Phytogenic silver nanoparticles from tissue-cultured Kaempferia angustifolia - an underutilized medicinal herb: a comparative antibacterial study on urinary pathogens.

BACKGROUND: Ethnomedicinally important Kaempferia angustifolia is a rhizomatous aromatic herb belonging to the family Zingiberaceae. The present manuscript deals with the green synthesis of silver nanoparticles through a rapid reduction process mediated by the rhizome extract of tissue culture-raised plants. The present study was conducted to evaluate the antimicrobial activity of the bio-nanoparticles, and the plant extracts themselves against seven multidrug-resistant urinary tract infecting (MDR-UTI) pathogens. RESULT: The ethanolic extracts of the rhizomes of the plant executed a very rapid synthesis of silver bio-nanoparticles, and the generation of the nanoparticles was confirmed through UV-vis spectrophotometry, dynamic light scattering (DLS), and electron dispersion spectroscopic (EDS) analysis. Finally, the precise shapes and dimensions of these nanoparticles were confirmed under the transmission electron microscope (TEM). The shapes of the nanoparticles obtained were diverse in nature and varied from rod, triangular, spherical, to oval shaped, with the size, ranging from 10-60 nm. Silver nanoparticles exhibited a maximum zone of inhibition (ZI) of 16.93 ± 0.04 mm against isolate no. 42332. The ex vitro and in vivo extracts exhibited ZI 14.03 ± 0.04 mm and 11.56 ± 0.04 mm, respectively, against the same strain, which are comparatively lower than the nanoparticles but unignorable. CONCLUSION: Although the pathogens used in the present study are resistant to at least three or more types of pharmacologically important antibiotics, nanoparticles, as well as the plant extracts, exhibited significant inhibition to all the seven MDR-UTI pathogens, which confirms that they are highly antimicrobic. Hence, this underutilized medicinal plant extracts of K. angustifolia and the bio-nanoparticles synthesized from these can be explored in pharmaceutical industries to treat multidrug-resistant human pathogenic bacteria. Furthermore, their broad-spectrum activity leads to the opportunity for the synthesis of future generation drugs.

Silver nanoparticle synthesis and their potency against multidrug-resistant bacteria: a green approach from tissue-cultured Coleus forskohlii.

Drug resistance is a major concern nowadays, and finding alternatives of the well-known antibiotic is necessary. Green nanoparticles are emerging as a tenable alternative to this with a large spectrum of activity. The present manuscript describes an eco-friendly approach for green synthesis of silver nanoparticles from both in vitro and in vivo leaf extract of Coleus forskohlii. Leaf extracts were used in synthesis of nanoparticles which were further analyzed through UV-Vis, dynamic light scattering, energy-dispersive spectroscopy, and transmission electron microscopy. Antimicrobial activity of silver nanoparticles alone, as well as crude extract of the plant itself, was carried out against eight multidrug-resistant respiratory tract infecting pathogenic strains. Satisfactory antimicrobial activities were found with nanoparticles, in vitro and in vivo leaf extracts. However, gradually higher to lower inhibition potential against pathogenic bacterial strains was found in silver nanoparticles, in vitro and in vivo leaf extracts. Seven bioactive compounds were detected in the crude extract through gas chromatography-mass spectroscopy analysis. Results revealed that nanoparticle formation occurred in a wide range of sizes (10-50 nm) and shapes (trigonal, hexagonal, spherical, rod). The diversity in size and shape of the nanoparticles makes them biologically active. Silver nanoparticle exhibits significantly better antimicrobial activities as compared to the plant extract in case of nearly all pathogens with a maximum zone of inhibition of 15.33 ± 0.94 mm where more than 12 well-known antibiotics failed to respond. Because of this broad-spectrum activity of nanoparticles as well as the leaf extracts against life-threatening microbes, it can be used as future generation drugs.

Callus mediated shoot organogenesis and regeneration of cytologically stable plants of Ledebouria revoluta: An ethnomedicinal plant with promising antimicrobial potency.

Ledebouria revoluta are important ethnomedicinal plant found in India and South Africa. Micropropagation via indirect shoot organogenesis had been established from three types of explant (i.e. scale leaf, leaf lamina and root) of L. revoluta. Scale leaf was found superior as compared to leaf lamina and root explant with respect to their organogenic callus induction potentiality. Murashige and Skoog (1962) [MS] media supplemented with 3.0 mg L-1 2,4-dichlorophenoxyacetic acid, 0.75 mg L-1 ß-naphthoxyacetic acid were best effective for inducing organogenic callus. Maximum 17.0 ±â€¯0.52 bulblets were induced from about 500 mg of callus within 42-46 days sub-culturing on a medium containing 0.75 mg L-1 kinetin. The bulblets were matured (86.7% success) after one month culture on the same medium composition. The best result of in vitro root induction with 100% response and 8.4 ±â€¯0.31 roots per bulb was achieved after 18 days of implantation on MS medium containing 2.0 mg L-1 indole-3-butyric acid. Plantlets were acclimatized with a 96.0% survival rate. Chromosomal studies revealed cytological stability of callus cells and all regenerants containing 2n = 30 chromosomes, same as parental plants. Antimicrobial activity of L. revoluta was tested against two Gram-positive bacteria, three Gram-negative bacteria and two fungi. The methanol and ethanol extract proved more effective against bacteria, whereas acetone and chloroform extract shows potential anti-fungal activities. Present protocol can be applied reliably to produce uniform planting materials in large scale. In addition, this efficient indirect regeneration pathway via callus culture opens a way for improvement through genetic transformation.

