Rugosa Var Research Articles

Camptothecin and its Analogs: High-Yielding Ophiorrhiza Species from Sri Lanka for Sustainable Anticancer Compound Production

Introduction: Camptothecin (CPT) is pivotal in cancer treatment, derived from various CPT-producing plant species, and is a fundamental component in synthesizing valuable cancer drugs like Irnotican and Topotecan. Sourcing from nature poses conservation issues, fostering interest in the herbaceous Ophiorrhiza plant as a more sustainable alternative. Ophiorrhiza species in Sri Lanka lack comprehensive study, warranting exploration for echo-friendly anticancer compound production. Objectives: This study examines CPT and analog content in Ophiorrhiza mungos, O. pectinata, and O. rugosa across diverse Sri Lankan regions. Methods: The study employs Thin Layer Chromatography (TLC), High-Performance Liquid Chromatography-DAD (HPLC-DAD), and Liquid Chromatography-Mass Spectrometry (LC-MS) to quantify and confirm CPT and its analogs. Results: Significant variations in the content of CPT and its analogs were observed among plant parts and regions. O. mungos from Deraniyagala and Bibile regions notably exhibited elevated CPT levels in fruits and roots. O. rugosa var. Angustifolia and O. pectinata also exhibited a considerable content of CPT in their roots, though it was significantly lower (p < 0.005) than O. mungos. Conclusion: The study validates analytical methods for specificity, linearity, precision, accuracy, and sensitivity per ICH guidelines. The results indicate that Ophiorrhiza species, especially O. mungos and O. rugosa var. Angustifolia holds the potential to be a sustainable source of CPT. Optimizing cultivation practices offers an eco-friendly solution for anticancer compound production, alleviating species threats and conserving biodiversity.

Comparative study on metabolite variations of two rose teas by plant metabolomics and revealing their skin-whitening candidates by spectrum-effect relationship analysis.

Rosa rugosa var. plena Rehd (CBR) and Rosa chinensis cv. "JinBian" (JBR) are two common species used in rose tea among different original species. CBR, the officially documented original plant of the rose species for food and medicinal purposes, is more costly than JBR. With increasing demand for different rose teas, it is meaningful to compare the chemical constituents for their quality control and reveal their skin-whitening components that will provide in-depth insights for the expansion of the rose tea industry. This study aims to reveal the chemical variances between CBR and JBR and determine their skin-whitening components. A strategy obtained by combining MS-based plant-metabolomics with spectrum-effect relationship analysis has been proposed for unveiling chemical differences between CBR and JBR and further exploring their potential skin-whitening components. A total of 2030 metabolites were found that revealed considerable differences between CBR and JBR. The results of bioactivity assay demonstrated that JBR exhibited stronger tyrosinase inhibition activity than CBR. Six potential skin-whitening compounds (di-O-galloyl-HHDP-glucoside, tri-O-galloyl-HHDP-glucoside, spiraeoside, quinic acid, rugosin A, and 1,2,3,6-tetra-O-galloyl-glucose) were discovered as potential tyrosinase inhibitors, via spectrum-effect relationship analysis. This is the first time that di-O-galloyl-HHDP-glucoside, tri-O-galloyl-HHDP-glucoside, rugosin A, and 1,2,3,6-tetra-O-galloyl-glucose have been reported with tyrosinase inhibitory activity. Additionally, molecular docking analysis was used to reveal the inhibition mechanism of these compounds toward tyrosinase. The finding of this study will be of great importance for the quality control of the two types of rose teas, and the revealed active ingredients will provide in-depth insights for the expansion of the rose tea industry.

Molecular phylogeny and taxonomy of four Remanella species (Protozoa, Ciliophora): A flagship genus of karyorelictean ciliates, with descriptions of two new species.

During faunal studies of psammophilic ciliates along the coast of Qingdao, China, several marine karyorelictean species were isolated. Among them, four species within the genus Remanella were investigated, including two species new to science: i.e., R. rugosa, Remanella elongata sp. nov., Remanella aposinica sp. nov., and R. unicorpusculata. Remanella rugosa has been reported several times, but this study is the first to provide detailed morphological characters and phylogenetics. Remanella elongata sp. nov. can be distinguished from its congeners by the presence of complex cortical granules, fewer macronuclei, and longer body size. Remanella aposinica sp. nov. differs from its congeners by having 14–17 right lateral ciliary rows and 24–37 dikinetids of intrabuccal kinety. Poorly known Remanella rugosa var. unicorpusculata (Kahl, 1933) Foissner, 1996 should be elevated from subspecies to species level, Remanella unicorpusculata (Foissner, 1996) stat. nov., based on detailed redescriptions with statistical data, living morphology, infraciliature, and species definitions. Small subunit (SSU) rDNA was sequenced for the four species, and phylogenetic analysis revealed that all known taxa in Remanella formed the outline branch to the genus Loxodes with moderate to high bootstrap support among Remanella lineages.

