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Insights into phylogenetic relationships of Veronica species (Plantaginaceae) based on comparative chloroplast genomics.

INTRODUCTION: Veronica L. is one of the most species-rich genera in Plantaginaceae and several species have medicinal, horticultural, or ecological value. METHODS: In this study, the complete chloroplast genomes of three Veronica species were assembled and annotated using Illumina sequencing data. RESULTS: The plastomes exhibited a typical quadripartite structures, with total lengths of 150,202 bp for Veronica biloba L., 151,159 bp for Veronica ciliata Fisch. and 151,098 bp for Veronica vandellioides Maxim. Each genome contained 130-132 unique genes, including 86-87 protein-coding genes, 36-37 tRNA genes, and 8 rRNA genes. Comparative analyses of 24 Veronica plastomes indicated that the IR/SC junctions were largely conserved, although slight boundary shifts occurred around rps19, ndhF, and ycf1. Forward, palindromic, complement, and reverse repeats were detected, and A/T mononucleotide repeats were the dominant SSR type. Nucleotide diversity analysis identified rpl32-trnL, trnK-rps16, rpl32, ycf1, ndhF, accD, matK, and rpoB as highly variable regions. Phylogenetic analyses recovered Veronica as a well-supported monophyletic lineage and clarified the plastid positions of the three newly sequenced species. Divergence time estimation suggested that the estimation suggested of Veronica was around 14.9 Ma, with V. biloba, V. ciliata and V. vandellioides diverging approximately 3.9 Ma, 0.6 Ma, and 6.9 Ma, respectively. DISCUSSION: Because the analyses were based on plastid genomes, the inferred topology should be interpreted as chloroplast phylogenetic evidence rather than a complete species-history reconstruction. These results provide plastome resources and molecular evidence for taxonomy, species identification, and future evolutionary studies of Veronica.

Plantaginaceae↗

Chemotaxonomic markers in Digitalideae (Plantaginaceae).

In a chemosystematic investigation of Digitalideae (Plantaginaceae), the water-soluble part of extracts of two species of Digitalis, two species of Isoplexis, as well as Erinus alpinus and Lafuentea rotundifolia were studied with regard to their content of main carbohydrates, iridoids and caffeoyl phenylethanoid glycosides (CPGs). Digitalis and Isoplexis contained sorbitol, cornoside and a number of other phenylethanoid glycosides including the new tyrosol beta-D-mannopyranoside, sceptroside but were found to lack iridoid glucosides. Erinus contained mainly glucose, the new 8,9-double bond iridoid, erinoside, and a number of known iridoid glucosides including two esters of 6-rhamnopyranosylcatalpol, as well as the CPG poliumoside. Finally, Lafuentea was characterized by the presence of glucose, aucubin and cryptamygin B but apparently lacked CPGs. The chemosystematic significance of the isolated compounds is discussed.

Carbohydrates↗

Chemotaxonomy of Veroniceae and its allies in the Plantaginaceae.

