Initiation, growth and cryopreservation of plant cell suspension cultures

Mustafa, Natali Rianika; Winter, Ward de; Iren, F. van; Verpoorte, Robert

doi:10.1038/nprot.2010.144

Cited by 185 publications

(123 citation statements)

References 58 publications

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“…An early improvement on the way to overcome protoplast recalcitrance was to exploit actively dividing cultured cells for protoplast isolation. Although very powerful this invention has an apparent limitation-it requires prior establishment of a suspension culture, a process which takes months and itself for many plant species is far from a routine (Mustafa et al 2011). This approach was successfully used also in case of sugar beet allowing to reach plating efficiency of about 40% (Majewska-Sawka et al 1994.…”

Section: Discussionmentioning

confidence: 99%

Phytosulfokine stimulates cell divisions in sugar beet (Beta vulgaris L.) mesophyll protoplast cultures

et al. 2012

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The aim of this work was to improve plating efficiency of sugar beet mesophyll protoplast cultures. Preliminary experiments showed that cultures of good quality, viable protoplasts were obtained in rich media based on the Kao and Michayluk formulation and with the calcium alginate as an embedding matrix. Nevertheless, in these cultures cell divisions were either not observed or very seldom confirming earlier reported recalcitrance of sugar beet protoplasts. The recalcitrant status of these cultures was reversed upon application of exogenous phytosulfokine (PSK)-a peptidyl plant growth factor. The highest effectiveness of PSK was observed at 100 nM concentration. Plating efficiencies obtained in the presence of PSK reached approximately 20% of the total cultured cells. The stimulatory effect of phytosulfokine was observed for all tested breeding stocks of sugar beet. Our data indicate that PSK is a powerful agent able to overcome recalcitrance of plant protoplast cultures.

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Section: Discussionmentioning

confidence: 99%

Phytosulfokine stimulates cell divisions in sugar beet (Beta vulgaris L.) mesophyll protoplast cultures

et al. 2012

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“…In vitro plant cell suspension culture facilitates large-scale production of fine chemicals in industrial bioreactors and for the study of cellular and molecular processes as it offers the advantage of a simplified model system for studying physiological effect of salt at the cellular level under a controlled environment [18] and the effect of heavy metal stress on growth, enzymes activities and altered biochemical parameters in cultured cells [19]. Cell suspension cultures contain a relatively homogeneous cell population, allowing rapid and uniform access to nutrition, precursors, growth hormones and signal compounds in the cells [20].…”

Section: Introductionmentioning

confidence: 99%

Enhanced Biosynthesis of Withanolides by Elicitation and Precursor Feeding in Cell Suspension Culture of Withania somnifera (L.) Dunal in Shake-Flask Culture and Bioreactor

Selvaraj²,

et al. 2014

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The present study investigated the biosynthesis of major and minor withanolides of Withania somnifera in cell suspension culture using shake-flask culture and bioreactor by exploiting elicitation and precursor feeding strategies. Elicitors like cadmium chloride, aluminium chloride and chitosan, precursors such as cholesterol, mevalonic acid and squalene were examined. Maximum total withanolides detected [withanolide A (7606.75 mg), withanolide B (4826.05 mg), withaferin A (3732.81 mg), withanone (6538.65 mg), 12 deoxy withanstramonolide (3176.63 mg), withanoside IV (2623.21 mg) and withanoside V (2861.18 mg)] were achieved in the combined treatment of chitosan (100 mg/l) and squalene (6 mM) along with 1 mg/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose in culture at 4 h and 48 h exposure times respectively on 28th day of culture in bioreactor. We obtained higher concentrations of total withanolides in shake-flask culture (2.13-fold) as well as bioreactor (1.66-fold) when compared to control treatments. This optimized protocol can be utilized for commercial level production of withanolides from suspension culture using industrial bioreactors in a short culture period.

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“…italica [15]. Such an approach might be beneficial; however, it is laborious and time consuming and requires the establishment of actively dividing suspension cultures [16]. Favorable effect of nurse cultures, feeder layers, or conditioned media suggested the presence of division-inducing factors that are secreted into the medium with cultured cells [17,18].…”

Section: Digital Signaturementioning

confidence: 99%

Early studies on the effect of peptide growth factor phytosulfokine-α on Brassica oleracea var. capitata L. protoplasts

Kiełkowska

Adamus

2017

Acta Soc Bot Pol

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Phytosulfokines (PSK) are peptidyl growth factors with the potential of inducing cell proliferation. We examined the effect of supplementation of liquid culture medium with 0.1 µM phytosulfokine-α (PSK-α) on protoplast viability and division frequencies in seven accessions of Brassica oleracea var. capitata L., including cultivars and breeding lines. Protoplasts were isolated from leaves and hypocotyls of in vitro grown plants and immobilized in calcium-alginate layers. Cabbage protoplast-derived cells cultured in medium supplemented with 0.1 µM of PSK-α had higher viability and division frequencies compared to cells cultured in PSK-α-free control medium. The effect of PSK-α was more pronounced in low-responding accessions ('Sława z Gołębiewa' , 'Ramkila F1' , LM, and LM98); however, in two cultivars with very low response ('Badger Shipper' and 'Oregon 123'), although the division frequencies in the media supplemented with PSK-α were increased over the control, the differences were not significant. Obtained callus colonies were subjected to regeneration. PSK-α supplemented into the liquid culture medium had an indirect effect on shoot regeneration by inducing sustained cell divisions leading to an increase in shoot regeneration in Sława z Gołębiewa and both breeding lines.

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Initiation, growth and cryopreservation of plant cell suspension cultures

Cited by 185 publications

References 58 publications

Phytosulfokine stimulates cell divisions in sugar beet (Beta vulgaris L.) mesophyll protoplast cultures

Phytosulfokine stimulates cell divisions in sugar beet (Beta vulgaris L.) mesophyll protoplast cultures

Enhanced Biosynthesis of Withanolides by Elicitation and Precursor Feeding in Cell Suspension Culture of Withania somnifera (L.) Dunal in Shake-Flask Culture and Bioreactor

Early studies on the effect of peptide growth factor phytosulfokine-α on Brassica oleracea var. capitata L. protoplasts

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