Abstract
Rauvolfia serpentina (Linn.) Benth. Ex Kurz belongs to the family Apocynaceae. The objective of the present study was to establish an effective protocol for the regeneration of leaf explants from R. serpentina and to study the α-amylase expression and phytochemical profiling by gas chromatography and mass spectrometry (GC-MS). Further antibacterial activity was studied using silver, gold, and copper nanoparticles. The leaf explants were cultured on Murashige and Skoog (MS) medium containing Benzyl amino purine (BAP) (2.0 mg/L-1) and Naphthalene acetic acid (NAA) (1.0 mg/L-1) induced the formation of callus and expressed α-amylase with underexposed to Cyclodextrin. Explants growing on MS medium fortified with 2,4-Dichlorophenoxy acetic acid (2,4-D) (1.0 mg/L-1), NAA (1.0 mg/L-1) with and without BAP (2.0 mg/L-1) and showed a maximum concentration of protein on the 75th day. Synthesis of a-amylase enzyme was expressed 40 days old culture and were confirmatic by western, further Silver, gold, and copper nanoparticles were synthesized using the ethyl acetate extract of callus tissue and subjected to thin-layer chromatography (TLC), which resolved 5 bands. These five bands were characterized by Fourier Transform Infrared (FTIR) Spectroscopy and screened for antimicrobial and antioxidant activities. TLC band 4 alone showed inhibitory activity against both Gram-negative and positive bacteria and potent antioxidant activity. For the first time, α-amylase was found in the callus extract by SDS-PAGE and confirmed by Western blot. The fourth band of TLC from the ethyl acetate extract as well as silver and gold nanoparticles synthesized using this extract revealed pronounced antimicrobial and antioxidant activities. GC-MS analysis revealed 26 compounds, which included mainly the phytosterols and fatty acid esters.
Highlights
Induction of callus The current study provided a protocol for large scale callus propagation of R. serpentina leaf explants
This is consistent with the report of Pandey et al (2010) who have reported higher frequency of callus formation when Murashige and Skoog (MS) media was fortified with Benzyl amino purine (BAP) and Naphthalene acetic acid (NAA)
Rashmi and Trivedi (2016) studied the effects of different combinations and concentrations of plant growth regulators on callus growth and shown maximum callusing response (75% in stem and 77% in leaf) when MS media was supplemented with a combination of BAP (0.5 mg/L-1) and NAA (1 mg/L-1)
Summary
Ex Kurz, popularly known as Sarpagandha, Indian snake root or devil pepper belongs to the family Apocynaceae. More than 100 species are included in the Rauwolfia genus, and they are native to tropical and subtropical regions of the world, including Europe, Africa, Asia, Australia and the Central and South America. R. serpentina is native to the moist, deciduous forests of Southeast Asia, including India, Burma, Bangladesh, Srilanka and Malaysia. R. serpentine is an ever green glaborous shrub which grows up to 60 m of height. Corresponding author: N Hariram Department of Biotechnology, Kalasingam Academy of Research and Education, Krishnankoil-626126, Tamil Nadu, India
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