Multitrophic Interactions Between Arbuscular Mycorrhizal Fungi, Foliar Endophytic Fungi and Aphids.

Almost all living plants can be simultaneously colonised by arbuscular mycorrhizal fungi in the roots and endophytes in the shoots, while also being attacked by insect herbivores. However, to date, no study has ever examined the multitrophic interactions between these two different fungal groups and insects on any species of forb. Here, we examined the effects of two commercial species mixtures of arbuscular mycorrhizal fungi (AMF) and two foliar endophytes (Colletotrichum acutatum and Cladosporium oxysporum) on the growth of an invasive weed, Impatiens glandulifera, and the aphids that attack it. AMF reduced plant biomass, which was most evident when C. oxysporum was inoculated. Mycorrhizal fungi had few effects on aphids, and these depended on the identity of the endophytes present. Meanwhile, endophytes tended to increase aphid numbers, but this depended on the identity of the AMF inoculum. Throughout, there were differences in the responses of the plant to the two mycorrhizal mixtures, demonstrating clear AMF specificity in this plant. These specific effects were also strongly affected by the endophytes, with a greater number of interactions found between the AMF and endophytes than between the endophytes themselves. In particular, AMF reduced infection levels by the endophytes, while some endophyte inoculations reduced mycorrhizal colonisation. We suggest that both AMF and endophytes could play an important part in future biological control programmes of weeds, but further multitrophic experiments are required to unravel the complexity of interactions between spatially separated parts of the plant microbiome.

Earlier onset of flowering and increased reproductive allocation of an annual invasive plant in the north of its novel range.

BACKGROUND AND AIMS: It remains unclear whether invasive species can maintain both high biomass and reproductive output across their invaded range. Along latitudinal gradients, allocation theory predicts that faster flowering onset at high latitudes results in maturation at smaller size and thus reduced reproductive output. For annual invasive plants, more favourable environmental conditions at low latitudes probably result in stronger competition of co-occurring species, potentially driving selection for higher investment in vegetative biomass, while harsher climatic conditions and associated reproductive uncertainty at higher latitudes could reduce selection for vegetative biomass and increased selection for high reproductive investment (stress-gradient hypothesis). Combined, these drivers could result in increased or constant reproductive allocation with increasing latitude. METHODS: We quantified life-history traits in the invasive annual plant Impatiens glandulifera along a latitudinal gradient in Europe. By growing two successive glasshouse generations, we assessed genetic differentiation in vegetative growth and reproductive output across six populations, and tested whether onset of flowering drives this divergence. KEY RESULTS: Trait variation was mainly caused by genetic differentiation. As expected, flowering onset was progressively earlier in populations from higher latitudes. Plant height and vegetative biomass also decreased in populations from higher latitudes, as predicted by allocation theory, but their variation was independent of the variation in flowering onset. Reproductive output remained constant across latitudes, resulting in increased reproductive allocation towards higher latitudes, supporting the stress-gradient hypothesis. We also observed trait genetic differentiation among populations that was independent of latitude. CONCLUSIONS: We show that an annual invasive plant evolved several life-history traits across its invaded range in ~150 years. The evolution of vegetative and reproductive traits seems unconstrained by evolution of flowering onset. This genetic decoupling between vegetative and reproductive traits possibly contributes to the invasion success of this species.

Biotic and abiotic drivers of intraspecific trait variation within plant populations of three herbaceous plant species along a latitudinal gradient.