Central and peripheral pain intervention by Ophiorrhizarugosa leaves: Potential underlying mechanisms and insight into the role of pain modulators

Ethnopharmacological relevanceOphiorrhiza rugosa var. prostrata is a traditional medicinal plant used by the indigenous and local tribes (Chakma, Marma and Tanchangya) of Bangladesh for the management of chest pain, body ache, and earache. However, the knowledge of anti-nociceptive and anti-inflammatory potentials of this plant is scarce. Aim of the studyTherefore, we scrutinized the anti-nociceptive and anti-inflammatory properties of O. rugosa leaves along with its possible mechanism(s) of action using chemical and heat-induced pain models. Methods and materialsO. rugosa was extracted using 100% ethanol (EEOR) followed by exploring phytochemicals and assessing acute toxicity. To determine anti-nociceptive potentials, chemical-induced (acetic acid and formalin) and heat-induced (hot plate and tail immersion) nociceptive models were followed. To investigate the possible involvement of opioid receptors during formalin, hot plate, and tail immersion tests, naltrexone was administered whereas methylene blue and glibenclamide were used to explore cGMP involvement and ATP-sensitive K+ channel pathways, respectively. Moreover, the anti-inflammatory potential was assessed using the carrageenan-induced paw edema test model. Motor behaviours of EEOR were assessed by the open-field test. Finally, bioactive constituents (identified by GC-MS) from O. rugosa were subjected to molecular docking and ADME/t analysis to evaluate its potency and safety. ResultsDuring chemical-induced and heat-induced pain models, EEOR exhibited significant and effective nociception suppression at all experimental doses (200 and 400 mg/kg). Also, the administration of naltrexone corroborated the association of opioid receptors with the anti-nociceptive activity by EEOR. Similarly, cGMP and ATP-sensitive K+ channel pathways were also found to be involved in the anti-nociceptive mechanism. Furthermore, significant and dose-dependent inhibition of inflammation induced by carrageenan was recorded for EEOR. Both doses of EEOR did not affect the animal's locomotor capacity in the open-field test. Besides, in silico test identified the key compounds (loliolide, harman, squalene, vitamin E, and gamma-sitosterol) that inhibited some particular receptors regarding pain and inflammation. ConclusionThis research exposes central and peripheral pain intervention as well as anti-inflammatory activity of O. rugosa. Also, the identified compounds from this plant support its activities by effectively inhibiting anti-nociceptive and anti-inflammatory receptors. Overall, these outcomes valorize the ethnomedicinal efficacy of O. rugosa in managing various painful conditions.

Investigation of the Biological Activities and Characterization of Bioactive Constituents of Ophiorrhiza rugosa var. prostrata (D.Don) & Mondal Leaves through In Vivo, In Vitro, and In Silico Approaches.

Ophiorrhiza rugosa var. prostrata is one of the most frequently used ethnomedicinal plants by the indigenous communities of Bangladesh. This study was designed to investigate the antidiarrheal, anti-inflammatory, anthelmintic and antibacterial activities of the ethanol extract of O. rugosa leaves (EEOR). The leaves were extracted with ethanol and subjected to in vivo antidiarrheal screening using the castor oil-induced diarrhea, enteropooling, and gastrointestinal transit models. Anti-inflammatory efficacy was evaluated using the histamine-induced paw edema test. In parallel, in vitro anthelmintic and antibacterial activities were evaluated using the aquatic worm and disc diffusion assays respectively. In all three diarrheal models, EEOR (100, 200 and 400 mg/kg) showed obvious inhibition of diarrheal stool frequency, reduction of the volume and weight of the intestinal contents, and significant inhibition of intestinal motility. Also, EEOR manifested dose-dependent anti-inflammatory activity. Anthelmintic action was deemed significant (P < 0.001) with respect to the onset of paralysis and helminth death. EEOR also resulted in strong zones of inhibition when tested against both Gram-positive and Gram-negative bacteria. GC-MS analysis identified 30 compounds within EEOR, and of these, 13 compounds documented as bioactive showed good binding affinities to M3 muscarinic acetylcholine, 5-HT3, tubulin and GlcN-6-P synthase protein targets in molecular docking experiments. Additionally, ADME/T and PASS analyses revealed their drug-likeness, likely safety upon consumption and possible pharmacological activities. In conclusion, our findings scientifically support the ethnomedicinal use and value of this plant, which may provide a potential source for future development of medicines.

