Synthesis and Identification of Epoxy Derivatives of 5-Methylhexahydroisoindole-1,3-dione and Biological Evaluation.

Cyclic imides belong to a well-known class of organic compounds with various biological activities, promoting a great interest in compounds with this functional group. Due to the structural complexity of some molecules and their spectra, it is necessary to use several spectrometric methods associated with auxiliary tools, such as the theoretical calculation for the structural elucidation of complex structures. In this work, the synthesis of epoxy derivatives of 5-methylhexahydroisoindole-1,3-diones was carried out in five steps. Diels-Alder reaction of isoprene and maleic anhydride followed by reaction with m-anisidine afforded the amide (2). Esterification of amide (2) with methanol in the presence of sulfuric acid provided the ester (3) that cyclized in situ to give imides 4 and 4-ent. Epoxidation of 4 and 4-ent with meta-chloroperbenzoic acid (MCPBA) afforded 5a and 5b. The diastereomers were separated by silica gel flash column chromatography, and their structures were determined by analyses of the spectrometric methods. Their structures were confirmed by matching the calculated 1H and 13C NMR chemical shifts of (5a and 5b) with the experimental data of the diastereomers using MAE, CP3, and DP4 statistical analyses. Biological assays were carried out to evaluate the potential herbicide activity of the imides. Compounds 5a and 5b inhibited root growth of the weed Bidens pilosa by more than 70% at all the concentrations evaluated.

Seed germination of Bidens subalternans DC. exposed to different environmental factors.

Bidens subalternans DC. is a weed found in several tropical countries such as Brazil. Large number of produced seeds and easy dispersion favor the colonization of agricultural fields by this species. To know the factors that affect the germination of B. subalternans can help to understand its ecology, permitting to develop control strategies. Laboratory experiments were carried out to evaluate how the temperature, photoperiod, burial depth, water deficit, and salt stress affect the seed germination of B. subalternans. The means of the treatments of each experiment were shown in scatter plots with the bars indicating the least significant difference (LSD, p≤0.05). The results showed a germination percentage above 77% for a wide alternating temperature (15/20 C to 30/35 C night/day). The highest germination and uniformity occurred at 25/30°C night/day. Only 11% of the seeds germinated at a temperature of 35/40°C night/day. The deeper burial of seeds reduced their germination. Only 17% of the seeds germinated in darkness conditions. However, in constant light and 12 hours of light/dark conditions the germination percentage was over 96%, confirming the light dependence of the B. subalternans during germination. In constant light and 12 hours of light/dark, the germination was over 96%. B. subalternans seeds showed sensitivity to water and salt stress, and their germination was inhibited under a water potential of -0.4 MPa and 100.09 mM, respectively. The sensitivity of B. subalternans seeds to high temperatures, water stress, and salt stress explains the high frequency of this weed in south-central Brazil. The light and sowing depth showed that burial of seeds by mechanical control is a strategy to reduce the high infestation of B. subalternans.

Chemical Characterization and Phytotoxic Effects of the Aerial Parts of Ruzigrass (Urochloa ruziziensis).

Studies of the phytotoxic effects between plants can be a crucial tool in the discovery of innovative compounds with herbicide potential. In this sense, we can highlight ruzigrass (Urochloa ruziziensis), which is traditionally used in the crop rotation system in order to reduce weed emergence. The aim of this work was to characterize the secondary metabolites of ruzigrass and to evaluate its phytotoxic effects. In total, eight compounds were isolated: friedelin, oleanolic acid, α-amyrin, 1-dehydrodiosgenone, sitosterol and stigmasterol glycosides, tricin and p-coumaric acid. Phytotoxic effects of the crude methanolic extract and fractions of ruzigrass were assessed using germination rate, initial seedling growth, and biomass of Bidens pilosa, Euphorbia heterophylla and Ipomoea grandifolia. Chemometric analysis discriminated the weed species into three groups, and B. pilosa was the most affected by fractions of ruzigrass. The phytotoxic activities of 1-dehydrodiosgenone, tricin, and p-coumaric acid are also reported, and p-coumaric acid and 1-dehydrodiosgenone were active against B. pilosa.

