Root exudate-assisted phytoremediation of copper and lead contamination using Rumex acetosa L. and Rumex K-1.

Heavy metal pollutants can be effectively removed from soil through phytoremediation using root exudates. Herein, experiments were conducted to assess the phytoremediation capabilities of Rumex acetosa L. and Rumex K-1 root exudates for copper (Cu) and lead (Pb) contamination. Results indicated that these root exudates effectively adsorbed Cu and Pb. Furthermore, the optimal adsorption conditions of Cu by the root exudates of both plants were as follows: light duration of 36 h, light intensity of 8000 Lx, temperature of 25 °C and CO(NH2)2 concentration of 0 %. Moreover, the optimal adsorption conditions of Pb by Rumex acetosa L. and Rumex K-1 root exudates were light duration of 48 h and 24 h, respectively, light intensity of 8000 Lx, temperature of 25 °C and CO(NH2)2 concentration of 0 %. In addition, the root exudates from both plants enhanced the enrichment and transport of Cu and Pb. Moreover, the root was found to be the main accumulation site of Pb, while the stems and leaves were the main accumulation sites of Cu. With the application of root exudates, plant growth increased, with growth indices in Rumex acetosa L. and Rumex K-1 groups treated with exudates being 1.08-1.81-fold and 1.06-1.9-fold higher, respectively, compared with the untreated ones; physiological indexes showed 1.14-2.62-fold and 1.14-2.71-fold improvements, respectively. Remediation efficiency indexes showed 1.05-1.62-fold and 1.10-1.89-fold improvements, respectively. Rumex acetosa L. and Rumex K-1 exhibited promising potential for the phytoremediation of Cu and Pb, with root exudates playing a critical role in metal adsorption and stabilisation, suggesting their potential for enhancing remediation capabilities. This study sheds light on the mechanisms of root exudate-assisted phytoremediation and provides insights into alleviating heavy metal pollution.

Allelopathic effect of the methanol extract of the weed species-red sorrel (Rumex acetosella L.) on the growth, phytohormone content and antioxidant activity of the cover crop - white clover (Trifolium repens L.).

Allelopathy is a biological process in which one organism releases biochemicals that affect the growth and development of other organisms. The current investigation sought to determine the allelopathic effect of Rumex acetosella on white clover (Trifolium repens) growth and development by using its shoot extract (lower IC50 value) as a foliar treatment. Here, different concentrations (25, 50, 100, and 200 g/L) of shoot extract from Rumex acetosella were used as treatments. With increasing concentrations of shoot extract, the plant growth parameters, chlorophyll and total protein content of Trifolium repens decreased. On the other hand, ROS, such as O2.- and H2O2, and antioxidant enzymes, including SOD, CAT, and POD, increased with increasing shoot extract concentration. A phytohormonal study indicated that increased treatment concentrations increased ABA and SA levels while JA levels were reduced. For the identification of allelochemicals, liquid‒liquid extraction, thin-layer chromatography, and open-column chromatography were conducted using R. acetosella shoot extracts, followed by a seed bioassay on the separated layer. A lower IC50 value was obtained through GC/MS analysis. gammaSitosterol was identified as the most abundant component. The shoot extract of Rumex acetosella has strong allelochemical properties that may significantly impede the growth and development of Trifolium repens. This approach could help to understand the competitive abilities of this weed species and in further research provide an alternate weed management strategy.

Study on the purification mechanism for ammonia nitrogen in micro-polluted rivers by herbaceous plant - Rumex japonicus Houtt.

