Evaluation of antidepressant activity of methanolic and hydroalcoholic extracts of Acorus calamus L.rhizome through tail suspension test and forced swimming test of mice

Objective:Acorus calamus(AC)L.(Araceae)is an annual semi-aquatic and aromatic plant found in Europe,North America and Asia.Its rhizomes are often used by Native Americans,Americans,and Chinese as well as by other cultures.Ethnobotanical studies and documents have shown their use in various disease treatments,such as insomnia,mental disorders,diabetes mellitus,epilepsy,inflammation,asthma,neuropathic pain,and diarrhea.In this study,the antidepressant activity of methanolic and hydroalcoholic extracts of the AC rhizome part in mice was investigated.Methods:Three doses of methanolic extract of AC rhizome(MEACR)(25,50 and 100 mg/kg b.wt),three doses of hydroalcoholic extract of AC rhizome(HAACR)(100,200 and 400 mg/kg b.wt),and standards(imipramine,15 mg/kg b.wt and fluoxetine,20 mg/kg b.wt)was daily oral administration to the mice for consecutive 14 days.The extract effect on the immobility time was monitored by a tail suspension test(TST)and a forced swimming test(FST).Monoamine oxidase(MAO)levels were also analyzed using standard methods.Results:The optimum antidepressant activity was viewed at 100 mg/kg b.wt of MEACR extract and400 mg/kg b.wt of HAACR extract with 23.82%and 20.59%immobility period reduction,respectively.Besides,the extracts weakened the FST-induced elevation of MAO activity significantly and returned to near-normal levels of neurotransmitters in the brain.100 mg/kg b.wt or above of MEACR extract significantly prevented the MAO-A and MAO-B activities in mice brain at a dose-dependent fashion.But,just 400 mg/kg b.wt of HAACR extract prevented the activity of MAO-A and MAO-B.Fluoxetine and imipramine showed a tendency to prevent the activity of MAO-A and MAO-B.Conclusion:This study suggests that AC rhizome extract mediated antidepressant activity by modulating the central neurochemical and hypothalamic-pituitary-adrenal(HPA)axis in response to FST and TSTinduced stress.Therefore,AC rhizome extract can be used as a valuable plant supplement to treat depressive disorders.

Protective effect of rhizome extracts of the herb, vacha (Acorus calamus) against oxidative damage: An in vivo and in vitro study

The rhizome of Acorus calamus,an herb widely used in Indian system of medicine for many ailments including epilepsy,mental illness and rheumatism,was subjected to soxhlet extraction to elucidate antioxidant property of different solvent extracts using in vitro assays.The benzene extract was most potent in scavenging hydroxyl and superoxide radicals and in reducing 1,1-diphenyl-2-picryl hydrazyl and ferric reducing antioxidant power.In addition the benzene extract prevented oxidative damage to DNA and mitochondria.It was also effective in preventing stress-induced decrease in total plasma anti-oxidant activity as determined in vivo using rat model wherein stress was induced by exposing to restraint and forced swimming.The minimum effective dose of the benzene extract was 5 mg/kg body weight(oral),and at this dose,its effect was similar to the same dose of a standard anti-oxidant,ascorbic acid.The study for the first time,clearly demonstrates a potent anti-oxidant activity of A.calamus combining in vitro and in vivo results.Hence,the therapeutic value of this herb maybe due to its anti-oxidant property.©2016 Beijing Academy of Food Sciences.Production and hosting by Elsevier B.V.All rights reserved.

