EFFICIENT in vitro PROPAGATION OF Amaranthus viridis L. USING NODE EXPLANTS
Hyperhydricity is a frequently problem in plants during in vitro culture and affected micropropagation of
plants. To develop an efficient in vitro regenerated system without hyperdydricity, we demonstrated the effect
of different disinfected agents (mercuric chlorite and hypochlorite), growth regulators, their concentrations
and combinations, Agar, pH, ammonium nitrate (NH4NO3) and number of subcultures. Mercuric chlorite at
0.07% and exposing time (9–10 min) was appropriate for hygienic culture. The shoots induced by Benzyladnine
(BA) alone or in combination with α-Naphthaleneacetic acid (NAA) exhibited maximum multiplication
with symptoms of hyperhydricity than those induced by Kinetin alone or in combination with NAA. Hyperhydricity
was also reduced by increasing the concentration of agar, pH and elimination of NH4NO3 from the
macroelements of Murashig and Skoog (MS) medium. Repeated subcultures affected both multiplication and
hyperhydricity. The multiplication of shoots increased from parental culture up to 5th subculture and thereafter
declined in 6th subculture. Although shoot hyperhydricity were observed from 1st subculture (19%) and
then increased up to 85% in 6th subculture. This increased in hyperhydricity could be due to the remaining
influence of hormones. In shoots of 5th subculture the content of chlorophyll (dark green) were higher than
shoots of 6th subculture.
agar; Amarinthus viridis; HgCl2; hyperhydricity; pH; subcultures
Bennici, A., Schiff, S., Bovelli, R. (1992). In vitro culture of species and verities of four Amaranthus L. species. Euphytica, 62, 181–186. DOI: 10.1007/BF00041752
Casanova, E., Moysset, L., Trillas, M.I. (2008). Effect of agar concentration and vessel closure on the organogenesis and hyperhydricity of adventitious carnation shoots. Biol. Plant., 52, 1–8. DOI: 10.1007/s10535-008-0001-z
Cheepala, S.B., Sharma, N.C., Sahi, S.V. (2004). Rapid in vitro regeneration of Sesbania drummondii. Plant Biol., 48, 13–18. DOI: 10.1023/B:BIOP.0000024269.72171.42
Faisal, M., Naseem, A., Mohammad, A., Abdulrahman, A.A., Ahmad, A.Q. (2018). Auxin-cytokinin synergism in vitro for producing genetically stable plants of Ruta graveolens using shoot tip meristems. Saudi J. Biol. Sci., 25(2), 273–277. DOI: 10.1016/j.sjbs.2017.09.009
Hussain, T.M., Chandrasekhar, T., Gopal, G.R. (2007). High frequency shoot regeneration of Sterculia urens Roxb. an endangered tree species through cotyledonary node cultures. Afr. J. Biotech., 6, 1643–1649.
Ivanova, M., Novak, O., Strnad, M., Van-Stadan, J. (2006). Endogenous cytokinins in shoots of Aloe polyphylla cultured in vitro in relation to hyperhydricity, exogenous cytokinins and gelling agents. Plant Growth Regul., 50, 219–230. DOI: 10.1007/s10725-006-9139-x
Ivanova, M., Johannes, V.S. (2011). Influence of gelling agent and cytokinins on the control of hyperhydricity in Aloe polyphylla. Plant Cell Tiss. Org. Cult., 104(1), 13–21. DOI: 10.1007/s11240-010-9794-5
Ivanova, M., Van-Staden, J. (2008). Effect of ammonium ions and cytokinins on hyperhydricity and multiplication rate of in vitro regenerated shoots of Aloe polyphylla. Plant Cell Tiss. Org. Cult., 92, 227–231. DOI: 10.1007/s11240-007-9311-7
Johnson, M., Berhanu, A., Mulugeta, K., Eyayu, M., Manickam, V.S. (2005). Regeneration from callus cultures of Rhinacanthus nasutus L. Kurtz. Ethnomed. J. Sci. Tech., 3(1), 17–24.
