Evaluation of the pathogenicity of endophytic fungi isolated from spines of rattan (Calamus castaneus) against other plant hosts.

AIMS: In this study, we sought to determine the pathogenic ability of endophytic fungi recovered from the spines of Calamus castaneus, a common rattan palm growing in the forests of Peninsula Malaysia. METHODS AND RESULTS: Ten endophytic fungal isolates were selected for pathogenicity tests: Colletotrichum boninense, Colletotrichum fructicola, Colletotrichum cliviae, Diaporthe hongkongensis, Diaporthe arengae, Diaporthe cf. nobilis, Neopestalotiopsis saprophytica, Neopestalotiopsis formicarum, Fusarium solani and Fusarium oxysporum. These endophytes were tested against leaves of bertam (Eugeissona sp.), oil palm (Elaeis guineensis) and mango (Mangiferae indica) and the fruits of chilli (Capsicum annum), tomato (Solanum lycopersicum) and banana (Musa acuminata). The fungal isolates showed infectivity against bertam, oil palm and mango leaves with degrees of virulence ranging from low to moderate, whereas infectivity against chilli, tomato and banana ranged from low to very high. CONCLUSIONS: Fungal endophytes isolated from the spines of C. castaneus are pathogenic to different crop plants with differing degrees of virulence or aggressiveness. SIGNIFICANCE AND IMPACT OF THE STUDY: Spines of C. castaneus can harbour fungal pathogens of a number of different crops as endophytes. The ability of the fungal endophytes to colonize and infect different crops demonstrate their importance towards agricultural crops. There is a possibility the endophytes behave as latent pathogen. When conditions become favourable, the fungal endophytes transform to pathogenic form and potentially infect other plants.

Euryhaliotrema dolichodrilus n. sp. and Euryhaliotrema mimulus n. sp. (Monogenoidea: Dactylogyridae) infecting the gill lamellae of porgies, Calamus spp. (Perciformes: Sparidae) from the Gulf of Mexico off Florida, U.S.A.

Two new species of Euryhaliotrema Kritsky & Boeger, 2002 (Monogenoidea: Dactylogyridae) were described from the gill lamellae of porgies, Calamus spp. (Perciformes, Sparidae), collected from the Gulf of Mexico off Florida: Euryhaliotrema dolichodrilus n. sp. from littlehead porgy Calamus proridens Jordan & Gilbert (type host), knobbed porgy Calamus nodosus Randall and Caldwell, grass porgy Calamus arctifrons Good & Bean, sheepshead porgy Calamus penna (Valenciennes), and jolthead porgy Calamus bajonado (Bloch & Schneider); and Euryhaliotrema mimulus n. sp. from jolthead porgy. The two new species along with Euryhaliotrema amydrum Kritsky & Bakenhaster, 2011, Euryhaliotrema carbuncularium Kritsky & Bakenhaster, 2011, Euryhaliotrema carbunculus (Hargis, 1955) Kritsky & Boeger, 2002, Euryhaliotrema luisae Cruces, Chero & Luque, 2018, Euryhaliotrema magnopharyngis Cruces, Chero & Luque, 2018, and Euryhaliotrema spirulum Kritsky & Bakenhaster, 2011 appeared to form a subgroup of species that developed secondarily within Euryhaliotrema.

Mammalian Arginase Inhibitory Activity of Methanolic Extracts and Isolated Compounds from Cyperus Species.

Polyphenolic enriched extracts from two species of Cyperus, Cyperus glomeratus and Cyperus thunbergii, possess mammalian arginase inhibitory capacities, with the percentage inhibition ranging from 80% to 95% at 100 µg/mL and 40% to 64% at 10 µg/mL. Phytochemical investigation of these species led to the isolation and identification of two new natural stilbene oligomers named thunbergin A-B (1-2), together with three other stilbenes, trans-resveratrol (3), trans-scirpusin A (4), trans-cyperusphenol A (6), and two flavonoids, aureusidin (5) and luteolin (7), which were isolated for the first time from C.thunbergii and C. glomeratus. Structures were established on the basis of the spectroscopic data from MS and NMR experiments. The arginase inhibitory activity of compounds 1-7 was evaluated through an in vitro arginase inhibitory assay using purified liver bovine arginase. As a result, five compounds (1, 4-7) showed significant inhibition of arginase, with IC50 values between 17.6 and 60.6 µM, in the range of those of the natural arginase inhibitor piceatannol (12.6 µM). In addition, methanolic extract from Cyperus thunbergii exhibited an endothelium and NO-dependent vasorelaxant effect on thoracic aortic rings from rats and improved endothelial dysfunction in an adjuvant-induced arthritis rat model.