In a chemosystematic investigation of tribe Veroniceae (Plantaginaceae), representatives of Camptoloma, Sibthorpia, Veronica subg. Pentasepalae and subg. Hebe, Veronicastrum, Wulfenia, and the related Ellisiophyllum and Globularia were examined for non-flavonoid glycosides. From the 14 species studied, 28 different iridoid glucosides and 10 caffeoyl phenylethanoid glucosides (CPGs), as well as salidroside and arbutin were isolated and characterized by NMR; of these, five compounds were previously unknown. It was found that the representatives of Veroniceae, as well as Globularia, were characterized by mannitol, aucubin, catalpol and catalpol esters. Each of the three studied species of Veronica subg. Hebe contained at least one of the 6-O-catalpol esters typical for Veronica s. str. (verminoside), supporting the inclusion of Hebe in Veronica. However, their main constituents were esters of 6-O-rhamnopyranosylcatalpol; a CPG, hebeoside (2'-beta-xylopyranosyl-verbascoside) was isolated from V. (Hebe) salicifolia. The two species of Veronicastrum also contained 6-O-rhamnopyranosylcatalpol esters, including the previously unknown 2'',3''- and 3'',4''-dicinnamoyl derivatives and, in contrast to the earlier reports, they lacked 6-O-catalpol esters. The main iridoid constituents in the three investigated species of Wulfenia were 10-O-aucubin and 10-O-catalpol esters (isoscrophularioside or globularin) while baldaccioside (10-O-cinnamoyl asystasioside E) was isolated from W. baldaccii. Globularia vulgaris contained 10-O-catalpol esters (e.g., globularin) and, in addition, asperuloside together with its benzoyl analogue named besperuloside. The representatives of Sibthorpia and Ellisiophyllum were almost completely devoid of iridoids; this, however, together with the CPGs present implied a close relationship between the two genera. Camptoloma lyperiiflorum lacked hexitols but contained esters of 6-O-rhamnopyranosylcatalpol different from those found in Veroniceae but known from Buddleja, Scrophularia and Verbascum (Scrophulariaceae s. str.).

Esters↗

Incongruence in Veroniceae (Plantaginaceae): evidence from two plastid and a nuclear ribosomal DNA region.

Nucleotide data from nuclear ribosomal and plastid DNA did not agree on all relationships within Veroniceae (Plantaginaceae) in previous analyses. We present here additional data from a second plastid DNA marker, the rps16 intron, and evaluate incongruence between the ribosomal and plastid DNA matrices. Based on these evaluations and analysis of secondary structure in ITS, we conclude that there is a biological cause for incongruence among the genera of Veroniceae. The sequence data imply a history of hybridization and polyploidization, in which ancestors of Wulfeniopsis and Veronica+Paederota gave rise to Picrorhiza and ancestors of Wulfeniopsis and Wulfenia gave rise to Veronicastrum.

Cell Nucleus↗

The genome sequence of Veronica verna L., 1753 (Lamiales: Plantaginaceae).

We present a genome assembly of Veronica verna (Spring Speedwell; Streptophyta; Magnoliopsida; Lamiales; Plantaginaceae). The genome sequence has a total length of 463.88 megabases. Most of the assembly (98.85%) is scaffolded into 8 chromosomal pseudomolecules. The mitochondrial sequence has a length of 311.81 kilobases and the plastid genome assembly has a length of 149.82 kilobases. Gene annotation of this assembly on Ensembl identified 22 903 protein-coding genes. This assembly was generated as part of the Darwin Tree of Life project, which produces reference genomes for eukaryotic species found in Britain and Ireland.

Asterales↗

Influence of restoration on arbuscular mycorrhiza of Biscutella laevigata L. (Brassicaceae) and Plantago lanceolata L. (Plantaginaceae) from calamine spoil mounds.

The arbuscular mycorrhizal status of two plant species, Biscutella laevigata L. and Plantago lanceolata L., was investigated on calamine spoil mounds in Bolesław (southern Poland). Although B. laevigata is a member of the Brassicaceae, a family generally accepted as non-mycorrhizal, this species formed AM symbioses on both heavy metal-contaminated and non-contaminated sites. Besides vesicles and coils, arbuscules were also observed, especially in roots collected prior to seed maturity. Relative mycorrhizal root length and relative arbuscular richness were usually much higher in P. lanceolata than in B. laevigata but not absolute arbuscule richness. Roots of P. lanceolata showed higher colonisation than B. laevigata. Although roots were collected from plants in close proximity, no correlation in mycorrhizal parameters was found between the two species.

Biodegradation, Environmental↗

Systematic significance of seed morphology in veronica (plantaginaceae): a phylogenetic perspective.

