The present investigation was carried out to develop an efficient regeneration protocol for Tecoma stans L., a valuable antidiabetic plant, using shoot tip explants. The effect of different concentrations (1.0–10.0 µM) of benzyl adenine (BA), kinetin (Kin) or isopentyl adenine (2-iP) either alone or in combination with different concentrations (0.1–2.0 µM) of α-naphthalene acetic acid (NAA) was evaluated for in vitro shoot bud induction and proliferation. Of the tested concentrations, MS medium containing 7.5 µM BA + 0.5 µM NAA proved to be optimal for the maximum (91%) regeneration with mean shoot number (9.8 ± 0.48) and length (4.36 ± 0.33 cm) after 8 weeks of incubation. The best in vitro rooting in regenerated microshoots was achieved on MS medium supplemented with 1.0 µM IBA, which produced a mean root number (6.8 ± 0.58) and length (4.24 ± 0.29 cm) after 4 weeks of incubation. The regenerated plantlets with well-developed roots and shoots were successfully acclimatized in thermocol cups filled with soilrite, for 8 weeks, in growth chamber. Thereafter transfered to field conditions where they grew well in garden soil with 80% survival rate. During the acclimatization period (0–56 days), a subsequent increase in the content of photosynthetic pigments (chlorophyll a, b and carotenoid) was observed up to 56th day. On assessing genetic fidelity among regenerated plants through RAPD analysis, no polymorphism in banding pattern was observed with that of donor plant, thus indicating clonal stability among micropropagated plants.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Anonymous (2003) The wealth of India. A Dictionary of Indian Raw Materials and Industrial Products Council of Scientific and Industrial Research, New Delhi, vol 10, pp 134–136
Andrade-Cetto A, Heinrich M (2005) Mexican plants with hypoglycaemic effect used in the treatment of diabetes. J Ethnopharmacol 99:325–348
Costantino L, Raimondi L, Parisino R, Brunetti T, Pessotto P, Giannessi F, Lins AP, Barlocco D, Antolini L, El-Abady SA (2003) Isolation and pharmacological activities of the Tecoma stans alkaloids. Il Farmaco 58:781–785
Marzouk M, Gamal-Eldeen A, Mohamed M, El-Sayed M (2006) Anti-proliferative and antioxidant constituents from Tecoma stans. Z Naturforsch 61(11–12):783–791
Robaszkiewicz A, Balcerczyk A, Bartosz G (2007) Antioxidative and prooxidative effects of quercetin on A549 cells. Cell Biol I 31:1245–1250
Kumar R, Sing G (2014) Investigations into the cause of sterility. Tecoma stans L. Bull Soc Bot France 135(2):131–135
Anis M, Ahmad N (2016) Plant tissue culture: propagation conservation and crop improvement. Springer, Singapore
Hussain SA, Ahmad N, Anis M (2018) Synergetic effect of TDZ and BA on minimizing the post-exposure effects on axillary shoot proliferation and assessment of genetic fidelity in Rauvolfia tetraphylla (L.). Rend Lincei Fis Nat 29:109–115
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Mac Kinney G (1941) Absorption of light by chlorophyll solution. J Biol Chem 140:315–322
Maclachlan S, Zalik S (1963) Plastid structure, chlorophyll concentration and free amino acid composition of chlorophyll mutant barely. Can J Bot 41:1053–1062
Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15
Fatima N, Anis M (2012) Role of growth regulators on in vitro regeneration and histological analysis in Indian ginseng (Withania somnifera L.) Dunal. Physiol Mol Biol Plants 18:59–67
Perveen S, Javed SB, Anis M, Aref IM (2013) Rapid in vitro multiplication and ex vitro establishment of Caribbean copper plant (Euphorbia cotinifolia L.): an important medicinal shrub. Acta Physiol Plant 35:3391–3400
Liu Z, Li Z (2001) Micropropagation of Camptotheca acuminate Decaisne from axillary buds, shoot tips, and seed embryos in a tissue culture system. Vitro Cell Dev Plant 37:84–88
