A Plant Model of α-Synucleinopathy: Expression of α-Synuclein A53T Variant in Hairy Root Cultures Leads to Proteostatic Stress and Dysregulation of Iron Metabolism.

Synucleinopathies, typified by Parkinson's disease (PD), entail the accumulation of α-synuclein (αSyn) aggregates in nerve cells. Various αSyn mutants, including the αSyn A53T variant linked to early-onset PD, increase the propensity for αSyn aggregate formation. In addition to disrupting protein homeostasis and inducing proteostatic stress, the aggregation of αSyn in PD is associated with an imbalance in iron metabolism, which increases the generation of reactive oxygen species and causes oxidative stress. This study explored the impact of αSyn A53T expression in transgenic hairy roots of four medicinal plants (Lobelia cardinalis, Artemisia annua, Salvia miltiorrhiza, and Polygonum multiflorum). In all tested plants, αSyn A53T expression triggered proteotoxic stress and perturbed iron homeostasis, mirroring the molecular profile observed in human and animal nerve cells. In addition to the common eukaryotic defense mechanisms against proteostatic and oxidative stresses, a plant stress response generally includes the biosynthesis of a diverse set of protective secondary metabolites. Therefore, the hairy root cultures expressing αSyn A53T offer a platform for identifying secondary metabolites that can ameliorate the effects of αSyn, thereby aiding in the development of possible PD treatments and/or treatments of synucleinopathies.

Latitudinal trends in mating system traits in the highly self-fertilizing Lobelia inflata revealed by community science.

Mating systems in angiosperms range from obligate outcrossing to highly self-fertilizing. The belief that obligate selfing does not exist is contradicted by genetic evidence in several populations of L. inflata, in which selfing is enforced by the anthers enclosing the style. However, whether the mating systems of these populations are typical, or an extreme across the species range is unknown. Such trends are hypothesized to result from selection for reproductive assurance under mate limitation at range margins. Here, we use ~7500 iNaturalist community science images, in which stylar exsertion can be observed, to test this hypothesis in L. inflata and, for comparison, in four typical congeneric Lobelias that express a staminate, then a pistillate phase (protandry). Specifically, we analyzed the effects of latitude and range marginality on the frequency of stylar exsertion and number of exserted flowers. Outcrossing capacity in L. inflata increased at low latitudes and near the overall range center, supporting our hypothesis, with exsertion frequencies significantly lower than in congenerics. Interestingly, in outcrossing capable individuals, the number of style-exserted flowers was consistent across the species range and among species, indicating outcrossing capable L. inflata individuals resemble congenerics. These findings suggest that variation in stylar exsertion is expressed among individuals rather than by all individuals within populations. However, whether this is a result of differences in exsertion allele frequencies or of differentiation in the induction of a threshold trait requires further study. Moreover, the trends in outcrossing capability revealed here imply the potential for geographic variation in L. inflata mating system.

In vitro anti-plasmodial activity of three selected medicinal plants that are used in local traditional medicine in Amhara region of Ethiopia.

BACKGROUND: The plants Aloe weloensis, Lepidium sativum, and Lobelia gibberoa have been used in Ethiopian folklore medicine to treat various diseases including malaria. METHOD: The in vitro anti-plasmodial activity of the three crude extracts was evaluated using parasite lactate dehydrogenase assay against the chloroquine (CQ)-sensitive D10 and the chloroquine (CQ)-resistant W2 strains. RESULT: The methanolic extract of L. gibberoa roots showed the highest in vitro anti-plasmodial effect against both D10 and W2 Plasmodium falciparum strains with IC50 value of 103.83 ± 26.17 µg/mL and 47.11 ± 12.46 µg/mL, respectively. However, the methanolic extract of L. sativum seeds and the leaf latex of A. weloensis were not active with an IC50 value > 200 µg/mL against both D10 and W2 strains. CONCLUSION: The methanolic extract of L. gibberoa roots showed a promising in vitro anti-plasmodial activity against the CQ-sensitive (D10) and CQ-resistant (W2) strains of P. falciparum. Thus, the anti-plasmodial activity of this plant partly justifies and may also support the traditional use against malaria. However, the methanolic extract of L. sativum seeds and the leaf latex of A. weloensis did not exert suppressive activity on the growth of P. falciparum strains.

