Regeneration of pea (Pisum sativum L.) by a cyclic organogenic system

• Published in: Plant cell reports Vol. 23; no. 7; pp. 453 - 460
• Main Authors: Tzitzikas, E.N, Bergervoet, M, Raemakers, K, Vincken, J.P, Lammeren, A. van, Visser, R.G.F
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.12.2004; Springer Nature B.V
• Subjects: acclimation; Acclimatization; Acclimatization - physiology; Acetic acid; Acids; Agriculture - methods; benzyladenine; Biological and medical sciences; Biotechnology; Cultivars; culture media; Culture Media - chemistry; Culture Media - pharmacology; cultures; embryos; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; genetic variation; gibberellic acid; growth; in vitro regeneration; indole acetic acid; Meristem - drug effects; Meristem - growth & development; Meristem - metabolism; meristems; Methods. Procedures. Technologies; micropropagation; Miscellaneous; Naphthalene; naphthaleneacetic acid; peas; Pisum sativum; Pisum sativum - drug effects; Pisum sativum - growth & development; Pisum sativum - metabolism; Plant cells and fungal cells; Plant Growth Regulators - pharmacology; plant growth substances; plant morphology; Plant Shoots - drug effects; Plant Shoots - growth & development; Plant Shoots - metabolism; Plant tissues; Regeneration - drug effects; Regeneration - physiology; Shoots; somatic embryogenesis; somatic embryos; stem nodes; thiazoles; thidiazuron; transformation; Vegetals; Thidiazuron; Adventitious regeneration; Meristem; Regeneration; Meristems; Shoot regeneration; Leguminosae; Cyclic; Dicotyledones; Pisum sativum; Angiospermae; Spermatophyta; Shoot
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-004-0865-0

In a five-step procedure, plants were regenerated from meristematic tissue initiated from nodal tissue in four pea cultivars ('Espace', 'Classic', 'Solara', and 'Puget'). In step 1, stem tissue with one node (1-cm size) was subcultured on medium containing thidiazuron. As a result multiple shoots were produced, appearing normal or swollen at their bases. The multiple shoots were subcultured in the same medium, resulting in the formation of a green hyperhydric tissue in the swollen bases of the multiple shoots, which is fully covered with small buds [bud-containing tissue (BCT)]. In step 2, BCT fragments were isolated and subcultured in the same medium and, as a result, they were able to reproduce themselves in a cyclic fashion. In step 3, subculture of BCT on medium supplemented with a combination of gibberelic acid, 6-benzyladenine and alpha-naphthalene acetic acid (NAA), resulted in the formation of shoots, which were rooted in step 4 on medium supplemented with 0.5 mg/l NAA, indole-3-acetic acid (IAA) or indole-3-butyric acid. In step 5, in vitro plants were transferred to the greenhouse for acclimatisation and further development. The four varieties tested were all able to produce meristematic tissue, suggesting that its production is genotype independent.