Intraspecific genetic variability,differentiation and evolutionary relationships revealed through microsatellite loci in seven economically important Calamus species

Population density,species richness and critical population parameters are crucial in determining the levels of gene diversity in dioecious species of the genus Calamus.The extent of intraspecific and intrageneric genetic variability in Calamus from the southern Western Ghats of India was studied using 26 microsatellite markers by sampling 227 individuals belonging to seven economically important species.The heterozygosity of microsatellite loci ranged from zero to 0.78.Average gene diversity within species was 0.13;in all species it was 0.18 and amongst species was 0.06.The Shannon Information Index was the lowest for Calamus metzianus(0.11),whereas it ranged from 0.16 to 0.26 for other species.The expected heterozygosity varied from 0.08 to 0.18.Calamus hookerianus and Calamu travancoricus showed the highest genetic differentiation(44%)revealed through Fst values,whereas the lowest(22%)was observed between Calamus gamblei and Calamu thwaitesii.Population structuring and phylogenetic analysis differentiated the seven species.Due to overexploitation and loss of rare alleles,small populations could lead to fertilization between closely related individuals,resulting in inbreeding and increasing the risk of extinction.This could be important for species such as C.metzianus where allelic polymorphism was 23%,whereas for all other species it was 38%to 46%.Genetic diversity“micro-hotspots”were identified from the protected area network of the southern and central Western Ghats with highest observed heterozygosity.Four microhotspots from the Agasthyamalai Biosphere Reserve and the Pushpagiri Wildlife Sanctuary may be possible for long-term conservation programs.The findings of this study lay a strong foundation for strengthening protected area networks,especially areas with intermediate levels of disturbance.

Population genetic structure and diversity of high value vulnerable medicinal plant Acorus calamus in India using RAPD and chloroplast microsatellite markers

Acorus calamus is a highly valued medicinal plant with global distribution used in several drugs of health care systems. We evaluated the genetic diversity and population structure of 50 populations of A. calamus from different geographical regions in India through RAPD and chloroplast microsatellite markers. From the total screened 82 RAPD primers and 18 cpSSR primers, 10 RAPD and nine cpSSRs were found polymorphic. The selected 10 RAPD primers produced a total of 96 reproducible bands, out of which 65 were polymorphic （67.70%）. Whereas, the selected nine cpSSR markers produced 26 alleles and all of them were polymorphic. The mean genetic diversity （H T ） among populations using RAPD （H T = 0.263） and cpSSR （H T = 0.530） markers was higher in comparison to the mean genetic diversity within populations. Mean coefficient of gene differentiation （G ST ） between the populations was also high for both RAPD （G ST = 0.830） and cpSSR markers （G ST = 0.735）, whereas the estimated gene flow was very low for RAPD （Nm = 0.102） and for cpSSR （Nm = 0.179）. AMOVA analysis revealed that more genetic variation resided among the populations than within populations. Significant differences （p 0.001） were observed between the populations and individuals within the populations. Cluster analysis of RAPD and cpSSR data using UPGMA algorithm based on Nei’s genetic similarity matrix placed the 50 populations into two main clusters. The implication of the results of this study in devising strategy for conservation of A. calamus is discussed.

Shoot drying and its cause in Calamus simplicifolius at Nanmeiling, Hainan, China

Many stems are subject to shoot drying in the plantations of Calamus simplicifolius Wei at the Nanmeiling Base of the Forestry Bureau of Baisha County, Hainan Province. As a palm, stems of C simplicifolius stop growing and later die if the shoots are dry. Generally, shoot drying means reduced cane production and reduced profit for a rattan plantation. We aimed to quantify the incidence of shoot drying and determine its reason. We sampled eight 20 m × 20 m plots in the C simplicifolius plantations. In each plot, we counted stems and categorized them as drying shoot or normal shoot, and mother stem or sucker stem. We measured stem length and categorized them as drying shoot and normal shoot for mother stems. We cut open each shoot-drying stem to determine the cause of drying from drying part to fresh part. The frequency of mother stems （259 stem·ha^-1）, was significantly lower than sucker stems （588 stem·ha^-1）. The percentage of shoot drying was 33.8% for mother stems, not significantly higher than that for sucker stems （18,9%）. The mother shoot stems were significantly shorter if they were drying than if normal. 80% of stems with a drying shoot were hollow within the leaf sheath, whereas 10% hosted lively mature larvae （one larvae per shoot） and 10% hosted several lively young larvae. Mature larvae were identified as the Red Palm Weevil, Rhynchophorus ferrugineus Oliver. To confirm the present of the weevil, three pheromone traps for the weevil were set up in the plantations and adult weevils were trapped on the next day and identified as the Red Palm Weevil. We conclude that shoot drying of C. simplicifolius is probably caused by the Red Palm Weevil.

