Clonal propagation, callus formation and plant regeneration of lavandin

doi:10.1016/0304-4238(88)90158-6

Scientia Horticulturae

Volume 37, Issues 1–2, November 1988, Pages 157-163

https://doi.org/10.1016/0304-4238(88)90158-6 Get rights and content

Abstract

Methods for in vitro propagation of lavandin cultivar ‘Grosso’ (Lavandula officinalis Chaix × Lavandula latifolia Villars) are described. The culture medium was Linsmaier and Skoog medium supplemented with 30 g l⁻¹ sucrose, 7.5 g l⁻¹ agar and growth regulators. Micropropagation was performed using stem node explants. Various hormonal treatments were compared: 0.2 mg l⁻¹ benzylaminopurine (BAP) (Medium A); 0.2 mg l⁻¹BAP + 0.5 mg l⁻¹gibberellic acid (GA₃) (Medium B); 15 days in culture on 10 mg l⁻¹ BAP medium (Medium C) followed by transfer to either Medium A or Medium B (C/A; C/B). Proliferation rates were similar under all culture conditions. Rooting was promoted on Medium F, supplied with 1 mg l⁻¹ naphthaleneacetic acid (NAA). Plants propagated on Medium A showed the highest rooting rate. Flower parts (bud and calyx), leaf, stem node and shoot tip were used as callus sources. Callus formation was promoted on Medium E, supplemented with 1 mg l⁻¹ 2,4-dichlorophenoxyacetic acid + 0.5 mg l⁻¹ kinetin. Callus formation percentages were recorded and the minimum value was observed in stem node explants. Callus growth and organogenesis were also investigated. Shoot regeneration was induced on Medium C and shoot elongation on Medium D, provided with 1 mg l⁻¹BAP + 0.5 mg l⁻¹GA₃. Explants from various sources exhibited different proliferation rates. Rooting of regenerated plants was promoted on Medium G, supplied with 0.5 mg l⁻¹ NAA. Stem node callus exhibited the highest number of shoots with the maximum rooting rate. Both micropropagated and regenerated plants were easily acclimatized to outdoor conditions.

References (11)

D.V. Banthorpe et al.
Ability of plant callus cultures to synthesize and accumulate lower terpenoids
Phytochemistry
(1986)
J. Du Manoir et al.
In vitro propagation of fennel (Foeniculum vulgare Miller)
Scientia Hortic.
(1985)
M. Khosh-Khui et al.
Callus induction and culture of Rosa
Scientia Hortic.
(1982)
K.T. Sebastian et al.
A micropropagation system for carob (Ceratonia siliqua L.)
Scientia Hortic.
(1986)
J.K. Webb et al.
Monoterpene synthesis in shoots regenerated from callus cultures
Phytochemistry
(1984)

There are more references available in the full text version of this article.

