Organogenesis and efficient in vitro plantlet regeneration from nodal segments of Allamanda cathartica L. using TDZ and ultrasound assisted extraction of quercetin
The present investigation was carried out to evaluate the instigative effect of thidiazuron (TDZ) on multiple shoot induction from nodal segments of Allamanda cathartica and estimated the flavonoid yield among the regenerants. High rate of shoot bud induction was achieved on Murashige and Skoog (MS) medium augmented with 0.3 µM TDZ from nodal segments exposed for 30 days. However, for shoot proliferation and elongation, TDZ exposed cultures were further cultured on MS medium devoid of TDZ and/or supplemented with different concentration of 6-benzyladenine (BA) and Kinetin (Kn). BA at 2.5 µM gave the maximum mean number of shoots (44.00 ± 1.30) and shoot length (7.50 ± 0.21 cm) per explant after 12 weeks of incubation in the secondary medium. The response of explant was influenced by the collection time. The highest rooting in the microshoots (5 cm) was achieved on 1/2 MS liquid medium supplemented with 0.5 µM Indole-3 butyric acid (IBA) which produced 4.50 ± 0.16 mean roots/shoot with 4.05 ± 0.17 cm mean root length. The leaves of 30 day old acclimatized plantlets were used for phytochemical screening. Ultrasonication mediated extraction and quantification of bioactive flavonoid namely quercetin through colorimetry and mass spectrometry analysis from the leaves of regenerants. Extraction was processed in methanol using 2 g leaf sample through sonication. Total yield of flavonoids and quercetin content was found to be maximum in 2.5 µM BA treated plants with respect to control and other treated samples. The concentration of total flavonoids was estimated to be 172.90 mg QE/g which yielded 51.39 mg/g quercetin. The study ensures a rapid cultivation of plantlets, thus enhancing the biomass production which may be utilized in the isolation and quantification of other biological potential compound for the use in treatment of various ailments.
KeywordsMicropropagation Cytokinins Colorimetry Flavonoid Ultrasonication Mass spectrometry
Award of UGC-BSR Faculty fellowship (2017) by the University Grants Commission, New Delhi to MA is gratefully acknowledged.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Deb CR, Deb MS, Jamir NS (2014) Effect of different factors on in vitro axillary shoot proliferation and plant regeneration of Cinnamomum tamala Nees: a spice yielding plant. Indian J Biotechnol 13:520–526Google Scholar
- Essiett AU, Udo ES (2015) Comparative phytochemical screening and nutritional potentials of the stems, leaves and flowers of Allamanda cathartica (Apocynaceae). Int J Sci Technol 4:248–253Google Scholar
- Fiola JA, Hassan MA, Swartz HJ, Bors RH, McNicols R (1990) Effect of thidiazuron, light fluence rates and kanamycin on in vitro shoot organogenesis from excised Rubus cotyledons and leaves. Plant Cell Tiss Org Cult 20:223–228Google Scholar
- Kostic DA, Dimitrijevic DS, Mitic SS, Mitic MN, Stojanovic GS, Zivanovic AV (2013) Phenolic content and antioxidant activities of fruit extracts of Morus nigra L (Moraceae) from Southeast Serbia. Trop J Pharmac Res 12:105–110Google Scholar
- Sambandam B, Thiyagarajan D, Ayyaswamy A, Raman P (2016) Extraction and isolation of flavonoid quercetin from the leaves of trigonella foenum-graecum and their anti-oxidant activity. Int J Pharm Sci 8(6):120–124Google Scholar
- Savithramma N, Linga Rao M, Suhrulatha D (2013) Qualitative and quantification analysis of phytochemicals from leaf aqueous extract of Allamanda cathartica L. and Terminalia paniculata Roth An Int J 1(8):821–825Google Scholar
- Wong KF, Taha RM, 2012. The effect of 2,4-dichlorophenoxyacetic Acid and 6-benzylaminopurine on callus induction and plant regeneration of Allamanda cathartica—a valuable medicinal plant. Res J Biotech 7(3):75–80Google Scholar