Influence of Post Space Preparation on the Apical Leakage of Calamus, Single-Cone and Cold Lateral Condensation Obturation Techniques: A Computerized Fluid Filtration Study.

PURPOSE: Apical microleakage between the root canal and periapical tissues is considered a common cause of endodontic failure. The aim of this study was to evaluate the apical microleakage of various obturation techniques after immediate post space preparations with different drills. MATERIALS AND METHODS: One hundred twenty human single-rooted teeth were selected for this study. The coronal portions were removed to achieve a uniform length of 15 mm. All root canals were instrumented with ProTaper Next rotary files using crown-down technique. The 120 roots were randomly assigned to 3 experimental groups according to obturation technique (n = 40). The root canals were obturated with lateral condensation in group 1, single-cone technique in group 2, and Calamus 3D obturation system in group 3. Each group was divided into two subgroups (n = 20) according to drill types (Gates Glidden and Peeso drills) leaving 5 mm of apical gutta-percha. The apical microleakage was measured using the computerized fluid filtration meter method. The results were evaluated using Kruskal-Wallis test with Bonferroni Corrected Mann-Whitney U multiple comparison post hoc tests. RESULTS: There was no statistically significant difference between obturation techniques after post space preparation with Peeso drills (p > 0.05). In Gates Glidden drill groups, single-cone technique demonstrated higher leakage (0.0051 ± 0.0037 µl x min-1 × cmH2 O-1 ) than Calamus techniques (0.0019 ± 0.0012 µl x min-1 × cmH2 O-1 ) (p < 0.017). CONCLUSIONS: Gates Glidden drills caused the highest apical leakage in teeth obturated with single-cone technique.

Screening of leaves extracts from Calamus aromaticus for their antimicrobial activity by disc diffusion assay.

The current research investigates the anti-microbial activities of different solvent extracted samples from the leaves of Calamus aromaticus against Gram positive, Gram negative bacteria and fungi using 500, 1000 and 2000 µg disc-1 concentrations. Escherichia coli, Citorbacter freundii and Candida albicans showed resistivity to crude methanolic extract and the same microbes were more susceptible to water extracted fractions. Maximum activity was measured by hexane extracted fractions against Pseudomonas aeruginosa, Bacillus subtilis and Xanthomonas campestris and minimum growth inhibition by water extracted fractions. Maximum growth of Klebsiella pneumoniae and Staphylococcus aureus was measured by ethyl acetate fraction. Majority of the tested microbes were resistant to water and butanol extracted fractions. Staphylococcus aureus revealed maximum susceptibility among gram positive bacteria and Bacillus subtilis showed minimum. Among Gram negative bacteria, Citorbacter freundii was more susceptibile while Xanthomonas campestris revealed resistively.

Cytotoxic Steroidal Glycosides from the Whole Plant of Calamus acanthophyllus.

A new steroidal glycoside, callaphylloside (1), together with seven known glycosides (2-8), was isolated from the whole plant of Calamus acanthophyllus. The structure of the new compound was elucidated by spectral data analyses and chemical transformations. Compounds 5 and 8 exhibited strong cytotoxic activity against four cancer cell lines (0.7 ≤ IC50 ≤ 3.4 µM). Evaluation of the structure-activity relationship among steroidal glycosides revealed that the structure of spirostanol with an α-L-rhamnopyranosyl linked to C-2 of the inner glucopyranosyl residue both play a critical role in the effects of these compounds on the cancer cell lines.

ZYGOTIC EMBRYO CRYOPRESERVATION OF Calamus vattayila RENUKA, AN ENDANGERED RATTAN PALM OF INDIA, AS INFLUENCED BY SEED MATURITY.