BACKGROUND: The importance of intraspecific trait variation (ITV) is increasingly acknowledged among plant ecologists. However, our understanding of what drives ITV between individual plants (ITVBI) at the population level is still limited. Contrasting theoretical hypotheses state that ITVBI can be either suppressed (stress-reduced plasticity hypothesis) or enhanced (stress-induced variability hypothesis) under high abiotic stress. Similarly, other hypotheses predict either suppressed (niche packing hypothesis) or enhanced ITVBI (individual variation hypothesis) under high niche packing in species rich communities. In this study we assess the relative effects of both abiotic and biotic niche effects on ITVBI of four functional traits (leaf area, specific leaf area, plant height and seed mass), for three herbaceous plant species across a 2300 km long gradient in Europe. The study species were the slow colonizing Anemone nemorosa, a species with intermediate colonization rates, Milium effusum, and the fast colonizing, non-native Impatiens glandulifera. RESULTS: Climatic stress consistently increased ITVBI across species and traits. Soil nutrient stress, on the other hand, reduced ITVBI for A. nemorosa and I. glandulifera, but had a reversed effect for M. effusum. We furthermore observed a reversed effect of high niche packing on ITVBI for the fast colonizing non-native I. glandulifera (increased ITVBI), as compared to the slow colonizing native A. nemorosa and M. effusum (reduced ITVBI). Additionally, ITVBI in the fast colonizing species tended to be highest for the vegetative traits plant height and leaf area, but lowest for the measured generative trait seed mass. CONCLUSIONS: This study shows that stress can both reduce and increase ITVBI, seemingly supporting both the stress-reduced plasticity and stress-induced variability hypotheses. Similarly, niche packing effects on ITVBI supported both the niche packing hypothesis and the individual variation hypothesis. These results clearly illustrates the importance of simultaneously evaluating both abiotic and biotic factors on ITVBI. This study adds to the growing realization that within-population trait variation should not be ignored and can provide valuable ecological insights.

Flavonoids from the flowers of Impatiens glandulifera Royle isolated by high performance countercurrent chromatography.

INTRODUCTION: Impatiens glandulifera Royle (Balsaminaceae) is an annual herb from the Himalaya region, currently widespread along European river systems and one of the most important neophyte invading plants in Germany. Exploring the effects of allelopathic plant chemicals is important for the understanding of its ecological impacts in the process of suppression of indigenous plant species. OBJECTIVE: To investigate the chemical composition of Impatiens glandulifera flowers (IGFs) using high performance countercurrent chromatography (HPCCC). METHODS: The flowers of Impatiens glandulifera were manually separated and extracted with ethanol. LC-ESI-MS/MS was used to characterise the crude extract of IGF. The various flavonoids detected were isolated by HPCCC using of methyl tert-butyl ether-acetonitrile-water (2:2:3, v/v/v). The combination of the data provided by preparative ESI-MS/MS metabolite profiling, LC-ESI-MS/MS, UV-vis and 1D/2D-NMR spectroscopic analysis was used to elucidate the structures of the isolated compounds. RESULTS: HPCCC runs led to the direct isolation of pure dihydromyricetin (ampelopsin), eriodictyol-7-O-glucoside, kaempferol-3-O-glucoside (astragalin) and kaempferol-3-O-6"-malonyl-glucoside, as well as the pre-purification of kaempferol-3-O-rhamno-rhamnosyldiglucoside, quercetin-3-O-galactoside (hyperoside), quercetin and kaempferol in a single step. CONCLUSION: This is the first report on the flavonoid composition of the species Impatiens glandulifera. The developed protocol was successfully used to isolate the main flavonoids from the crude extract of IGFs. This combined HPCCC and HPLC procedure could be applied to the fast fractionation and recovery of flavonoid derivatives of other plant extracts.

An invasive plant alters pollinator-mediated phenotypic selection on a native congener.

UNLABELLED: • PREMISE OF STUDY: Recent studies suggest that invasive plants compete reproductively with native plants by reducing the quantity or quality of pollinator visits. Although these studies have revealed ecological consequences of pollinator-mediated competition between invasive and native plants, the evolutionary outcomes of these interactions remain largely unexplored.• METHODS: We studied the ecological and evolutionary impact of pollinator-mediated competition with an invasive jewelweed, Impatiens glandulifera, on a co-occurring native congener, I. capensis. Using a pollinator choice experiment, a hand pollination experiment, and a selection analysis, we addressed the following questions: (1) Do native pollinators show preference for the invasive or native jewelweed, and do they move between the two species? (2) Does invasive jewelweed pollen inhibit seed production in the native plant? (3) Does the invasive jewelweed alter phenotypic selection on the native plant's floral traits?• KEY RESULTS: The pollinator choice experiment showed that pollinators strongly preferred the invasive jewelweed. The hand pollination experiment demonstrated that invasive pollen inhibited seed production in the native plant. The selection analysis showed that the presence of the invasive jewelweed altered phenotypic selection on corolla height in the native plant.• CONCLUSIONS: Invasive plants have the potential to alter phenotypic selection on floral traits in native plant populations. If native plants can evolve in response to this altered selection pressure, the evolution of floral traits may play an important role in permitting long-term coexistence of native and invasive plants.

