Anti-bacterial studies on Hemigraphis colorata(Blume) H.G.Hallier and Elephantopus scaber L.

Objective:To examine the ethanol,aqueous,chloroform,benzene,acetone and petroleum ether extracts of Hemigraphis colorata(H.colorata) leaves and stem and Elephantopus scaber (E.scaber) leaves,root and flower for the presence of phyto-constituents and screened the antibacterial activity against the selected pathogens.Methods:The fresh materials were shade dried and powdered using the tissue blender.The dried and powered materials(50 g) were extracted successively with 200 mL of aqueous,acetone,benzene,chloroform,etganol.and petroleum ether by using Soxhlet extractor for 8 h at a temperature not exceeding the boiling point of the solvent.Aqueous,acetone,benzene,chloroform,ethanol.and petroleum ether extracts were prepared from powdered materials were used for preliminary phytnehemical and antimicrobial studies using standard methods.Results:The crude aqueous,acetone,benzene,chloroform, ethauol.and petroleum ether extracts E.scaber leaves,flower and root and H.colorata leaves and stem demonstrated that out of(5×6×12 = 360) tests for the presence or absence of the above compounds.188 tests gave positive results and the remaining 172 gave negative results. The results of the phvtochemical screening revealed that phenol(12/12).carbohydrates(9/12). steroids(8/12).saponins and coumarins(7/12).tannins(6/12),proteins(5/12).earboxylic acid and flavonoids(4/12).xanthoproteins(3/12) and alkaloids(2/12) presence in the crude aqueous, acetone,benzene,chloroform,ethanol.and petroleum ether extracts of H.colorata leaves and stem.The crude aqueous,acetone,benzene,chloroform,ethanol.and petroleum ether extracts E.scaber leaves,flower and root displayed the presence of phenol(18/18).tannin(17/18). carbohydrates(16/18).steroids(14/18),oarboxylic acid and coumarins(12/18).saponins(10/18), xanthoprotein(9/18).flavonoids(7/18).protein(4/18) and alkaloids(2/18).The root ethanolic extracts of E.scaber illustrated the highest zone of inhibition against three pathogens viz.. Staphylococcus aureus(S.aureus)(24 mm).Escherichia coli(E.coli)(16 mm) and Psemlomonas aeruginosa {P.aeruginosa)(13 mm).The chlorofrom extracts of E.scaber showed the highest zone of inhibition against Bacillus cereus(B.ceretus)(12 mm).The leaves ethanolic extracts of E.scaber demonstrated the highest zone of inhibition against three pathogens viz.,Enterococcus faecalis (E.faecalis)(18 mm).Proteus mirabilis(P.mirabilis)(17 mm).Salmonella Typhi(S.typhi)(14 mm) and Enterobacter sp.(11 mm) While the benzene extracts of H.colorata demonstrated maximum zone of inhibition against the pathogen Acinetobucter sp.(14 mm) and S.aureus (12 mm).Conclusions:It is hoped that this study would direct to the establishment of some compounds that could be used to invent new and more potent antimicrobial drugs of natural origin.

In vitro clonal propagation of Achyranthes aspera L. and Achyranthes bidentata Blume using nodal explants

Objective:To develop the reproducible in vitro propagation protocols for the medicinally important plants viz.,Achyranthes aspera(A.aspera)L.and Achyranthes bidentata(A.bidentata)Blume using nodal segments as explants.Methods:Young shoots of A.aspera and A.bidentata were harvested and washed with running tap water and treated with 0.1%bavistin and rinsed twice with distilled water.Then the explants were surface sterilized with 0.1%(w/v)HgCl_2 solutions for I min.After rinsing with sterile distilled water for 3-4 times,nodal segments were cut into smaller segments(1 cm)and used as the explants.The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog(MS)medium supplemented with 3%sucrose,0.6%(w/v)agar(HiMedia,Mumbai)and different concentration and combination of 6-benzyl amino purine(BAP),kinetin(Kin),naphthalene acetic acid(NAA)and indole acetic acid(IAA)for direct regeneration.Results:Adventitious proliferation was obtained from A.aspera and A.bidentata nodal segments inoculated on MS basal medium with 3%sucrose and augmented with BAP and Kin with varied frequency.MS medium augmented with 3.0 mg/L of BAP showed the highest percentage(93.60±0.71)of shootlets formation for A.aspera and(94.70±0.53)percentages for A.bidentata.Maximum number of shoots/explants(10.60±0.36)for A.aspera and(9.50±0.56)for A.bidentata was observed in MS medium fortified with 5.0 mg/L of BAP.For A.aspera,maximum mean length(5.50±0.34)of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A.bidentata(5.40±0.61)was observed in the very same concentration.The highest percentage,maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of 1BA.Seventy percentages of plants were successfully established in polycups.Sixty eight percentages of plants were well established in the green house condition.Sixty five percentages of plants were established in the field.Conclusions:The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A.aspera and A.bidentata.The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can he easily adopted for commercial large scale cultivation.

Studies on isozymic variation among the South Indian species of Sphaerostephanos

Objective:To explore the identity and phylogenetic relationships among the three medicinally important species of Sphaerostephanos from South India using isozymic profile.Methods:The young fronds were homogenized with 3.5 mL of ice-cold homogenizing buffer in a pre-chilled pestle and mortar.The supernatant was subjected to electrophoresis as described by Anbalagan poly acrylamide gel electrophoresis.Staining solutions for isoperoxidase was prepared as per Smila method for the detection of isoenzymes.Results:A total of six different bands in five different positions with different molecular weight/Rf values and four active zones have been observed in the isoperoxidase enzyme system of Sphaerostephanos.Only one band with MW/Rf 0.399 is common to two different species i.e.Sphaerostephanos arbuscula(S.arbuscula) and Sphaerostephanos unitus(S.unitus).Among the remaining four bands,two bands(R_f.0.23,0.47)are present in Sphaerostephanos subtruncatus(S.subtruncatus) and one distinct band has been observed individually in S.arbuscula(R_f.0.507) and S.unitus(R_f.0.56).Conclusions:The present preliminary molecular study through isozymic analysis shows the identity of all the three species and the present results confirm distinctness of these three species based on macromicromorphology,phytochemistry and cytology.