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antidiabtic medicinal plants

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Information about antidiabtic medicinal plants
Science-Technology

Published on September 29, 2007

Author: vadivel007

Source: authorstream.com

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IN VITRO CULTURE OF ANTIDIABETIC MEDICINAL PLANTS:  IN VITRO CULTURE OF ANTIDIABETIC MEDICINAL PLANTS ANTIDIABETIC MEDICINAL PLANTS:  ANTIDIABETIC MEDICINAL PLANTS Allium ceba Allium sativum Andrographis paniculata Stevia rebaudiana Morinda citrifolia Gymnema sylvestre Pterocarpus marsupium Centella asiatica Slide3:  Houttuynia cordata Chlorophytum borivilianum Trigonella foenum graecum Cassia auriculata Taraxacum officinale Syzygium cumini Vaccinium spp. Nigella sativa In vitro regeneration of Pterocarpus marsupium:  In vitro regeneration of Pterocarpus marsupium Conventional propagation method: The winged fruit is the only propagating material, but its germinationis low (only 30%). Hard fruitcoat, less germinability. Slide5:  IN VITRO PROPAGATION: Fruit coats were removed manually with the help of a cutter and the seeds were kept in running tap water for 30 min,then soaked for 24 h in distilled water. Seeds were then treated with liquid detergent (Teepol 5%) for 5 min. surface sterilized with 0.1% HgCl2 for 5 min, washed 4–5 times in sterile water. Implanted the explants aseptically on half strength MS medium containing 3% sucrose and gelled with 0.8% agar (Qualigens,India). Slide6:  When the seedling grew up to 5 cm, the cotyledons, nodal segments, cotyledonary nodes and shoot tips derived from 18-day-old aseptic seedlings were excised and used to initiate culture. The MS medium supplemented with cytokinins or auxins (IAA,IBA, NAA), either singly or in combination. pH of the medium adjusted to 5.8 . All cultures were incubated at 25 ± 2°C in a photoperiod of 16 h for a day under fluorescent light (about 1200 lux) and with the RH 55–60%. For rooting, half and full strength MS medium augmented with various auxins (IAA, IBA, NAA) along with a phenolic acid were used. Slide7:  a, Induction and proliferation of multiple shoots from cotyledonary node explant on MS + BA b, Shoot elongation on MS + BA+ IAA . c, Multiple shoot formation on MS + BA after 6 weeks. d,Growth and proliferation of shoots on the same medium after 8 weeks. e, Normal rooted shoot developed on ½ MS + IBA through a two-step culture procedure. In vitro multiplication of Centella asiatica:  In vitro multiplication of Centella asiatica Leaf explants, collected from 5–6 months old plants of C. Asiatica Washed with a detergent solution (1% teepol) for 2–3 min, followed by thorough washing under running tap water for 30 min. The explants are surface sterilized with 0.1% HgCl2 for 2–3 min, followed by a final 5–6 rinses with sterile water. The sterilized leaf explants, dissected into two halves with or without petioles, were cultured on MS medium containing different concentrations and combinations of cytokinins [6-furfuryl benzylaminopurine] and kinetin and auxins [indole butyric acid] and naphthaleneacetic acid. Slide9:  The cultures are maintained at 25 ± 2° C, under light of 3000 lux and with RH 55–60%. BAP (2 mg/l) along with IBA (0.1 mg/l) produced maximum sprouting within two weeks. Differentiation of shoots and their growth required transfer of cultures to a medium with a relatively higher concentration of BAP (3.0 mg/l) and NAA at a lower concentration (0.05 mg/l) instead of IBA. Slide10:  the average number of roots and root length was better on half strength MS medium containing 1.0 mg/l IBA . The rooted plants were transferred into earthen pots containing a soil and sand mixture (2:1), with 89 ± 3% survival under glasshouse conditions. Slide11:  The average number of roots and root length was better on half strength MS medium containing 1.0 mg/l IBA . The rooted plants were transferred into earthen pots containing a soil and sand mixture (2:1), with RH 89 ± 3% survival under glasshouse conditions. a and b, Initial response of leaf segments on MS medium c, Multiple shoot regeneration on MS + 3.0 mg/l BAP and 0.05 mg/l NAA from pre-cultured leaf segments; d, Root induction in half strength MS medium. e, In vitro