Water eutrophication caused by nitrogen pollution is an urgent global issue that requires attention. The Qingyi River is a typical micro-polluted river in China. In this study, we took this river as the research object to investigate the nitrogen pollution purification capacity of a herbaceous plant, Rumex japonicus Houtt. (RJH). Compared to nitrate nitrogen (NO3--N) and nitrite nitrogen (NO2--N), RJH showed better purification performance on total nitrogen (TN), total phosphorus (TP) and ammonia nitrogen (NH4+-N), with a highest removal rate of 37.22%, 52.13%, and 100%, respectively. RJH could completely remove ammonia nitrogen and exhibit excellent resistance to pollutant interference when the initial concentration of ammonia nitrogen in the cultivation devices increased from 1 mg/L to 10 mg/L or in the actual river. This indicated the great application potential of RJH in ammonia nitrogen removal from natural micro-polluted rivers. In addition, combined effects of nitrification of roots, absorption of self-growth, stripping, and others contributed to nitrogen removal by RJH. Particularly, the nitrification of roots played a dominant role, accounting for 73.85% ± 8.79%. High-throughput sequencing results indicate that nitrifying bacteria accounted for over 75% of all bacterial species in RJH. Furthermore, RJH showed good growth status and strong adaptability. The correlation coefficients of its relative growth rate with chlorophyll A and the degradation rate of absorption were 0.9677 and 0.9594, respectively. Our research demonstrates that RJH is one of the excellent varieties for ammonia removal. This provides a very promising and sustainable method for purifying micro-polluted rivers.

Variance of soil bacterial community and metabolic profile in the rhizosphere vs. non-rhizosphere of native plant Rumex acetosa L. from a Sb/As co-contaminated area in China.

Heavy metals (HMs) contamination poses a serious threat to soil health. However, the rhizosphere effect of native pioneer plants on the soil ecosystem remains unclear. Herein, how the rhizosphere (Rumex acetosa L.) influenced the process of HMs threatening soil micro-ecology was investigated by coupling various fractions of HMs, soil microorganisms and soil metabolism. The rhizosphere effect alleviated the HMs' stress by absorbing and reducing HMs' direct bioavailability, and the accumulation of ammonium nitrogen increased in the rhizosphere soil. Meanwhile, severe HMs contamination covered the rhizosphere effect on the richness, diversity, structure and predicted function pathways of soil bacterial community, but the relative abundance of Gemmatimonadota decreased and Verrucomicrobiota increased. The content of total HMs and physicochemical properties played a more important role than rhizosphere effect in shaping soil bacterial community. Furthermore, As was observed to have a more significant impact compared to Sb. Moreover, plant roots improved the stability of bacterial co-occurrence network, and significantly changed the critical genera. The process influenced bacterial life activity and nutrient cycling in soil, and the conclusion was further supported by the significant difference in metabolic profiles. This study illustrated that in Sb/As co-contaminated area, rhizosphere effect significantly changed soil HMs content and fraction, soil properties, and microbial community and metabolic profiles.

Anti-cancer effect of Rumex obtusifolius in combination with arginase/nitric oxide synthase inhibitors via downregulation of oxidative stress, inflammation, and polyamine synthesis.

Cancer continues to be a leading cause of death worldwide, making the development of new treatment methods crucial in the fight against it. With cancer incidence rates increasing worldwide, ongoing research must focus on identifying new and effective ways to prevent and treat the disease. The combination of herbal extracts with chemotherapeutic agents has gained much interest as a novel strategy to combat cancer. Rumex obtusifolius L. is a wild plant known for its medicinal properties and is widely distributed worldwide. Our preclinical evaluations suggested that R. obtusifolius seed extracts possessed cancer-inhibiting properties and we also evaluated the beneficial effects of the arginase inhibitor NG-hydroxy-nor-L-arginine and nitric oxide inhibitor NG-nitro-L-arginine methyl ester in the treatment of breast cancer. The current study aimed to combine these observations and evaluate the antioxidant and antitumor properties of R. obtusifolius extracts alone and in combination with the arginase and nitric oxide synthase inhibitors. Metabolic characterization of the plant extract using a liquid chromatography/high-resolution mass spectrometry advanced system revealed the presence of 240 phenolic compounds many of which possess anticancer properties, according to the literature. In vitro studies revealed a significant cytotoxic effect of the R. obtusifolius extracts on the human colon (HT29) and breast cancer (MCF-7) cell lines. Thus, a new treatment approach of combining R. obtusifolius bioactive phytochemicals with the arginase and nitric oxide synthase inhibitors NG-nitro-L-arginine methyl ester and/or NG-hydroxy-nor-L-arginine, respectively, was proposed and could potentially be an effective way to treat breast cancer. Indeed, these combinations showed immunostimulatory, antiproliferative, antioxidant, anti-inflammatory, and antiangiogenic properties in a rat breast cancer model.