Sharma S, Sharma R, Sharma P, Thakur K, Dutt B (2018) Direct shoot organogenesis from seedling derived shoot tip explants of endangered medicinal plant Saussurea costus (Falc.) Lipsch. Proc Natl Acad Sci India Sect B Biol Sci 1:1. https://doi.org/10.1007/s40011-018-0983-z
Lavanya M, Venkateshwarlu B, Devi BP (2009) Acclimatization of neem microshoots adaptable to semi-sterile conditions. Indian J Biotechnol 8:218–222
Kant T, Tomar Prajapati UK, Parmar AK (2010) In vitro propagation as a viable conservation strategy for Commiphora wightii, an endangered medicinally important desert tree. Conserv Evid 7:94–99
Borkowska B (2001) Morphological and physiological characteristics of micropropagated strawberry plants rooted in vitro or ex vitro. Sci Hortic 89:195–206
Jeon WW, Ali MB, Hahn EJ, Paek KY (2005) Effects of photon flux density on the morphology, photosynthesis and growth of a CAM orchid, doritaenopsis during post micropropagation acclimatization. Plant Growth Regul 45:139–147
Kenneth E, Pallet KE, Young AJ (2000) Carotenoids. In: Ruth GA, Hess JL (eds) Antioxidants in higher plants. CRC Press, Boca Raton, pp 60–81
Kadlecek P, Ticha I, Capkova V, Schafer C (1998) Acclimatization of micropropagated tobacco plantlets. In: Garab G (ed) Photosynthesis: mechanisms and effects, vol V. Kluwer, Dordrecht, pp 3853–3856
Pospisilova J, Ticha I, Kadlecek P, Haisel D, Plzakova S (1999) Acclimatization of micropropagated plants to ex vitro conditions. Biol Plant 42:481–497
Faisal M, Anis M (2010) Effect of light irradiations on photosynthetic machinery and oxidative enzymes during ex vitro acclimatization of Tylophora indica plantlets. J Plant Int 5:21–27
Ahmad N, Javed SB, Khan MI, Anis M (2013) Rapid plant regeneration and analysis of genetic fidelity in micropropagated plants of Vitex trifolia: an important medicinal plant. Acta Physiol Plant 35:2493–2500
Larkin PJ, Scowcroft W (1981) Somaclonal variation—a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60:197–214
Fatima N, Ahmad N, Anis M, Ahmad I (2013) An improved in vitro encapsulation protocol, biochemical analysis and genetic integrity using DNA based molecular markers in regenerated plants of Withania somnifera L. Ind Crop Prod 50:468–477
Saha S, Adhikari S, Dey T, Gosh P (2016) RAPD and ISSR based evaluation of genetic stability of micropropagated plantlets of Morus alba L. variety S-1. Meta Gene 7:7–13
The authors extend their appreciation to the international Scientific Partnership Program (ISPP# 0082), at King Saud University for funding this research. The award of UGC-BSR Faculty Fellowship (2017) to one of the authors by the University Grants Commission, New Delhi [F.18-1/2011(BSR)], is duly acknowledged. One of the authors is thankful to the University Grants Commission, New Delhi (MANF-2013-14-MUS-JAM-22128), for the award of SRF under MANF.
Conflict of interest
The authors declare that there is no conflict of interest to publish this manuscript.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
No plant tissue culture report is available for micropropagation of this important antidiabetic plant using shoot tip explants and thus, an attempt has been made to generate an efficient and reliable protocol. Through this protocol, T. stans plants can be reproduced on large scale without any genetic variation and can survive well during the ex vitro establishment. This regeneration method is simple, efficient and amenable for commercialization of this important medicinal shrub.
About this article
Cite this article
Hussain, S.A., Anis, M. & Alatar, A.A. Efficient In Vitro Regeneration System for Tecoma stans L., Using Shoot Tip and Assessment of Genetic Fidelity Among Regenerants. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 90, 171–178 (2020). https://doi.org/10.1007/s40011-019-01093-x
- Benzyl adenine