regenerated plantlets showing prostrate stem with rooting at the nodes f, Ex vitro establishment of regenerated plants. :  a and b, Initial response of leaf segments on MS medium c, Multiple shoot regeneration on MS + 3.0 mg/l BAP and 0.05 mg/l NAA from pre-cultured leaf segments; d, Root induction in half strength MS medium. e, In vitro regenerated plantlets showing prostrate stem with rooting at the nodes f, Ex vitro establishment of regenerated plants. In vitro regeneration of – Houttuynia cordata :  In vitro regeneration of – Houttuynia cordata Healthy shoots are collected from the plants grown in the greenhouse and washed with tap water several times. The shoot segments were then washed several times with sterile distilled water in a laminar airflow. Then surface sterilized in 0.1% mercuric chloride solution for about 3 min followed by a thorough rinsing in SDW. These surface sterilized shoot segments were cut into 6–8 mm pieces. The shoot pieces which consisted of a single node containing a small portion of the internode on either side were used as explants. The explants were cultured on MS medium with 0.8% agar and 3% sugar, and with or without N-6 benzyladenine . The pH of the medium was adjusted to 5.8 prior to autoclaving. Slide14:  The media autoclaved at a pressure of 15 psi for 15 min. The culture are maintained under 2000 lux light intensity at 25 ± 2oC. The growing explants are subcultured only once after 30 days. After 4 weeks of subculture, the growing shoots are cultured for rooting on MS medium (pH 5.8) The nodal explants started growing after about one week of culture on MS medium with BA ,producing green shoot buds. Maximum number of shoots were obtained in the medium days of culture. The shoots were subcultured after 30 days. After 4 weeks of subculture, the shoots were cultured on MS medium supplemented with IAA. Slide15:  Adventitious roots developed from the base of the shoots after 21 days of culture and maximum number of roots developed in the presence of 1.0 mg/l IAA . The rooted plants were allowed to grow on the MS medium containing IAA up to 30 days and then transferred to liquid MS medium without hormone for 10 days . Thereafter, the plantlets were transferred into pots containing a sterilized mixture of garden soil and vermiculite (2:1). The plantlets were kept covered with a polythene bag for 10 days to check excessive transpiration. The plantlets were then successfully established in field condition and in big earthen pots . Slide16:  a, Initiation of rooting of H. cordata Thunb. in in vitro condition. b, In vitro growth of plantlet. c, Fully grown plant after 4 months of transfer. In vitro culture of Chlorophytum borivilianum:  In vitro culture of Chlorophytum borivilianum In vitro cultures were established from young shoot apices obtained from the tuberous roots of these field-grown plants. Each shoot of a cluster was separated and trimmed from the top leaving behind about 1–1.5 cm portion along with shoot apex. MS medium supplemented with 3% sucrose, 6- benzylaminopurine and with 0.8% agar or without agar was used. The pH of the medium adjusted to 5.80 ± 0.1 using 0.10 N HCl and/or 0.10 N NaOH prior to autoclaving at 121°C and 15 lb pressure for 20 min. Slide18:  Two explants are inoculated per culture flask All aseptic cultures were maintained under 16 h photoperiod at 25 ± 2°C temperature. For root induction in in vitro regenerated shoots, three-fourths strength MS liquid medium was supplemented with 9.8 ml indole 3butyric acid (IBA). Liquid culture medium not only supported multiplication of shoots but shoot growth was also better compared to solid medium. Slide19:  Agitated liquid culture supported better shoot growth and multiplication than static liquid culture. In vitro-regenerated shoots exhibited optimal root induction and growth in three fourths strength basal MS liquid medium supplemented with 9.8 mM IBA In vitro-regenerated plants exhibited 80% survival rate . Slide20:  a) solid (control), liquid agitated and liquid static (from left to right); (b–c) rooting in shoots regenerated in agitated liquid medium; and (d) In vitro raised C. borivilianum plants in earthen pots after 8 weeks. Slide21:  By… N.SUDHAKAR R.SURENDAR

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