An invasive plant alters phenotypic selection on the vegetative growth of a native congener.

PREMISE OF THE STUDY: The ecological consequences of plant competition have frequently been tested, but the evolutionary outcomes of these interactions have gone largely unexplored. The study of species invasions can make an important contribution to this field of research by allowing us to watch ecological and evolutionary processes unfold as a novel species is integrated into a plant community. We explored the ecological and evolutionary impact of an invasive jewelweed, Impatiens glandulifera, on a closely related native congener, I. capensis and asked: (1) Does the presence of the invasive jewelweed alter the fitness of native jewelweed populations? (2) Does the invasive jewelweed affect the vegetative growth of the native congener? and (3) Does the invasive jewelweed alter phenotypic selection on the vegetative traits of the native congener? METHODS: We used a greenhouse competition experiment, an invasive species removal field experiment, and a survey of natural populations. KEY RESULTS: We show that when the invasive jewelweed is present, phenotypic selection favors native jewelweed individuals investing less in rapid upward growth and more in branching and fruiting potential through the production of nodes. CONCLUSIONS: This research demonstrates that invasive plants have the potential to greatly alter natural selection on native competitors. Studies investigating altered selection in invaded communities can reveal the potential evolutionary impact of invasive competitors, while deepening our understanding of the more general role of competition in driving plant evolution and permitting species coexistence.

Interference of field evidence, morphology, and DNA analyses of three related Lysiphlebus aphid parasitoids (Hymenoptera: Braconidae: Aphidiinae).

This study provides evidence on integrating the morphological, field, and laboratory data, and application of the cytochrome oxidase subunit I (COI) barcoding gene to the three asexual or sexual Lysiphlebus spp., i.e., Lysiphlebus cardui (Marshall), Lysiphlebus confusus Tremblay and Eady and Lysiphlebus fabarum (Marshall) (Hymenoptera: Braconidae: Aphidiinae). New aphid- invasive plant association, Aphis fabae Scopoli (Hemipreta: Aphididae) on Impatiens glandulifera Royle, has been used in the same model area in the Czech Republic under the same sampling and rearing method for several consecutive years and throughout the season. For molecular identification of these three species, we used DNA sequences of the barcoding region of the mitochondrial COI gene. Although our results confirmed ecological and morphological differences among L. cardui, L. confusus, and L. fabarum, genetic analysis on the basis of COI mitochondrial barcoding gene does not support species status of the mentioned Lysiphlebus taxa. The level of morphological differentiation in these Lysiphlebus Förster species is in accordance with the usual species variability within subfamily Aphidiinae. However, it should be examined how appearance of asexual lineages affects the morphological or genetical variability.

Pollen wall development in Impatiens glandulifera: exine substructure and underlying mechanisms.

The aim of this study was to investigate in detail the pollen wall ontogeny in Impatiens glandulifera, with emphasis on the substructure and the underlying mechanisms of development. Sporopollenin-containing pollen wall, the exine, consists of two parts, ectexine and endexine. By determining the sequence of developing substructures with TEM, we have in mind to understand in which way the exine substructure is connected with function. We have shown earlier that physical processes of self-assembly and phase separation are universally involved in ectexine development; currently, we try to clear up whether these processes participate in endexine development. The data received were compared with those on other species. The ectexine ontogeny of I. glandulifera followed the main stages observed in many other species, including the late tetrad stage named "Golden gates". It turned out that the same physico-chemical processes act in endexine development, especially expressed in aperture sites. Another peculiar phenomenon observed in exine development was the recurrency of micellar sequence at near-aperture and aperture sites where the periplasmic space is widened. It should be noted that, in the whole, the developmental substructures observed during the tetrad and early post-tetrad period are similar in species with columellate exines. Evidently, these basic physical processes proceed, reiterating again and again in different species, resulting in an enormous variety of exine structures on the base of a relatively modest number of genes. Granular and alveolar exines emerge on the base of the same basic processes but are arrested at spherical and cylindrical micelle mesophases correspondingly.

