International Journal of Noni Research - Noni Family

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J. Subramani S. Antony Selvaraj D. Vijay M. Sakthivel Authors’ affiliation : J. Subramani S. Antony Selvaraj D. Vijay M. Sakthivel Crop Research division of WNRF, INRF, Chennai, Tamilnadu, India - 600 119. Correspondence to : J. Subramani S. Antony Selvaraj D. Vijay M. Sakthivel M. Umashanthi Crop Research division of WNRF, INRF, Chennai, Tamilnadu, India - 600 119. santony@nonifamily.net Micropropagation of Morinda citrifolia L. Keywords : Morinda sp, Nodal explant, multiple shoots, Rhizogenic calli Abstract : The varying effect of Cytokinin and Auxin combinations on Morinda citrifolia L. for effective in – vitro induction of shoots from nodal explants was studied by using both WP and MS basal media. BAP alone with 2 mg/ l concentration was found effective in both the basal media, to initiate auxillary shoot induction. Further BAP along with 1mg/l Kinetin combination gave 4-5 multiple shoots from a single node within a period of two weeks. Further by using leaf ex–plants, callus was induced. Actively growing yellowish callus was induced by using 2 mg /l-2,4-D. Further the initiated calli were subcultured in MS basal media containing 0.5 mg/l NAA to produce rhizogenic calli with lots of roots at periphery. These calli mass with roots in BAP and KN media combination, became organogenic and resulted in producing shoot (s). Introduction Morinda citrifolia L. commercially known as Noni, grows widely throughout Asia and Pacific, is one of the most significant sources of traditional medicines among Pacifiic Islands Societies. (Clatchey, 2002; Nelson, 2001). The botanical name for the genus was derived from two Latin word Morus, mulberry and indicus, Indian, in reference to the similarity of the fruit of Noni to that of true mulberry (Morus alba), belongs to the family Rubiaceae (Morton, 1992). Noni is relatively easy to propogate from seeds, stem or root cuttings and air layering. The preferred methods of propagation are by seeds and by cuttings made from stem verticals (Nelson, 2001). Till date, not much work on in vitro propagation by using tissue culture techniques were carried out in this plant. Preliminary studies on in vitro propagation of Morinda citrifolia L. was attempted at this center for the past one year. Both the nodal ex-plants and rhizogenic calli have given shoot(s) in both the WP and MS media with the combination of Cytokinin and Auxin. Materials and Methods Plant Material Seeds of Morinda citrifolia L. brought from the Western Ghats, India were sown in the gardens of Health India Laboratories, Sholinganallur. Two leaf Intl. J. Noni Res. 2007, 2(1-2) 38
39 Intl. J. Noni Res. 2007, 2(1-2) stage plants were brought to Tissue Culture Laboratory and used for explant collection. Apical regions and nodal explants were used. Young leaves were used for callus initiation. Sterilization details a. Nodal explant The nodal explants were collected from 6-8 months old seedlings. Top 3 nodes were taken for tissue culture purpose by leaving the basal single node in the mother plant. Due to the removal of upper axillary buds, the axillary region sprouts very efficiently by producing two axillary sprouts in vivo within 8-10 days. b. Procedure Top 3 nodes were taken and then washed with sterile water and kept in 1: 2 (Sodium hypochlorite : Sterile water) with constant stirring for 30 minutes. The ex-plants were washed 3-4 times with sterile water, then treated with 0.1% mercuric chloride for 5 minutes and rinsed with sterile water before inoculation. c. Leaf ex–plant The above sterilization treatment was given for leaf ex plant also to initiate callus. Media details J. Subramani et al. Micropropagation of Morinda citrifolia L. Fig.1 Fig.2 Following basal media with various combinations of Cytokinin and Auxin were tried in our in-vitro studies. The results are shown in Table 1, 2 & 3. Table 1. Various combinations of media for in-vitro Multiplication Media BAP (mg/I) KN (mg/I) IBA (mg/I) 2.0 - - 4.0 - - MS 1.0 1.0 - 2.0 0.75 2.0 1.0 0.25 -
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