Seed germination ecology of Bidens pilosa and its implications for weed management.

It is now widely recognized that Bidens pilosa has become a problematic broadleaf weed in many ecosystems across the world and, particularly in the light of recent climate change conditions, closer management strategies are required to curtail its impact on agricultural cropping. In this investigation, experiments were conducted to evaluate the effect of environmental factors on the germination and emergence of B. pilosa, and also on the response of this weed to commonly available post-emergence herbicides in Australia. The environmental factors of particular interest to this current work were the effect of light and temperature, salinity, burial depth and moisture on B. pilosa since these are key management issues in Australian agriculture. In addition, the effects of a number of commonly used herbicides were examined, because of concerns regarding emerging herbicide resistance. In the tested light/dark regimes, germination was found to be higher at fluctuating day/night temperatures of 25/15 °C and 30/20 °C (92-93%) than at 35/25 °C (79%), whilst across the different temperature ranges, germination was higher in the light/dark regime (79-93%) than in complete darkness (22-38%). The standard five-minute temperature pretreatment required for 50% inhibition of maximum germination was found to be 160 °C, and it was further shown that no seeds germinated at temperatures higher than 240 °C. With regard to salinity, some B. pilosa seeds germinated (3%) in 200 mM sodium chloride (NaCl) but all failed to germinate at 250 mM NaCl. Germination declined from 89% to 2% as the external osmotic potential decreased from 0 to -0.6 MPa, and germination ceased at -0.8 MPa. Seeding emergence of B. pilosa was maximum (71%) for seeds placed on the soil surface and it was found that no seedlings emerged from a depth of 8 cm or greater. A depth of 3.75 cm was required to inhibit the seeds to 50% of the maximum emergence. In this study, application of glufosinate, glyphosate and paraquat provided commercially acceptable control levels (generally accepted as >90%) when applied at the four-leaf stage of B. pilosa. However, none of the herbicide treatments involved in this study provided this level of control when applied at the six-leaf stage. In summary, B. pilosa germination has been clearly shown to be stimulated by light and thus its emergence was greatest from the soil surface. This suggests that infestation from this weed will remain as a problem in no-till conservation agriculture systems, the use of which is increasing now throughout the world. It is intended that information generated from this study be used to develop more effective integrated management programs for B. pilosa and similar weeds in commercial agricultural environments which are tending toward conservation approaches.

Preponderance of Oxygenated Sesquiterpenes and Diterpenes in the Volatile Oil Constituents of Lactuca serriola L. Revealed Antioxidant and Allelopathic Activity.

Using synthetic chemicals in industry and agriculture has led to several environmental problems. Thus, plant products derived from volatile oils (VOs) could be a potential green source for bioherbicides. Little is known about the VOs of Lactuca serriola. Hence, the present study aimed to characterize the VOs chemical composition from the aerial parts of L. serriola, assessment of antioxidant activity, and evaluate allelopathic potential against the noxious weed Bidens pilosa. The VOs were extracted by hydrodistillation and analyzed by GC/MS. The VOs from the Egyptian ecospecies of L. serriola were found to comprise 34 compounds mainly oxygenated sesquiterpenes and diterpenes. The major compounds were isoshyobunone (64.22 %), isocembrol (17.35 %), and alloaromadendrene oxide-1 (7.32 %). So, L. serriola can be considered as a good source for isoshyobunone, considering that it has a much higher concentration than any other plants. Also, this plant has a high content of the oxygenated diterpene compound, isocembrol, which is rarely found in the VOs of most plants. The VOs expressed strong antioxidant activity. Also, for the first time, our results showed a strong allelopathic activity of VOs from L. serriola on germination and seedling growth of the noxious weed, B. pilosa. We suppose that the activity of the VOs from L. serriola could be attributed to these previously mentioned major compounds, as they represent about 89 % of the total identified oil constituents. Nevertheless, to evaluate these compounds as new allelochemicals, further study is needed to test the allelopathic activity of authenticated standard of these compounds either singular or in combination on several weeds as well as evaluate the safety, and improve the efficacy and stability at the field scale.

Soil lead pollution modifies the structure of arbuscular mycorrhizal fungal communities.

