Establishment of embryogenic cell suspension cultures of garlic (Allium sativum L.), plant regeneration and biochemical analyses

Abstract

Embryogenic cell suspension cultures of garlic (Allium sativum L.) were initiated in liquid medium from friable embryogenic tissue. The optimal parameters for culture maintenance were: (1) an initial cell density of 1-4% (v/v); (2) medium renewal every 14 days and subculturing every 28 days; (3) a low 2,4-dichlorophenoxyacetic acid concentration (0.1-0.3 mg/l). Cultures regenerated during a 14-month period. The cell suspension cultures differentiated embryos following transfer to a semi-solid embryo induction medium, with histological studies confirming and characterising the embryogenic nature of the process. Forty percent of these embryos converted into plantlets, which produced micro bulbs in vitro. The composition of the sulphur compounds of the micro bulbs obtained from cell suspension embryo-derived plantlets differed slightly from those produced by in vitro shoot proliferation-derived plantlets, but after two cycles of multiplication in the field these differences had disappeared.

Effect of plant growth regulators, temperature and sucrose on shoot proliferation from the stem disc of Chinese jiaotou (Allium chinense) and in vitro bulblet formation

“…In vitro regeneration has been successfully achieved for many Allium species from a range of explant sources, including the stem disc (Mukhopadhyay et al 2005;Luciani et al 2006), young leaves (Nagasawa and Finer 1988), shoot tips (Kehr and Schaeffer 1976), basal plates (Koch et al 1995;Luciani et al 2006) and flower organs (Luthar and Bohanec 1999;Luciani et al 2006), root tips (Haque et al 1997;Myers and Simon 1998;Barandiaran et al 1999;Kim et al 2003;Martín-Urdíroz et al 2004;Luciani et al 2006), and immature umbels (Luciani et al 2006) through a number of methods like cell suspension culture (Fereol et al 2005) and protoplast culture (Ayabe et al 1995). Culture temperature and sucrose are known to exert a great influence on in vitro bulblet formation and growth of in vitro bulblets in various bulbous crops (Al-aghabary et al 2001;Lian et al 2003;Kim et al 2003;Kumar et al 2005;Maesato et al 1994;Niimi 1995;Stimart and Ascher 1978).…”

Somatic embryogenesis and plant regeneration from root sections of Allium schoenoprasum L.

“…By contrast, many members of the genus Allium have been successfully cultivated and propagated in vitro, either by caulogenesis or somatic embryogenesis. The most well-studied species of the genus Allium are the commercially important A. cepa (Dunstan and Short 1978;Zheng et al 1998Zheng et al , 1999Eady et al 1998;Luthar and Bohanec 1999;Zhang et al 2004) and A. sativum Simon 1998, 1999;Sumi 1998, 2001; Barandiaran et al 1999a, b;Fereol et al 2002Fereol et al , 2005Martín-Urdíroz et al 2004;Luciani et al 2006). However, protocols for other Allium species are also available: A. ampeloprasum (Buiteveld et al 1993;Buiteveld and Creemers-Molenaar 1994;Yasseen et al 1995), A. chinense (Xu et al 2008), A. porrum (Debergh and StandaertDe Metsenaere 1976; Van der Valk et al 1992;Hong and Debergh 1995), A. fistulosum (Van der Valk et al 1992;Song and Peffley 1994;Kim and Soh 1996), A. fistulosum 9 A. cepa (Van der Valk et al 1992;Song and Peffley 1994) and A. tuberosum (Matsuda and Adachi 1996).…”

Biotechnological Tools for Garlic Propagation and Improvement

“…Thirty percent of the somatic embryos developed into plants which acclimated successfully to greenhouse conditions. Later, Fereol et al (2005b) established a protocol for embryo regeneration through suspension cultures by using young leaf sections from cloves of the variety Morasol. Embryogenic calli were obtained when explants were grown on B5 medium with 4.5µM 2,4-D and 0.47µM kinetin, then transferred to a modified B5 medium with 2.2, 1.1, 1.1, 0.4µM 2,4-D, IAA, NAA and kinetin, respectively, plus 175mM sucrose and 2mM proline.…”