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Salinity is one of the major environmental threats to plants because a large number of plants are intolerant to NaCl stress. One such plant, Justicia vasica, which is the subject of our study, is said to have great medicinal properties. However, the natural occurrence and cultivation of this species has been constrained due to various factors including its poor seed germination rate under its normal conditions of habitat, and a low availability of seeds. This study was undertaken to examine the efficiency of callus formation and regenerative tendencies of Justicia vasica under saline conditions. The leaf, shoot tips and inter nodal cuttings were employed for this study using MS media formulated with several concentrations of different kinds of hormonal combinations. This study also tested the effects of different concentrations of NaCl on callus induction and growth. Each experiment was conducted thrice with 12 replicates. A superior level of callus formation was achieved by treating shoot tips with 2.0 mg/l α-naphthalene acetic acid (NAA) and 0.5 mg/l 6-benzyladenine purine (BAP) in combination. From this study, it was concluded that there was no inhibitory action from the medium supplemented with 0.05 mg/l NaCl. The optimal hormonal combination also exhibited highest rate of callus induction and growth with 0.05 mg/l NaCl added. There was no callus necrosis at this saline concentration after 60 days of inoculation. Furthermore, some somatic embryogenic callus displayed the organogenesis of leaf like structures after 75 days of inoculation. The development of salinity tolerance of plants by genetic application has been a vital area on modern agriculture as well as medicinal plant research because more land has been salinized through poor local irrigation practices. The present investigation assumed that the NaCl tolerance of this plant could be effectively obtained in vitro.
Keywords: callus necrosis; explant browning; NaCl stress; salinity tolerance; somatic embryo; tissue culture
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 Chakrabarty, A. and Brantner, A.H., 2001. Study of alkaloids from Adhatoda vasica Nees on their anti-inflammatory activity. Phytotherapy Research, 15, 532-534.
 Shinwari, Z.K., Khan, I., Naz, S. and Hussain, A., 2009. Assessment of antibacterial activity of three plants used in Pakistan to cure respiratory diseases. African Journal of Biotechnology, 8(24), 7082-7086.
 Bimal, R. and Shahnawaz, M.D., 2012. Plant regeneration from nodal explants of Adhatoda vasica Nees. Journal of Medicinal Plants Research, 6(7), 1229-1233.
 Prajila, P. and Indulekha, P., 2016. Shoot organogenesis in Justicia adhatoda (L). Nees.
International Journal of Botany Studies, 1(7), 21-22.
 Grange, J.M. and Snell, N.J., 1996. Activity of bromehexine and ambroxol, semisynthetic derivatives of vasicine from the Indian shrub Adhatoda vasica against Mycobacterium tuberculosis in vitro. Journal of Ethnopharmacology, 50(1), 49-53.
 Maurya, S. and Singh, D., 2010. In vitro callus culture of Adhatoda vasica: A medicinal plant. Annals of Biological Research, 1(4), 57-60.
 Anand, Y. and Bansal, Y.K., 2002. Synthetic seeds: a novel approach of in vitro plantlet formation in vasaka (Adhatoda vasica Nees.). Plant Biotechnology, 19(3), 159-162.
 Lone, S.A., Wani, S.H. and Tafazul, M., 2017. Cost effective protocol for in vitro propagation of Adhatoda vasica Nees. using modified tissue culture media. International Journal of Advanced Science and Research, 2(2), 67-69.
 Dagar, J.C., 2005. Ecology, management and utilization of halophytes. Bulletin of the NIE, 15, 81-97.
 Khenifi, M.L., Boudjeniba, M. and Kameli, A., 2011. Effects of salt stress on micropropagation of potato (Solanum tuberosum L.). African Journal of Biotechnology, 10(40), 7840-7845.
 Atabaki, N., Nulit, R., Kalhori, N., Lasumin, N., Sahebi, M. and Abiri, R., 2018. In vitro selection and development of Malaysian salt-tolerant rice (Oryza sativa L. cv. MR263) under salinity. Acta Scientific Agriculture, 2(8), 8-17.
 Saputro, T.B., Dianawati, S., Sholihah, N.F. and Ermavitalini, D., 2017. Genetic diversity of improved salt tolerant calli of maize (Zea mays L.) using RAPD. AIP Conference Proceedings, 1854, 020033, https://doi.org/10.1063/1.4985424.
 Benderradji, L., Brini, F., Kellou, K., Ykhlef, N., Djekoun, A., Masmoudi, K. and Bouzerzour, H., 2012. Callus induction, proliferation, and plantlets regeneration of two bread wheat (Triticum aestivum L.) genotypes under saline and heat stress conditions. ISRN Agronomy, 2012, https://doi.org/10.5402/2012/367851.
 Nasser, S., Iwan, D.T. and Paul, J.K., 1980. Effect of repeated application of water stress on water status and growth of wheat. Physiologia Plantarum, 50(1), 11-15.
 Murashige, T. and Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15(3), 473-497.
 Duncan, D.B., 1955. Multiple Range and Multiple F Test. Biometrics, 11(1), 1-42.
 Khalekuzzaman, M., Rahman, M.S., Rashid, M.H. and Hossain, M.S., 2008. High frequency in vitro propagation of Adhatoda vasica Nees through shoot tip and nodal explants culture. Journal of Bio-Science, 16, 35-39.
