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Micropropagation of snapdragon

by J.M. Iriondo and E. Torres

Dpto. Biología Vegetal
E.U.I.T. Agrícola
Universidad Politécnica de Madrid
E-28040 Madrid, Spain


Regenerated plantlet
Cultivar "Floral Carpet"
Full size picture (99855 Bytes)

Introduction

Literature review

Conclusions

Bibliography

Abbreviations

Introduction

Multiple shoot formation,
Cultivar "Floral Carpet",
Full size picture (109270 Bytes)
Snapdragons can be propagated wih relative ease through sexual and conventional vegetative multiplication. Nevertheless, in vitro propagation has the advantage of rapidly cloning a selected plant. Fortunately, snapdragons can be included among the group of species that are highly adaptable and responsive to in vitro culture conditions.

Several works have been carried out on the in vitro culture of snapdragon cultivars. A large part of them deal with callus formation, protoplast isolation and plant regeneration from callus cultures, whereas others report on a direct way of micropropagation based on multiple shoot formation and the rooting of the formed shoots. The different developmental responses are obtained culturing the explants in nutritious media with different combinations and concentrations of auxins and cytokinins.

Literature review

Explant material, methods and results of published works on the in vitro culture of Antirrhinum majus can be summarized as follows:



Explant


Medium


pH


Sucrose

(g/l)


Agar

(g/l)


Plant Growth Regulators (mg/l)


Results


Author
Leaf and stem segments Murashige & Skoog (1962) 5.830 81 (2,4-D) Callus formationRao et al. (1976)
Stem internodeSangwan & Harada (1975) 5.520 60.25 NOA Callus formationPfister & Widholm (1984)
Seedling rootSangwan & Harada (1975) 5.520 60.25 (NOA) Callus formationPfister & Widholm (1984)
Shoot tipsMurashige & Skoog (1962) 5.730 71 (BAP) Callus formationAtkinson et al. (1989)
Seedling rootSangwan & Harada (1975) 5.520 60.25 (NOA) Shoot regeneration via callus Pfister & Widholm (1984)
Root callusSyono & Furuya (1972) 5.830 Liquid0.4 (2,4-D) Shoot regeneration via callus Pfister & Widholm (1984)
Stem internodePoirier-Hamon et al. (1974) 5.520 60.5-2 (2,4-D) Embryoids via callus Poirier-Hamon et al. (1974)
ProtoplastsPoirier-Hamon et al. (1974) ---20.5 Liquid1 (2,4-D) + 0.5 (BAP) Embryoids via colonies Poirier-Hamon et al. (1974)
Internode stem segments Nitsch et al (1967) 5.520 70.25-0.5 (NOA) or

0.5 (2,4-D)

Friable green callus embryogenesis Sangwan & Harada (1975)
Callus Murashige & Skoog (1962) 5.830 81 (2,4-D) + 0.5 (BAP) Indirect embryogenesis Rao et al. (1976)
Internode stem segments Nitsch et al. (1967) 5.520 72 (IAA) + 4 (Kin) Roots & multiple shoots Sangwan & Harada (1975)
Shoot tipsMurashige & Skoog (1962) 5.730 71 (BAP) Multiple shootsAtkinson et al. (1989)
Nodal segmentsMurashige & Skoog (1962) 5.830 80.5 (NAA) + 1 (BAP) Multiple shootsGonzález-Benito et al.

(1996)

Shoots Bourgin & Nitsch (1967) 5.510 8None Rooted plantletsPfister & Widholm (1984)
ShootsMurashige & Skoog (1962) 5.830 8None Rooted plantletsGonzález-Benito et al.

(1996)


Conclusions

Root development,
Cultivar "Floral Carpet"
Full size picture (127311 Bytes)
Published results show that snapdragon can be grown on a wide selection of media. In general terms, callus formation and shoot regeneration from callus can be induced at low auxin concentrations (e.g. Pfister & Widholm, 1984) and embryoids can also be formed in the presence of 2,4-D (Poirier-Hamon et al., 1974). Nevertheless, for clonal plant production, a micropropagation system which avoids callus formation is highly desirable to reduce the chances of obtaining somaclonal variants.

The presence of cytokinins (kinetin or BAP) in the medium seems to be favourable for multiple shoot formation (Atkinson et al., 1989; González-Benito et al., 1996). According to Newbury et al. (1992) less callus was formed when shoot tips cultured on 1mg/l BAP, were excised from older plants (greenhouse grown plants vs. 48 day old seedlings). In González-Benito et al. (1996), no callus growth was observed on most media tested.