In Vitro Antioxidant and Free Radical Scavenging Activity of Curcuma longa, Acorus calamus and Camellia sinensis

The purpose of the current study was to determine the total phenolic and flavonoid content and total antioxidant activity of the Curcuma longa, Acorus calamus, and Camellia sinensis ethanolic extracts and their free radical scavenging activity. The study concluded that the Curcuma longa, Acorus calamus, and Camellia sinensis ethanolic extracts have a good source of phenolics, flavonoids, and antioxidant sources in turn which opens the high possibility of the extracts being used as food preservatives. The DPPH assay for scavenging free radicals showed that the IC50 value was above 123% of Curcuma longa ethanolic extract, 129.9% μg/ml of Acorus calamus ethanolic extract and 25% of Camellia sinensis ethanolic extracts shows very strong inhibition of the free radicals. Thus, comparing the DPPH assay for scavenging free radicals of Curcuma longa, Acorus calamus and Camellia sinensis ethanolic extracts with the positive control ascorbic acid, Curcuma longa and Camellia sinensis ethanolic extracts showed strong inhibition of the free radicals.

Influence of the Repeated Flooding on Growth and Development of Acorus calamus Vegetation in Water-level-fluctuation Zone of the Three Gorges Reservoir

[ Objective] The research aimed to study influence of the repeated flooding on growth and development of A. calamus in water-level-fluc- tuation zone of the Three Gorges Reservoir. [ Method] A. calamus plants were exposed to water under the dark conditions respectively in Septem- ber 2009 and September 2010. Then, they were taken away from the water, and grew in natural conditions in the following March and April respec- tively （ marked as S1, S2, S1 and S2）. The plant number was conducted statistics respectively. On May 7, the leaf number was made statistics. Length, width and rapid light responding curve of the leaf were determined. [ Result] Repeated flooding restrained significantly plant germination un- der the dark condition. Plant number in S1 and S2 groups decreased by 38.9% and 33.3% respectively compared with the control. It also reduced survival rate of the plant. Plant number in S2 group decreased by 16.7% （P 〈 0.05） compared with that of S2 group when they were taken away from the water. Both of flooding promoted elongation of the leaves, restrained leaves to grow wider, and also restrained leaf formation of the plants except for S1 and S1 groups. Total leaf length of a plant decreased markedly after the second flooding which in S1 and S2 groups were 50.6% and 36.9% respectively less than that after the first flooding. Total leaf number of the plant in S1and S2 groups reduced significantly, and they were only 63.1% and 38.5% respectively of the control. Compared with the first flooding, total leaf length of a plant in the control increased signifi- cantly and decreased remarkably in S2 group after the second flooding. Furthermore, light response ability of the relative electronic transfer rate （rETR） in S1 group wasn＇t significantly different from the control, and rETRmax, in S2 group was significantly less than the control. Moreover, non- photochemical quenching （NPQ） decreased remarkably in S1 and S2 groups. It indicated that A. calamus had good restore ability of the light re- sponse, but restoring of its heat dissipation capacity was slower. [ Conclusion] Repeated flooding inhibited plant growth and population recovery of A. calamus under the dark condition.

Isolation and characterization of insecticidal activity of （Z）- asarone from Acorus calamus L.

