Micropropagation ofFagus sylvatica L.

Viéitez, A. M.; Ferro, Enrique; Ballester, A.

doi:10.1007/bf02632033

Cited by 13 publications

(11 citation statements)

References 18 publications

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“…Changes of medium within subculture cycles have also proved necessary for the stabilization of other Fagaceae, including Fagus sp. (Vieitez et al 1993b) and Q. robur (Vieitez et al 1994).…”

Section: Discussionmentioning

confidence: 95%

In vitro regeneration of the important North American oak species Quercus alba, Quercus bicolor and Quercus rubra

Viéitez

Corredoira

Ballester

et al. 2009

Plant Cell Tiss Organ Cult

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North American oak species, with their characteristic strong episodic seasonal shoot growth, are highly problematic for clonal micropropagation, resulting in the inability to achieve a stabilized shoot multiplication stage. The potential for initiating and proliferating shoot cultures derived from Quercus alba, Q. bicolor and Q. rubra explants was investigated, and a micropropagation method for these species was developed. Branch segments from 6 to 7-year-old trees were forced-flushed and the forced shoots were used as source of explants for culture initiation. A consistent shoot multiplication stage was achieved, in 13 of the 15 genotypes established in vitro, although marked differences occurred in explants from different genotypes/species. The control of efficient shoot multiplication involved the culture of decapitated shoots in a stressful horizontal position on cytokinin-containing medium with a sequence of transfers within a 6-week subculture cycle, which was beneficial to overcoming the episodic character of shoot growth. During each subculture cycle, the horizontally placed explants were cultured on media containing 0.2 mg l -1 benzyladenine (BA) for 2 weeks with two successive transfers (2 weeks each) to fresh medium with 0.1 mg l -1 BA, giving a 6-week subculture cycle. The general appearance and vigor of Q. alba and Q. bicolor shoot cultures were improved by the inclusion of both 0.1 mg l -1 BA and 0.5 mg l -1 zeatin in the medium used for the second transfer within the 6-week subculture cycle. Addition of AgNO 3 (3 mg l -1 ) to the shoot proliferation medium of Q. rubra had a significant positive effect on shoot development pattern by reducing deleterious symptoms, including shoot tip necrosis and early senescence of leaves. The three species showed acceptable in vitro rooting rates by culturing microcuttings in medium containing 25 mg l -1 indolebutyric acid for 48 h with subsequent transfer to auxin-free medium supplemented with 0.4% activated charcoal. Although an initial 5-day dark period generally improved the rooting response, it was detrimental to the quality of regenerated plantlets. However, activated charcoal stimulated not only the rooting frequencies, but it also enhanced plant quality, as evidenced by root, shoot and leaf growth.

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“…Changes of medium within subculture cycles have also proved necessary for the stabilization of other Fagaceae, including Fagus sp. (Vieitez et al 1993b) and Q. robur (Vieitez et al 1994).…”

Section: Discussionmentioning

confidence: 95%

In vitro regeneration of the important North American oak species Quercus alba, Quercus bicolor and Quercus rubra

Viéitez

Corredoira

Ballester

et al. 2009

Plant Cell Tiss Organ Cult

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“…In vitro shoots of E orientalis established from a 2-mo.-old seedling (clone 3) and a 4-yr-old plant (Bulgarian selection) were maintained on a shoot multiplication medium and subcultured every 6 wk, following the protocol previously reported for £ sylvatica (Vieitez et al, 1993). The shoot multiplication medium consisted of Woody Plant Medium (WPM; Lloyd and McCown, 1980), 2.2 p,M benzyladenine (BA), 9.1 I.tM zeatin, 2.9 ~M indole-3-acetic acid (IAA), 30 g sucrose per L, and 7 g agar (Sigma No.…”

Section: Methodsmentioning

confidence: 99%

Histological study of in vitro development of adventitious buds on leaf explants of oriental beech (Fagus orientalis lipsky)

