Micropropagation of Helleborus orientalis Lam. and Aconitum uncinatum Linn. (Ranunulaceae)

Lim, Chon C.; Kitto, S.L.

doi:10.21273/hortsci.30.4.871d

Cited by 5 publications

(7 citation statements)

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“…For H. niger, a satisfactory survival rate for rooting in vitro (87%) and ex vitro (85%) was obtained after chilling (7°C) microshoots in an IBA and NAA-containing solution for 7 days [18]. A low survival percentage (50-56%) was found for ex vitro rooting of H. orientalis [15]. Dhooghe and van Labeke [17] suggested ex vitro rooting for four Helleborus species which were induced for one week by drenching them in a solution of IBA and NAA at 5°C, but no data on the rooting rate and survival percentage was provided.…”

Section: Discussionmentioning

confidence: 97%

“…Helleborus species has been investigated in vitro and ex vitro [11,[15][16][17][18]. Studies on in vitro rooting have shown that cultivation of H. niger shoots on IBA and NAA-containing media resulted in a higher proportion of rooted shoots than when cultivation took place on auxin-free medium.…”

Section: Discussionmentioning

confidence: 99%

“…In vitro propagation of many plants has played a very important role in rapid multiplication of cultivars and the production of healthy plants. Relatively little tissue culture research has been published on the micropropagation of hellebores [11,[15][16][17][18][19][20]. The shoot multiplication efficacy of hellebores is influenced by several factors, such as: type of initial explants, genotype, growth regulators, and environmental factors [11,16,17].…”

Section: Introductionmentioning

confidence: 99%

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Effect of different sucrose and nitrogen salt levels in the medium and temperature on in vitro propagation of Helleborus niger L.

Gabryszewska

2015

Acta Agrobot

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Helleborus niger L. is a rhizomatous, herbaceous perennial with overwintering, divided, basal leaves. The objective of the study was to investigate the influence of different levels of sucrose (10, 20, 30, 40, 50, 60, 70, and 80 g l−1) and nitrogen salts (25%, 50%, and 100% according to MS medium) as well as temperature (15°C, 20°C) on in vitro multiplication and rooting and ex vitro acclimatization of H. niger. The growth and multiplication of axillary shoots were performed on modified MS medium supplemented with various growth regulators (2iP, BAP and kinetin – each at a concentration of 1.0 mg l−1, GA3 2.5 mg l−1). For the induction of roots, the medium was supplemented with IBA 1 mg l−1 and NAA 0.1 mg l−1. Rooted plants were transplanted in a peat–perlite substrate (4:1) in a heated greenhouse for ex vitro acclimatization. The multiplication rate of H. niger shoots, in vitro rooting, and ex vitro acclimatization were strongly dependent on the sucrose/nitrogen salt relationship in the medium. The highest multiplication rate of axillary shoots (3.7) was found at a temperature of 15°C or 20°C, on the medium with cytokinins and GA3 supplemented with sucrose 20–30 g l−1 and nitrogen salts at 50%. Sucrose at a concentration of 50 g l−1 strongly stimulated the number of roots per microplant (5.8–6.0) on the media with a reduced level of nitrogen salts (25% and 50%) when the temperatures were 20°C and 15°C, respectively. The plants rooted on the media with a high sucrose/nitrogen salt ratio showed acclimatization rates which ranged from 82% to 100%. Morphological observation of plantlets revealed obvious differences in leaf shape and size and the architecture of the root system as well as differences in the developmental stages of shoots grown on media with different sucrose and nitrogen salt concentrations.

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Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effect of different sucrose and nitrogen salt levels in the medium and temperature on in vitro propagation of Helleborus niger L.

Gabryszewska

2015

Acta Agrobot

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“…developed by Lim and Kitto (1995) is suitable for H. niger. The advantage of this system is that the rooting could be carried out ex vitro.…”

Section: Discussionmentioning

confidence: 99%

“…Springer (1995) reported a successful attempt at in vitro propagation without, however, giving detailed information about either the Helleborus species propagated or the cultivation method used. Lim and Kitto (1995) presented a system used for H. orientalis Lam. in which the propagation of shoots was carried out in vitro and the rooting was done ex vitro.…”

Section: Introductionmentioning

confidence: 99%

In vitro cloning of Helleborus niger

Seyring

2002

Plant Cell Rep

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This study shows that it is possible to propagate Helleborus niger by means of in vitro cloning. Universal (U) medium has been used for the in vitro culture. This medium was created by Haensch as a universal medium to meet the average nutritional requirements of many different plants. Apical buds of H. niger seedlings were established on U-medium supplemented with 8.9 µmol/l 6-benzylaminopurine (BAP) and 2.7 µmol/l α-naphthaleneacetic acid. Propagation was carried out on U-medium supplemented with 2.2 µmol/l BAP and 2.9 µmol/l gibberellic acid. The highest rooting success rate of 96.7% was achieved by adding 4.9 µmol/l indole-3-butyric acid to the medium. Shooting and rooting were dependent on the seedling cloned. More than 80% of the in vitro plants survived and thrived in the greenhouse.

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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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Micropropagation of Helleborus orientalis Lam. and Aconitum uncinatum Linn. (Ranunulaceae)

Cited by 5 publications

References 0 publications

Effect of different sucrose and nitrogen salt levels in the medium and temperature on in vitro propagation of Helleborus niger L.

Effect of different sucrose and nitrogen salt levels in the medium and temperature on in vitro propagation of Helleborus niger L.

In vitro cloning of Helleborus niger

Micropropagation of Helleborus through Axillary Budding

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