The impact of lead (Pb) pollution on native communities of arbuscular mycorrhizal fungi (AMF) was assessed in soil samples from the surroundings of an abandoned Pb smelting factory. To consider the influence of host identity, bulk soil surrounding plant roots soil samples of predominant plant species (Sorghum halepense, Bidens pilosa, and Tagetes minuta) growing in Pb-polluted soils and in an uncontaminated site were selected. Molecular diversity was assessed by sequencing the 18S rDNA region with primers specific to AMF (AMV4.5NF/AMDGR) using Illumina MiSeq. A total of 115 virtual taxa (VT) of AMF were identified in this survey. Plant species did not affect AMF diversity patterns. However, soil Pb content was negatively correlated with VT richness per sample. Paraglomeraceae and Glomeraceae were the predominant families while Acaulosporaceae, Ambisporaceae, Archaeosporaceae, Claroideoglomeraceae, Diversisporaceae, and Gigasporaceae were less abundant. Acaulosporaceae and Glomeraceae were negatively affected by soil Pb, but Paraglomeraceae relative abundance increased under increasing soil Pb content. Overall, 26 indicator taxa were identified; four of them were previously reported in Pb-polluted soils (VT060; VT222; VT004; VT380); and five corresponded to cultured spores of Scutellospora castaneae (VT041), Diversispora spp. and Tricispora nevadensis (VT060), Diversispora epigaea (VT061), Glomus proliferum (VT099), and Gl. indicum (VT222). Even though AMF were present in Pb-polluted soils, community structure was strongly altered via the differential responses of taxonomic groups of AMF to Pb pollution. These taxon-specific differences in tolerance to soil Pb content should be considered for future phytoremediation strategies based on the selection and utilization of native Glomeromycota.

Evaluation of Growth, Photosynthetic Pigments and Genotoxicity in the Wetland Macrophyte Bidens laevis Exposed to Tebuconazole.

The fungicide tebuconazole (TBZ) has been used to prevent terrestrial fungi in agroecosystems, but it can also induce negative effects to non-targeted aquatic organisms, such as plants. The aim of the present work was to evaluate the potential cyto- and genotoxicity of TBZ in the aquatic macrophyte Bidens laevis, exposed to a range of concentrations of 0.1-100 µg/L. Mitosis in root tips were analyzed showing decreased mitotic index and an increase of chromosomal aberrations at 10 and 100 µg/L. The regression of TBZ concentration vs. aneugenic aberrations was significant, indicating the mechanism of genotoxicity. The specific growth rate (Gr) for total length decreased in plants exposed to 0.1, 10 and 100 µg/L. Gr for root decreased in plants exposed at 0.1 and 10 µg/L, reaching a maximum percent inhibition root growth rate (Ir) of 68.8%. These results show that TBZ resulted cyto- and genotoxic to B. laevis at environmentally relevant levels.

Responses of the weed Bidens pilosa L. to exogenous application of the steroidal saponin protodioscin and plant growth regulators 24-epibrassinolide, indol-3-acetic acid and abscisic acid.

The exogenous application of plant hormones and their analogues has been exploited to improve crop performance in the field. Protodioscin is a saponin whose steroidal moiety has some similarities to plant steroidal hormones, brassinosteroids. To test the possibility that protodioscin acts as an agonist or antagonist of brassinosteroids or other plant growth regulators, we compared responses of the weed species Bidens pilosa L. to treatment with protodioscin, brassinosteroids, auxins (IAA) and abscisic acid (ABA). Seeds were germinated and grown in agar containing protodioscin, dioscin, brassinolides, IAA and ABA. Root apex respiratory activity was measured with an oxygen electrode. Malondialdehyde (MDA) and antioxidant enzymes activities were assessed. Protodioscin at 48-240 µm inhibited growth of B. pilosa seedlings. The steroidal hormone 24-epibrassinolide (0.1-5 µm) also inhibited growth of primary roots, but brassicasterol was inactive. IAA at higher concentrations (0.5-10.0 µm) strongly inhibited primary root length and fresh weight of stems. ABA inhibited all parameters of seedling growth and also seed germination. Respiratory activity of primary roots (KCN-sensitive and KCN-insensitive) was activated by protodioscin. IAA and ABA reduced KCN-insensitive respiration. The content of MDA in primary roots increased only after protodioscin treatment. All assayed compounds increased APx and POD activity, with 24-epibrassinolide being most active. The activity of CAT was stimulated by protodioscin and 24-epibrassinolide. The results revealed that protodioscin was toxic to B. pilosa through a mechanism not related to plant growth regulator signalling. Protodioscin caused a disturbance in mitochondrial respiratory activity, which could be related to overproduction of ROS and consequent cell membrane damage.