Multiple shoot cultures of Ophiorrhiza rugosa var. decumbens Deb and Mondal – A viable renewable source for the continuous production of bioactive Camptotheca alkaloids apart from stems of the parent plant of Nothapodytes foetida (Wight) Sleumer

Camptotheca alkaloids were isolated from multiple shoot cultures of O. decumbens (0.056% dry weight) and stems of N. foetida. The cytotoxicity of the extracts and products were tested in a panel of five cell lines. Crude extract from O. decumbens (Cr-Od) and N. foetida (Cr-Nf) showed more potent cytotoxic activity as compared to the isolated camptothecin from O. decumbens (CPT-Od) and N. foetida (CPT-Nf). CPT isolated from shoot cultures contained biological activity suggesting the possibility of using this system of O. decumbens as a renewable source for the production of camptotheca alkaloids. 9-Methoxy camptothecin (9-mCPT), isolated from N. foetida, was a very effective cytotoxic agent as compared to Cr-Nf or CPT-Nf. The IC50 of 9-mCPT was 0.84, 0.32, and 0.35μg/ml for A549, MCF7 and Jurkat cell lines and >3μg/ml for U937. Viability assays using MTT dye were further confirmed by assessing extent of apoptosis in these cells. These findings suggest that shoot cultures of O. decumbens offer a rich alternative plant source for the anticancer compound, CPT and 9-mCPT is a more potent compound in N. foetida as compared to CPT.

High-frequency Plant Regeneration and Accumulation of the Anticancer Alkaloid Camptothecin in Ophiorrhiza rugosa var. decumbens Deb & Mondal

An efficient protocol for obtaining high-frequency regeneration and camptothecin production from Ophiorrhiza rugosa var. decumbens (Rubiaceae) was developed. Leaves, stem, petiole, and root explants from in vitro grown plants of O. decumbens were selected, cultured on Murashige and Skoog's (MS) basal medium supplemented with benzyl adenine (5 mg.L−1) and indole acetic acid (0.1 mg.L−1). Root explants produced maximum shoot regeneration. The regenerated shoots were rooted on MS medium containing α- naphthalene acetic acid (0.1 mg.L−1) and transferred to soil. The regenerated and potted plants indicated the accumulation of camptothecin. Whole regenerated plant of O. decumbens grown in a pot for 1 month showed maximum camptothecin (0.132% d.w) compared to the regenerated shoots from other explants (0.013-0.020 % d.w).

Search for Camptothecin-yielding Ophiorrhiza species from southern Western Ghats in India: A HPTLC-densitometry study

Camptothecin (CPT) content in 44 accessions belonging to nine Ophiorrhiza species and three varieties collected from their natural habitats in the Kerala region of Western Ghats were estimated by HPTLC-densitometry. Ophiorrhiza mungos 170.11–503.15 (μg/g, dr. wt.) and O. mungos var. angustifolia 127.86–476.89 (μg/g, dr. wt.) accessions showed the highest CPT contents. O. trichocarpon (19.50–28.31μg/g) and O. pectinata (0.28–38.65μg/g) showed moderate contents of CPT. Among the nine Ophiorrhiza species and three varieties screened O. barberi, O. caudata, O. nairii and O. rugosa var. decumbens showed zero or non-detectable levels of CPT. O. grandiflora (1.07–1.34μg/g), O. eriantha (0.00–0.30μg/g), O. rugosa var. prostrata (0.16μg/g) and O. shendurunii (0.05μg/g) showed only very low contents of CPT. O. mungos var. angustifolia is an established variant and is a small herb which could be relatively easily cultivated or multiplied by tissue culture compared to O. mungos. Significant variations in CPT contents were found between the ecotypes/genotypes of the same Ophiorrhiza species or variety collected from different geographic locations. This revealed the importance of choosing high CPT yielding Ophiorrhiza genotypes or ecotypes for commercial purposes.