Lobelia chinensis Extract and Its Active Compound, Diosmetin, Improve Atopic Dermatitis by Reinforcing Skin Barrier Function through SPINK5/LEKTI Regulation.

The skin acts as a mechanical barrier that protects the body from the exterior environment, and skin barrier function is attributed to the stratum corneum (SC), which is composed of keratinocytes and skin lipids. Skin barrier homeostasis is maintained by a delicate balance between the differentiation and exfoliation of keratinocytes, and keratinocyte desquamation is regulated by members of the serine protease kalikrein (KLK) family and their endogenous inhibitor SPINK5/LEKTI (serine protease inhibitor Kazal type 5/lympho-epithelial Kazal-type-related inhibitor). Furthermore, SPINK5/LEKTI deficiency is involved in impaired skin barrier function caused by KLK over-activation. We sought to determine whether increased SPINK5/LEKTI expression ameliorates atopic dermatitis (AD) by strengthening skin barrier function using the ethanol extract of Lobelia chinensis (LCE) and its active compound, diosmetin, by treating human keratinocytes with UVB and using a DNCB-induced murine model of atopic dermatitis. LCE or diosmetin dose-dependently increased the transcriptional activation of SPINK5 promoter and prevented DNCB-induced skin barrier damage by modulating events downstream of SPINK5, that is, KLK, PAR2 (protease activated receptor 2), and TSLP (thymic stromal lymphopoietin). LCE or diosmetin normalized immune response in DNCB treated SKH-1 hairless mice as determined by reductions in serum immunoglobulin E and interleukin-4 levels and numbers of lesion-infiltrating mast cells. Our results suggest that LCE and diosmetin are good candidates for the treatment of skin barrier-disrupting diseases such as Netherton syndrome or AD, and that they do so by regulating SPINK5/LEKTI.

Plasticity in floral longevity and sex-phase duration of Lobelia siphilitica in response to simulated pollinator declines.

PREMISE: Pollinator declines can reduce the quantity and quality of pollination services, resulting in less pollen deposited on flowers and lower seed production by plants. In response to these reductions, plant species that cannot autonomously self-pollinate and thus are dependent on pollinators to set seed could plastically adjust their floral traits. Such plasticity could increase the opportunity for outcross pollination directly, as well as indirectly by affecting inflorescence traits. METHODS: To test whether plants can respond to pollinator declines by plastically adjusting their floral traits, we simulated declines by experimentally reducing pollinator access to Lobelia siphilitica plants and measuring two traits of early- and late-season flowers: (1) floral longevity; and (2) sex-phase duration. To test whether plasticity in these floral traits affected inflorescence traits, we measured daily display size and phenotypic gender. RESULTS: We found that experimentally reducing pollination did not affect female-phase duration, but did extend the male-phase duration of early-season flowers by 13% and the longevity of late-season flowers by 12.8%. However, plants with an extended male phase did not have a more male-biased phenotypic gender, and plants with an extended floral longevity did not have a larger daily display. CONCLUSIONS: Our results suggest that plants can respond to pollinator declines by plastically adjusting both the longevity and sex-phase duration of their flowers. If this plasticity increases the opportunity for outcross pollination, then it could be one mechanism by which pollinator-dependent plant species maintain seed production as pollinators decline.

Do Genetic Drift and Gene Flow Affect the Geographic Distribution of Female Plants in Gynodioecious Lobelia siphilitica?

Variation in population sex ratio is particularly pronounced in gynodioecious angiosperms. Extremely high female frequencies in gynodioecious populations cannot be readily explained by selective forces alone. To assess the contributions of drift and gene flow to extreme sex-ratio variation, we documented sex ratio and population size in 92 populations of Lobelia siphilitica across its range and genotyped plants using plastid and nuclear genetic markers. Similarity in spatial patterns of genetic and demographic variables may suggest that drift and/or gene flow have contributed to population sex-ratio variation in L. siphilitica. We found strong spatial structuring of extremely high female frequencies: populations with >50% female plants are restricted to the south−central portion of the range. However, we did not detect any spatial structuring in population size nor metrics of genetic diversity, suggesting that extreme variation in female frequency is not strongly affected by drift or gene flow. Extreme sex-ratio variation is frequently observed in gynodioecious plants, but its causes are difficult to identify. Further investigation into mechanisms that create or maintain the spatial structure of sex ratios in gynodioecious species will provide much needed insight.