Rumex acetosella Inhibits Platelet Function via Impaired MAPK and Phosphoinositide 3-Kinase Signaling.

OBJECTIVE: To examine the antiplatelet and antithrombotic activity of Rumex acetosella extract. METHODS: Standard light aggregometry was used for platelet aggregation, intracellular calcium mobilization assessed using Fura-2/AM, granule secretion (ATP release) by luminometer, and fibrinogen binding to integrin αIIbß3 detected using flow cytometry. Western blotting is carried out to determine the phosphorylation of mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K)/Akt signaling. RESULTS: Rumex acetosella displayed the ability to inhibit platelet aggregation, calcium mobilization, granule secretion, and fibrinogen binding to integrin αIIbß3. Rumex acetosella has also down-regulated MAPK and PI3K/Akt phosphorylation (all P<0.01). CONCLUSION: Rumex acetosella extract exhibits antiplatelet activity via modulating GPVI signaling, and it may protect against the development of platelet-related cardiovascular diseases.

The impact of aqueous and N-hexane extracts of three Fabaceae species on seed germination and seedling growth of some broadleaved weed species.

Weed infestation is a persistent problem for centuries and continues to be major yield reducing issue in modern agriculture. Chemical weed control through herbicides results in numerous ecological, environmental, and health-related issues. Moreover, numerous herbicides have evolved resistance against available herbicides. Plant extracts are regarded as an alternative to herbicides and a good weed management option. The use of plant extracts is environmentally safe and could solve the problem of herbicide resistance. Therefore, laboratory and wire house experiments were conducted to evaluate the phytotoxic potential of three Fabaceae species, i.e., Cassia occidentalis L. (Coffee senna), Sesbania sesban (L.) Merr. (Common sesban) and Melilotus alba Medik. (White sweetclover) against seed germination and seedling growth of some broadleaved weed species. Firstly, N-hexane and aqueous extracts of these species were assessed for their phytotoxic effect against lettuce (Lactuca sativa L.). The extracts found more potent were further tested against germination and seedling growth of four broadleaved weed species, i.e., Parthenium hysterophorus L. (Santa-Maria), Trianthema portulacastrum L. (Pigweed), Melilotus indica L (Indian sweetclover). and Rumex dentatus L. (Toothed dock) in Petri dish and pot experiments. Aqueous extracts of all species were more toxic than their N-hexane forms for seed germination and seedling growth of lettuce; therefore, aqueous extracts were assessed for their phytotoxic potential against four broadleaved weed species. Aqueous extracts of all species proved phytotoxic against T. portulacastrum, P. hysterophorus, M. indica and R. dentatus and retarder their germination by 57, 90, 100 and 58%, respectively. Nevertheless, foliar spray of C. occidentalis extract was the most effective against T. portulacastrum as it reduced its dry biomass by 72%, while M. alba was effective against P. hysterophorus, R. dentatus and M. indica and reduced their dry biomass by 55, 68 and 81%, respectively. It is concluded that aqueous extracts of M. alba, S. sesban and C. occidentalis could be used to retard seed germination of T. portulacastrum, P. hysterophorus, M. indica and R. dentatus. Similarly, aqueous extracts of C. occidentalis can be used to suppress dry biomass of T. portulacastrum, and those of M. alba against P. hysterophorus, R. dentatus. However, use of these extracts needs their thorough testing under field conditions.

Secondary Metabolites, Ferulic Acid and p-Hydroxybenzoic Acid Induced Toxic Effects on Photosynthetic Process in Rumex acetosa L.