The insecticidal activity of Acorus calamus L. rhizome-derived material against adults of Sitophilus zeamais Motschulsky was examined by using repellency method and contact toxicity. The biologically active constituent of the A. calamus rhizome was separated and identified. The results showed that the ethanol extract of A. calamus had strong repellency and contact effect to S. zeamais and the active constituent of the A. calamus was characterized as （Z）-asarone by spectroscopic analysis. Responses from the tests varied with exposure times and doses. In the repellency test, ethanol extract of A. calamus had 93.92% repellency at 629.08 μg/cm^2 but only 71.38% at 157.27 μg/cm^2 12 h after treatment. As a contrast, （Z）-asarone showed 84.50% repellency at 314.54μg/cm^2 and 77.02% at 78.63 μg/cm^2 12 h after treatment. In the filter paper diffusion test, ethanol extract of A. calamus caused 95.56% and 17.78% mortality to S. zeamais at 314.54 μg/cm^2 and 78.63 μg/ cm^2 4 days after treatment, while （Z）-asarone brought about 100.00% and 15.56% mortality at 40.89 μg/cm^2 and 15.73 μg/cm^2 respectively. These results indicate that the insecticidal activity of the A. calamus extract may be due to （Z）-asarone.

A new anti-diabetic sesquiterpenoid from Acorus calamus

A new sesquiterpenoid, 1β,5α-guaiane-4β,10α-diol-6-one （1）, was isolated from 70% EtOH extract of the rhizomes of Acorus calamus. The structure was determined on spectroscopic methods, especially 2D NMR techniques. The absolute configuration of 1 was confirmed by TDDFT quantum chemical calculation of its ECD spectrum. Compound 1 showed promising anti-diabetic activity on a insulin-mediated glucose consumption model of HepG2 cells.

Characterization of natural fiber from manau rattan(Calamus manan)as a potential reinforcement for polymer-based composites

Researches on novel natural fibers in polymer-based composites will help promote the invention of novel reinforcement and expand their possible applications.Herein,in this study,novel cellulosic fibers were extracted from the stem of manau rattan(Calamus manan)by mechanical separation.The chemical,thermal,mechanical and morphological properties of manau rattan fibers were comprehensively analyzed and studied by Fourier transform infrared spectroscopy(FT-IR),X-ray photoelectron spectroscopy(XPS),X-ray diffraction(XRD)analysis,thermogravimetric analysis(TGA),single fiber tensile test and scanning electron microscopy(SEM).Component analysis re-sults showed that the cellulose,hemicellulose and lignin contents of C.manan fibers were 42wt%,20wt%,and 27wt%,respectively.The surface of the rattan fiber was hydrophilic according to the oxygen/carbon ratio of 0.49.The C.manan has a crystalline index of 48.28%,inducing a max-imum degradation temperature of 332.8°C.This reveals that it can be used as a reinforcement for thermoplastic composites whose operating temperature is below 300°C.The average ten-sile strength can reach(273.28±52.88)MPa,which is beneficial to improve the mechanical properties of rattan fiber reinforced composites.The SEM images displayed the rough surface of the fiber,which helped to enhance the interfacial adhesion between the fibers and matrices in composites.These results indicate the great potential of C.manan fibers as the reinforcement in polymer-based composites.

Identification of Random Amplified Polymorphic DNA Markers Linked to Sex Determination in Calamus simplicifolius C. F. Wci

The random amplified polymorphic DNA （RAPD） molecular marker technique was used to determine the sex of Calamus simplicifolius C. F. Wei In the present study, DNA samples were extracted individually from 10 male and 10 female plants. After a total of 1 040 decamer primers had been tested, an approximate 500-bp male-specific DNA fragment was generated with the S 1443 primer. It is feasible to identify sex at the early stages of plant life, which is beneficial for improving breeding programs of this dioecious species. In addition, we have obtained a proper RAPD protocol that is useful for other species of rattan.