Phytoremediation potential of invasive plant species for potentially toxic elements along the Sava River upstream.

Invasive plant species (IPS) have many characteristics that are necessary for successful phytoremediation and the accumulation of large amounts of potentially toxic elements (PTEs). The most common IPS from the source of the Sava River are Reynoutria japonica, Solidago canadensis and Impatiens glandulifera. Considering that the riparian soils of the Sava River are classified as moderately polluted, this study investigated their enrichment with PTEs (As, B, Cd, Cr, Cu, Ni, Pb, Zn) and the potential for accumulation of these elements in roots and leaves of the most common IPS. The soil and plant samples were prepared using the wet digestion method in CEM Mars 6 microwave oven. The content of PTEs in soil and plant samples was determined by ICP-OES. The results showed a moderate to very high enrichment of Cu in the soils. Impatiens glandulifera has the highest uptake capacity in leaves and phytoextraction ability of B, Cd, Cu and Zn. Reynoutria japonica has some potential for phytoextraction of Pb, Zn and especially B, while Solidago canadensis has potential for phytoextraction of B and Zn, while excluding Cr and Cu. The analyzes have shown that the studied species are not suitable for bioindication of PTEs in the riparian soils of the Sava River. Considering the enrichment of soils with Cu, Impatiens glandulifera is the most suitable species for phytoremediation of these elements among the studied species.

Genetic evidence for predominantly hydrochoric gene flow in the invasive riparian plant Impatiens glandulifera (Himalayan balsam).

BACKGROUND AND AIMS: Riparian systems are prone to invasion by alien plant species. The spread of invasive riparian plants may be facilitated by hydrochory, the transport of seeds by water, but while ecological studies have highlighted the possible role of upstream source populations in the establishment and persistence of stands of invasive riparian plant species, population genetic studies have as yet not fully addressed the potential role of hydrochoric dispersal in such systems. METHODS: A population genetics approach based on a replicated bifurcate sampling design is used to test hypotheses consistent with patterns of unidirectional, linear gene flow expected under hydrochoric dispersal of the invasive riparian plant Impatiens glandulifera in two contrasting river systems. KEY RESULTS: A significant increase in levels of genetic diversity downstream was observed, consistent with the accumulation of propagules from upstream source populations, and strong evidence was found for organization of this diversity between different tributaries, reflecting the dendritic organization of the river systems studied. CONCLUSIONS: These findings indicate that hydrochory, rather than anthropogenic dispersal, is primarily responsible for the spread of I. glandulifera in these river systems, and this is relevant to potential approaches to the control of invasive riparian plant species.

No evidence for local adaptation in an invasive alien plant: field and greenhouse experiments tracing a colonization sequence.

BACKGROUND AND AIMS: Local adaptation enables plant species to persist under different environmental conditions. Evolutionary change can occur rapidly in invasive annual species and has been shown to lead to local adaptation. However, the patterns and mechanisms of local adaptation in invasive species along colonization sequences are not yet understood. Thus, in this study the alien annual Impatiens glandulifera was used to investigate local adaptation to distinct habitats that have been consecutively invaded in central Europe. METHODS: A reciprocal transplant experiment was performed using 15 populations from alluvial deciduous forests, fallow meadows and coniferous upland forests, and a greenhouse experiment was performed in which plants from these habitats were grown under treatments reflecting the main habitat differentiators (shade, soil acidity, competition). KEY RESULTS: Biomass production, specific leaf area, plant height and relative growth rate differed between habitats in the field experiment and between treatments in the greenhouse, but not between seed origins. Overall, there was no indication of local adaptation in either experiment. CONCLUSIONS: Since I. glandulifera is a successful invader in many habitats without showing local adaptation, it is suggested that the species is coping with environmental variation by means of high phenotypic plasticity. The species seems to follow a 'jack-and-master' strategy, i.e. it is able to maintain high fitness under a wide range of environmental conditions, but performs particularly well in favourable habitats. Therefore, the proposed colonization sequence is likely to be based primarily on changes in propagule pressure. It is concluded that invasive alien plants can become dominant in distinct habitats without local adaptation.