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Furthermore, the efficacy of growth substances to induce organogenesis of explants is influenced by the presence of various concentrations of mineral salts in the nutrient medium [42]. In this study, callogenesis and caulogenesis were obtained independently after three months with low regeneration rates, while previous studies have recorded callus and shoots formation after a very short period and with high success rates [15,21,26,35,36,42–44]. Our data can be attributed to the type and size of explants used to propagate the target plants.
The overuse of natural medicinal plants forces the search for alternative production techniques to meet the growing needs, compensate for habitat degradation and conserve natural populations of plants. This study investigated the macropropagation and micropropagation of four medicinal plants from the Lamiaceae family growing in Algeria: Thymus algeriensis Boiss & Reut., Rosmarinus officinalis L., Marrubium vulgare L. and Salvia officinalis L. This baseline study aimed to describe biotechnology techniques that will help in restoring these important species and the developed callus culture can be a source of pharmaceutically important metabolites. It included two experiments: In Experiment 1, macrocuttings of 10−15 cm were cultivated in plastic pots containing soil and submitted in open greenhouse conditions. The rates of developed cuttings into whole plants were estimated. Experiment 2: segments of 5 mm taken from young stems were washed, sterilized using 0.1 % HgCl₂ and then cultured on semi-solid Murashige and Skoog (MS) medium supplemented with various combined concentrations of plant growth regulators (PGRs): indole-3-acetic acid (IAA) and Kinetin or N-6-furfuryladenine (Kin). The explants were incubated in a photoperiod (16 h light/25 °C, 8 h dark/15 °C) in a phytotron. The rates of callogenesis and caulogenesis were evaluated for different IAA/Kin combinations. The obtained results from macropropagation experiment showed significant differences (P < 0.001) in regeneration rates between the plant species. Salvia officinalis presented the highest success rate of regeneration (100 %) compared to other plants that showed low success rates. The combinations between IAA and Kin exhibited significant effects (P < 0.05 and P < 0.001) on callogenesis and caulogenesis. The best results of callus formation in T. algeriensis were observed at the combination of (5 mg/L IAA, 5 mg/L Kin) and (5 mg/L IAA, 10 mg/L Kin) where regeneration rates reached 23.3 % and 20 % respectively. In R. officinalis, the greatest callus production was obtained at (2.5 mg/L IAA, 5 mg/L Kin) with a rate of 56.7 %. In M. vulgare, the highest callus formation was marked at (2.5 mg/L IAA, 2.5 mg/L Kin), (2.5 mg/L IAA, 5 mg/L Kin) and (5 mg/L IAA, 5 mg/L Kin) with success rates varied between 33.3 % and 36.7 %. While S. officinalis can be significantly regenerated through macrocutting, the application of IAA and Kin combinations influenced greatly callus and shoots proliferation in T. algeriensis, R. officinalis and M. vulgare in in vitro micropropagation. The caulogenesis scores were low in the three above plants. Whereas, S. officinalis did not show any organogenic changes with all PGR combinations.
Trichomes, essential oils and in vitro propagation of Lavandula pedunculata (Lamiaceae)
2010, Industrial Crops and Products
Citation Excerpt :
Explants showing callus formation were not used for further experiments since genetic variability may occur in the plantlets obtained (Kaepler et al., 2000). Nevertheless, shoot organogenesis from basal callus is likely to occur and has been reported in several Lavandula species such as L. angustifolia (Quazi, 1980), L. x intermedia (Dronne et al., 1999), L. latifolia (Calvo and Segura, 1988; Jordan et al., 1990), L. officinalis x L. latifolia (Panizza and Tognoni, 1988) and L. vera (Tsuro et al., 2000). Organogenesis may be an alternative to shoot proliferation but a careful analysis of the propagated material must be performed to confirm the uniformity of the regenerated plants.
Lavanders (Lavandula spp.) are aromatic plants with high industrial and commercial value, used in food industry, perfumery and pharmaceutical preparations. Micropropagation can be used for large-scale multiplication of essential oil producing plants thus avoiding an overexploitation of natural resources. This work aims to develop a reliable protocol for the in vitro propagation of Lavandula pedunculata and to evaluate the potential of these plants for essential oil production with industrial application.
In vitro cultures were established on media with different concentrations of benzyladenine. Trichome morphology was examined by SEM and their secretion analyzed by histochemical tests. Essential oils were isolated by hydrodistillation and analyzed by GC and GC-MS. The essential oils and trichomes of in vitro plants were compared with those of field-growing plants.
Best propagation rates were achieved with 0.25 mg/l BA. Rooting occurred without treatment with auxins thus preventing callus growth. Trichomes and essential oils of the in vitro propagated plants were similar to those of the parent plants. Two chemotypes were characterized: 1,8-cineole/camphor type and fenchone type.
