Biochemical Changes during Plantlet Regeneration in Two Accessions of Mucuna pruriens

S. Raghavendra, C. K. Ramesh, V. Kumar, M. H. M. Khan, B. S. Harish

Abstract


The genus Mucuna is an important medicinal herb and is extensively used in traditional Indian systems of medicine for various ailments. In vitro culture technique provides an alternative to plant propagation and germplasm conservation. Our aim was to study the biochemical changes occurring during regeneration of shoots (plantlets) from explants of two accessions of Mucuna pruriens, by monitoring the efficiency of nitrogen utilization and changes in levels of some hydrolytic enzymes. A rapid micropropagation system was developed using Murashige and Skoog's (MS) medium supplemented with BAP and IAA combined. In both the accessions, 3.0mg l-1 6-BAP, in combination with 0.2mg l-1 IAA, induced shoot buds and shoot elongation; however for multiple-shoot induction, a slightly higher concentration of cytokinin, i.e., 3.5mg l-1 6-BAP, in combination with 0.2mg l-1 IAA, was required. Results of the present study confirm an active growth of explants revealed by nitrate assimilation enzymes and hydrolytic enzymes. It is concluded that medium composition, growth regulator combination and culture incubation conditions are all vital in both the accessions of Mucuna pruriens for induction of in vitro plant regeneration.

Keywords


Mucuna, in vitro, Biochemical Changes, Regeneration, Enzymes.

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References


Altaf Ahmad and Abdin, M.Z. 1999. NADH: nitrate reductase and NAD(P)H: nitrate reductase activities in mustard seedlings. Pl. Sci., 143:1-8

Angosto, T., Gonzalez, F. and Matilla, A. 1988. Partial purification and some biochemical properties of acid phsophatase in germinating chick pea (Cicer arietinum L.) seeds. Physiol. Pl., 74:715-719

Bradley, D., Kjellborn, P. and Lamb, C. 1992. Elicitor- and wound-induced oxidative cross-linking of a prolinerich plant cell wall protein: a novel, rapid defense response. Cell., 70:21-30

Brown, D.C.W. and Thorpe, T.A. 1980. Adenosine phosphate and nicotinamide adenine dinucleotide pool sizes during shoot initiation in tobacco callus. Pl. Physiol., 65:587-590

Campbell, W.H. and Smarrelli, Jr. J. 1978. Purification and kinetics of higher plant nitrate reductase. Pl. Physiol., 61:611-616

Christopher, T., Prolaram, B. and Subhash, K. 1991. Differential in vitro morphogenetic response in hypocotyl segments of Capsicum annuum. Indian J. Expt’l. Biol., 29:68-69

Kavi Kishor, P.B. and Mehta, A.R. 1988. Changes in some enzyme activities during growth and organogenesis in dark-grown tobacco callus cultures. Pl. Cell Physiol., 29:255-259

Kumar, V. 1998. Biochemical and in vitro plantlet regeneration studies in fig (Ficus carica L. cv. Gular). M. Phil. dissertation submitted to Sri Krishnadevaraya University, Anantapur, Andhra Pradesh, India

Lagrimini, L.M., Gingas, V., Finger, F., Rothstein, S. and Liu, T-T.Y. 1997a. Characterization of antisense transformed plants deficient in the tobacco anionic peroxidase. Pl. Physiol., 114:1187-1196

Lagrimini, L.M., Joly, R.J., Dunlap, J.R., and Liu, T-T.Y. 1997b. The consequence of peroxidase overexpression in transgenic plants on root growth and development. Pl. Mol. Biol., 33:887-895

Lam, H.M., Coshigano, K., Oliveira, I., Melo-Oliveira, R. and Coruzzi, G. 1996. The molecular genetics of nitrogen assimilation into amino acids in higher plants. Pl. Mol. Biol., 47:569-593

Lochab, S., Pathak, R.R. and Raghuram, N. 2007. Molecular approaches for enhancement of nitrogen use efficiency in plants. In: Agricultural Nitrogen use & its Environmental Implications (Eds. Abrol, Y.P., Raghuram, N. and Sachdev, M.S.). IK International, Delhi, India, pp. 327-350

Lorenza M. Bellani, Massimo Guarnieri and Anna Scialabba. 2002. Differences in the activity and distribution of peroxidases from three different portions of germinating Brassica oleracea seeds. Physiol. Pl., 114:102-108

Malik, C.P. and Kumari, U. 1977. Histochemical studies on the localization of metabolic reserves and enzymes during the initiation and formation of adventitious roots inImpatiens balsamina L. New Bot., 4:113-115

Miller, W.B. and Ranwala, A.P. 1994. Characterization and localization of three soluble forms of invertase from Lilium longiflorum flower buds. Physiol. Pl., 92:247 -253

Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Pl., 15:473-497

Naidu, K.R. and Kavi Kishor, P.B. 1995. Activities of hydrolytic enzymes in callus cultures of tobacco during organogenesis. J. Biosci., 20:629-636

Naik, M.S., Abrol, Y.P., Nair, T.V.R., and Ramaras, C.S. 1982. Nitrate assimilation – its regulation and relationship to reduced nitrogen in higher plants. Phytochem., 21:495-504

Narender Singh, Kuldeep Yadav, Suman Kumari, and Renu. 2011. Metabolic changes during differentiation in callus cultures of Stevia rebaudiana (Bertoni). J. Phytol., 3:63-67

Crawford, N.M. 1995. Nitrate: Nutrient and signal for plant growth. Pl. Cell, 7:859-868

Oskar J. Sanchez, Alberto Pan, Gregorio Nicolas and Emilia Labrador. 1989. Relation of cell wall peroxidase activity with growth in epicotyls of Cicer arietinum. Physiol. Pl., 75:275-279

Philippe Lenee and Yves Chupeau. 1989. Development of nitrogen assimilating enzymes during growth of cells derived from protoplasts of sunflower and tobacco. Pl. Sci., 59:109-117

Riley, P.A. 1997. Melanin. Int’l. J. Biochem. Cell Biol., 29:1235-1239

Sadasivam, S. and Manickam, A. 2008. Biochemical Methods. New Age International (P) Limited Publishers, New Delhi, India, pp. 137-139

Singh, S.R., Singh, R. and Dhawan, A.K. 2009. Biochemical changes related to shoot differentiation in callus cultures of Tylophora indica Wight and Arn. J. Indian Bot. Soc., 88:49-53

Srivastava, H.S. 1980. Regulation of nitrate reductase activity in higher plants. Phytochem., 19:725-733

Sujatha, M., Sivaraj, N. and Satya Prasad, M. 2000. Biochemical and histological changes during in vitro organogenesis in Jatropha integerrima. Biologia Plant., 40:167-171

Suzuki, A., Audet, C. and Oaks, A. 1987. Influence of light on the ferredoxin-dependent glutamate synthase in maize leaves. Pl. Physiol., 95:384-389

Yolanda Cuadrado, Hilario Guerra, Ana Belen Martin, Piedad Gallego, Oscar Hita, Ana Dorado and Nieves Villalobos. 2001. Differences in invertase activity in embryogenic and non-embryogenic calli from Medicago arborea. Pl. Cell Tiss. Org. Cult., 67: 145-151






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