A Modeling Framework to Frame a Biological Invasion: Impatiens glandulifera in North America.

Biological invasions are a major component of global environmental change with severe ecological and economic consequences. Since eradicating biological invaders is costly and even futile in many cases, predicting the areas under risk to take preventive measures is crucial. Impatiens glandulifera is a very aggressive and prolific invasive species and has been expanding its invasive range all across the Northern hemisphere, primarily in Europe. Although it is currently spread in the east and west of North America (in Canada and USA), studies on its fate under climate change are quite limited compared to the vast literature in Europe. Hybrid models, which integrate multiple modeling approaches, are promising tools for making projections to identify the areas under invasion risk. We developed a hybrid and spatially explicit framework by utilizing MaxEnt, one of the most preferred species distribution modeling (SDM) methods, and we developed an agent-based model (ABM) with the statistical language R. We projected the I. glandulifera invasion in North America, for the 2020-2050 period, under the RCP 4.5 scenario. Our results showed a predominant northward progression of the invasive range alongside an aggressive expansion in both currently invaded areas and interior regions. Our projections will provide valuable insights for risk assessment before the potentially irreversible outcomes emerge, considering the severity of the current state of the invasion in Europe.

Characterization of Rare Himalayan Balsam (Impatiens glandulifera Royle) Honey from Croatia.

Himalayan balsam (Impatiens glandulifera Royle) is an invasive garden ornamental plant species originating from Asia, which produces significant amounts of nectar. In Croatia, it is widely distributed along the banks of the Mura River. Although this plant species is widespread in Europe, there are still no available scientific data about this unifloral honey type. The results showed that Himalayan balsam honey is characterized by the high presence of pollen grains in the pollen spectrum (59−85%), natural higher diastase activity (39.1 ± 7.98 DN), negative specific rotation (−21.2° ± 6.89) and an extra light amber color (48.5 ± 12.69 mm Pfund). The carbohydrate profile is characterized by monosaccharides fructose (39.34 ± 0.65 g/100 g) and glucose (31.91 ± 1.42 g/100 g) with a ratio >1.23, while the most commonly represented disaccharide was maltose (3.04 ± 0.79 g/100 g). The average total phenolic content was 130.97 ± 11.17 mg gallic acid/kg honey, and the average antioxidant capacity value was 225.38 ± 29.58 µM Fe(II). The major mineral element was K, with an average of 533.92 ± 139.70 mg/kg. The sensory profile was characteristic with a light orange color and medium-intensity odor and aroma. The crystallization rate was moderate and characterized by the appearance of opalescence and gelatinous forms of crystals. The results of this study provide the first insight into the melissopalynological, physico-chemical and sensory profile of Himalayan balsam honey.

Fast and Reproducible 96-Well Plate-Based Method for the Evaluation of the Antigerminative Potential of Plant Extracts and Phytotoxic Compounds.

With the aim of evaluating the antigerminative activity of plant extracts, a miniaturized assay using 96-well plates (WP assay) was developed and compared to the long-established assay using Petri dishes (PD assay). The WP assay yielded results comparable to those of the PD assay using an ethanolic extract of the Himalayan balsam and lawsone as a standard. It also allowed the needed volume of the test solution to be cut by half and the number of required cress seeds to be cut by more than 1.5. The WP assay was then successfully applied to various extracts of Himalayan balsam, molecules (2,4-dichlorophenoxyacetic acid (2,4-D), glyphosate, and 2-methoxy-1,4-naphthoquinone (2-MNQ)) and target seeds (radish, lettuce, and wheat). By being adapted to a 96-well plate format, the antigerminative WP assay is a promising alternative to the PD assay. Besides, its convenience and low resource consumption make it ready for accelerated and high-throughput screening, as well as automation.

Impacts of the Invasive Impatiens glandulifera: Lessons Learned from One of Europe's Top Invasive Species.