Production of camptothecin by hairy roots and regenerated transformed shoots of Ophiorrhiza rugosa var. decumbens

Camptothecin (CPT), the derivatives of which are used clinically for the treatment of metastatic colon cancer, is isolated from intact plants that can be subjected to environmental fluctuations. In vitro cultures may be an alternate and continuous source for year-round production of CPT. Since CPT production by undifferentiated cell cultures is low, differentiated tissues such as root cultures may be a viable alternate source for CPT production. Hairy roots were induced in Ophiorriza rugosa, a source of CPT, using Agrobacterium rhizogenes strain LBA9402. The hairy roots, when cultured in light, showed spontaneous regeneration of shoots. Analysis of CPT levels in the hairy roots and in vitro-grown transformed shoots revealed 0.009% d.w. and 0.012% d.w., respectively.

Frankia Populations in Soil and Root Nodules of Sympatrically Grown Alnus Taxa

The genetic diversity of Frankia populations in soil and in root nodules of sympatrically grown Alnus taxa was evaluated by rep-polymerase chain reaction (PCR) and nifH gene sequence analyses. Rep-PCR analyses of uncultured Frankia populations in root nodules of 12 Alnus taxa (n=10 nodules each) growing sympatrically in the Morton Arboretum near Chicago revealed identical patterns for nodules from each Alnus taxon, including replicate trees of the same host taxon, and low diversity overall with only three profiles retrieved. One profile was retrieved from all nodules of nine taxa (Alnus incana subsp. incana, Alnus japonica, Alnus glutinosa, Alnus incana subsp. tenuifolia, Alnus incana subsp. rugosa, Alnus rhombifolia, Alnus mandshurica, Alnus maritima, and Alnus serrulata), the second was found in all nodules of two plant taxa (A. incana subsp. hirsuta and A. glutinosa var. pyramidalis), and the third was unique for all Frankia populations in nodules of A. incana subsp. rugosa var. americana. Comparative sequence analyses of nifH gene fragments in nodules representing these three profiles assigned these frankiae to different subgroups within the Alnus host infection group. None of these sequences, however, represented frankiae detectable in soil as determined by sequence analysis of 73 clones from a Frankia-specific nifH gene clone library. Additional analyses of nodule populations from selected alders growing on different soils demonstrated the presence of different Frankia populations in nodules for each soil, with populations showing identical sequences in nodules from the same soil, but differences between plant taxa. These results suggest that soil environmental conditions and host plant genotype both have a role in the selection of Frankia strains by a host plant for root nodule formation, and that this selection is not merely a function of the abundance of a Frankia strain in soil.

Micropropagation and production of Camptothecin from in vitro plants of Ophiorrhiza rugosa var. decumbens

Camptothecin and its derivatives are used as anticancer alkaloids. At present, they are obtained by the extraction from the Chinese tree Camptotheca acuminata and the indigenous tree Nothapodytes foetida. Identification of alternate species of plants like Ophiorrhiza species and development of tissue culture methods may be a suitable alternative for large-scale micropropagation as well as for the production of camptothecin. Plantlets were successfully regenerated from shoot cultures of Ophiorrhiza rugosa initiated from axillary meristems on medium containing benzyladenine (BA) (4.0 mg L−1) + α-naphthalene acetic acid (NAA) (0.05 mg L−1). Rooting was initiated in half concentration of Murashige and Skoog's (MS) basal medium devoid of growth hormones. Tissue culture derived plants of O. rugosa were similar to the normal plants in their morphological characteristics and chemical constitution. Ten O. rugosa plants that were obtained through micropropagation showed when analysed a higher alkaloid content compared to the normal plant. Chemical analysis of the different organs of the tissue culture plant of O. rugosa established in soil indicated 0.002% dry weight of camptothecin in the roots, 0.011% dry weight in the stems, 0.090% dry weight in the leaves and 0.015% in the floral parts.