Cuenca

Viéitez

1999

In Vitro Cell.Dev.Biol.-Plant

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Adventitious shoots were induced on the proximal portion of leaves excised from Fagus orientalis shoot cultures derived from a 2-mo.-old or a 4-yr-old seedling. Up to 90% of the explants formed between 13 and 19 buds after culture on Woody Plant Medium containing 2.9 pM indole-3-acetic acid and 4.5 pM thidiazuron. Adventitious buds developed mostly on petiole stub callus, but also on the midvein. The histological events leading to shoot organogenesis were examined. Some shoots developed directly from subepidermis or epidermis, but most originated indirectly from cell file proliferation produced by periclinally dividing cells subadjacent to the epidermis. Some cells in the outermost layers of these files became meristematic and divided extensively, resulting in the formation of meristemoids after 16 d of culture. Dedifferentiation into meristematic cells, which exhibited a large, prominent nucleus, densely-stained cytoplasm, and a high nucleus-to-cell area ratio, was generally associated with anticlinal divisions in cells previously originated by periclinal division. Subepidermal cell proliferation occurred mainly in the abaxial surface of the explant, which initially formed a diffuse cambium and later evolved to a phellogenic cambium. Some meristemoids were also differentiated by lenticel phellogen. Organized cell divisions in meristemoids gave rise to bud primordia that emerged from the explant surface and differentiated a protoderm. The progressive structural differentiation of the apical meristem, leaf primordia, and procambial strands led, after about 28 d of culture, to shoots with vascular connections with treachery elements previously differentiated in adjacent tissues.

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“…There are several reports concerning in vitro propagation of Fagus sylvatica based on organogenesis (Vieitez et al 1993;Vieitez and San-José 1996;Cuenca et al 2000). The regeneration of adventitious buds of this beech species was achieved from intact hypocotyls of whole plantlets obtained from embryonic axes, which were initial explants, but also from hypocotyl and cotyledon segments isolated from in vitro growing seedlings derived from embryos (Vieitez et al 1993), and from leaf and internode explants derived from in vitro grown shoots (Vieitez and SanJosé 1996;Cuenca et al 2000).…”

Section: Discussionmentioning

confidence: 99%

“…The regeneration of adventitious buds of this beech species was achieved from intact hypocotyls of whole plantlets obtained from embryonic axes, which were initial explants, but also from hypocotyl and cotyledon segments isolated from in vitro growing seedlings derived from embryos (Vieitez et al 1993), and from leaf and internode explants derived from in vitro grown shoots (Vieitez and SanJosé 1996;Cuenca et al 2000). Cuenca and coworkers (2000) succeeded in induction of adventitious buds from internode explants on WPM supplemented with 4.5 µM thidiazuron (TDZ) or 17.8 µM BA, although the medium containing TDZ was more efficient.…”

Section: Discussionmentioning

confidence: 99%

In vitro responses of various explants of Fagus sylvatica

Hazubska-Przybył¹,

Chmielarz²,

Bojarczuk³

2015

Dendrobiology

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There are limited published data on in vitro reproduction of Fagus sylvatica L. (European beech).This study was aimed to determine the efficiency of induction of somatic embryogenesis or organogenesis of beech from different types of explants in various culture conditions. Explants derived from immature, fresh seeds (collected in 2011 and 2013) and from mature seeds, stored at -10ºC and some stratified at 3ºC, were placed on induction media with various combinations of plant growth regulators: zeatin, 2,4-dichlorophenoxyacetic acid (2,4-D) and/or benzyladenine (BA). Initial cultures were kept in darkness or weak light (white fluorescent or blue-red LED). Limited success has been achieved in initiation of somatic embryogenesis. We obtained friable, yellow-white callus with characteristic PEM-like structures (cPEM-ls, from embryonic axes or fragments of immature embryos with embryonic axes), which may be an early developmental stage of embryogenic callus of Fagus sylvatica. This type of callus regenerated from explants incubated in darkness, mainly on WPM medium with addition of 6.8 µM zeatin or WPM and MSG media with 9.1 µM 2,4-D and 2.2 µM BA. The highest frequency of regeneration of callus with cPEM-ls was 5%. Instead, we succeeded to induce organogenesis from both immature and mature zygotic embryos and from embryonic axes. The best results were obtained for mature zygotic embryos incubated on ½WPM medium (half-strength Woody Plant Medium) with 9.1 µM 2,4-D and 2.2 µM BA. Adventitious buds were regenerated on up to 15% of the explants. The induced buds developed into shoots, enabling us to establish tissue cultures of beech. Induction of organogenesis from the tested explants was more efficient than induction of somatic embryogenesis.

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Micropropagation ofFagus sylvatica L.

Cited by 13 publications

References 18 publications

In vitro regeneration of the important North American oak species Quercus alba, Quercus bicolor and Quercus rubra

In vitro regeneration of the important North American oak species Quercus alba, Quercus bicolor and Quercus rubra

Histological study of in vitro development of adventitious buds on leaf explants of oriental beech (Fagus orientalis lipsky)

In vitro responses of various explants of Fagus sylvatica

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