The elimination of broadleaf weeds from agricultural fields has become an urgent task in plant and environment protection. Allelopathic control is considered a potential approach because of its exclusive and ecological safety measures. Plant secondary metabolites also called allelochemicals are released from plant leaves, roots, stem, bark, flowers and play significant roles in soil rhizosphere signaling, chemical ecology, and plant defense. The present study was carried out to evaluate the impact of two allelochemicals; ferulic acid (FA) and p-hydroxybenzoic acid (pHBA) on photosynthetic characteristics; Fv/Fm: efficiency of photosystem II photochemistry in the dark-adapted state; ΦPSII: photosynthetic quantum yield; NPQ, non-photochemical quenching; qP, photochemical quenching, and photon energy dissipation (1-qP)/NPQ in Rumex acetosa following 6 days exposure. R. acetosa seedlings were grown in perlite culture, irrigated with Hoagland solution and treated with allelopathic compounds FA and pHBA and were evaluated against the photosynthetic attributes. Both compounds behaved as potent inhibitors of photosynthetic traits such as Fv/Fm, ΦPSII, qP, and NPQ in R. acetosa. Photon energy dissipation (1-qP)/NPQ increased significantly from days 3 to 6. Higher dissipation of absorbed energy indicates the inactivation state of reaction centers and their inability to effectively use the absorbed energy in photosynthesis. These results indicated the potential allelopathic application of FA and pHBA for control of broadleaf weed, Rumex acetosa.

Anti-inflammatory effects of Beopje curly dock (Rumex crispus L.) in LPS-induced RAW 264.7 cells and its active compounds.

Inflammation is a defense response of the body to stimuli. Curly dock (CD) is an herbal food with anti-inflammatory effects. Beopje is an herbal food processing method that reduces toxicity and enhances beneficial effects. This study investigated the effects of CD and Beopje curly dock (CD-B) extracts on lipopolysaccharide (LPS)-induced inflammatory damage in RAW 264.7 cells. Cell survival rate and nitrite concentration were determined using the MTT assay and Griess method, respectively. Enzyme-linked immunosorbent assay was used to detect the inflammatory cytokine levels. The mRNA and protein expression levels of inflammatory associated genes were detected by qPCR and Western blot, respectively. CD and CD-B extracts compositions were assessed by UPLC-Q-TOF MS analysis. Our results indicate that CD-B has a more significant inhibitory effect on the LPS-induced inflammatory response in RAW 264.7 cells than CD, suggesting that the Beopje process potentially enhances the anti-inflammatory effect of CD. PRACTICAL APPLICATIONS: Long-term inflammation can cause a variety of chronic diseases. Therefore, it is necessary to suppress the occurrence of body inflammation in time. This study preliminarily clarified the mechanism of herbal foods to alleviate inflammation by regulating the immune response, and further confirms that applying the Beopje process enhances the anti-inflammatory effect. This research can serve as a significant reference for future research, prevention and treatment of inflammation-related diseases, and the development of functional foods with anti-inflammatory activity. It also provides a theoretical basis for the further reasonable application of Beopje processing method.

Growth performance and emergence of invasive alien Rumex confertus in different soil types.

Rumex confertus is known to be one of the most serious invasive weed species infesting meadows, pastures and ruderal lands throughout the Central Europe. Rumex confertus was grown in pot experiments using 8 soil types at 5 concentrations of nitrogen (N) and phosphorus (P). Based on harvest data, the variables, seedlings emergence, root: shoot (R: S) ratio, N and P concentration, chlorophyll content, Relative Colimitation Index (RCI) and dry matter allocation to plant components, were determined. N and P addition stimulated the growth of plants in different soils, as reflected by a significant increase in seedling growth parameters such as total plant biomass and shoot biomass. Across all soil types, emergence of seedlings was negatively affected by very high N, but positively affected by increased P availability. This study indicates that Dystric Arenosol, Entic Podzol, Brunic Arenosol and Calcaric Leptosol are unfavorable for R. confertus growth, excluding  R. confertus completely. Moreover, evidence suggests that plant growth is limited by both N and P, therefore R. confertus could be controlled by reducing available N and P content in the soil.