 Tejavathi, D.H., Manjula, B.L. and Anitha, P., 2010. Multiple Shoot Regeneration from the Cultures of Adhatoda vasica Nees. Acta horticulturae, 865, 367-370.
 Adesoye, A.I., Okooboh, G.O., Akande, S.R., Balogun, M.O. and Odu, B.O., 2012. Effect of phytohormones and genotype on meristem and shoot tip culture of Telfairia occidentalis Hook F. Journal of Applied Biosciences, 49, 3415- 3424.
 Isah, T., 2019. De novo in vitro shoot morphogenesis from shoot tip-induced callus cultures of Gymnema sylvestre (Retz.) R.Br. ex Sm. Biological Research, 52, 3, https://doi.org/ 10.1186/ s40659-019-0211-1.
 Al-Mayahi, A.M.W., Ali, A.H. and Shareef, H.J., 2018. Influence of cold pretreatment on shoot regeneration from callus in date palm (Phoenix dactylifera L.) cv. ‘Barhee’. Journal of Genetic Engineering and Biotechnology, 16(2), 607-612.
 Altan, F., Bürün, B. and Sahin, N., 2010. Fungal contaminants observed during micropropagation of Lilium candidum L. and the effect of chemotherapeutic substances applied after sterilization. African Journal of Biotechnology, 9(7), 991-995.
 Singh, C.R., 2018. Review on problems and its remedy in plant tissue culture. Asian Journal of Biological Sciences, 11(4), 165-172.
 Garcia, C., Almeida, A.F.D., Costa, M., Britto, D., Valle, R., Royaert, S. and Marelli, J.P., 2019. Abnormalities in somatic embryogenesis caused by 2, 4-D: an overview. Plant Cell, Tissue and Organ Culture, 137, 193-212.
 Peixoto, F.P., Gomes-Laranjo, J., Vicente, J.A. and Madeira, V.M.C., 2007. Comparative effects of the herbicides dicamba, 2,4-D and paraquat on non-green potato tuber calli. Journal of Plant Physiology, 165, 1125-1133.
 Lukas, A.M., Christoper, D.G., Rebecca, A.S. and Virgnia, W., 2000. AN9-a petunia glutathione s-transferase required for anthocyanin sequestration, is a flavonoid binding protein. Plant Physiology, 123, 1561-1570.
 Sridhar, T.M. and Naidu, C.V., 2011. An efficient callus induction and plant regeneration of Solanum nigrum (L.) - An important antiulcer medicinal plant. Journal of Phytology, 3(5), 23-28.
 Seyyedyousefi, S.R, Kaviani, B., Dehkaei, N.P. and Salehzadeh, A., 2013. Callus induction in Alstroemeria using NAA and BAP. European Journal of Experimental Biology, 3(5), 137-140.
 Khan, N., Ahmed, M., Hafiz, I., Abbasi, N., Ejaz, S. and Anjum, M., 2015. Optimizing the concentrations of plant growth regulators for in vitro shoot cultures, callus induction and shoot regeneration from calluses of grapes. Journal International des Sciences de la Vigne et du Vin, 49, 37-45.
 Michel, Z., Hilaire, K.T., Mongomaké, K., Georges, A.N. and Justin, K.Y., 2008. Effect of genotype, explants, growth regulators and sugars on callus induction in cotton (Gossypium hirsutum L.). Australian Journal of Crop Science, 2(1), 1-9.
 Sundarasekar, J., Anthony, J.J.J., Murugaiyah, V. and Subramaniam, V., 2012. Preliminary responses of 2, 4-D and BAP on callus initiation of an important medicinal-ornamental Hymenocallis littoralis plants. Journal of Medicinal Plants Research, 6(11), 2088-2093.
 Ahmad, S. and Spoor, W., 1999. Effects of NAA and BAP on callus culture and plant regeneration in Curly Kale (Brassica oleraces L.). Pakistan Journal of Biological Sciences, 2 (1), 109-112.
 Rahman, M.H., Islam, R., Hossain, M. and Haider, S.A., 2009. Differential response of potato under sodium chloride stress conditions in vitro. Journal of Bio-Science, 16, 79-83.
 Hasegawa, P.M., Bressan, R.A., Zhu, J.K. and Bohnert, H.J., 2000. Plant cellular and molecular responses to high salinity. Annual Review of Plant Physiology and Plant Molecular Biology, 51, 463-499.
 Ashraf, M. and Foolad. M.R., 2007. Roles of glycinebetaine and proline in improving plant abiotic stress resistance. Environmental and Experimental Botany, 59(2), 206-216.
 Efferth, T., 2019. Biotechnology applications of plant callus cultures. Engineering, 5, 50-59.
 Ahuja, M.R., 1991. Application of biotechnology to preservation of forest tree germplasm. In: M.R. Ahuja, ed. Woody Plant Biotechnology. New York: Plenum Press, pp. 307-313.
 Ahmad, N., Faisal, M., Anis, M. and Aref, I.M., 2010. In vitro callus induction and plant regeneration from leaf explants of Ruta graveolens L. South African Journal of Botany, 76(3), 597-600.
 Winicov, I. and Bastola, D.R., 1997. Salt tolerance in crop plants: new approaches through tissue culture and gene regulation. Acta Physiologiae Plantarum, 19, 435-449.