BACKGROUND: Calamus vattayila Renuka is an endemic and endangered rattan palm of the Western Ghats, India where the development of a protocol for cryopreservation is important for their ex situ conservation in gene banks. OBJECTIVE: The objective of the study is to devise an efficient protocol for cryopreservation of the species, comparing the relative efficiency of seeds and zygotic embryos as storage material. MATERIALS AND METHODS: Mature seeds extracted from ripened fruits were subjected to cryopreservation through a simple dehydration method and zygotic embryos isolated from seeds of three different maturity stages were cryopreserved through an encapsulation-dehydration method. RESULTS: The mature seeds did not tolerate desiccation and freezing but the isolated zygotic embryos tolerated both desiccation and freezing. Embryos isolated from immature, partially mature and mature seeds harvested respectively after 160-170, 210-220 and 250-260 days after flowering possessed 82 - 86% moisture content (m.c.) and tolerated desiccation down to 9% m.c. with 80% viability. Embryos from immature seeds gave a maximum 63+/-26% regeneration after LN exposure, which is nearly equal to the corresponding desiccated control (73%). LN tolerance declined with maturity of seeds with a maximum of 49% of embryos from partially mature seeds and 25% from mature seeds subjected to LN exposure showing post-freeze recovery and regeneration. CONCLUSION: Although immature seeds collected during February-March were recalcitrant (desiccation sensitivity), their embryos tolerated cryopreservation through encapsulation-dehydration better than embryos isolated from more mature seeds.

Identification of floral genes for sex determination in Calamus palustris Griff. by using suppression subtractive hybridization.

Calamus palustris Griff. is an economically important dioecious rattan species in Southeast Asia. However, dioecy and onset of flowering at 3-4 years old render uncertainties in desired female:male seedling ratios to establish a productive seed orchard for this rattan species. We constructed a subtractive library for male floral tissue to understand the genetic mechanism for gender determination in C. palustris. The subtractive library produced 1536 clones with 1419 clones of high quality. Reverse Northern screening showed 313 clones with differential expression, and sequence analyses clustered them into 205 unigenes, including 32 contigs and 173 singletons. The subtractive library was further validated with reverse transcription-quantitative polymerase chain reaction analysis. Homology identification classified the unigenes into 12 putative functional proteins with 83% unigenes showing significant match to proteins in databases. Functional annotations of these unigenes revealed genes involved in male flower development, including MADS-box genes, pollen-related genes, phytohormones for flower development, and male flower organ development. Our results showed that the male floral genes may play a vital role in sex determination in C. palustris. The identified genes can be exploited to understand the molecular basis of sex determination in C. palustris.

Removal of fluoride and arsenic by pilot vertical-flow constructed wetlands using soil and coal cinder as substrate.

This study evaluated the performance of soil and coal cinder used as substrate in vertical-flow constructed wetlands for removal of fluoride and arsenic. Two duplicate pilot-scale artificial wetlands were set up, planted respectively with cannas, calamus and no plant as blank, fed with a synthetic sewage solution. Laboratory (batch) incubation experiments were also carried out separately to ascertain the fluoride and arsenic adsorption capacity of the two materials (i.e. soil and coal cinder). The results showed that both soil and coal cinder had quite high fluoride and arsenic adsorption capacity. The wetlands were operated for two months. The concentrations of fluoride and arsenic in the effluent of the blank wetlands were obviously higher than in the other wetlands planted with cannas and calamus. Fluoride and arsenic accumulation in the wetlands body at the end of the operation period was in range of 14.07-37.24% and 32.43-90.04%, respectively, as compared with the unused media.

[Chemical constituents of Acori Calami Rhizoma from Hunan Province].

OBJECTIVE: To study the chemical components from the ethanol extract of Acori Calami Rhizoma from Hunan Province. METHODS: Components were isolated and purified through various chromatographic methods and recrystallization, and identified by spectroscopic data. RESULTS: Ten compounds were isolated and identified as follows: heptadecanoic acid(1), monopentadecanoin(2), syringic acid(3), aurantiamide acetate(4), monononadecanoin(5),tatarine A(6),tatanan C(7),cerevisterol(8),2 ,6-dioxopiperidin-3-yl acetate(9) and palmatine(10). CONCLUSION: Compounds 1-5 and 8-10 are isolated from Acorus genus for the first time, and compounds 1-5 and 7-10 are isolated from this plant for the first time.