Identification of lobetyolin as a major antimalarial constituent of the roots of Lobelia giberroa Hemsl.

Lobelia giberroa Hemsl. is an endogenous Ethiopian medicinal plant with a long history of use in the treatment of malaria, bacterial and fungal diseases, and cancer. Here, we present the in vivo bioassay-guided fractionation of the 80% methanol extract of L. giberroa roots, which led to the isolation of lobetyolin. L. giberroa roots were extracted with 80% methanol, and the dried 80% methanol extract was fractionated with hexane, ethyl acetate, methanol, and water. Acute oral toxicity study was conducted according to the Organisation for Economic Co-operation and Development Guideline 425 by using female Swiss albino mice. Antimalarial activity was assessed in Plasmodium berghei-infected Swiss albino mice. Through in vivo bioassay-guided fractionation processes lobetyolin, a C14-polyacetylene glucoside, was isolated from the methanol fraction by silica gel column chromatography as the main active ingredient from the plant. The chemical structure of lobetyolin was elucidated by interpretation of spectroscopic data (1HNMR, 13CNMR, IR. MS) including two dimensional NMR. The plant extract was considered safe for administration up to 2000 mg/kg. In the four-day suppressive test, the 80% methanol extract (400 mg/kg), methanol fraction (400 mg/kg), and lobetyolin (100 mg/kg) exhibited antimalarial activity, with chemosuppression values of 73.05, 64.37, and 68.21%, respectively. Compared to the negative control, which had a mean survival time of 7 days, the lobetyolin (100 mg/kg) and methanol fraction (400 mg/kg) treated groups had mean survival times of 18 and 19 days, respectively. The current study supports the traditional use of the plant for the treatment of malaria. The structural differences between lobetyolin and existing antimalarials, as well as its previously unknown antimalarial activity, make it of interest as an early lead compound for further chemical optimization.

Pleistocene origin and colonization history of Lobelia columnaris Hook. f. (Campanulaceae: Lobelioideae) across sky islands of West Central Africa.

We aimed to infer ancestral area and historical colonization of Lobelia columnaris in the sky islands of Bioko and Cameroon through dated phylogeny using chloroplast genomes. Specifically, we aim to answer the following questions: (1) What are the phylogenetic relationships among Bioko Island and Cameroon populations? (2) Are the older populations found in the older sky islands? We assembled novel plastomes from 20 individuals of L. columnaris from 5 mountain systems. The plastome data were explored with phylogenetic analyses using Maximum Likelihood and Bayesian Inference. The populations of L. columnaris have a monophyletic origin, subdivided into three plastomes-geographic clades. The plastid phylogenomic results and age of the sky islands indicate that L. columnaris colonized first along with the Cameroon Volcanic Line's young sky islands of Bioko. The crown group (1.54 Ma) split the population in Bioko and mainland Cameroon. It is possible that Bioko was the ancestral area and likely isolated during cold and dry conditions in forest refugia. Presumably, the colonization history occurred during the middle-late Pleistocene from South Bioko's young sky island to North Bioko and the northern old sky islands in Cameroon. Furthermore, the central depression with lowland forest between North and South Bioko is a current geographic barrier that keeps separating the populations of Bioko from each other. Also, the shallow sea channel keeps isolated the populations of Bioko and the mainland populations. The Pleistocene climatic oscillations led to the divergence of the Cameroon and Bioko populations into three clades. L. columnaris colonized the older sky islands in mainland Cameroon after establishment in Bioko's younger sky islands. Contrary to expectations, the biogeography history was an inverse progression with respect to the age of the Afromontane sky islands.