Riverine wetland vegetation: importance of small‐scale and large‐scale environmental variation

Abstract. The purpose of this study was to evaluate the relative importance of small‐scale variation in abiotic factors and large‐scale spatio‐temporal variation on the distribution of wetland vegetation of a section of the Upper St. Lawrence River in Québec. Vegetation data have been classified with agglomerative clustering into 11 community types, from Acer rubrum, Acer saccharinum and Fraxinus swamps, to scrubs dominated by Salix petiolaris, Alnus rugosa var. americana or Myrica gale, to Typha, Typha/Lythrum, Carex lacustris and Calamagrostis canadensis marshes. Canonical Correspondence Analysis (CANOCO) suggested that peat thickness and water level are the most important abiotic variables correlated with plant community composition. As a whole, small‐scale variation accounts for 25.6 % of the species variation. Another 21.6% is explained by large‐scale variation based on location data summarizing large‐scale spatial distribution, and historical landscape dynamics differentiated into (a) no net loss of wetlands, (b) net loss of wetland, and (c) changes within wetlands, which are correlated with the actual variation in herbaceous and scrubby vegetation. The interaction between small‐scale and large‐scale variations explains another 1.7 %. In total, 48.9% of the species variation is explained by the two data sets, leaving 51.1% unexplained. Whereas the omission of some abiotic variables is possible, it is hypothesized that the abiotic conditions measured in this study play an important role, especially in the distribution of forested swamps. In addition, past history, particularly that of human interventions, becomes another important factor leading to the observed importance of large‐scale spatio‐temporal variables. This is particularly true for Alnus rugosa var. americana shrublands. Time lag between a relative stabilization of species distribution and the reduction of natural disturbances (water level fluctuations and fires) could be a possible cause of the importance of spatio‐temporal variables and the undetermined portion of species variation.

New Species and Varieties of Passiflora (Passifloraceae) from the Andes of South America

Three new species and two new varieties of Passiflora in the subgenera Tacsonia and Rathea are described. All are endemic to Andean habitats at altitudes above 2600 meters, with Passiflora huamachucoensis, P. runa, and P. trifoliata var. tarmensis known only from Peru. Passiflora rugosa var. venezolana is found in the Cordillera de Merida, Venezuela and northeastern Colombia while P. colombiana is confined to the southernmost Andes of Colombia.

Chromosome Numbers in Compositae, XV: Liabeae

Chromosome Numbers in Compositae, XV: Liabeae

Levels of Endogenous Indole-3-acetic Acid in Achenes of Rosa rugosa during Dormancy Release and Germination

Indole-3-acetic acid (IAA) in highly purified extracts of rose achenes (Rosa rugosa var rubra) was quantified by means of ion-pair reversephase high performance liquid chromatography with spectrofluorimetric detection. Changes in IAA content were determined during a 14-week 4 degrees C stratification, which leads to dormancy breakage, and during subsequent germination at 17 degrees C. IAA was also determined in achenes stratified in parallel at 17 degrees C, which does not induce release from dormancy. IAA decreased during the first 2 weeks of stratification both at 4 degrees C and at 17 degrees C. IAA remained low during the remaining 12 weeks of stratification at 4 degrees C, whereas it continued to decrease in achenes kept at 17 degrees C. An immediate increase in IAA during germination was followed by transients in the IAA level. The results suggest that IAA is without a regulating role in dormancy release although it seems to be involved in the germination process.

Levels of Endogenous Cytokinin-like Substances in Rosa rugosa Achenes During Dormancy Release and Early Germination

Summary Rosa rugosa var. rubra possess not only a coat-induced dormancy but are also embryo dormant and need a period of chilling to germinate. Achenes stored under moist, cold conditions, showed marked but transient peaks of free and bound cytokinin-like substances detected by the Amaranthus test. In achenes stored at 17°C no such changes were observed. Germinating achenes, previously stored at 4°C for 14 weeks, showed a transient peak of bound cytokinin-like substances and no changes in free forms. The early changes in cytokinin-like substances during cold stratification are considered to be related to the breakage of dormancy while the second increase in cytokinins seems to be more closely involved with germination. Most of the cytokinin-like activity detected in the Amaranthus test co-chromatographed with zeatin and ribosylzeatin on paper-chromatography. Scarification of achenes with 70% sulphuric acid did not improve germination or reduce the time needed for chilling. The germination was, however, increased two-fold in partly after-ripened achenes by removing leached inhibitors from the imbibition water. The role of the seed coat is discussed.

Notes on Acrobasis rubrifasciella Pack. (Lepidoptera: Phycitidae), with a List of its Parasites

In the Ottawa region about the middle of June, frequently certain leaves of the common alder, Alnus rugosa var. americana (Regel) Fern., are drawn together by strands of silk, concealing an elongate silken tube or case. The latter is occupied by an active brownish-grey larva that, when unmolested, extends itself from the broadly open end of the rube to feed on the foliage. This insect is the phycitid Acrobasis rubrifasciella Pack., a species that occors in northeastern United States and adjacent parts of Canada.