Efficient and stable regeneration from protoplasts of Cyclamen coum Miller via somatic embryogenesis

Abstract

Embryogenic cultures of Cyclamen coum were established on solid media and in suspension, and their growth characteristics in response to different concentrations of plant growth regulators (PGRs) were evaluated. Embryogenic cultures exhibited a high regeneration capacity of 876 somatic embryos per gram fresh mass. Up to 4.24 9 10 5 protoplasts per gram of fresh mass were isolated from somatic embryos and embryogenic suspension cultures. Protoplasts derived from both embryos and suspension cultures were successfully cultured in vitro and regenerated into plants via somatic embryogenesis. Phenotypic analyses and flow cytometric measurements revealed that some regenerated plants were tetraploid. About 20% of the protoplast-derived calluses used for regeneration were tetraploid, while tetraploidy was found in 0.9% of the plants regenerated from the embryogenic cultures.

Flow cytometric analysis of somatic embryos, shoots, and calli of the cactus Copiapoa tenuissima Ritt. forma monstruosa

“…The measurement of the DNA content and defining the size of the genome could be precisely and rapidly made with the flow cytometry (FCM) (Doležel 1991;Thiem and Ś liwińska 2003;Sliwinska et al 2009;Obae and West 2010). Flow cytometric analysis is useful for the study of genetic stability in plants regenerated in vitro, especially via somatic embryogenesis (Loureiro et al 2005;Mallón et al 2010;Pinto et al 2010;Prado et al 2010;Prange et al 2010;Yang et al 2010).…”

Somatic embryogenesis in the medicinal legume Desmodium motorium (Houtt.) Merr.

“…Appropriate types, concentration and combination of plant growth regulators have enabled manipulation of response and recovery of plants in a number of species (Ammirato 1983b). Perusal of the literature reveals that, in most plants, the auxin most commonly used to induce somatic embryogenesis is 2,4-D (Hofmann et al 2004;Korbes and Droste 2005;Hiraga et al 2007;Pacheco et al 2009;Viba et al 2009;Chen et al 2010;Pan et al 2010;Palmer and Keller 2010;Prange et al 2010;Chai et al 2011;Ram et al 2011). Induction of somatic embryogenesis by cytokinin alone is relatively rare among legumes (Maheshwaran and Williams 1986;Malik and Saxena 1992).…”

Protoplast isolation and plant regeneration of different doubled haploid lines of cauliflower (Brassica oleracea var. botrytis)

“…Moreover, plant regeneration from hypocotyls protoplasts of different DH lines of cauliflower was reproducibly achieved and the frequency of shoot regeneration is higher than previously reported for cauliflower, which gave a maximum regeneration rate of 72% (Veera et al 2009). Plant regeneration from protoplasts is influenced by many factors, including the methods of protoplast isolation, inoculation density, culture system, nurse cells, and medium compositions (Meyer et al 2009;Eeckhaut et al 2009;Castelblanque et al 2010;Prange et al 2010;Tiwari et al 2010). The use of feeder layer-culture systems with protoplasts of other species as nurse cells provided a simple and reliable technique for enhancing cell division and plating efficiency of isolated protoplasts (Walters and Earle 1990;Karamian and Ebrahimzadeh 2001).…”