A sit-and-wait predator, but not an active-pursuit predator, alters pollinator-mediated selection on floral traits.

Indirect species interactions are ubiquitous in nature, often outnumbering direct species interactions. Yet despite evidence that indirect interactions have strong ecological effects, relatively little is known about whether they can shape adaptive evolution by altering the strength and/or direction of natural selection. We tested whether indirect interactions affect the strength and direction of pollinator-mediated selection on floral traits of the bumble-bee pollinated wildflower Lobelia siphilitica. We estimated the indirect effects of two pollinator predators with contrasting hunting modes: dragonflies (Aeshnidae and Corduliidae) and ambush bugs (Phymata americana, Reduviidae). Because dragonflies are active pursuit predators, we hypothesized that they would strengthen pollinator-mediated selection by weakening plant-pollinator interactions (i.e., a density-mediated indirect effect). In contrast, because ambush bugs are sit-and-wait predators, we hypothesized that they would weaken or reverse the direction of pollinator-mediated selection by altering pollinator foraging behavior (i.e., a trait-mediated indirect effect). Specifically, if ambush bugs hunt from plants with traits that attract pollinators (i.e., prey), then pollinators will spend less time visiting those plants, weakening or reversing the direction of selection on attractive floral traits. We did not find evidence that high dragonfly abundance strengthened selection on floral traits via a density-mediated indirect effect: neither pollen limitation (a proxy for the strength of plant-pollinator interactions) nor directional selection on floral traits of L. siphilitica differed significantly between high- and low-dragonfly abundance treatments. In contrast, we did find evidence that ambush bug presence affected selection on floral traits via a trait-mediated indirect effect: ambush bugs hunted from L. siphilitica plants with larger daily floral displays, reversing the direction of pollinator-mediated selection on daily display size. These results suggest that indirect species interactions have the potential to shape adaptive evolution by altering natural selection.

Beyond Alkaloids: Novel Bioactive Natural Products From Lobelia Species.

In this work, we reviewed the progress in the phytochemical and biological investigations of bioactive components derived from medicinally valuable Lobelia species. In the last 60 years, Lobelia has garnered significant attention from the phytochemist from around the world, majorly due to the discovery of bioactive piperidine alkaloids (e.g., lobinaline and lobeline) in the early 1950s. Later, lobeline underwent clinical trials for several indications including the treatment of attention deficit hyperactivity disorder and a multicenter phase three trial for smoking cessation. Subsequently, several other alkaloids derived from different species of Lobelia were also investigated for their pharmacological characteristics. However, in the last few years, the research focus has started shifting to the characterization of the other novel chemical classes. The major shift has been noticed due to the structurally similar alkaloid components, which essentially share similar pharmacological, physicochemical, and toxicological profiles. In this review, we present an up-to-date overview of their progress with special attention to understanding the molecular mechanisms of the novel bioactive components.

Distribution pattern and habitat preference for Lobelia species (Campanulaceae) in five countries of East Africa.

East Africa is one of the centres of distribution and diversity for Lobelia L. (Campanulaceae, sub-family Lobelioideae). Lobelia habitats in East Africa have been facing habitat fragmentation and loss, which are recognised as a major threat to biodiversity. However, previous plant conservation studies in East Africa only focused on protected areas and ignored unprotected areas. Future conservation strategies of plants, such as Lobelia, will depend on knowledge of their distribution patterns and habitat preference in East Africa. To understand the distribution pattern and the habitat preference of Lobelia in five countries (Kenya, Uganda, Tanzania, Rwanda and Burundi) of East Africa, we conducted a literature review in the seven major vegetation regions (afro-alpine, afro-montane forest, drier savannah, grasslands, wetter savannah, Zambezian woodland and semi-desert and desert). We also employed meander and patterned searches, which allowed greater opportunities for recording Lobelia species. Our results showed that the genus is distributed in all of the seven regions of the five countries with 54 taxa. The afro-montane forest region, with 41 taxa, is the richest in species diversity, followed by the Zambezian woodland region with 18 taxa. The semi-desert and desert region has the lowest number with only four taxa. The afro-alpine region has 15 taxa, although the region is the smallest by area. The herbaceous type was found in all regions, while the giant type has a clear preference for the afro-alpine and afro-montane forest regions. Future conservation for Lobelia should consider its habitat preference by, for example, focusing on the afro-alpine and afro-montane forest regions. This study will facilitate the setting of future conservation strategies for Lobelia.

