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Synergistic effect of ethylene inhibitors and putrescine on shoot regeneration from hypocotyl explants of Chinese radish (Raphanus sativus L. var. longipinnatus Bailey) in vitro
Authors:Eng-Chong Pua  Guek-Eng Sim  Gek-Lan Chi  Lan-Feng Kong
Institution:(1) Department of Botany, Faculty of Science, National University of Singapore, Lower Kent Ridge Road, 119260 Singapore, Republic of Singapore;(2) Present address: Department of Biotechnology, Ngee Ann Polytechnic, 535 Clementi Road, 599489 Singapore, Republic of Singapore;(3) Present address: Parks & Recreation Department, 5th storey, National Development Building, Annexe B, 7 Maxwell Road, 069111 Singapore, Republic of Singapore
Abstract:Summary The role of ethylene and putrescine on shoot regeneration from hypocotyl explants of Chinese radish (Raphanus sativus L. var. longipinnatus Bailey cv. Red Coat) was investigated. Explants were recalcitrant in culture, but exogenous application of ethylene inhibitor 20–30 mgrM aminoethoxyvinylglycine (AVG) or AgNO3] enhanced shoot regeneration of explants grown on medium supplemented with 2 mg/l N6-benzyladenine and 1 mg/l 1-naphthaleneacetic acid. The best regeneration occurred in the medium containing AgNO3 in combination with AVG. Culture medium solidified with agarose in the presence of AgNO3 but not AVG was also beneficial to shoot regeneration. Exogenous putrescine, 2-chloroethylphosphonic acid and 1-aminocyclopropane-1-carboxylate had no effect on shoot regeneration. However, regeneration was greatly promoted by 10–25 mM putrescine in combination with 30 mgrM AgNO3 or AVG. Explants with high regenerability grown in the presence of AgNO3 or in combination with putrescine emanated high levels of ethylene throughout the 21-d culture period. By contrast, AVG or putrescine alone resulted in a decrease in ethylene production. For rooting of shoot cuttings, IAA and IBA at 1–5 mg/l were more effective than NAA.Abbreviations ACC 1-aminocyclopropane-1-carboxylate - AVG aminoethoxyvinylglycine - BA N6-benzyladenine - CEPA 2-chloroethylphosphonic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - MS Murashige and Skoog (1962) medium - NAA 1-naphthaleneacetic acid - PAs polyamines - SAM S-adenosyl-L-methionine
Keywords:AgNO3  aminoethoxyvinylglycine  Chinese radish  putrescine  ethylene  Raphanus sativus  var  longipinnatus  shoot regeneration
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