Rooting is achieved when shoots are cultured in the absence of plant growth regulators (Sangwan & Harada, 1975; Pfister & Widholm, 1984; González-Benito et al., 1996). In González-Benito et al. (1996), the use of IBA at concentrations of 2 mg/l and higher was detrimental to root formation.

The last stage of micropropagation, that is the acclimatization to ex vitro conditions and transfer to the greenhouse, which is critical in many species, can be successfully achieved in snapdragon. For instance, in González-Benito et al. (1996), a 90% survival was obtained when rooted vitroplants were transferred to pots with an autoclaved 3 peat : 1 vermiculite v/v mixture. The potted plants were kept in fully enclosed propagators with transparent lids and the windows of the micropropagators were opened for increasing periods of time each day for 15 days, before being transferred to the greenhouse.

A similar general pattern of response has been found when different snapdragon cultivars have been cultured in the same media, although some differences occur (Newbury et al. 1992). Moreover, the same can be said with regard to the micropropagation of wild types of snapdragon (Antirrhinum majus ssp. barrelieri and A. microphyllum) (González-Benito et al., (1996).

A general protocol for the in vitro propagation of snapdragon would consist of the following: Nodal segments or shoot tips from "mother plants” can be used as explants. The explants can then be cultured on MS + 1 mg/l BAP for multiple shoot formation. Root formation of isolated shoots can take place on MS medium without growth regulators. Initially high moisture conditions are needed for the acclimatization of vitroplants before transfer to greenhouse.

The use of microproopagation techniques in snapdragon may be especially best suited for the propagation of triploid cultivars where sexual propagation is not possible.

Bibliography

Atkinson, N.J. et al. (1989) Regeneration of plants from Antirrhinum majus L. callus. Plant Cell Tiss. Organ. Cult. 17:59-70

Atkinson, N.J. et al. (1991) In vitro adventitious root induction in Antirrhinum majus L. Plant Cell Tiss. Organ. Cult. 27:77-79

Bourgin, J.P. & Nitsch, J.P. (1967) Production of haploid Nicotiana from excised stamens. Ann. Physiol. Vég. 9:377-382.

González-Benito et al. (1996) Micropropagation of comercial and wild genotypes of snapdragon (Antirrhinum spp.) Journal of Horticultural Science 71:11-15

Murashige, T. & Skoog, F. (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol. Plant. 15:473-497

Newbury, H.J. et al. (1992) Micropropagation of snapdragon (Antirrhinum majus L.). In: Biotechnology in agriculture and forestry. High tech and micropropagation . IV. (Bajaj, Y.P.S., Ed.). Springer Verlag, Berlin, 19-33

Nitsch, J.P., Nitsch, C. Rossini, L.M.E. and Bui Dang Ha, D. (1967) The role of adenine in bud differentiation. Phytomorph. 17:446-453

Pfister, J. M. & Widholm, J.M. (1984). Plan regeneration from snapdragon tissue cultures. Hort Science, 19:852-854

Poirier-Hammon, S. et al. (1974). Culture of mesophyll protoplasts and stem segments of Antirrhinum majus (snapdragon): growth and organization of embryoids. Journal of Experimental Botany. 87: 752-60

Rao, P. S. etal. (1976) Gamma radiation and hormonal factors controlling morphogenesis in organ cultures of Antirrhinum majus L. cv. Red. Majestic Chief. Z. Pflanzenphysiol., 80:144-152

Sangwan, R.S. & Harada, H. (1975) Chemical regulation of callus growth, organogenesis, plant regeneration, and somatic embryogenesis in Antirrhinum majus tissue and cell cultures. Journal of Experimental Botany.26:868-81

Sangwan, R. S. et al. (1987) In vitro culture of shoot-tip meristems in some higher plants. Acta Hort. 212: 661-666

Sangwan, R.S. & Sangwan, J.S. (1990). Snapdragon. In: Handbook of plant cell culture. V. Ornamental species. (Ammirato, P.V., Evans, D.A., Sharp, W.R., Bajaj, Y.P.S., Eds). Springer Verlag, New York, 744-62

Syono, K. & Furuya, T. (1972) The differentiation of Coptis plants in vitro from callus cultures. Experientia 28:236

Abbreviations

2,4-D: 2,4 dichlorophenoxyacetic acid

BAP: 6-benzylaminopurine

IAA: 3-indoleacetic acid

IBA: 3-indolebutiric acid

Kin: kinetin; 6-furfurylaminopurine

NAA: 1-naphtylacetic acid

NOA: 2-naphthoxyacetic acid


This file has been submitted on April 15, 1997 by J. M. Iriondo.
It is part of the Snapdragon Home Page.
The URL of this page is http://www.mpiz-koeln.mpg.de/~stueber/snapdragon/methods/micropropagation.html