Efficacy of Lobelia alsinoides Lam ethanolic extract on a third-degree burn: An experimental study on rats.

This is the first study to evaluate the topical application of Lobelia alsinoides Lam (LT) ethanolic extract on burns in rats. A deep third-degree burn wound was inflicted in adult male Wistar rats and the burns were dressed daily with a topical ointment formulation (Patent filed) comprising of Lobelia alsinoides Lam (5% and 10% w/w). The wound had noteworthy contraction and quicker eschar removal in 10% w/w LT-treated groups followed by 5% w/w treated groups on comparing with the commonly prescribed ointment (SilverexTM containing 1% w/w Silver sulfadiazine). Histopathological analysis showed that ointment containing 10% w/w LT ethanolic extract significantly increased fibroblast growth, which plays a major role in anatomic integrity, collagen synthesis, and accelerated the rate of the healing process. This study shows that the ethanolic extract of Lobelia alsinoides Lam, a previously pharmacologically unreported traditional medicinal plant, possesses wound contraction and eschar removal properties on burn wounds.

Chemical structure elucidation of an inulin-type fructan isolated from Lobelia chinensis lour with anti-obesity activity on diet-induced mice.

A neutral polysaccharide (LCPS) was obtained from Lobelia chinensis via hot water extraction and ethanol precipitation. After separation and purification, the homogeneous polysaccharide was obtained with a molecular weight of 2.6 × 103 Da. The chemical composition of the extracted polysaccharide contains fructose and glucose with protein-free identified by gas chromatography-mass spectrometer (GC-MS) method. Chemical structure of LCPS was indicated by nuclear magnetic resonance (NMR) spectroscopy. Data indicated that LCPS was an inulin-type fructan with α-D-Glcp-(1→[1)-ß-D-Fruf-(2]15 linkage. LCPS intake reduced at significant level high fat diet (HFD)-induced body weight gain, liver weight, total cholesterol and triglyceride in serum and hepatic tissue, respectively. The lipid droplet accumulation in hepatic tissue was similar between lean and LCPS groups lower than in HFD-induced mice from tissue section staining results. Together, polysaccharides from Lobelia chinensis could be a new source of natural anti-obesity agent against obesity with potential value in the manufacturing supplements and drugs.

Network Pharmacology-Based Dissection of the Anti-diabetic Mechanism of Lobelia chinensis.

Diabetes mellitus (DM) is a chronic inflammatory disease, and the rapidly increasing DM is becoming a major problem of global public health. Traditional Chinese medicine (TCM) has a long history of treating diabetes. It has been developed and utilized because of its good efficacy and no toxic side effects. Lobelia chinensis is a traditional whole grass herbal. With the continuous deepening of pharmacological research on TCM, the active ingredients of L. chinensis are continuously revealed, which contained the alkaloids, flavonoids, flavonoid glycosides and amino acids that have the good effects of anti-inflammatory, anti-viral and anti-diabetic. In order to further explore the targets of active ingredients and its anti-diabetic mechanism, a feasible network pharmacology analysis model based on chemical, pharmacokinetic and pharmacological data was developed by network construction method to clarify the anti-diabetic mechanism of L. chinensis. The present study conducted by gas chromatography-mass spectrometer (GC/MS), which identified 208 metabolites of L. chinensis, of which 23 ingredients may have effective pharmacological effects after absorption, distribution, metabolism, and excretion (ADME) screening. Network pharmacological analysis on the active ingredients revealed that 5-hydroxymethylfurfural in L. chinensis affects the insulin resistance signaling pathway by acting on GSK3B, TNF, and MAPK1, acacetin affects the diabetic pathway by acting on INSR, DPP4, and GSK3B, that regulate type 2 diabetes, non-insulin-dependent DM, and inflammatory diseases. These results successfully indicated the potential anti-diabetic mechanism of the active ingredients of L. chinensis.