Since trichomes and essential oils of L. pedunculata micropropagated plantlets are identical to those produced by field-growing plants, in vitro cultures can be used for essential oil production without affecting natural resources. Moreover, the quality of the essential oils is assured and this protocol can be further applied to the propagation of selected chemotypes for industrial purposes.
Efficient plant regeneration from multiple shoots formed in the leaf-derived callus of Lavandula vera, using the 'open culture system'
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To develop an efficient procedure for plant regeneration from leaf-derived callus of Lavandula vera DC, the production of multiple shoots and the formation of roots from these shoots were studied. When calli were cultured in a medium with 4.0×10⁻⁷ M N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU), urea-type cytokinin, multiple shoots were obtained from the greenish surface of the callus efficiently at a rate of 52.2%. For multiple shoot development, the shoots were divided into 7–20 shoot clusters with 2–3 shoots and subcultured every 2 weeks. Two modified culture-systems were employed for subculture; a “closed culture system” and an “open culture system”. In the open-system, shoot clusters grew and elongated vigorously. In the closed-system, most of the clusters showed vitrification and did not elongate. In root induction culture, shoot clusters in the open-system were well rooted and grew vigorously, the highest rate of root formation being 74.0%, in the $12$ MS medium supplemented with 1.0×10⁻⁶ M indoleacetic acid (IAA). On the other hand, shoot clusters in the closed-system showed vitrification and did not root well. About seven times more regenerants were obtained from the multiple shoots than from normal shoots treated with 4.4×10⁻⁶ M 6-benzylamino purine (BA), a purine-type cytokinin. These results indicate that efficient plant regeneration in L. vera from callus using multiple shoots is possible by means of an open-system.
Comparative effect of different types of cytokinin for shoot formation and plant regeneration in leaf-derived callus of lavender (Lavandula vera DC)
1999, Scientia Horticulturae
Leaf-derived calli of Lavandula vera DC were cultured on Murashige and Skoog (MS) solid medium supplemented with purine or urea-type-cytokinins at 25°C under continuous light conditions. When calli were cultured on the medium with 6-benzylamino purine (BA) at 4.4 × 10⁻⁷–4.4 × 10⁻⁵ M, shoots were formed on green spots appearing on the callus surface; the highest value for shoot formation rate was 55.3% at 4.4 × 10⁻⁶ M. In the medium with thidiazuron (TDZ) or N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU), green spots were not observed, but callus-surface turned compact and greenish. Only multiple shoots were formed at all the concentration regimes. CPPU at 4.0 × 10⁻⁷ M was most effective for multiple shoot formation, 52.0%. Most shoots obtained in the presence of BA formed vigorous roots in 1/2 MS medium with or without 3-indolebutyric acid (IBA) at 9.9 × 10⁻⁷ M. Plantlets could be easily acclimatized in a greenhouse. On the other hand, multiple shoots obtained when using TDZ or CPPU could not form any root and died during the culture period.
Involvement of activated charcoal in the modulation of abiotic and biotic ethylene levels in tissue cultures
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Activated charcoal in the culture medium improved axillary bud proliferation of lavandin and growth of Anemone seedlings, but it negatively influenced root formation. In Anemone, severe medium browning was prevented by activated charcoal (AC). Abiotic ethylene released by the agarized medium was adsorbed by AC, depending upon the culture vessel and the medium volume. Biotic ethylene produced by lavandin nodes, laying directly on the medium surface, was easily adsorbed by AC. On the contrary, Anemone seedlings, 5 cm long, looked healthier in the presence of AC and evolved higher ethylene levels than the control. Therefore, the positive effect of AC on both species cannot be related to ethylene adsorption but probably to the removal of unidentified inhibitory substances from the culture medium.
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Scientia Horticulturae

Abstract

References (11)

Ability of plant callus cultures to synthesize and accumulate lower terpenoids

Phytochemistry

In vitro propagation of fennel (Foeniculum vulgare Miller)

Scientia Hortic.

Callus induction and culture of Rosa

Scientia Hortic.

A micropropagation system for carob (Ceratonia siliqua L.)

Scientia Hortic.

Monoterpene synthesis in shoots regenerated from callus cultures

Phytochemistry

Cited by (18)

New insights for the production of medicinal plant materials: Ex vitro and in vitro propagation of valuable Lamiaceae species from northern Africa

Trichomes, essential oils and in vitro propagation of Lavandula pedunculata (Lamiaceae)

Efficient plant regeneration from multiple shoots formed in the leaf-derived callus of Lavandula vera, using the 'open culture system'

Comparative effect of different types of cytokinin for shoot formation and plant regeneration in leaf-derived callus of lavender (Lavandula vera DC)

Involvement of activated charcoal in the modulation of abiotic and biotic ethylene levels in tissue cultures

Micropropagation and cryopreservation by vitrification of the Spanish endemic medicinal plant Sideritis leucantha Cav. subsp. leucantha (Lamiaceae)