Chloroplast genome of Calamus tetradactylus revealed rattan phylogeny.

BACKGROUND: Calamus tetradactylus, a species primarily distributed in Vietnam, Laos, and southern China, is highly valued for its utilization as a small-diameter rattan material. While its physical and mechanical properties have been extensively studied, the genomic characteristics of C. tetradactylus remain largely unexplored. RESULTS: To gain a better understanding of its chloroplast genomic features and evolutionary relationships, we conducted sequencing and assembly of the chloroplast genome of C. tetradactylus. The complete chloroplast genome exhibited the typical highly conserved quartile structure, with specific variable regions identified in the single-copy region (like psbF-psbE, π = 0.10327, ndhF-rpl32, π = 0.10195), as well as genes such as trnT-GGU (π = 0.05764) and ycf1 (π = 0.03345) and others. We propose that these regions and genes hold potential as markers for species identification. Furthermore, phylogenetic analysis revealed that C. tetradactylus formed a distinct clade within the phylogenetic tree, alongside other Calamus species, and C. tetradactylus was most closely related to C. walkeri, providing support for the monophyly of the genus. CONCLUSION: The analysis of the chloroplast genome conducted in this study provides valuable insights that can contribute to the improvement of rattan breeding programs and facilitate sustainable development in the future.

New flavan trimer from Daemonorops draco as osteoclastogenesis inhibitor.

Dragon's blood is a red resin obtained from different plants and is considered highly efficacious and used in medicine owing its wound healing function. Two new compounds (7 and 8) were isolated from the dragon's blood of Daemonorops draco fruits, along with eight known compounds (1-6, 9, and 10). Their structures, including their absolute configurations, were elucidated by nuclear magnetic resonance (NMR), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electronic circular dichroism (ECD) analysis. According to the spectroscopic data, 8 was determined to be a quinone methide derivative of flavan and 7 was deduced to be a flavan trimer. All compounds were evaluated for their anti-osteoclastogenesis activity, compound 1 and 7 exhibited anti-osteoclastogenesis activity with IC50 values of 31.3 and 36.8 µM, respectively.

Phytotoxicity and Phytotoxic Substances in Calamus tenuis Roxb.

Calamus tenuis is a shrub species distributed across South Asia. It grows well in diversified habitats and tends to dominate plants in the surrounding environment. The phytotoxicity of C. tenuis and the action of its phytochemicals against other plant species could explain its dominant behavior. Compounds with phytotoxic activity are in high demand as prospective sources of ecofriendly bioherbicides. Therefore, we investigated the phytotoxicity of C. tenuis. Aqueous methanol extracts of this plant species significantly limited the growth of four test plant species, two monocots (barnyard grass and timothy), and two dicots (alfalfa and cress), in a dose- and species-dependent manner. Bio-directed chromatographic isolation of the C. tenuis extracts yielded two major active substances: a novel compound, calamulactone {(S)-methyl 8-(5-oxo-2,5-dihydrofuran-2-yl) octanoate}, and 3-oxo-α-ionone. Both of the identified compounds exerted strong growth inhibitory effects on cress and timothy seedlings. The concentrations of 3-oxo-α-ionone and calamulactone required to limit the growth of the cress seedlings by 50% (I50) were 281.6-199.5 and 141.1-105.5 µM, respectively, indicating that the effect of calamulactone was stronger with lower I50 values. Similarly, the seedlings of timothy also showed a considerably higher sensitivity to calamulactone (I50: 40.5-84.4 µM) than to 3-oxo-α-ionone (I50: 107.8-144.7 µM). The findings indicated that the leaves of C. tenuis have marked growth-inhibitory potential, and could affect surrounding plants to exert dominance over the surrounding plant community. Moreover, the two identified phytotoxic substances might play a key role in the phytotoxicity of C. tenuis, and could be a template for bioherbicide development. This paper was the first to report calamulactone and its phytotoxicity.

Potent hepatoprotective activity of common rattan (Calamus rotang L.) leaf extract and its molecular mechanism.