Impact of prairie fragment size on proportion of females and reproductive success of Lobelia spicata Lam., a gynodioecious species.

Due to ongoing human impacts, plant species increasingly occur in landscapes that are highly fragmented, with remaining natural habitats occupying small areas, resulting in populations that are smaller and more isolated than in previous time periods. This changed metapopulation structure is expected to have negative impacts on seed production. For example, the proportion of female plants within gynodioecious populations may be more volatile due to genetic drift in small populations associated with small habitat fragments, with concomitant impacts on seed production. My aims were to determine: (i) if variation in proportion of females is larger in smaller fragments; and (ii) if such changes in female frequency in small fragments result in reduced seed production. Thirty-two populations of Lobelia spicata Lam., a gynodioecious species, were surveyed in 2000, 2001 and 2009 in the tallgrass prairie region of Midwestern North America (Illinois and Indiana, USA). Data were collected for: proportion of female plants, total number of flowering plants (measure of population size), seed set per plant and prairie fragment size (another measure of population size). The proportion of females is more variable in smaller prairie fragments. Seed number per fruit decreases as the proportion of females increases in a population, but only significantly for female plants. The number of flowering plants is positively associated with fruit production for both genders. Populations within larger prairie fragments have higher seed production. The reproductive consequences of habitat fragmentation depend on the plant breeding system. While both sexes were negatively impacted, females were more adversely affected.

Hepatoprotective effect of Lobelia alsinoides Lam. in Wistar rats.

BACKGROUND: Traditional healing practitioners of South India use fine paste (an Ayurvedic dosage form known as 'kalka') of Lobelia alsinoides Lam., an ethno medicinal plant for curing hepatic diseases. OBJECTIVE: To evaluate in-vivo hepatoprotective effect of a candidate formulation viz. kalka containing whole plant (L. alsinoides Lam.) in rat model of Carbon-tetrachloride (CCl4) induced hepatotoxicity. MATERIALS & METHODS: Hepatotoxicity was induced in Wistar albino rats by oral administration of 1.25 ml/kg CCl4 once every day for 7 consecutive days. A candidate kalka formulation (fine paste) was prepared and administered to rats at different dose rates of 0.54 g/kg, 1.08 g/kg and 2.16 g/kg daily. At the end of the study-period, the serum levels of aspartate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase (ALP), total bilirubin, total protein, albumin and total cholesterol were monitored. Further, the hepatic pathology was evaluated for assessing the extent of hepatotoxicity in the control and hepatoprotective effect in treatment groups. Meanwhile in-vitro antioxidant activity of kalka was evaluated by hydroxy radical, nitric oxide and DPPH (2, 2 diphenyl-1-picrylhydrazil) radical scavenging assays. Further, a 'limit test' was done in accordance with OECD Guidelines 425 (acute toxicity). RESULTS: The animals treated with the fine paste of L. alsinoides did not show an elevation in the biochemical values compared to CCl4 treated rats and during histomorphologic evaluation, hepatoprotective effect was evident with scattered mitotic figures in the parenchyma. Acute toxicity evaluation indicated that doses up to 2500 mg/kg are not toxic to rats. It has a good anti-oxidant activity also. CONCLUSIONS: From the study, it was obvious that L. alsinoides had significant hepatoprotective effect in CCl4 induced liver toxicity in rats. This ethno medicinal plant is certainly a promising hepatoprotective drug in liver disorders.

Characterization of microsatellite loci in Brighamia insignis and transferability to other genera in the Hawai'ian lobelioid group.

PREMISE: Microsatellite markers were developed to measure genetic diversity and relatedness of ex situ collections of Brighamia insignis (Campanulaceae). METHODS AND RESULTS: Potential microsatellite markers were identified from two sources; 28 were developed for B. insignis and an additional 12 markers from a previously published study of Lobelia villosa. Primer pairs were tested on 30 individuals of B. insignis and 24 individuals of B. rockii to provide measures of genetic diversity and inbreeding. We assessed cross-species amplification in an additional 13 taxa that represented all six genera within the Hawai'ian lobelioid group to determine the broader applicability of the markers. CONCLUSIONS: Results indicate that these primers will provide useful estimates of genetic diversity and relatedness of ex situ collections of both Brighamia species. In addition, we have also demonstrated the widespread applicability of these markers for use in population genetic studies of several species within the Hawai'ian lobelioid group.