BACKGROUND AND AIMS: A new infrageneric rearrangement for Veronica has been proposed based on the most recent evidence from DNA sequence data, morphological evidence, and biogeographical considerations. Looking for morphological synapomorphies for each monophyletic subgenus has been problematic, due to difficulties arising from widespread homoplasy (mainly parallel evolution). In an attempt to overcome these difficulties, previously underexplored morphological characters are starting to be studied in more depth. METHODS: A molecular phylogenetic hypothesis was used based on sequences of ITS (nuclear ribosomal DNA) and plastid trnL-F regions, as a framework to test the use of seed coat ultrastructure (studied under scanning electron microscope) in the systematics of the genus. A sample of 132 taxa representing ten of the 13 subgenera in Veronica, excluding the species of the southern hemisphere Hebe complex and the exclusively North American subgenus Synthyris, was studied. KEY RESULTS AND CONCLUSIONS: The results demonstrate that, in many cases, the ultrastucture of the testa can be employed to assess relationships of taxa within the genus, and the character provides additional support for molecular trees. Further characters relevant for the classification of Veronica, i.e. base chromosome number, iridoid chemical data, life cycle, inflorescence position, have been taken into consideration in a discussion where an attempt is made to highlight the best traits to characterize each subgenus investigated.

Classification↗

Family- and population-level responses to atmospheric CO2 concentration: gas exchange and the allocation of C, N, and biomass in Plantago lanceolata (Plantaginaceae).

To ascertain the inheritance of responses to changing atmospheric CO(2) content, we partitioned response to elevated CO(2) in Plantago lanceolata between families and populations in 18 families in two populations. Plants were grown in 35 Pa and 71 Pa partial pressure of CO(2) (pCO(2)) in open-top chambers. We measured above- and belowground mass, carbon (C), nitrogen (N), hexose sugar, and gas exchange properties in both CO(2) treatments. Families within populations differed in mass, mass allocation, root : shoot ratios, aboveground percentage N, C : N ratio, and gas exchange properties. The CO(2) × family interaction is the main indicator of potential evolutionary responses to changing CO(2). Significant CO(2) × family interactions were observed for N content, C : N ratio, and photosynthetic rate (A: instantaneous light-saturated carbon assimilation capacity), intercellular CO(2) concentration, transpiration rate (E), and water use efficiency (WUE = A/E), but not for stomatal conductance. Families differed significantly in acclimation across time. The ratio of A in elevated vs. ambient growth CO(2), when measured at a common internal CO(2) partial pressure was 0.79, indicating down-regulation of A under CO(2) enrichment. Mass, C : N ratio, percentage, C (%C), and soluble sugar all increased significantly but overall %N did not change. Increases in %C and sugar were significant and were coincident with redistribution of N aboveground. The observed variation among populations and families in response to CO(2) is evidence of genetic variation in response and therefore of the potential for novel evolutionary trajectories with rising atmospheric CO(2).

Journal Article↗

[Sub-chronic toxicity and test of eye irritability of leaf aqueous extract from Plantago major (plantaginaceae)].

For the sub-chronic toxicity an aqueous preparation of Plantago major leaves was tested in 20 male NGP mice, with an average weight of 20.15 g and separated in two groups of ten individuals each. The dose used was 2000 mg/kg and the control group received 0.5 ml of distilled water. The extract administration was done daily during five days at week for a total period of 40 days. Signs of sub-chronic toxicity were observed in the days two and 12 of treatment. No significant change in corporal weight was observed. The ocular irritation was tested in five New Zeland male rabbits, with an average weight of 3.640 kg. The dose used was a 200 microliters the preparation (100 mg/ml) of Plantago major leaves, instill into the right eye and the control was used the left eye instill 200 microliters of distilled water. The administration was done daily during five days. The extract shows no significant irritation during the observation period.

Animals↗

In vitro anti-hepatoma activity of fifteen natural medicines from Canada.