Simulated warming enhances biological invasion of Solidago canadensis and Bidens frondosa by increasing reproductive investment and altering flowering phenology pattern.

Phenological and reproductive shifts of plants due to climate change may have important influences on population dynamics. Climate change may also affect invasive species by changing their phenology and reproduction, but few studies have explored this possibility. Here, we investigated the impact of climate change on the phenology, reproduction and invasion potential of two alien Solidago canadensis and Bidens frondosa and one native weed, Pterocypsela laciniata, all of which are in the Asteraceae family. The three species responded to simulated climate change by increasing reproductive investments and root/leaf ratio, prolonging flowering duration, and while the two alien species also displayed a mass-flowering pattern. Moreover, our experimental results indicated that the alien invasive species may have greater phenological plasticity in response to simulated warming than that of the native species (P. laciniata). As such, climate change may enhance the invasion and accelerate the invasive process of these alien plant species.

AM fungi facilitate the competitive growth of two invasive plant species, Ambrosia artemisiifolia and Bidens pilosa.

Invasive species often cause enormous economic and ecological damage, and this is especially true for invasive plants in the Asteraceae family. Arbuscular mycorrhizal fungi (AMF) play an important role in the successful invasion by exotic plant species because of their ability to promote growth and influence interspecific competition. However, few studies have evaluated the effects of invasive Asteraceae species on AMF diversity and how feedback mechanisms during competition with native species subsequently affect the accumulation of nutrient resources. Two exotic Asteraceae, Ambrosia artemisiifolia and Bidens pilosa, were monitored during competition with a native grass species, Setaria viridis, which is being replaced by these exotic species in natural areas around the study site. From these species continuously maintained in a field plot for 5 years, we collected the rhizosphere soil and cloned and identified soil AMF. Furthermore, AM fungal spores were isolated from rhizosphere soil of the two invasive species and used as inoculum in greenhouse experiments, to compare growth and nutrient accumulation during competition. The results indicate that although the AMF diversity in the rhizosphere soil of A. artemisiifolia and B. pilosa differed, the three most abundant species (Septoglomus viscosum, Septoglomus constrictum, Glomus perpusillum) were identical. The addition of AMF inoculum changed the competition between the plants, increasing the competitive ability of the invasives and decreasing that of the native. The results show a similar AMF community composition between A. artemisiifolia and B. pilosa, increased AMF root colonization of the invasive species during competition, AMF-enhanced N accumulation, and AMF-facilitated competitive growth of the invasive species.

A greater foraging scale, not a higher foraging precision, may facilitate invasion by exotic plants in nutrient-heterogeneous conditions.

Background and Aims: Soil nutrient heterogeneity has been proposed to influence competitive outcomes among different plant species. Thus, it is crucial to understand the effects of environmental heterogeneity on competition between exotic invasive and native species. However, the effects of soil nutrient heterogeneity on the competition between invasive and native plants have rarely been linked to root foraging behaviour. Methods: In this study, a competition experiment was performed with two invasive-native species pairs (BP-VC, Bidens pilosa vs. Vernonia cinerea; MM-PS, Mikania micrantha vs. Paederia scandens) grown under homogeneous and heterogeneous conditions in a common greenhouse environment. Root activity was assessed by determining the amount of strontium (Sr) taken up by the shoot of each species. Key Results: The invasive species exhibited a greater foraging scale, whereas the native species exhibited a higher foraging precision. A trade-off between foraging scale and precision was observed within each pair of invasive-native species. Compared with soil homogeneity, soil heterogeneity significantly increased the biomass of the two invasive species, B. pilosa and M. micrantha, under competitive conditions. Within each pair, the invasive species exhibited greater relative competitive ability with respect to shoot mass, and considerably more Sr taken up by the invasive species compared with the native species. The Sr acquisition results indicate that nutrient-poor conditions may facilitate the competitive ability of the native species V. cinerea, whereas M. micrantha may possess a stronger competitive ability regardless of soil nutrient conditions. Conclusion: Soil nutrient heterogeneity has the potential to promote the invasion of these two exotic species due to their larger foraging scale, stronger competitive ability and greater root activity relative to their counterpart native species. The present work highlights the importance of soil heterogeneity in plant invasion, particularly with regards to root foraging traits and competition between invasive and native plants.

