce Moc. & Sessé than to the two South American genera Poecilanthe Benth. and Cyclolobium Benth., which are more similar morphologically. Optimisation of selected morphological characters on one of the most parsimonious trees indicates that the sessile ovary and the explosive pollen presentation are putative synapomorphies of the Tabaroa-Harpalyce clade. The genus Tabaroa may be diagnosed by the absence of peltate glandular trichomes; leaves imparipinnate, exstipellate and with opposite leaflets; flowers sessile, not resupinate, grouped in panicles; anthers apiculate; and fruit indehiscent. The only known species inhabits areas of arboreal caatinga on sandy soil in southwestern Bahia, near the boundaries of Dom Basílio and Livramento de Nossa Senhora municipalities.     

Growth and Reproductive Strategy in VeronicaSpp.

Seed production, germination and plant growth form were studiedin several species of Veronica. The objectives were to comparethe reproductive strategies of annual and perennial speciesof Veronica which varied in their habitats and aggressivenessas weeds. ‘Reproductive effort’ was fairly similar for bothannuals and perennials, falling within the commonly observedrange for annual species. Seed number per capsule and the 100-seedweight were unaffected by the growth conditions but plant growthform was substantially modified in V. agrestis. The small-seeded Veronica species required light for germinationand the winter annual V. hederifolia germinated much more vigorouslyafter a low temperature treatment. The growth form, vigour andextent of adventitious root production and rapid seed germinationof V. persica must be significant factors in the spread andestablishment of this species. Veronica spp., germination, growth form, reproductive strategy