Study of the Climbing Behavior of the Flagella of Calamus Simplicifolius Based on Micro-CT and Nanoindentation

Rattan is a typical tropical climbing plant that uses flagella to climb supports to grow.A comprehensive understanding of the anatomic structure and micromechanics of rattan flagella might inspire more research on biomimetic climbing materials.Here,the structure and micromechanical properties of flagella in calamus simplicifolius were examined by Micro-Computed Tomography(Micro-CT)and nanoindentation techniques,respectively.The results showed that the rachis of the flagella mainly comprised vascular bundles surrounded by basic tissues,which had a gradient density decreasing from outsides to insides.The prickles are derived from the epidermis or the epidermis and cortical tissue of the flagellum,which do not possess vascular tissue.The entire tip of the prickle was composed almost of fibrous cells.The indentation modulus of elasticity of the prickle was 17.03 GPa,which was 17.93%higher in comparison with the rachis.The hardness of the prickle was 539.27 MPa and was slightly higher than that of the rachis.The results indicated that the discrepancy of micromechanical strengths in different parts of flagella reflects on their unique roles in the process of climbing.

Phytochemical screening, antioxidant and cytotoxic activity of fruit extracts of Calamus tenuis Roxb

Objective:To investigate the antioxidant and cytotoxic activity of the fruits of Calamus tenuis Roxb.Methods:The preliminary phytochemical group tests were done,which revealed the presence of alkaloid,tannin,flavonoid and steroid.The dried fruit was extracted in soxhlet apparatus using petroleum ether,ethyl acetate and methanol.Antioxidant potential of each extract was evaluated using total phenol content,total flavonoid content,cupric reducing antioxidant capacity,1,1-diphenyl-2-picrylhydrazyl radical scavenging activity,and total antioxidant capacity determinations.Results:The extracts were found to possess moderate to high amounts of phenolic and flavonoid contents.In cupric reducing antioxidant capacity assay the extracts showed moderate reducing power which increases with concentration.Scavenging of 1,1-diphenyl-2-picrylhydrazyl radical was found to rise with concentration with lowest IC_(50)value for methanol extract,which was confirmed by total antioxidant activity test that shows highest(95 mg/g of extract)in ascorbic acid equivalent for methanol extract.In Brine shrimp lethality bioassay the methanol and petroleum ether extracts were found to be toxic to Brine shrimp nauplii,with LC_(50)of 25.53μg/mL and 28.07μg/mL respectively while the LC_(50)of the reference vincristine sulphate was 1.32μg/mL.Ethyl acetate extract was found to be moderately cytotoxic showing LC_(50)of 47.79μg/mL.Conclusions:The results of the present study suggest that the fruits of Calamus tenuis Roxb possess antioxidant and cytotoxic potential.Moreover,phytochemical screening reveals the presence of alkaloid,tannin,flavonoid and steroid,which may be responsible for the observed bioactivities.

菖蒲对不同程度富营养化水体的适应性研究

对5种不同程度富营养化水体中的菖蒲(Acorus calamusL.)在静态培养过程中的叶片长度变化、叶片数目变化进行了观测,比较了培养后各水体中菖蒲根长、根数、鲜重、根活力、叶绿素含量、叶片可溶性总糖含量、鲜重增量和根系活力的差异。研究发现,菖蒲在5种不同程度富营养化水体中均能正常生长。在非极度富营养化的水体组中,水体富营养化程度越高,菖蒲生长越好。但在极度富营养水体中,菖蒲根系生长受到抑制,其根短且须根多,且叶片生长对此水体有一段适应期,试验前8 d,其中菖蒲叶片长度增长明显小于重度富营养,8 d后,叶片生长速度高于其他处理组。结果表明,菖蒲具有较好的耐污性、耐贫瘠性,较适合于在中、高度富营养水体中生长,在水体生态修复工程中具有良好的应用前景。

水菖蒲药学研究概况

在广泛文献检索基础上,对水菖蒲的成分、药理、临床应用、代表性膳食及应用注意事项的研究进展进行综述,为进一步药物开发利用提供资料。

Assigning conservation value and identifying hotspots of endemic rattan diversity in the Western Ghats, India