Optimization of plant hormonal balance by microorganisms prevents plant heavy metal accumulation.

Heavy metal contamination is a threat to global food safety. Reducing heavy metal uptake in plants is a promising way to make plants safer, yet breeding the right set of traits can be tedious. We test whether microorganisms are able to impact the plant's hormonal balance hereby helping to manage plant heavy metal uptake. We focus on ethylene, a plant hormone regulating plant stress tolerance and nutrition. We grew three phylogenetically distinct plants, Rumex palustris, Alcea aucheri and Arabidopsis thaliana, on a cadmium-spiked soil. Plants roots were coated with the bacterium Pseudomonas putida UW4, which degrades the precursor of ethylene, or an isogenic ACC deaminase-deficient mutant lacking this ability. We followed ethylene concentrations, plant growth and cadmium uptake. Wildtype bacteria reduced shoot cadmium concentration by up to 35% compared to the control, while the mutant increased cadmium concentration. This effect was linked to ethylene, which was consistently positively correlated with cadmium concentration. We therefore propose that bacteria modulating plant hormonal balance may offer new possibilities to improve specific aspects of plant phenotype, in the present context reducing heavy metal. They may thus pave the way for new strategies to improve food safety in a context of the widespread soil contamination.

The effect of biogenic manufactured silver nanoparticles on human endothelial cells and zebrafish model.

Human health and environment have been continuously getting exposure to toxic chemicals including nanomaterial; therefore, nontoxicity has recently attracted huge amount of attention. In this study, RU-AgNPs were synthesized by a green synthesis procedure and evaluated for their toxicity in human umbilical vein endothelial cells (HUVECs) as well as on zebrafish embryos via apoptotic pathway. The synthesized RU-AgNPs were average in size (20-25 nm) with a negative surface charge of -13.43 mV. As a result, RU-AgNPs potentiated the formation of reactive oxygen species (ROS) in HUVECs as confirmed by the results of immunoblotting analysis using apoptotic markers, such as Bax, Bcl2, and cytochrome C. Moreover, the induction of apoptosis in HUVECs was also authenticated in a dose-dependent manner after the treatment with RU-AgNPs by the Incucyte analysis. In vivo trials conducted on zebrafish visualized the mortality, malformation, and imbalanced in the heart rate, and cell death of the whole embryo, including severe morphological changes in the yolk sac and the tail of zebrafish. Furthermore, the results of western blot analysis demonstrated the increasing intensity of apoptotic biomarkers such as Bax, Bcl2, and Cyto C, including enhanced production of ROS, validating the cell death in zebrafish larvae. In addition, chemically functionalized silver nanoparticles found to be more cytotoxic than biogenic functionalized silver nanoparticles. Above-mentioned findings clearly demonstrate that Ru-AgNPs cause the toxicity via ROS-induced apoptotic pathway. Therefore, it is necessary to decide RU-AgNPs toxicity levels before being used in any biomedical application.

Use of ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry system as valuable tool for an untargeted metabolomic profiling of Rumex tunetanus flowers and stems and contribution to the antioxidant activity.

Rumex tunetanus is an untapped resource with no available information about their chemical composition. That is why, the present study has conducted comprehensive metabolites profiling of the hydromethanol extracts of the Rumex tunetanus flowers and stems using RP-UHPLC-ESI-QTOF-MS. As results, 60 compounds were characterized for the first time in Rumex tunetanus between them, 18 photochemical were firstly identified in the Polygonaceae family in negative ionization mode. In this regard, quercetin-3-O-glucuronide and (-)-epicatechin gallate were the most abundant phenolic compounds in flowers and stems, respectively. Moreover, positive correlations were found between the antioxidant activity measured by DPPH and FRAP assays with the total phenolic compounds (r = 0.98; r = 0.99, respectively) and the abundance of some phenolic subfamilies such as hydroxycinnamic acids, hydroxybenzoic acids, flavonols and flavones with r > 0.86. The compounds displaying significant (P < 0.01) and good correlations with the antioxidant activity (r > 0.93) were hydroxybenzoic acid, rutin, quercetin-3-O-glucuronide, quercetin-3-O-glucoside, quercetin and luteolin-7-O-rutinoside. In conclusion, Rumex tunetanus flowers and stems showed different bioactive compound profiles and significant antioxidant properties and could be further used in food and nutraceutical industries.