Changes in gametophyte physiology of Pteris multifida induced by the leaf leachate treatment of the invasive Bidens pilosa.

In recent years, the response of fern gametophytes to environment has raised much attention. However, studies on the influence of plant invasion to fern gametophytes are scarce. Allelopathy plays an important role in biological invasion. Hence, it is necessary to study the allelopathic effects of invasive plants on fern gametophytes and elucidate the mechanisms by which invasive plants cause phytotoxicity. As one of the main invasive plants in China, Bidens pilosa exhibits allelopathic effects on spermatophyte growth. Field investigation shows that many ferns are threatened by the invasion of B. pilosa. The distribution of Pteris multifida overlaps with that of B. pilosa in China. To examine the potential involvement of allelopathic mechanisms of B. pilosa leaves, changes in the physiology in P. multifida gametophytes are analyzed. We found that cell membrane and antioxidant enzyme activities as well as photosynthesis pigment contents of the gametophytes were affected by B. pilosa leachates. Gametophytes of P. multifida exposed to B. pilosa had increased damages to cell membranes, expressed in thiobarbituric acid reacting substance (TBARS) concentrations, malondialdehyde (MDA), electrolyte leakage (membrane permeability), and degree of injury. Enzyme activities, assessed by superoxide dismutase (SOD) and catalase (CAT) as well as guaiacol peroxidase (GPX) enhanced with the increase in leachate concentration after 2-day exposure. Meanwhile, lower chlorophyll a (Chl a), chlorophyll b (Chl b), carotenoid (Car), and the total chlorophyll were measured as leachate concentrations increased. At day 10, leaf leachates of B. pilosa exhibited the greatest inhibition. These results suggest that the observed inhibitory or stimulatory effects on the physiology studied can have an adverse effect on P. multifida and that allelopathic interference seems to have involved in this process.

Assessment of the root system of Brassica juncea (L.) czern. and Bidens pilosa L. exposed to lead polluted soils using rhizobox systems.

The purpose of this study was to compare the behavior of the root system of one of the most frequently cited species in phytoremediation Indian mustard [Brassica juncea (L.) Czern.] and a representative perennial herb (Bidens pilosa L.) native of Argentina, for different concentrations of lead in soils through chemical and visualization techniques of the rhizosphere. Lead polluted soils from the vicinity of a lead recycling plant in the locality of Bouwer, were used in juxtaposed rhizobox systems planted with seedlings of B. juncea and B. pilosa with homogeneous and heterogeneous soil treatments. Root development, pH changes in the rhizosphere, dry weight biomass, lead content of root and aerial parts and potential extraction of lead by rhizosphere exudates were determined. In both species lead was mainly accumulated in roots. However, although B. juncea accumulated more lead than B. pilosa at elevated concentrations in soils, the latter achieved greater root and aerial development. No changes in the pH of the rhizosphere associated to lead were observed, despite different extractive potentials of lead in the exudates of the species analyzed. Our results indicated that Indian mustard did not behave as a hyperaccumulator in the conditions of the present study.

Evaluation of two Brazilian indigenous plants for phytostabilization and phytoremediation of copper-contaminated soils.