Rattans, or canes, are one of the most important non-timber forest products supporting the livelihood of many forest-dwelling communities in South and North-eastern India. Due to increased demand for rattan products, rattans have been extracted indiscriminately from the Western Ghats, a 1600-km mountain chain running parallel to the west coast of India. Extensive harvesting, loss of habitat and poor regeneration has resulted in dwindling rattan populations, necessitating an urgent attempt to conserve existing rattan resources. In this study, using niche-modelling tools, an attempt has been made to identify areas of high species richness of rattans in the Western Ghats, one of the mega-diversity regions of the world. We have also developed conservation values for 21 economically important and endemic rattans of the Western Ghats. We identified at least two to three sites of extremely high species richness outside the existing protected area network that should be prioritized for in situ conservation.This study emphasizes the need to develop strategies for the long-term conservation of rattans in the Western Ghats, India.

Contact Activity of Combination Use of Three Kinds of Plant Extracts on Helicoverpa armigera

[Objectives] The aim was to study the contact activity of combination use of garlic,onion and stone calamus extracts on Helicoverpa armigera and their co-toxicity coefficient( CTC) and mixture ratio. [Methods] Leaf dipping method was used to study the contact activity of the three kinds of plant extracts against H. armigera. Based on this,the co-toxicity coefficient of the compound of the three kinds of extracts was determined. Orthogonal test was used to explore the optimal mixture ratio of the three kinds of extract for combination use. [Results]The extracts of garlic,onion and stone calamus had good inhibition and contract activity on H. armigera larvae. The LC50 values of the three kinds of plant extracts against H. armigera were 175. 0,217. 0 and 79. 2 mg/L,respectively. When the mixture ratios of stone calamus to garlic,and stone calamus to onion were both 2:1,the extract showed a synergistic effect( CTC > 120); when the mixture ratios of stone calamus to garlic,and stone calamus to onion were 1:1 and 1:2,respectively,the extracts showed an additive effect( 80 < CTC < 120); and then garlic extract and onion extract were mixed according to the ratio of 1:1,the additive effect was slight( CTC < 90). Among the three kinds of extracts,the extract of stone calamus showed the strongest contact activity,followed by that of garlic. The results of co-toxicity coefficient determination and orthogonal test showed that the contact activity of combination use of the three kinds of extracts was stronger than that of the single extract overall. When the extracts of garlic,onion and stone calamus were mixed according to the ratio of 2:3:1( mass concentration ratio),the contact activity of the compound against H. armigera reached 95. 3%. [Conclusions]The study results provide a scientific basis and theoretical reference for the development of new plant-derived pesticides based on the pharmacodynamics of garlic,onion and stone calamus.

菖蒲:上得案头,入得香囊

菖蒲是江南地区比较常见的一种水生湿地植物,属天南星科植物,为多年生草本。常见有两个种：一种生活在湿地淤泥中,通常称之为菖蒲,Acorus calamus;一种多野生于山涧溪流和水石缝隙处,称为石菖蒲Acorus tatarinowii。前者有着高大剑形叶片,后者叶片矮小密集成丛,二者都有匍匐蜿蜒的根茎。植物界中中文名字带“菖蒲”有不少,唐菖蒲和黄菖蒲。唐葛蒲是重要的鲜切花植物,属于“世界四大切花”之一。

施肥、解淀粉芽孢杆菌对高地省藤苗生长的影响

采用正交设计试验,研究施肥、解淀粉芽孢杆菌对高地省藤(Calamus nambariensis Becc.var.alpinus S.J.Pei et)苗期生长的影响。结果表明:(1)解淀粉芽孢杆菌(Y2菌株)和施肥(尿素、过磷酸钙、复合肥)对高地省藤苗生长具有显著影响;(2)综合各因素水平对高地省藤苗地径和苗高生长指标的影响以及节省生产成本的原则,确定的最佳组合为A4B1C2D1,即8×10~6 CFU/MLY2菌株,不施尿素(0g)、施用过磷酸钙20g,不施复合肥(0g)时,高地省藤苗生长最好。