Abstract
Here we report the establishment of a simple protocol for the micropropagation and acclimatization of U. minor. Branches with dormant buds were collected from mature elms and sprouted in a greenhouse. Tip and node segments were used as starting material for in vitro proliferation in a medium (designated here as DKW1) already used for the micropropagation of a clone of the English Elm (U. procera SR4). In the first assay, in which explants from nine different trees were used, 88.5% of the tip segments produced new axillary shoots thus proving to be the best explant type. Afterwards, material from four different trees (F4, F7, F13, F14), that had the highest sprouting rate in the greenhouse, was used to test for genotype influence. F14 proved to be the best genotype in culture and it was used for all the subsequent experiments. Shoots from F14 were used to assay in vitro rooting using five DKW based media. Rooting percentages were high for all media and varied between 80% and 100%. For acclimatization two approaches were assayed: the use of previously rooted in vitro plants and the direct acclimatization of shoots from cultures in DKW1. After 6 weeks, 86.4% of the in vitro rooted plants were successfully acclimatized and a slightly higher value, 88.6%, was attained by direct acclimatization of shoots with thick stems and hard leaves. These results proved that there is no need for a previous in vitro rooting step and that direct acclimatization can effectively reduce time and costs. Thus U. minor micropropagation and acclimatization can be divided into only two steps: proliferation of shoots in DKW1 and direct acclimatization of these shoots in a sterile soil mixture.
This is a preview of subscription content, log in via an institution to check access.
Abbreviations
- BAP:
-
Benzylaminopurine
- DED:
-
Dutch elm disease
- DKW:
-
Driver and Kuniyuki (1984) medium
- DKW1:
-
Medium by Fenning et al. (1993) with 4.4 μM BAP and 0.05 μM IBA
- IBA:
-
Indole-3-butyric acid
- NAA:
-
1-Naphtalene acetic acid
- Wgr:
-
Without growth regulators
References
Ben Jouira H, Hassairi A, Bigot C, Dorion N (1998) Adventitious shoot production from strips of stem in the Dutch elm hybrid ‘Commelin’: plantlet regeneration and neomycin sensitivity. Plant Cell Tissue Organ Cult 53:153–160
Ben Jouira H, Bigot C, Dorion N (2000) Plant regeneration from leaves of Ulmus × ‘Commelin’. ISHS Acta Horticulturae 520: XXV International Horticultural Congress, Part 10: application of biotechnology and molecular biology and breeding – in vitro culture. Brussels, Belgium
Biroscíková M, Spisáková K, Lipták S, Pichler V, Durkovic J (2004) Micropropagation of mature wych elm (Ulmus glabra Huds.). Plant Cell Rep 22:640–644
Bolyard MG, Hajela RK, Sticklen MB (1991a) Microprojectile and Agrobacterium-mediated transformation of ‘Pioneer’ elm. J Arboricult 17:34–37
Bolyard MG, Srinivasan C, Cheng J, Sticklen M (1991b) Shoot regeneration from leaf explants of American and Chinese elm. HortScience 26:1554–1555
Bonga JM, Von Aderkas P (1992) Clonal propagation. In: Bonga JM, Von Aderkas P (eds) In vitro culture of trees. Kluwer Academic Publishers, Boston, pp 72–125
Cheng Z-M, Shi N-Q (1995) Micropropagation of mature Siberian elm in two steps. Plant Cell Tissue Organ Cult 41:197–199
Conde P, Santos C (2006) An efficient protocol for Ulmus minor Mill. protoplast isolation and culture in agarose droplets. Plant Cell Tissue Organ Cult 86:359–366
Conde P, Loureiro J, Santos C (2004) Somatic embryogenesis and plant regeneration from leaves of Ulmus minor Mill Plant Cell Rep 22:632–639
Corchete MP, Fenning T, Gartland JS, Valle T (1997) Micropropagation of Ulmus species (elms). In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, high tech and micropropagation, vol 39. Springer-Verlag, Berlin, pp 381–392
Corredoira E, Vieitz AM, Ballester A (2002) Somatic embryogenesis in elm. Ann Bot 89:637–644
Dorion N, Danthu P, Bigot C (1987) Multiplication végétative in vitro de quelques espèces d’ ormes. Ann Sci For 44:103–118
Dorion N, Hassairi A, Guyon P, Godin B, Bigot C (1995) In vitro budding ability of woody internode and Agrobacterium susceptibility as prerequisites for elm genetic transformation. J Plant Physiol 146:699–703
Driver J, Kuniyuki A (1984) In vitro propagation of paradox walnut rootstock. HortScience 19:507–509
Dytham C (1999) Choosing and using statistics, a biologist’s guide. Blackwell Science Ltd, Oxford
Fenning TM, Gartland KMA, Brasier CM (1993) Micropropagation and regeneration of English elm, Ulmus procera Salisbury. J Exp Bot 44:1211–1217
Fink CVM, Sticklen MB, Lineberger RD, Domir SC (1986) In vitro organogenesis from shoot tip, internode, and leaf explants of Ulmus × ‘Pioneer’. Plant Cell Tissue Organ Cult 7:237–245
Gartland KMA, McHugh AT, Crow RM, Garg A, Gartland JS (2005) 2004 SIVB congress symposium proceeding: biotechnological progress in dealing with Dutch elm disease. In Vitro Cell Dev Biol Plant 41:364–367
Gautheret RJ (1940) Nouvelles recherches sur le bourgeonnement du tissue cambial d’ Ulmus campestris cultivé in vitro. C R Acad Sci 210:744–746
George MW, Tripepi RR (1994) Citokinins, donor plants and time in culture affect shoot regenerative capacity of American elm leaves. Plant Cell Tissue Organ Cult 39:27–36
GM trees fight DED (2001) BBC news. http://www.news.bbc.co.uk/1/hi/scotland/1512210.stm. Cited 27 August 2001
Harvengt L, Dumas E, Meier-Dinkel A, Collin E (2004) Establishment of a cryopreserved gene bank of European elms. Can J For Res 34:43–55
Kapaun J, Cheng Z (1997) Plant regeneration from leaf tissues of Siberian elm. HortScience 32:301–303
Karnosky DF, Podila GK (1993) Elm improvement via biotechnological methods. In: Sticklen MB Sherald JL (eds) Dutch elm disease research, cellular and molecular approaches. Springer-Verlag, New York, pp 75–80
Sticklen MB, Hajela RK, Bolyard MG, Graham LS, Sherald JL (1994) Genetic transformation in Ulmus species (elms). In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, plant protoplasts and genetic engineering V, vol 29. Springer-Verlag, Berlin, pp 401–410
Stipes RJ (2000) The management of Dutch elm disease. In: Dunn CP (ed) The elms: breeding, conservation and disease management. Kluwer Academic Publishers, Boston, pp 157–172
Acknowledgements
The authors thank Engineer Quadros, and his group, for the technical support during plant material harvest in “Mata Nacional do Choupal”. This work was financed by Project Praxis XXI (Praxis XXI/BD/3152/94) and Project FCT AGR/60672/2004.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Conde, P., Sousa, A., Costa, A. et al. A protocol for Ulmus minor Mill. micropropagation and acclimatization. Plant Cell Tiss Organ Cult 92, 113–119 (2008). https://doi.org/10.1007/s11240-007-9310-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-007-9310-8