Indigenous plants have been grown naturally and vigorously in copper contaminated soils. Thus, the aim of this study was to evaluate the phytoremediation ability of two indigenous plants naturally grown in two vineyard soils copper contaminated, and in a copper mining waste. However, it was evaluated the macro and micronutrient uptake and the potential of phytoremediation. So, a greenhouse study was carried out with Bidens pilosa and Plantago lanceolata in samples of vineyard soils (Inceptisol and Mollisol) copper contaminated, and in a copper mining waste. Plant growth, macro and micronutrient up take, tolerance index (TI), translocation factor (TF), metal extraction ratio (MER), bioaccumulation factor (BCF), plant effective number of the shoots (PENs), and plant effective number of the total plant (PENt) were analyzed. Both plants grown in vineyard soils showed high phytomass production and TI. P. lanceolata plants cultivated in the Inceptisol showed the highest copper concentrations in the shoots (142 mg kg-1), roots (964 mg kg-1) and entire plants (1,106 mg kg-1). High levels of copper were phytoaccumulated from the Inceptisol by B. pilosa and P. lanceolata with 3,500 and 2,200 g ha-1 respectively. Both B. pilosa and P. lanceolata plants showed characteristics of high copper hyperaccumulator. Results showed that both species play an important role in the natural copper phytoaccumulation in both vineyard soils contaminated with copper, being important to its phytoremediation.

Evaluation of two Brazilian indigenous plants for phytostabilization and phytoremediation of copper-contaminated soils / Avaliação de duas plantas nativas brasileiras para fitoestabilização e fitorremediação de solos contaminados com cobre

Abstract Indigenous plants have been grown naturally and vigorously in copper contaminated soils. Thus, the aim of this study was to evaluate the phytoremediation ability of two indigenous plants naturally grown in two vineyard soils copper contaminated, and in a copper mining waste. However, it was evaluated the macro and micronutrient uptake and the potential of phytoremediation. So, a greenhouse study was carried out with Bidens pilosa and Plantago lanceolata in samples of vineyard soils (Inceptisol and Mollisol) copper contaminated, and in a copper mining waste. Plant growth, macro and micronutrient up take, tolerance index (TI), translocation factor (TF), metal extraction ratio (MER), bioaccumulation factor (BCF), plant effective number of the shoots (PENs), and plant effective number of the total plant (PENt) were analyzed. Both plants grown in vineyard soils showed high phytomass production and TI. P. lanceolata plants cultivated in the Inceptisol showed the highest copper concentrations in the shoots (142 mg kg–1), roots (964 mg kg–1) and entire plants (1,106 mg kg–1). High levels of copper were phytoaccumulated from the Inceptisol by B. pilosa and P. lanceolata with 3,500 and 2,200 g ha–1 respectively. Both B. pilosa and P. lanceolata plants showed characteristics of high copper hyperaccumulator. Results showed that both species play an important role in the natural copper phytoaccumulation in both vineyard soils contaminated with copper, being important to its phytoremediation.

Resumo Plantas nativas crescem naturalmente e vigorosamente em solos contaminados com cobre. Assim, o objetivo deste estudo foi avaliar a capacidade de fitorremediação de duas plantas nativas, naturalmente encontradas em dois solos de vitivinicultura contaminados com cobre, e em rejeito de mineração de cobre. Foram avaliados os teores de macro e micronutrientes nos tecidos das plantas, e o potencial de fitorremediação. Assim, um estudo em casa de vegetação foi realizado com plantas de Bidens pilosa e Plantago lanceolata, com amostras de dois solos de vitivinicultura (Neossolos e Cambissolos) contaminados com cobre, e com rejeito de mineração de cobre. O crescimento das plantas, teores de macro e micronutrientes nos tecidos, índice de tolerância (TI), fator de translocação (TF), taxa de extração do metal (MER), fator de bioacumulação (BCF), número efetivo dos plantas da parte aérea (PENs) e número efetivo de plantas inteiras (PENt) foram analisados. Ambas as espécies cultivadas em solos vitivinicultura mostraram elevada produção de fitomassa e os TI. P. lanceolata cultivadas no Neossolo mostraram as concentrações de cobre mais elevados na parte aérea (142 mg kg–1), nas raízes (964 mg kg–1) e nas plantas inteiras (1.106 mg kg–1). Altos níveis de cobre foram fitoacumulados pelas plantas B. pilosa e P. lanceolata com 3.500 e 2.200 g ha–1, respectivamente, quando cultivadas em Neossolo. Ambas as espécies apresentaram características hiperacumuladoras de cobre. Os resultados mostraram que estas espécies desempenham um papel importante na fitoacumulação de cobre naturalmente em ambos os solos de vitivinicultura contaminados com cobre, sendo importantes para a fitorremediação.