Carbon (δ13C) and Nitrogen (δ15N) Stable Isotope Composition Provide New Insights into Phenotypic Plasticity in Broad Leaf Weed Rumex acetosa under Allelochemical Stress.

Phenolic compounds, hydroquinone and cinnamic acid derivatives have been identified as major allelochemicals with known phytotoxicity from allelopathic plant Acacia melanoxylon R. Br. Several phenolic compounds such as ferulic acid (FA), p-hydroxybenzoic acid (pHBA) and flavonoid (rutin, quercetin) constituents occur in the phyllodes and flowers of A. melanoxylon and have demonstrated inhibitory effects on germination and physiological characteristics of lettuce and perennial grasses. However, to date, little is known about the mechanisms of action of these secondary metabolites in broad-leaved weeds at ecophysiological level. The objective of this study was to determine the response of Rumex acetosa carbon isotope composition and other physiological parameters to the interaction of plant secondary metabolites (PSM) (FA and pHBA) stress and the usefulness of carbon isotope discrimination (Δ13C) as indicative of the functional performance of intrinsic water use efficiency (iWUE) at level of plant leaf. R. acetosa plant were grown under greenhouse condition and subjected to PSM stress (0, 0.1, 0.5, 1.0, and 1.5 mM) for six days. Here, we show that FA and pHBA are potent inhibitors of Δ13C that varied from 21.0‰ to 22.9‰. Higher pHBA and FA supply enhanced/retard the Nleaf and increased the Cleaf while ratio of intercellular CO2 concentration from leaf to air (Ci/Ca) was significantly decreased as compared to control. Leaf water content and leaf osmotic potential were decreased following treatment with both PSM. The Ci/Ca decreased rapidly with higher concentration of FA and pHBA. However, iWUE increased at all allelochemical concentrations. At the whole plant level, both PSM showed pronounced growth-inhibitory effects on PBM and C and N concentration, root fresh/dry weight, leaf fresh/dry weight, and root, shoot length of C3 broad leaf weed R. acetosa. Carbon isotope discrimination (Δ) was correlated with the dry matter to transpiration ratio (transpiration efficiency) in this C3 species, but its heritability and relationship to R. acetosa growth are less clear. Our FA and pHBA compounds are the potent and selective carbon isotope composition (δ13C) inhibitors known to date. These results confirm the phytotoxicity of FA and pHBA on R. acetosa seedlings, the reduction of relative water content and the induction of carbon isotope discrimination (Δ) with lower plant biomass.

The improvement of bioactive secondary metabolites accumulation in Rumex gmelini Turcz through co-culture with endophytic fungi

Abstract Aspergillus sp., Fusarium sp., and Ramularia sp. were endophytic fungi isolated from Rumex gmelini Turcz (RGT), all of these three strains could produce some similar bioactive secondary metabolites of their host. However the ability to produce active components degraded significantly after cultured these fungi alone for a long time, and were difficult to recover. In order to obtain more bioactive secondary metabolites, the co-culture of tissue culture seedlings of RGT and its endophytic fungi were established respectively, and RGT seedling was selected as producer. Among these fungi, Aspergillus sp. showed the most significant enhancement on bioactive components accumulation in RGT seedlings. When inoculated Aspergillus sp. spores into media of RGT seedlings that had taken root for 20 d, and made spore concentration in co-culture medium was 1 × 104 mL-1, after co-cultured for 12 d, the yield of chrysophaein, resveratrol, chrysophanol, emodin and physcion were 3.52-, 3.70-, 3.60-, 4.25-, 3.85-fold of the control group. The extreme value of musizin yield was 0.289 mg, which was not detected in the control groups. The results indicated that co-culture with endophytic fungi could significantly enhance bioactive secondary metabolites production of RGT seedlings.

