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Effect of TDZ on plant regeneration from mature seeds in pea (Pisum sativum)
Authors:S. Zhihui  M. Tzitzikas  K. Raemakers  M. Zhengqiang  R. Visser
Affiliation:(1) Plant Breeding Laboratory, Wageningen University and Research Center, 6708 PB Wageningen, Netherlands;(2) National Key Laboratory of Crops Genetics & Germplasm Enhancement, Nanjing Agriculture University, Nanjing, 210095, China;(3) Present address: Genetwister Technologies, Nieuwe Kanaal, 7b6709 PA Wageningen, Netherlands;
Abstract:Pea (Pisum sativum L. cv. Espace) seeds directly cultured on thidiazuron (TDZ)-containing medium formed high numbers of shoots. The number of shoots per seedling depended on the concentration and duration of the TDZ treatment. The best treatment was 12-wk incubation on MS medium supplemented with 4 mg/l TDZ followed by 4-wk culture on MS medium supplemented with 0.5 mg/l benzylaminopurine (BA) and produced more than 400 shoots/seedling. Isolated shoots rooted at a high frequency on MS medium containing 2–3 mg/l indole-3-butyric acid and 2 mg/l α-naphthalene acetic acid. In addition to the formation of shoots, bud-containing tissues (BCT) were formed at the cotyledonary nodes, shoot nodes, tendrils, stipules, and internodes. The BCT from the cotyledonary nodes and the shoot nodes was maintained in its pure state on MS medium supplemented with 4 mg/l TDZ by repeated culture. Shoot development was accomplished when the BCT were left on MS medium supplemented with 4 mg/l TDZ without subculture prior to transfer onto MS medium supplemented with 0.5 mg/l BA.
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