[Biomorphological Features and Microevolution of the Invasive Species Bidens L. in European Russia].

Species of the genus Bidens that have invaded natural communities in Europe were observed. Fourteen species have been introduced in European botanical gardens since the 18th century, but only two of them have become invasive in Russia-Bidensfrondosa and B. connata. B.frondosa demonstrates microevolutional ability in the second distribution range. Nevertheless, it has a low ability of hybridization. B. frondosa has higher competitiveness compared with that of B. connata.

Performance and responses to competition in two congeneric annual species: does seed heteromorphism matter?

Variations in seed characteristics observed in heteromorphic species may affect various stages of their life cycles, e.g. seed dormancy, germination characteristics or even adult plant performance. Highly specialised seed morphs - described as colonisers and maintainers - exhibit a trade-off between colonisation capacity and competitive traits. The performance of distinct seed morph progeny under competitive conditions, and especially in multi-species arrangements, had previously not been given much attention. In this study, we compared performance and response to competition among distinct seed morph progenies in two congeneric, co-occurring species: the invasive Bidens frondosa and the non-invasive Bidens tripartita. We hypothesised that maintainer seed morphs of both species would perform better under increased plant densities and within inter-morphic mixtures, while coloniser morphs would show stronger responses to increased densities and perform relatively poorly in inter-morphic mixtures. We conducted a growth trial and a greenhouse experiment, which revealed that seed morph progeny differed significantly in plant height when grown without competition, while under competitive conditions such differences became less apparent. The observed pattern was more strongly pronounced in B. frondosa, which showed a general predominance in stature and biomass over its non-invasive congener. Although seed morphs performed equally well under competitive conditions, increased plant height and more rapid germination can favour the maintainer seed morph on sites where vegetation is already present.

The role of pentacyclic triterpenoids in the allelopathic effects of Alstonia scholaris.

Alstonia scholaris is a tropical evergreen tree native to South and Southeast Asia. Alstonia forests frequently lack understory species. However, potential mechanisms-particularly the allelochemicals involved-remain unclear. In the present study, we identified allelochemicals of A. scholaris, and clarified the role of allelopathic substances from A. scholaris in interactions with neighboring plants. We showed that the leaves, litter, and soil from A. scholaris inhibited growth of Bidens pilosa-a weed found growing abundantly near A. scholaris forests. The allelochemicals were identified as pentacyclic triterpenoids, including betulinic acid, oleanolic acid, and ursolic acid by using (1)H and (13)C-NMR spectroscopy. The half-maximal inhibitory concentration (IC50) for radicle growth of B. pilosa and Lactuca sativa ranged from 78.8 µM to 735.2 µM, and ursolic acid inhibited seed germination of B. pilosa. The triterpenoid concentrations in the leaves, litter, and soil were quantified with liquid chromatography-electrospray ionization/tandem mass spectrometry. Ursolic acid was present in forest soil at a concentration of 3,095 µg/g, i.e., exceeding the IC50. In the field, ursolic acid accumulated abundantly in the soil in A. scholaris forests, and suppressed weed growth during summer and winter. Our results indicate that A. scholaris pentacyclic triterpenoids influence the growth of neighboring weeds by inhibiting seed germination, radicle growth, and functioning of photosystem II.