Beyond balancing selection: frequent mitochondrial recombination contributes to high-female frequencies in gynodioecious Lobelia siphilitica (Campanulaceae).

Gynodioecy is a sexual system in which females and hermaphrodites co-occur. In most gynodioecious angiosperms, sex is determined by an interaction between mitochondrial male-sterility genes (CMS) that arise via recombination and nuclear restorer alleles that evolve to suppress them. In theory, gynodioecy occurs when multiple CMS types are maintained at equilibrium frequencies by balancing selection. However, some gynodioecious populations contain very high frequencies of females. High female frequencies are not expected under balancing selection, but could be explained by the repeated introduction of novel CMS types. To test for balancing selection and/or the repeated introduction of novel CMS, we characterised cytoplasmic haplotypes from 61 populations of Lobelia siphilitica that vary widely in female frequency. We confirmed that mitotype diversity and female frequency were positively correlated across populations, consistent with balancing selection. However, while low-female populations hosted mostly common mitotypes, high-female populations and female plants hosted mostly rare, recombinant mitotypes likely to carry novel CMS types. Our results suggest that balancing selection maintains established CMS types across this species, but extreme female frequencies result from frequent invasion by novel CMS types. We conclude that balancing selection alone cannot account for extreme population sex-ratio variation within a gynodioecious species.

The Reappearance of An Extremely Rare and Critically Endangered Nitella translucens (Charophyceae) in Poland.

We report the reappearance of the rare charophyte Nitella translucens in Poland. It was identified in the soft-water lobelian Lake Jelen (North Poland) during 2013 and 2018 phytolittoral surveys. This species is considered critically endangered in various European countries and was previously classified as extinct in Poland. Its occurrence was confirmed using morphological and molecular data (ITS1-18S, ITS2-28S, rDNA, and rbcL). The N. translucens occupied ~20% of the lake bottom, at depths of 1.5-6.5 m, water pH 7.5-8.6, conductivity of 59-66 µS · cm-1 , and total nitrogen and phosphorus during growing season in the range of 1.1-1.4 mg · L-1 and 0.07-0.1 mg · L-1 , respectively. It co-occurred mainly with plant species typical for lobelia lakes: Isoetes lacustris, Littorella uniflora, and Myriophyllum alterniflorum, as well as Ceratophyllum demersum and Elodea canadensis, which are characteristic for eutrophicated waters. It appears that N. translucens may thrive in lobelia lakes during their transformation to more eutrophic states.

Chemical Profiling of Lobelia chinensis with High-Performance Liquid Chromatography/Quadrupole Time-of-Flight Mass Spectrometry (HPLC/Q-TOF MS) Reveals Absence of Lobeline in the Herb.

Lobelia chinensis is a kind of herbal medicine widely distributed and used in Asia. The chemical components of this herb, however, have not been well studied until now. Lobeline, as an essential and famous bioactive compound in Lobelia genus, has been assumed to be present in L. chinensis. In order to ascertain its presence and, more importantly, proper use of this herb, chemical profiling this herb with highly sensitive and high-resolution analytical mass spectrometry was applied. In this study, high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC/Q-TOF MS) method was employed to systematically profile the chemical constituents of L. chinensis for the first time. Comparative chemical profiling study of L. chinensis and Lobelia inflata was also conducted to provide evidence whether lobeline is present or not. Piperidine alkaloids except for lobeline, alkaloid-lignan hybrids, flavonoids, polyacetylenes, nonanedioic acid, and some new phytochemicals were successfully identified in L. chinensis simultaneously. Comparing to the chemical profiles of L. inflata, lobeline was found to be absent in L. chinensis. All of the secondary metabolites in L. chinensis were determined with the HPLC/Q-TOF MS method. The absence of lobeline in L. chinensis was confirmed after this extensive study.