Fifteen crude drugs, Stellaria media Cyrill. (Caryophyllaceae), Calendula officinalis L. (Compositae), Achillea millefolium L. (Compositae), Verbascum thapsus L. (Scrophulariaceae), Plantago major L. (Plantaginaceae), Borago officinalis L. (Boraginaceae), Satureja hortensis L. (Labiatae), Coptis groenlandica Salisb. (Ranunculaceae), Cassia angustifolia Vahl. (Leguminosae), Origanum majorana L. (Labiatae), Centella asiatica L. (Umbelliferae), Caulophyllum thalictroides Mich. (Berberidaceae), Picea rubens Sargent. (Pinaceae), Rhamnus purshiana D.C. (Rhamnaceae) and Hibiscus sabdariffa L. (Malvaceae), which have been used as folk medicine in Canada, were evaluated for their anti-hepatoma activity on five human liver-cancer cell lines, i.e. HepG2/C3A, SK-HEP-1, HA22T/VGH, Hep3B and PLC/PRF/5. The samples were examined by in vitro evaluation for their cytotoxicity. The results showed that the effects of crude drugs on hepatitis B virus genome-containing cell lines were different from those against non hepatitis B virus genome-containing cell lines. C. groenlandica was observed to be the most effective against the growth of all five cell lines and its chemotherapeutic values will be of interest for further studies.

Antineoplastic Agents, Phytogenic↗

Neighbor species differentially alter resistance phenotypes in Plantago.

In this study, we investigated how neighbors (i.e., competitors) altered resistance phenotypes, namely plant size and levels of secondary compounds (iridoid glycosides), of individual plants and specifically tested whether neighbor identity mattered. We conducted a greenhouse experiment with Plantago lanceolata and Plantago major (Plantaginaceae) in which each species served as focal plants as well as neighbors in a factorial design. In addition, we harvested plants six and nine weeks after transplantation to test whether effects changed as plants grew. In both species, competition reduced plant size, and this effect increased over time. Plantago lanceolata neighbors suppressed growth of both focal plant species more than P. major neighbors. Effects of competition on levels of secondary compounds were more complex. Concentrations of iridoid glycosides were increased by competition in both species at harvest one. By the second harvest, an effect of competition on iridoid glycosides was found only in P. major. Neighbor identity influenced levels of iridoid glycosides in P. lanceolata at harvest one; concentrations were higher in plants grown with P. lanceolata neighbors than in plants grown with P. major neighbors. We also tested whether there was a trade-off between growth (biomass) and defense (levels of iridoid glycosides). Biomass and iridoid glycoside content were significantly correlated only in plants grown with competition and harvested at nine weeks, and this relationship was positive in both species, indicating that there was no trade-off between growth and defense. This study suggests that neighbor identity could play an important role in interspecific interactions, including the interactions of plants with other trophic levels.

Biomass↗

Biology of weed pollen allergens.

Weeds represent a heterogeneous group of plants, usually defined by no commercial or aesthetic value. Important allergenic weeds belong to the plant families Asteraceae, Amaranthaceae, Urticaceae, Euphorbiaceae, and Plantaginaceae. Major allergens from ragweed, mugwort, feverfew, pellitory, goosefoot, Russian thistle, plantain, and Mercurialis pollen have been characterized to varying degrees. Four major families of proteins seem to be the major cause of allergic reactions to weed pollen: the ragweed Amb a 1 family of pectate lyases; the defensin-like Art v 1 family from mugwort, feverfew, and probably also from sunflower; the Ole e 1-like allergens Pla l 1 from plantain and Che a 1 from goosefoot; and the nonspecific lipid transfer proteins Par j 1 and Par j 2 from pellitory. As described for other pollens, weed pollen also contains the panallergens profilin and calcium-binding proteins, which are responsible for extensive cross-reactivity among pollen-sensitized patients.

Allergens↗

Screening of medicinal plants for induction of somatic segregation activity in Aspergillus nidulans.