BACKGROUND: Calamus rotang L. (CR) is an Indian shrub. The leaves and other organs of the plant are traditionally used in India for treatment of various diseases. The in vitro antioxidant property of the leaves extract was previously established. Thus, the current study aimed to evaluate the antioxidant and hepatoprotective effects of CR ethyl acetate extract at a dose of 350 mg/kg on CCl4 induced hepatotoxic rats through different mechanisms. METHODS: Histopathological examination of the treated rats' group in comparison with positive and negative controls were performed. Quantitative measuring of the proinflammatory cytokines (TNF α), inflammatory regulators (Arginase, PPAR α) and the antiapoptotic protein Bcl-2 in comparison with positive and negative control groups was achieved using immunohistochemical examination. HPLC profiling of the polyphenol contents and molecular docking of the identified compounds against BH3 proapoptotic protein were correspondingly studied to evaluate the potential antiapoptotic property. RESULTS: The CR extract greatly protects the liver tissue through the suppression of TNF α, arginase and PPAR α induced by CCl4 as well as its enhancement of the antiapoptotic Bcl-2 protein. Fourteen polyphenols of different classes were identified in CR extract and tested via molecular docking for their potential antiapoptotic activities against BH3 protein. Naringin, rutin, 7-hydroxy flavone, and ellagic acid compounds exhibit the highest affinity and potential inhibition of pro-apoptotic protein BH3 via molecular docking study. CONCLUSIONS: The ethyl acetate fraction of the leaves of C. rotang is rich in polyphenols that exhibited potent hepatoprotective effect on CCl4 induced hepatotoxic rats through its antioxidant, anti-inflammatory, anti-steatosis and antiapoptotic properties.

Advanced phosphorus removal for secondary effluent using a natural treatment system.

Mechanisms for low concentrations phosphorus removal in secondary effluent were studied, and a process was developed using limestone filters (LF), submerged macrophyte oxidation ponds (SMOPs) and a subsurface vertical flow wetland (SVFW). Pilot scale experimental models were applied in series to investigate the advanced purification of total phosphorus (TP) in secondary effluent at the Chengjiang sewage treatment plant. With a total hydraulic residence time (HRT) of 82.52 h, the average effluent TP dropped to 0.17 mg L(-1), meeting the standard for Class III surface waters. The major functions of the LF were adsorption and forced precipitation, with a particulate phosphorus (PP) removal of 82.93% and a total dissolved phosphorus (TDP) removal of 41.07%. Oxygen-releasing submerged macrophytes in the SMOPs resulted in maximum dissolved oxygen (DO) and pH values of 11.55 mg L(-1) and 8.10, respectively. This regime provided suitable conditions for chemical precipitation of TDP, which was reduced by a further 39.29%. In the SVFW, TDP was further reduced, and the TP removal in the final effluent reached 85.08%.

[Prediction of the lengths of fibers and vessels of rattans using near infrared spectroscopy].

The morphological characteristics of fibers and vessels of six rattan species in Southern China were investigated to study the feasibility of predicting the lengths of fibers and vessels of rattan species with application of analytical technologies of near infrared spectroscopy (NIR). The results showed that the average lengths of fibers and vessels of six rattan species were from 1 229 to 1 917 mm and from 1 035 to 2 129 mm, respectively. The models of length of fiber and vessel were constructed by combining partial least square (PLS) and full cross-validation, and a good correlation between the length of fibers and the spectrum transformed by the first derivative was found within the spectral range of 350 - 2 454 nm, and the correlation coefficient (r(c) and r(p)) and standard error (SEC and SEP ) of calibration model and prediction model are 0.98, 0.85 and 70, 178 respectively, while a good correlation between the length of vessels and the spectrum transformed by the first derivative was found within the spectral range of 350-2 500 nm, the correlation coefficient (r(c) and r(p)) and standard error (SEC and SEP) of calibration and prediction model is 0.97, 0.80 and 101, 261 respectively. Their model parameters showed that NIR spectroscopic technique can rapidly and accurately predict the lengths of fibers and vessels of the six rattan species.

Quiquelignan A-H, eight new lignoids from the rattan palm Calamus quiquesetinervius and their antiradical, anti-inflammatory and antiplatelet aggregation activities.