Biological invasions are renowned for their negative ecological and economic implications, however from studying invasions invaluable insights can be gained in the fields of ecology and evolution- potentially contributing towards conservation plans to deal, not only with biological invasion, but with other concerning issues, such as climate change. Impatiens glandulifera, or Himalayan balsam, is widely considered to be a highly problematic invasive, having spread across more than thirty countries during the past century. This paper will examine the findings which have arose from studying I. glandulifera and its impacts on the invaded ecosystem.

Assessment of genetic relationships among native and introduced Himalayan balsam (Impatiens glandulifera) plants based on genome profiling.

We conducted genomic characterization based on SNP and SilicoDArT markers on the invasive Himalayan balsam (Impatiens glandulifera) plants originating from native and non-native regions of their distribution. When genetic relationships were explored by PCoA using SNP and SilicoDArT marker data, the first, second, and third principal coordinates explained altogether 37.4% and 31.0% of the variability, respectively. Samples from the UK, Canada, and Pakistan were grouped together, while Indian plants were clearly distinct based on SNP markers but relatively close to the UK-Canada-Pakistan group based on SilicoDArT markers. Constructed trees differentiated individuals into clusters resembling the PCoA patterns. The Bayesian BAPS analysis performed for the SNP data revealed that the individuals were distributed in seven clusters, representing samples from each of the four Finnish populations, India, Pakistan, and the combination of the UK and Canada. Similar clustering was visible in the UPGMA tree. The Indian cluster did not display any ancestral gene flow with the others, while the Pakistani cluster showed ancestral gene flow only with the combined UK and Canada cluster. Furthermore, the latter cluster displayed ancestral gene flow with the Finnish populations varying from 0% to 3.1%. The BAPS analyses conducted for the SilicoDArT data differ slightly: The individuals were distributed in nine clusters, and the Indian cluster exhibited ancestral gene flow with the mixed cluster including Canadian, Pakistani, and UK samples, and one Finnish sample. The AMOVA showed that 45% and 26% of variation was present among the I. glandulifera groups/populations and the rest within them based on SNP and SilicoDArT markers, respectively. The Bayesian BAPS analyses and the gene flow networks were the most informative tools for resolving relationships among native and introduced plants. It is notable that the small sample sizes for non-Finnish plant materials may affect the accuracy of the gene flow and other estimates.

Impatiens glandulifera (Himalayan balsam) chloroplast genome sequence as a promising target for populations studies.

BACKGROUND: Himalayan balsam Impatiens glandulifera Royle (Balsaminaceae) is a highly invasive annual species native of the Himalayas. Biocontrol of the plant using the rust fungus Puccinia komarovii var. glanduliferae is currently being implemented, but issues have arisen with matching UK weed genotypes with compatible strains of the pathogen. To support successful biocontrol, a better understanding of the host weed population, including potential sources of introductions, of Himalayan balsam is required. METHODS: In this molecular study, two new complete chloroplast (cp) genomes of I. glandulifera were obtained with low coverage whole genome sequencing (genome skimming). A 125-year-old herbarium specimen (HB92) collected from the native range was sequenced and assembled and compared with a 2-year-old specimen from UK field plants (HB10). RESULTS: The complete cp genomes were double-stranded molecules of 152,260 bp (HB92) and 152,203 bp (HB10) in length and showed 97 variable sites: 27 intragenic and 70 intergenic. The two genomes were aligned and mapped with two closely related genomes used as references. Genome skimming generates complete organellar genomes with limited technical and financial efforts and produces large datasets compared to multi-locus sequence typing. This study demonstrates the suitability of genome skimming for generating complete cp genomes of historic herbarium material. It also shows that complete cp genomes are solid genetic markers for population studies that could be linked to plant evolution and aid with targeting native range and natural enemy surveys for biocontrol of invasive species.

Phenolic constituents of the aerial parts of Impatiens glandulifera Royle (Balsaminaceae) and their antioxidant activities.