Evaluation of metabolic changes in oxalate-rich plant Rumex obtusifolius L. caused by ion beam irradiation.

Some Rumex species such as sorrel are edible as baby leaf salad greens. On the other hand, Rumex plants accumulate soluble oxalate, a toxic metabolite which causes serious diseases such as renal syndrome. We attempted to produce low-oxalate plants of R. obtusifolius, a perennial weed which has higher vitamin C and amino acid content and higher tolerance to stress than many other Rumex species. Ion beams are ionising radiation with high linear energy transfer that causes a wide spectrum of mutations. Thus, in the present study we evaluated the effects of ion beams on oxalate and other primary metabolites in leaves of R. obtusifolius using CE-MS. The results showed that oxalate content was increased by irradiation with carbon ion beams. Metabolome analysis revealed that ion beams affected carbon flow to the isocitrate pathway, which is involved in oxalate synthesis. These observations suggested that modulation of carbon flow to the isocitrate pathway is important to regulate oxalate levels in plants.

Evaluation of raw nepodin extraction from Rumex japonicus and R. obtusifolius and their DNA polymorphisms.

Nepodin, found in the roots of Rumex japonicus Houtt. (Polygonaceae), inhibits osteoclast differentiation and has an antidiabetic effect. We propose nepodin as an ingredient of new functional foods or as a drug candidate for reducing the risk of reduced locomotion resulting from diseases such as osteoporosis. Although there are no previous reports of R. obtusifolius L., which is found throughout Japan, having roots containing nepodin, we found nepodin in the roots of this species. Therefore, R. obtusifolius as well as R. japonicus was considered a candidate raw material for nepodin extraction. We also discuss the suitability of R. japonicus and R. obtusifolius as sources of raw nepodin for cultivation on the Ryukyu Islands. In this study, all specimens on the Ryukyu Islands were identified as R. japonicus. Conversely, all specimens on mainland Japan were R. obtusifolius. The DNA sequence of the chloroplast trnL-trnF intergenic spacer region and partial nuclear internal transcribed spacer was consistent with the identification of R. japonicus and R. obtusifolius by morphological characteristics of the perianth segments. Therefore, to avoid erroneous identification and misuse of the plant species used for extraction of raw materials, it is preferable to develop DNA markers for these two regions. The content of nepodin varied from undetectable to 0.34% of the fresh weight (%FW) in R. japonicus and from undetectable to 0.21%FW in R. obtusifolius. From a pharmacological perspective, as plants that might be suitable as raw materials for nepodin extraction, it became clear that both R. japonicus and R. obtusifolius can be used with the same expected extraction efficiency. Based on our findings, R. obtusifolius could not be confirmed as inhabiting the Ryukyu Islands. For this reason, to conserve the endemic genetic characteristics of the Ryukyu Islands and to prevent genetic pollution by R. obtusifolius, only R. japonicus should be cultivated on the Ryukyu Islands.

The improvement of bioactive secondary metabolites accumulation in Rumex gmelini Turcz through co-culture with endophytic fungi.

Aspergillus sp., Fusarium sp., and Ramularia sp. were endophytic fungi isolated from Rumex gmelini Turcz (RGT), all of these three strains could produce some similar bioactive secondary metabolites of their host. However the ability to produce active components degraded significantly after cultured these fungi alone for a long time, and were difficult to recover. In order to obtain more bioactive secondary metabolites, the co-culture of tissue culture seedlings of RGT and its endophytic fungi were established respectively, and RGT seedling was selected as producer. Among these fungi, Aspergillus sp. showed the most significant enhancement on bioactive components accumulation in RGT seedlings. When inoculated Aspergillus sp. spores into media of RGT seedlings that had taken root for 20d, and made spore concentration in co-culture medium was 1×104mL-1, after co-cultured for 12d, the yield of chrysophaein, resveratrol, chrysophanol, emodin and physcion were 3.52-, 3.70-, 3.60-, 4.25-, 3.85-fold of the control group. The extreme value of musizin yield was 0.289mg, which was not detected in the control groups. The results indicated that co-culture with endophytic fungi could significantly enhance bioactive secondary metabolites production of RGT seedlings.