Eight new lignin derivatives, termed quiquelignan A-H (1-8), comprising three tricin-type flavonolignans (1-3) and five 8-O-4' neolignans (4-8), were isolated from the ethanol extract of Calamus quiquesetinervius stems. Structural elucidation of the new isolates was accomplished on the basis of spectroscopic data. Compounds 1-8 showed strong-to-moderate antioxidant activity against the hydroxy radical (()OH). Among them, compound 5 showed significantly higher hydroxy radical scavenging activity (IC(50) 4.4microg/mL). Compounds 2-4 and 6-8 dose-dependently suppressed the LPS-stimulated production of nitric oxide (NO) in RAW 264.7 cells. The anti-inflammatory potency of 4 and 6 was 2.7-4.5-fold higher compared with quercetin. Compounds 2-4, 6 and 8 also exhibited mild collagen-antagonistic activity, but were inactive with respect to thrombin-induced platelet aggregation.

Antioxidant and anti-inflammatory phenylpropanoid derivatives from Calamus quiquesetinervius.

Eight new phenylpropanoid derivatives [quiquesetinerviusides A (1), B (2), C (3), D (4), and E (5), as well as quiquesetinerviusins A (6), B (7), and C (8)] and seven known compounds (8-15), were isolated from an EtOAc extract of Calamus quiquesetinervius stems. The structures of 1-8 were elucidated on the basis of 1D- and 2D-NMR spectroscopic data analysis. Bioassay results showed that 1-5 possess weak DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging activity, but potent (·)OH radical scavenging activity (IC(50) 3.6-8.4 µM). Of the tested isolates, compounds 4-6 and 9 showed potent inhibition (IC(50) 9.2-29.5 µM) of LPS-stimulated NO production when compared with a positive control substance, quercetin (IC(50) 34.5 µM).

Biomorphous porous hydroxyapatite-ceramics from rattan (Calamus Rotang).

The three-dimensional, highly oriented pore channel anatomy of native rattan (Calamus rotang) was used as a template to fabricate biomorphous hydroxyapatite (Ca(5)(PO(4))(3)OH) ceramics designed for bone regeneration scaffolds. A low viscous hydroxyapatite-sol was prepared from triethyl phosphite and calcium nitrate tetrahydrate and repeatedly vacuum infiltrated into the native template. The template was subsequently pyrolysed at 800 degrees C to form a biocarbon replica of the native tissue. Heat treatment at 1,300 degrees C in air atmosphere caused oxidation of the carbon skeleton and sintering of the hydroxyapatite. SEM analysis confirmed detailed replication of rattan anatomy. Porosity of the samples measured by mercury porosimetry showed a multimodal pore size distribution in the range of 300 nm to 300 microm. Phase composition was determined by XRD and FT-IR revealing hydroxyapatite as the dominant phase with minimum fractions of CaO and Ca(3)(PO(4))(2). The biomorphous scaffolds with a total porosity of 70-80% obtained a compressive strength of 3-5 MPa in axial direction and 1-2 MPa in radial direction of the pore channel orientation. Bending strength was determined in a coaxial double ring test resulting in a maximum bending strength of approximately 2 MPa.

Rattan spines as deterrence? A spinescence study on different species of rattans.

Rattan spines are most often regarded as an identification trait and perhaps as a physical protection structure. In this study, we study the spinescence traits from five different species rattan: Daemonorops lewisiana, Daemonorops geniculata, Calamus castaneus, Plectomia griffithii, and Korthalsia scortechinii. We tested length, width, angle, strength, spine density, cross-section surface, spine color, and leaf trichomes (only for D. lewisiana, C. castaneus and D. geniculata). We also tested whether the spines were capable of deterring small climbing mammals (for Plectomia griffithii and Calamus castaneus) by using a choice selection experiment. Due to a variety of spine traits, we could not categorize whether any species is more or less spinescent than the others. We suggest that spines have a much more significant role than merely as a physical defense and work together with other rattan characteristics. This is also evidenced by our choice selection experiment, in which the spines on a single stem donot deter small climbing mammals. However, this is a work in progress, and we have outlined several alternative methods to be used in future work.