Somatic embryogenesis and synthetic seed production--a biotechnological approach for true-to-type propagation and in vitro conservation of an ornamental bulbaceous plant Drimiopsis kirkii Baker.

An efficient plant regeneration protocol through indirect somatic embryogenesis pathway via callus had been developed from the leaf explant of an ornamental bulbaceous plant Drimiopsis kirkii. Optimum friable calli were induced on Murashige and Skoog (MS) basal medium supplemented with 3.0 mg/l of 2,4-dichlorophenoxyacetic acid and 1.0 mg/l of α-naphthalene acetic acid (NAA). On subculturing the callus on MS medium supplemented with 2.5 mg/l of thidiazuron (TDZ), 73.3 % of the cultures responded with 20.4 ± 0.3 somatic embryos (SEs) per 500 mg callus at different stages of development after 6 weeks of culture. The highest response of 86.7 % with 28.3 ± 0.5 embryos per 500 mg callus was observed on MS medium supplemented with 2.5 mg/l TDZ and 1.0 mg/l NAA. SEs were encapsulated in calcium alginate beads for the production of synthetic seeds (SSs) and their storability was investigated. The highest SS germination (93.3 %) was observed in 1.0 % sodium alginate followed by 86.7 % germination with 2.5 % sodium alginate. The SSs were stored at three different temperatures (4, 15, and 24 ºC) up to 6 months. The SSs kept at 15 °C showed 64.4 % germinability even after 4 months of storage. Both nonencapsulated and encapsulated SE-derived plants were successfully transferred to soil with 93.3 and 88.3 % survival rate accordingly. Randomly amplified polymorphic DNA (RAPD) analysis revealed that there were no somaclonal variations among the plants produced via somatic embryogenesis and they are true-to-type to their parental plant. These results confirmed the most reliable methods, which can be further used for genetic transformation studies as well as for mass propagation of ornamental D. kirkii at a commercial level.

High frequency microcloning of Aloe vera and their true-to-type conformity by molecular cytogenetic assessment of two years old field growing regenerated plants.

BACKGROUND: Aloe vera (L.) Burm.f is an important industrial crop, which has enormous application in pharmaceutical, cosmetic and food industries. Thereby, the demand for quality planting material of A. vera is increasing worldwide. Micropropagation is the widely accepted practical application of plant biotechnology that has gained the status of a multibillion-dollar industry throughout the world and this techniques can be used to meet the industrial demand of A. vera. Present studies aim to develop a proficient methods of high-frequency true-to-type plantlet regeneration without intermediate callus phase for A. vera. RESULTS: Nodal portion of rhizomatous stem of A. vera were cultured on Murashige and Skoog (MS) medium (Physiol. Plant. 15:473 - 497, 1962) supplemented with various cytokinin and A. vera leaf gel (AvG) as organic supplement. Number of proliferated shoots per explant was increased along with the regeneration cycles and on MS medium supplemented with 2.5 mg/L 6-benzylaminopurine and 10.0% (v/v) AvG, only 17.8 ± 0.35 shoots per explant were induced on 1st regeneration cycle whereas on 3rd regeneration cycle these number increase to 38.5 ± 0.44 shoots per explant on the same medium composition. AvG have an encouraging role to increase the proliferation rate and on 3rd regeneration cycle 27.6 ± 0.53 shoot per explant induced on 2.5 mg/L BAP, but these number increase to 38.5 ± 0.44 shoots per explant when 10.0% (v/v) AvG was added along with 2.5 mg/L BAP. After transfer of individual excised shoots to a one-third strength MS medium containing 20.0% (v/v) AvG, all the shoots formed whole plantlets with maximum number (9.6 ± 0.29) of roots per shoot. 95.0% of the regenerated plantlets survived on poly-green house. Normal flower appeared in 84.2% field growing micropropagated plants after 18 to 20 months of field transfer. Further, clonal fidelity of the two years old micropropagated plants was established by studying mitotic and meiotic chromosomal behavior and also considered the chromosome number and structural organization. There were no alterations in chromosome phenotypes, somatic haploid (pollen mitosis) and diploid chromosome count (n = 7; 2n = 14), or meiotic behavior. Randomly amplified polymorphic DNA analyses revealed there were no somaclonal variations among these regenerants. CONCLUSIONS: These results confirm the very reliable method for large scale production of true-to-type plantlets of A. vera, which can be used for commercial purpose.