Efficient Plant Regeneration from Suspension-cultured Cells of Tall Bearded Iris

Abstract

<jats:p>A protocol was developed for efficient plant regeneration of <jats:italic>Iris germanica</jats:italic> L. `Skating Party' from suspension cultures. Suspension cultures were maintained in Murashige and Skoog (MS) basal medium (pH 5.9) supplemented with 290 mg·L<jats:sup>–1</jats:sup> proline, 50 g·L<jats:sup>–1</jats:sup> sucrose, 5.0 μ<jats:sc>m</jats:sc> 2,4-D, and 0.5 μ<jats:sc>m</jats:sc> Kin. Suspension-cultured cells were transferred to a shoot induction medium (MS basal medium supplemented with 10 mg·L<jats:sup>–1</jats:sup> pantothenic acid, 4.5 mg·L<jats:sup>–1</jats:sup> nicotinic acid, 1.9 mg·L<jats:sup>–1</jats:sup> thiamine, 250 mg·L<jats:sup>–1</jats:sup> casein hydrolysate, 250 mg·L<jats:sup>–1</jats:sup> proline, 50 g·L<jats:sup>–1</jats:sup> sucrose, 2.0 g·L<jats:sup>–1</jats:sup> Phytagel, 0.5 μ<jats:sc>m</jats:sc> NAA, and 12.5 μ<jats:sc>m</jats:sc> Kin). Cell clusters that proliferated on this medium differentiated and developed shoots and plantlets in about 5 weeks. Regeneration apparently occurred via both somatic embryogenesis and shoot organogenesis. A series of experiments was conducted to optimize conditions during suspension culture to maximize subsequent plant regeneration. Parameters included 2,4-D and Kin concentrations, the subculture interval, and the size of cell clusters. The highest regeneration rate was achieved with cell clusters ≤280 μm in diameter, derived from suspension cultures grown for 6 weeks without subculturing in liquid medium containing 5 μ<jats:sc>m</jats:sc> 2,4-D and 0.5 μ<jats:sc>m</jats:sc> Kin. Up to 4000 plantlets with normal vegetative growth and morphology could be generated from 1 g of suspension-cultured cells in about 3–4 months. Chemical names used: 2,4-dichlorophenoxyacetic acid (2,4-D); kinetin (Kin); 1-naphthaleneacetic acid (NAA).</jats:p>

Somatic embryogenesis in saffron (Crocus sativus L.). Histological differentiation and implication of some components of the antioxidant enzymatic system

“…7 Densitometric scan of gels stained for total SOD activity detected at the different stages of saffron somatic embryogenesis showing the different Mn-SOD and Cu, Zn-SOD isoenzymes comparisons from reports on other species related to Crocus sativus. The stages of differentiation during somatic embryogenesis in saffron reported in this work are in coincidence with the observations previously reported in several species of geophytes as the case of Iris (Radojevic and Subotic 1992;Wang et al 1999), Gladiolus (Stefaniak 1994), Narcissus (Sage et al 2000) and Allium sativum (Fereol et al 2002). Although the process of somatic embryogenesis must start earlier than it is actually visible at the microscope, the attainment of a globular appearance is regarded as one of the first key features of somatic embryo development (Thorpe and Stasolla 2001).…”

Improved Plant Regeneration from Suspension-cultured Cells of Iris germanica L. `Skating Party'

“…Iris suspension cultures and media. Suspension cultures of Iris germanica 'Skating Party' established from friable calli by Wang et al (1999) were maintained in MS-L medium (Table 1) in the dark on a gyrating shaker (100 rpm) at 23 °C. They were subcultured every 3 weeks (unless otherwise described) by decanting MS-L medium and transferring the cells into two 250-mL flasks, each containing 75 mL of MS-L medium.…”

“…An efficient and reproducible protocol for plant regeneration from cell suspension cultures of I. germanica 'Skating Party' was developed by Wang et al (1999). Several parameters were optimized in the cell suspension culture phase, including concentration of plant growth regulators, size of cell aggregates, and the length of subculture interval.…”

<p align="center">Endemik Kaba Navruz Bitkisinin (<em>Iris galatic</em>a Siehe) In Vitro Çoğaltımı</p>

“…Ascough et al (2009)'un bildirdiğine göre Iris türlerinde yapılan ilk çalışma 1945 yılında Randolph tarafından yürütülen embriyo kültürüdür. I. ensata (Yabuya et al 1991;Boltenkov et al 2007), I. germanica (Shimizu et al 1997;Wang et al 1999aWang et al ,1999b, I. nigricans (Shibli and Ajlouni 2000), I. pumila (Jevremovic and Radojevic 2002), I. ensata, I. setosa, I. sanguinea (Boltenkov and Zarebno 2005), I. hollandica (Fidalgo et al 2005), I. atrofusca, I. petrana, I. vartanii (Al-Gabbiesh et al 2006, I. adriatica (Keresa et al 2009), I. sari ve I. schachtii (Uzun et al 2014) gibi birçok Iris türünde kök, yaprak, çiçek organları, soğan segmentleri, olgunlaşmamış embriyo ve zigotik embriyo gibi birçok eksplant kullanılarak rejenerasyon çalışmaları yapılmıştır. Ancak Iris galatica ile ilgili herhangi bir çalışmaya rastlanmamıştır.…”

“…Değişik araştırıcılar tarafından da belirtildiği gibi İris türlerinin çoğunun, tohum ekiminden çiçek açacak zamana ulaşması ortalama 4-5 yıl süre alması ve vejetatif çoğaltım hızının da düşük olması nedeniyle bir takım hızlı çoğaltma tekniklerinin bu bitkiler üzerinde geliştirilmesi gerekmektedir (Boltenkov et al 2007;Shibli and Ajlouni 2000;Wang et al 1999a;Wang et al 1999b). Bu çalışmada da endemik ve az tehdit altında bulunan Iris galatica türünün doku kültürü teknikleri ile in vitro çoğaltımı amaçlanmıştır.…”