CO 2 accumulation in bioreactor suspension cultures of Cyclamen persicum Mill. and its effect on cell growth and regeneration of somatic embryos

Hohe, Annette; Winkelmann, Traud; Schwenkel, H.-G.

doi:10.1007/s002990050675

Cited by 36 publications

(15 citation statements)

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“…This result is particularly interesting, since the model genotype used in most previous studies (Hohe et al, 1999a(Hohe et al, , b, 2001Winkelmann et al, 2004a, b) was also a cultivar with purple flamed flowers. The reasons for the high regeneration response of cultivars with flamed flowers remain unclear and have to be verified in future analyses with cultivars of different genetic background.…”

Section: Discussionmentioning

confidence: 74%

“…While some protocols included seedling tissues as starting material (Kreuger et al, 1995;Takamura et al, 1995), we have concentrated on the method starting from somatic tissue in ovules . By applying this method, mass propagation was achievable, especially if liquid culture systems and bioreactors were used Hohe et al, 1999aHohe et al, , b, 2001. Stability of plants regenerated via somatic embryogenesis using this protocol was high .…”

Section: Introductionmentioning

confidence: 98%

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Genotypic differences in callus formation and regeneration of somatic embryos in Cyclamen persicum Mill

Winkelmann

Serek

2005

Euphytica

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This study was performed in order to check the applicability of a protocol for somatic embryogenesis in Cyclamen persicum and to compare the in vitro response of cultivars belonging to different cyclamen growth types and flower colours obtained from two breeders. One hundred ovules per plant were prepared from 32 tested F 1 hybrid cultivars represented by five plants each. Callus induction monitored after 8 weeks varied between 42 and 85%. Variation occurred between replications and single plants of a cultivar. Consistency and colour of the developing calluses were diverse. Only in 1 cultivar out of the 32 tested no regeneration of somatic embryos was obtained. The regeneration rates varied from 0 to 65% among cultivars. In the group of midi-type cultivars the regeneration response was lower than that in mini and giganteum types. High regeneration rates were observed in cultivars with flamed flowers.

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Section: Discussionmentioning

confidence: 74%

Section: Introductionmentioning

confidence: 98%

Genotypic differences in callus formation and regeneration of somatic embryos in Cyclamen persicum Mill

Winkelmann

Serek

2005

Euphytica

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“…Mass propagation using embryogenic suspension cultures was found to be even more effective if bioreactor culture was applied (Hohe et al 1999a(Hohe et al ,b, 2001. Although the potential to develop somatic embryos is present in many genotypes and the regenerative ability seems to be under the control of two dominant genes (Pueschel et al 2003), soft and suspendable embryogenic callus has been achieved in only a few genotypes and the establishment of stably growing embryogenic callus has been difficult and time-consuming.…”

Section: Introductionmentioning

confidence: 97%

Cryopreservation of embryogenic suspension cultures of Cyclamen persicum Mill.

2004

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We have developed a simple protocol for the cryopreservation of embryogenic suspension cultures of Cyclamen persicum. Embryogenic suspension cultures in the linear growth phase 7-10 days after subculture were used for cryopreservation. Of the different cryoprotectants tested during a 2-day pre-culture, 0.6 M sucrose resulted in the highest re-growth rates of 75%. An additional pre-treatment with 0.6 M sucrose and 10% DMSO (dimethylsulfoxide) for 1 h also positively affected re-growth. Microscopic studies on viability revealed that only few small embryogenic cells survived cryopreservation, while vacuolated single cells died. Experiments in which the duration of the pre-culture period--i.e. the length of time the embryogenic suspension cells were exposed to 0.6 M sucrose--was varied showed that 2-4 days was the most optimal exposure time to 0.6 M sucrose. Callus re-grown after cryopreservation showed growth rates similar to that of unfrozen callus and regenerated even higher numbers of somatic embryos than unfrozen callus.

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“…In suspension cultures of Picea glauca, Kumar et al (1989) found that the concentration of CO 2 accumulated around 25% on day 8 and depressed cell growth. Hohe et al (1999) reported that CO 2 accumulation was the growth-limiting factor in bioreactors. They stated that the CO 2 accumulation during suspension culture benefited the regeneration of cyclamen somatic embryos in subsequent culture stages.…”

Section: Effects Of Individual Gas Componentsmentioning

confidence: 98%

“…By dynamic DO control, the formation rate of torpedo-stage embryos was doubled compared to the control in which P O 2 was kept at 20%. Hohe et al (1999) reported that most embryos were generated from suspension cells in which the mole fraction of CO 2 in the gas phase of the reactor was higher than 10%. Kumar et al (1989) reported that embryogenic cultures of Picea glauca were inhibited by the accumulation of high levels of CO 2 and C 2 H 4 .…”

Section: Introductionmentioning

confidence: 99%

Induction of somatic embryos of celery by control of gaseous compositions and other physical conditions

2006

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The celery (Tall Utah, Apium graveolens var. dulce) was adopted as a model system to investigate effective methods for enhancing embryogenesis. The focus was placed on attaining the optimum gaseous composition and the mixing intensity and viscosity of culture systems. The mixing intensity was investigated by using creased flasks and evaluated by shear force index (SFI). By using different closures with different ventilation coefficients, Kw, to adjust the gaseous composition in the flasks, it was found that embryo frequency (EF) increased with high P O 2 or low ethylene concentration in the headspace of the culture flasks. Cultivating the embryogenic cells under controlled ethylene concentrations depressed EF at ethylene concentrations higher than 0.08 ppm. Moreover, 0.1 lM of CoCl 2 remarkably repressed the ethylene formation in the culture medium and enhanced EF. P CO 2 and P O 2 around 3% and 30%, respectively, favored EF. For non-creased flasks, addition of CMC (carboxymethyl cellulose) decreased the EF and DO level of the medium, due to lowered mass transfer rate attributed to high viscosity. For creased flasks, supplementation of 0.4 g/ 100 ml CMC reduced the ill-effects of high shear stress and maximized EF (7.22%). Our results are potentially applicable to other species and largescale embryo production systems.

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CO 2 accumulation in bioreactor suspension cultures of Cyclamen persicum Mill. and its effect on cell growth and regeneration of somatic embryos

Cited by 36 publications

References 0 publications

Genotypic differences in callus formation and regeneration of somatic embryos in Cyclamen persicum Mill

Genotypic differences in callus formation and regeneration of somatic embryos in Cyclamen persicum Mill

Cryopreservation of embryogenic suspension cultures of Cyclamen persicum Mill.

Induction of somatic embryos of celery by control of gaseous compositions and other physical conditions

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