Kataeva, N.V., Irena, G.A., Raisa, G.B., Elena, V.D.K.A. (1991). Effect of applied and internal hormones on vitrification and apical necrosis of different plants cultured in vitro. Plant Cell. Tiss. Org. Cult., 27(2), 149–154.
Kevers, C., Franck, T., Strasser, R.J., Dommes, J., Gaspar, T. (2004). Hyperhydricity of micropropagated shoots: a typically stress-induced change of physiological state. Plant Cell Tiss. Org. Cult., 77, 181–191. DOI: 10.1023/B:TICU.0000016825.18930.e4
Liu, M., Fangling, J., Xiangyu, K., Jie, T., Zexiu, W., Zhen, W. (2017). Effects of multiple factors on hyperhydricity of Allium sativum L. Sci. Hortic., 217, 285–296. DOI: 10.1016/j.scienta.2017.02.010
Masanori, K., Yoshiji, N. (2003). Effects of cytokinin types and their concentrations on shoot proliferation and hyperhydricity in in vitro pear cultivar shoots. Plant Cell Tiss. Org. Cult., 72, 261–265.
Martin, G., Geetha, S.P., Raja, S.S., Raghu, A.V., Balachandran, I., Ravindran, P.N. (2006). An efficient micropropagation system for Celastrus paniculatus Wild.: A vulnerable medicinal plant. J. Forest Res., 11, 461–465. DOI:10.1007/s10310-006-0237-4
Mikula, A., Rybczynski, J.J. (2006). Cryopreservation – a tool for long-term storage of cells, tissues and organs from in vitro culture derived. Biotechnologia, 4, 145–163.
Murashige, T., Skoog, F. (1962), A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant., 15(3), 473–497. DOI.org/10.1111/j.1399-3054.1962.tb08052.x
Naik, P.M., Patil, B.R., Jaggal L.G., Jangid, V.K. (2013). The Effect of Subculture on the Bacoside A Content in Adventitious Shoot Cultures of Bacopa monnieri (L). Res. J. Pharma. Biol. Chem. Sci., 4(4), 1111–1116.
Piatczak, E., Wysokinska, H. (2003). In vitro regeneration of Centaurium erythraea Rafn from shoot tips and other seedling explants. Acta Soc. Bot. Pol., 72(4), 283–288.
Remphrey, W.R., Palmer, C.E., Blouw, M.J. (1993). In vitro branching in relation to repeated subculture in two cultivars of Potentilla fruticosa. Plant Cell Tiss. Org. Cult., 32, 235–240. DOI: 10.1007/BF00029848
Saxena, R., Venkaiah, K., Anitha, P., Venu, L., Raghunath, M. (2007). Antioxidant activity of commonly used plant foods of India: Contribution of their phenolic content. Int. J. Food. Sci. Nutr., 58, 250–260. DOI: 10.1080/09637480601121953
Tisserat, B., Galleta, P.D. (1988). In vitro flowering in Amaranthus. HortScience, 23(1), 210–212.
Vijayalakshmi, G., Giri, C. (2003). Plant regeneration via organogenesis from shoot base derived callus of Arachis stenosperma, Arachis villosa. Curr. Sci., 85, 1624–1629.
Wesely, E.G., Johnson, M., Kavitha, M.S., Selvan, N. (2011). Micropropagation of Alternanthera sessilis (L.) using shoot tip and nodal segments. Iran. J. Biot., 9(3), 206–212.
Yaacob, J.S., Hwei, L.C., Taha, R.M., Mat Nor, N.A., Aziz, N. (2012). Pigment Analysis and Tissue Culture of Amaranthus cruentus L. Acta Hortic., 20, 171–178. DOI: 10.17660/ActaHortic.2012.958.20
Articles are made available under the CC BY-NC-ND 4.0 (recognition by authorship, non-commercial use, no dependent works).
The author signs a statement on the originality of the work and the contribution of individuals.
Submission of the paper implies that it has not been published previously, that it is not under consideration for publication elsewhere.