As a continuation of investigating Impatiens L. genus, eight flavonoids, eriodyctiol, eriodyctiol 7-O-ß-ᴅ-glucoside, kaempferol 3-O-ß-ᴅ-glucoside, kaempferol 3-O-ß-ᴅ-galactoside, kaempferol 3-rhamnosyl-di-glucoside, kaempferol 3-O-ß-ᴅ-rutinoside, quercetin 3-O-ß-ᴅ-glucoside and quercetin 3-O-ß-ᴅ-galactoside, two phenolic acids - p-hydroxybenzoic acid and protocatechuic acid, and 2-methoxynaphthalene-1,4-dione were isolated from the aerial parts of I. glandulifera collected in Poland. The structures of the compounds were established by analysis of their spectroscopic (1H and 13C NMR) and spectrometric (MS) data, as well as by comparison of these with those reported in the literature. Quercetin 3-O-ß-ᴅ-glucoside, kaempferol 3-O-ß-ᴅ-galactoside and kaempferol 3-O-ß-ᴅ-rutinoside were isolated for the first time from the investigated taxon. In addition, the antioxidant activities in different tests of all obtained compounds were evaluated. The results clearly showed that among analyzed constituents, quercetin 3-O-ß-ᴅ-glucoside exhibited antioxidant activity comparable or better than ascorbic acid and Trolox which were used as a positive control.

Specialized naphthoquinones present in Impatiens glandulifera nectaries inhibit the growth of fungal nectar microbes.

The invasion success of Impatiens glandulifera (Himalayan balsam) in certain parts of Europe and North America has been partially attributed to its ability to compete for bee pollinators with its rich nectar and due to its capacity to produce and release allelopathic 1,4-naphthoquinones (1,4-NQs) from its roots and leaves. Given that other 1,4-NQs present in the digestive fluids of certain carnivorous plants are proposed to control microbial colonization, we investigated the potential for the 1,4-NQs, 2-methoxy-1,4-naphthoquinone (2-MNQ) and lawsone, to fulfill an analogous role in the nectaries of I. glandulifera. Both 2-MNQ and lawsone were detected in the floral nectaries of I. glandulifera at levels comparable to leaves and roots, but were discovered to be at significantly higher levels in its extra-floral nectaries (EFNs) and to be present in EFN nectar itself. Nectar microbe inhibition assays revealed that the common nectar bacteria Gluconobacter oxydans and Asaia prunellae are not inhibited by 2-MNQ or lawsone, although both compounds were found to inhibit the growth of the common fungal nectar microbes Metschnikowia reukaufii and Aureobasidium pullulans. Taken together, these findings suggest that 2-MNQ and lawsone could serve to protect the rich nectar of I. glandulifera against fungal growth. The high abundance of 2-MNQ and lawsone in I. glandulifera EFNs may also point to an unsuspected mechanism for how allelopathic 1,4-NQs are leached into the soil where they exhibit their known allelopathic effects.

Glanduliferins A and B, two new glucosylated steroids from Impatiens glandulifera, with in vitro growth inhibitory activity in human cancer cells.

Impatiens glandulifera has been imported from Himalaya in Europe and is considered as an invasive alien plant whose spreading arouses increasing interest among scientific literature. Via anti-cancer bioguiding, two new glucosylated steroids, named glanduliferins A and B, were isolated from the dried stem of I. glandulifera plants, together with the well-known α-spinasterol and 2-methoxy-1,4-naphthoquinone, which are also isolated from roots and leaves. They were characterized as 17-(2-hydroxy-2-pentamethylcyclopropyl-ethyl)-10,13-dimethyl-2,3,4,5,6,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopents[a]phenathren-3-O-(4-O-acetyl)-α-D-glucopyranoside and 17-(4-ethyl-1,5-dimethyl-hex-2-enyl)-10,13-dimethyl-2,3,4,5,6,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopents[a]phenathren-3-O-(6-O-acetyl)-ß-D-glucopyranoside using various NMR and HRESIMS techniques and chemical methods. In vitro determination of the growth inhibitory activity of the four isolated compounds using the MTT colorimetric assay revealed mean IC50 growth inhibitory value of ~30 µM for glanduliferin A while glanduliferin B and α-spinasterol were poorly active till 100 µM. 2-methoxy-1,4-naphthoquinone revealed to be active in the single micromolar digit range as previously described. Quantitative videomicroscopy analyses of the effects of glanduliferins A and B suggested cytostatic rather than cytotoxic activity in U373 glioblastoma (GBM) cells.