Allelopathic effect of essential oils of medicinal plants in Bidens pilosa L / Efeito Alelopático de oleos essencias de plantas medicinais em Bidens pilosa L

We determined the inhibitory allelopathic effects of the volatile extracts of Cinnamomum zeylanicum Ness, Lippia sidoides Cham. and Cymbopogum nardus L. on seed germination and root growth of seedlings of Bidens pilosa. The experiment was conducted at the Seed Analysis Laboratory of the Department of Plant Science, Federal University of Ceará. For this end, we used oils at the concentrations of 0.01, 0.02, 0.04 and 0.08% (v/v). Five treatments were used for each of the oils arranged in a completely randomized design with four replications of 25 seeds. The seeds were sown in Petri dishes lined with filter paper moistened with distilled water and, aiming at the indirect contact with each oil, two sheets of filter paper were placed on top of the lid, in which three (3) mL of each oil solution were added. Then, the dishes were incubated in a germination chamber at 25°C. The pH did not contribute to alter the results; the volatile extracts of essential oils of C. zeylanicum, L. sidoides and C. nardus inhibited seed germination and root growth of seedlings of B. pilosa, which shows allelopathic potential; and the concentration of 0.08% of oils caused the overall deterioration of the roots and death of seedlings of B. pilosa.

Determinou-se o efeito alelopático inibitório dos extratos voláteis de Cinnamomum zeylanicum Ness, Lippia sidoides Cham. e Cymbopogum nardus L., sobre a germinação de sementes e o crescimento de raiz de plântulas de Bidens pilosa. O experimento foi conduzido no Laboratório de Análises de Sementes do Departamento de Fitotecnia da Universidade Federal do Ceará. Para isso, utilizaram-se óleos essenciais nas concentrações 0,01, 0,02, 0,04 e 0,08%. (v/v). Foram utilizados cinco tratamentos para cada óleo dispostos em delineamento inteiramente casualizado com quatro repetições de 25 sementes. As sementes foram semeadas em placas de Petri forradas com papel de filtro umedecido com água destilada e, visando o contato indireto com cada óleo, foram colocadas duas folhas de papel de filtro na parte superior interna da tampa, onde foi adicionado 3(três) mL da solução de cada óleo. Em seguida, as placas foram incubadas em câmara de germinação a 25ºC. O pH não contribuiu para alterar os resultados; os extratos voláteis de óleos essenciais de C. zeylanicum, L. sidoides and C. nardus inibiram a germinação de sementes e o crescimento da raiz de plântulas de B. pilosa, evidenciando potencial alelopático; a concentração de 0,08% dos óleos causaram a deterioração total das raízes e morte das plântulas de B. pilosa.

Effects of arbuscular mycorrhizal fungi on seedling growth and development of two wetland plants, Bidens frondosa L., and Eclipta prostrata (L.) L., grown under three levels of water availability.

To identify the importance of arbuscular mycorrhizal fungi (AMF) colonizing wetland seedlings following flooding, we assessed the effects of AMF on seedling establishment of two pioneer species, Bidens frondosa and Eclipta prostrata grown under three levels of water availability and ask: (1) Do inoculated seedlings differ in growth and development from non-inoculated plants? (2) Are the effects of inoculation and degree of colonization dependent on water availability? (3) Do plant responses to inoculation differ between two closely related species? Inoculation had no detectable effects on shoot height, or plant biomass but did affect biomass partitioning and root morphology in a species-specific manner. Shoot/root ratios were significantly lower in non-inoculated E. prostrata plants compared with inoculated plants (0.381 ± 0.066 vs. 0.683 ± 0.132). Root length and surface area were greater in non-inoculated E. prostrata (259.55 ± 33.78 cm vs. 194.64 ± 27.45 cm and 54.91 ± 7.628 cm(2) vs. 46.26 ± 6.8 cm(2), respectively). Inoculation had no detectable effect on B. frondosa root length, volume, or surface area. AMF associations formed at all levels of water availability. Hyphal, arbuscular, and vesicular colonization levels were greater in dry compared with intermediate and flooded treatments. Measures of mycorrhizal responsiveness were significantly depressed in E. prostrata compared with B. frondosa for total fresh weight (-0.3 ± 0.18 g vs. 0.06 ± 0.06 g), root length (-0.78 ± 0.28 cm vs.-0.11 ± 0.07 cm), root volume (-0.49 ± 0.22 cm(3) vs. 0.06 ± 0.07 cm(3)), and surface area (-0.59 ± 0.23 cm(2) vs.-0.03 ± 0.08 cm(2)). Given the disparity in species response to AMF inoculation, events that alter AMF prevalence in wetlands could significantly alter plant community structure by directly affecting seedling growth and development.