In vitro cloning of Helleborus niger

Seyring, M.

doi:10.1007/s00299-001-0420-1

Cited by 12 publications

(19 citation statements)

References 6 publications

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“…In our experiment, the main obstacle for effective micropropagation of Helleborus was too high temperature in the growth room. The cultures began to proliferate shoots when transferred to a growth room with a temperature of 15 o C. The beneficial effect of such a temperature for Helleborus micropropagation is known from the report of Seyring (2002). For rooting, Dhooghe and Van Labeke (2007), Beruto and Curir (2009) and Beruto et al (2013) applied pulse, 7 day-long chilling at 5-7 o C, together with drenching in auxin solution.…”

Section: Resultsmentioning

confidence: 99%

“…In a nursery environment, the chilling of young plants stimulated growth (Lowder et al, 2010) and accelerated flowering time (Christiaens et al, 2012). As it turned out, microshoot multiplication and rooting in vitro is more effective at a temperature of around 15 o C (Seyring, 2002;Poupet et al, 2006;Dhooghe and van Labeke, 2007;Beruto and Curir, 2009;Gabryszewska, 2014). Establishing of this condition can be a problem due to the cost of cooling.…”

Section: Introductionmentioning

confidence: 99%

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Burkholderia phytofirmansPsJN promotes in vitro rooting and acclimatization ofHelleborus

Orlikowska¹,

Nowak²,

Ogórek³

2017

Acta Hortic.

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Helleborus is a valuable ornamental plant, flowering in early spring or winter. Its hybrid cultivars can be propagated vegetatively, including in vitro via axillary shoots. One of the obstacles preventing effective micropropagation is the necessity for culture to be kept at a temperature of around 15 o C, which additionally increases costs. Knowledge obtained up to date shows that inoculating plants with bacteria Burkholderia phytofirmans PsJN can increase their tolerance to the non-optimal growth temperature. In this study, we tested whether the bacterium, which is known to produce a relatively high amount of IAA and demonstrates ACC deaminase activity, was able to stimulate Helleborus root formation at a temperature of 23 o C.In our experiment, the microshoots cultured without auxin and not inoculated with PsJN rooted in 83% with 1.7 roots of 0.6 cm long. The microshoots, which were induced to root with auxins IBA 3 mg/L and NAA 1 mg/L but not inoculated, were rooted in 94% with 2.0 roots of 1.1 cm long. A similar result was obtained for the microshoots not rooted on auxin containing medium but inoculated with PsJN -95% rooted shoots with 2.3 roots of 1.2 mm long. The microshoots that were induced to root on the auxins containing medium and then inoculated with PsJN were rooted in 100% with 6.9 roots of 2.1 cm long. Almost all microplants from this treatment acclimatized to greenhouse conditions and grew vigorously, then flowered after winter precooling in a mildly heated greenhouse.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Burkholderia phytofirmansPsJN promotes in vitro rooting and acclimatization ofHelleborus

Orlikowska¹,

Nowak²,

Ogórek³

2017

Acta Hortic.

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“…To explore these advantages an efficient propagation system for Helleborus species is crucial. Unfortunately, until now there is no easy way to multiply Helleborus (Seyring 2002). Methods of generative propagation are limited since the degree of variation in the offspring is often too high.…”

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confidence: 98%

“…The choice of the explants for initiation of an aseptic culture is important. Starting in vitro propagation with rhizome buds lead to a high degree of pathological contamination due to their subterranean origin (Seyring 2002). Successful in vitro protocols for H. niger were initiated from seedlings (Seyring 2002) or from meristem tip culture (Onesto et al 2004).…”

mentioning

confidence: 99%

“…Starting in vitro propagation with rhizome buds lead to a high degree of pathological contamination due to their subterranean origin (Seyring 2002). Successful in vitro protocols for H. niger were initiated from seedlings (Seyring 2002) or from meristem tip culture (Onesto et al 2004). In an attempt to increase the efficiency of propagation and to reduce the variation in the offspring we searched for an in vitro propagation system using axillary buds as initial explants.…”

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confidence: 99%

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In vitro propagation of Helleborus species

Dhooghe

Labeke

2007

Plant Cell Tiss Organ Cult

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A general in vitro cloning system was established for four Helleborus species: H. argutifolius, H. foetidus, H. niger and H. orientalis. The plant material was introduced in vitro from axillary buds. A Murashige and Skoog (MS)-based medium (Murashige and Skoog 1962) was used supplemented with 2% (w/v) sucrose, 2-isopentenyladenine (2-iP) and 6-benzylaminopurine (BA). Multiplication rates depended on the genotype and varied from 1.3 for H. foetidus till 3.8 for H. niger. The first results showed that the rooting phase could be done ex vitro. Rooting was induced by a drench for one week in a solution of indole-3-butyric acid (IBA -3 mg l -1 ) and 1-naphthaleneacetic acid (NAA-1 mg l -1 ) at 5°C.

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Micropropagation of Helleborus through Axillary Budding

Beruto

Viglione

Bisignano

2012

Methods in Molecular Biology

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Helleborus genus, belonging to the Ranunculaceae family, has 20 species of herbaceous perennial flowering plants. The commercial exploitation of this plant is dependent on the selection and propagation of appropriate lines. High propagation rate could be accomplished by using a suitable tissue culture method enabling the rapid introduction of valuable selections in the market. However, in vitro cultivation of Helleborus is still very difficult. Thereby the development of reliable in vitro propagation procedures is crucial for future production systems. Axillary buds cultured on agar-solidified Murashige and Skoog medium supplemented with 1 mg/L benzyladenine, 0.1 mg/L β-naphthoxyacetic acid, and 2 mg/L isopentenyl adenine develop shoots after 16 weeks of culture under 16 h light regime, 50-60 μmol/s/m(2), and 19 ± 1°C. The multiplication rate ranges from 1.4 to 2.1. However, the genotype and the number of subcultures affect the efficiency of the micropropagation process. The rooting of shoots is about 80% in solidified MS medium containing 1 mg/L 1-naphthaleneacetic acid and 3 mg/L indole-3-butyric acid. The described protocol provides information which can contribute to the commercial production of Helleborus plants.

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In vitro cloning of Helleborus niger

Cited by 12 publications

References 6 publications

Burkholderia phytofirmansPsJN promotes in vitro rooting and acclimatization ofHelleborus

Burkholderia phytofirmansPsJN promotes in vitro rooting and acclimatization ofHelleborus

In vitro propagation of Helleborus species

Micropropagation of Helleborus through Axillary Budding

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