Knowledge about mutagenic properties of plants commonly used in traditional medicine is limited. A screening for genotoxic activity was carried out in aqueous or alcoholic extracts prepared from 13 medicinal plants widely used as folk medicine in Cuba: Lepidium virginicum L. (Brassicaceae): Plantago major L. and Plantago lanceolata L. (Plantaginaceae); Ortosiphon aristatus Blume, Mentha x piperita L., Melissa officinalis L. and Plectranthus amboinicus (Lour.) Spreng. (Lamiaceae); Cymbopogon citratus (DC.) Stapf (Poaceae); Passiflora incarnata L. (Passifloraceae); Zingiber officinale Roscoe (Zingiberaceae); Piper auritum HBK. (Piperaceae); Schinus terebinthifolius Raddi (Anacardeaceae) and Momordica charantia L. (Cucurbitaceae). A plate incorporation assay with Aspergillus nidulans was employed, allowing detection of somatic segregation as a result of mitotic crossing-over, chromosome malsegregation or clastogenic effects. Aspergillus nidulans D-30, a well-marked strain carrying four recessive mutations for conidial color in heterozygosity, which permitted the direct visual detection of segregants, was used throughout this study. As a result, only in the aqueous extract of one of the plants screened (Momordica charantia) a statistical significant increase in the frequency of segregant sectors per colony was observed, and consequently, a genotoxic effect is postulated.

Aspergillus nidulans↗

Effect of Plantago major on cell proliferation in vitro.

Plantago major (Plantaginaceae) is popularly used to treat tumors, infections and as a blood purifier. Aqueous, methanol, chloroform and hexane extracts of the aerial parts (leaves and seeds) were added to CD(1) mice bone marrow and spleen cultures incubated at 37 degrees C for 72h, and also added to Escherichia coli, Bacillus subtilis and Candida albicans cultures, while methanol extract dilutions were added to HTC-15, OVCAR, UISO and KB cell line cultures. Doses of 0.4 and 0.2 mg/mL of aqueous and methanol extracts increased the bone marrow cell concentration by 2.70- and 3.15-fold, respectively, and increased the spleen cell concentration by 3.38- and 6.39-fold, respectively (p < 0.001). Aqueous extract inhibited Bacillus subtilis growth from 78 to 21%; hexane extract inhibited the growth of Escherichia coli, and methanol and chloroform extracts weakly inhibited the growth of Bacillus subtilis and Escherichia coli, respectively. Methanol extract (1 microg/mL) decreased the UISO and OVCAR cell concentrations to 59 and 82%, respectively. Data demonstrate for the first time that Plantago major has hematopoietic activity in vitro.

Animals↗

Chemotaxonomy of Plantago. Iridoid glucosides and caffeoyl phenylethanoid glycosides.

Data for 34 species of Plantago (Plantaginaceae), including subgen. Littorella (= Littorella uniflora), have been collected with regard to their content of iridoid glucosides and caffeoyl phenylethanoid glycosides (CPGs). In the present work, 21 species were investigated for the first time and many known compounds were found together with three new iridoid glucosides. Of these, arborescoside and arborescosidic acid, both of the uncommon type with an 8,9-double bond, were present in several species, while 6-deoxymelittoside was found only in P. subulata. The known compounds deoxyloganic acid, caryoptoside and rehmannioside D were isolated from the genus for the first time. The earlier reported occurrence of sorbitol in the family was confirmed, and this compound was shown by NMR spectroscopy to be the main sugar in the three species investigated for this. The combined data show that CPGs are present in all species investigated. With regard to the iridoids, the distribution patterns showed a good correlation with the classification of Rahn. Thus, aucubin is typical for the whole genus, while bartsioside and catalpol as well as 5-substituted iridoids are each characteristic for a subgenus in the family. Finally, the close relationship between Plantago and Veronica suggested by chloroplast DNA sequence analysis. could be corroborated by the common occurrence of the rare 8,9-unsaturated iridoids in these two genera.

Glucosides↗