Biological Activities and Secondary Metabolite Screening of Rumex hastatus Extract through Fourier Transform Infrared and Raman Spectroscopy.

BACKGROUND: Human infectious diseases are caused by various pathogens including bacteria, fungi, viruses, parasites, and protozoans. These infectious agents are controlled by using synthetic drugs as well as natural sources. OBJECTIVE: The aim of current study was to evaluate the antibacterial effect of Rumex hastatus against clinical bacterial pathogens. METHODS: In current research antibacterial effect of Rumex hastatus was analyzed against seven clinical pathogenic bacteria such as Escherichia coli, Serratia marcescens, Streptococcus pyogenes, Staphylococcus epidermidis, Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa through agar well diffusion method. The boiled extract was used for the phytochemical screening, antioxidant potential, thin layer chromatography, bio-autography, and spot screening. Genomic DNA was extracted to find the DNA protection effect of R. hastatus. RESULTS: Antibacterial results showed that diethyl ether extract has the maximum inhibition of S. pyogenes (9.66 ± 0.57 mm). Acetone and diethyl ether extracts showed moderate inhibition of K. pneumoniae (6.33 ± 1.52 mm and 5.66 ±1.15 mm) and S. aureus (6.33 ± 1.52 mm and 5.66 ± 0.57 mm). Similarly, chloroform extract indicated moderate inhibition of S. pyogenes (5.66 ± 1.15 mm). Ethanol extract had low or even no effect on the growth of bacteria. Genomic DNA extraction also encouraged the antibacterial effect of R. hastatus. Various phytochemical constituents such as ketoses, oligosaccharides, amino acids, amines, sugars, flavonoids, and antioxidant constituents were detected. TLC-Bioautography and spot screening results revealed the potential use of R. hustatus as an antibacterial agent. CONCLUSION: It was concluded that most of the tested fractions appeared as an important source for the discovery of new antimicrobial drugs.

Differential expression of vacuolar and defective cell wall invertase genes in roots and seeds of metalliferous and non-metalliferous populations of Rumex dentatus under copper stress.

Acid invertase activities in roots and young seeds of a metalliferous population (MP) of Rumex dentatus were previously observed to be significantly higher than those of a non-metalliferous population (NMP) under Cu stress. To date, no acid invertase gene has been cloned from R. dentatus. Here, we isolated four full-length cDNAs from the two populations of R. dentatus, presumably encoding cell wall (RdnCIN1 and RdmCIN1 from the NMP and MP, respectively) and vacuolar invertases (RdnVIN1 and RdmVIN1 from the NMP and MP, respectively). Unexpectedly, RdnCIN1 and RdmCIN1 most likely encode special defective invertases with highly attenuated sucrose-hydrolyzing capacity. The transcript levels of RdmCIN1 were significantly higher than those of RdnCIN1 in roots and young seeds under Cu stress, whereas under control conditions, the former was initially lower than the latter. Unexpected high correlations were observed between the transcript levels of RdnCIN1 and RdmCIN1 and the activity of cell wall invertase, even though RdnCIN1 and RdmCIN1 do not encode catalytically active invertases. Similarly, the transcript levels of RdmVIN1 in roots and young seeds were increased under Cu stress, whereas those of RdnVIN1 were decreased. The high correlations between the transcript levels of RdnVIN1 and RdmVIN1 and the activity of vacuolar invertase indicate that RdnVIN1 and RdmVIN1 might control distinct vacuolar invertase activities in the two populations. Moreover, a possible indirect role for acid invertases in Cu tolerance, mediated by generating a range of sugars used as nutrients and signaling molecules, is discussed.