INFLUENCE OF CULTURE MEDIUM, EXPLANT LENGTH AND GENOTYPE ON MICROPROPAGATION OF <i>Pinus taeda</i> L.
DOI:
https://doi.org/10.5902/1980509817440Keywords:
loblolly pine, <i>in vitro</i> culture, forestry species, agar-water.Abstract
http://dx.doi.org/10.5902/1980509817440
This work aimed to establish a micropropagation protocol for Pinus taeda L. Apical shoots from 5-day seedlings, of different genotypes (F27, B05 and PC), were cultured on WV5 medium supplemented with 44 μM 6-benzylaminopurine (BAP) and 0.05 μM α-naphtaleneacetic acid (NAA), for 14 days, followed by subcultures on growth regulator-free medium. Explants were sectioned into apical shoots and nodal segments for multiplication. Different lengths of explants, BAP concentrations and cultivation periods were tested. Rooting was induced on WV5/2, WV3/2, GDm/2 and agar-water culture media supplemented with 2.68 μM NAA and 0.44 μM BAP for nine days, followed by a 4-week subculture on growth regulator-free medium. It was verified that genotype influenced shoot formation and rooting. The length of 1.0 cm is recommended for nodal segment explants to obtain a high number of axillary shoots. For apical shoots, 0.5 cm explant and WV5 medium formulation allowed the best results for elongation. The best period of subculture was eight weeks, both for nodal segments and for apical shoots. The higher percentage of nodal segments with shoots (99.2%) and the higher average number of shoots per explant (4.0) were obtained with F27 genotype in medium culture containing 2.5 μM BAP. For apical shoots, the best result for elongation was observed for the shorter explants (0.5 cm) on WV5 culture medium (218.3%). The maintenance of in vitro clonal micro garden of vigorous shoots was obtained for two years of 8-week subcultures on WV5 culture medium. The best rooting rate (55.6%) was obtained when shoots were inoculated in agar-water medium with 2.68 μM NAA and 0.44 μM BAP for nine days. Plantlets were successfully acclimatized (85% of survival), so a micropropagation protocol was established.
Downloads
References
ÁLVAREZ, J. M.; MAJADA, J.; ORDÁS, J. An improved micropropagation protocol for maritime pine (Pinus pinaster Ait.) isolated cotyledons. Forestry, v. 82, p. 175-184, 2009.
AMERSON,H. V. et al. Loblolly pine tissue culture: laboratory, greenhouse and field studies. In: HENKE, R. R. (ed.) et al. Tissue culture in forestry and agriculture. Plenum Press: New York, p.271-287, 1985.
BAXTER, R. et al. Production of clonal plantlets of tropical pine in tissue culture via axillary shoot activation. Can J For Res, v. 19, p. 1338-1342, 1989.
COKE, J. E. Basal nutrient medium for in vitro cultures of Loblolly pines. US Patent 5534434. http://www.freepatentsonline.com/5534434.html, 1996a.
COKE, J. E. Basal nutrient medium for in vitro cultures of Loblolly pines. US Patent 5534433. http://www.freepatentsonline.com/5534433.html, 1996b.
CORTIZO, M. et al. Benzyladenine metabolism and temporal competence of Pinus pinea cotyledons to form buds in vitro. Journal of Plant Physiology, v. 166, p. 1069-1076, 2009.
DE KLER, K et al. Regeneration of roots, shootbs and embryos: physiological, biochemical and molecular aspects. Biol Plant, v. 39, n. 1, p. 53-66, 1997.
DIAZ-SALA C. et al. Maturation-related loss in rooting competence by loblolly pine stem cuttings: The role of auxin transport, metabolism and tissue sensitivity. Physiol Plant, v. 97, p. 481-490, 1996.
GEORGE, E. F.; HALL, M. H.; DE KLERK, G. J. Plant propagation by tissue culture. 3rd ed. Springer: Berlin, 1998., v. 1.
GRESSHOFF, P. M.; DOY, C. H. Development and differentiation of haploid Lycopersicum esculentum (tomato). Planta, v. 107, p. 473-497, 1972.
HANDLEY, L. W et al. Research and development of commercial tissue culture systems in loblolly pine. Tappi J, v. 78, n. 5, p. 169-175,1995.
JANG, J. C.; TAINTER, F. H. Micropropagation of shortleaf, Virginia and loblolly pine x shortleaf pine hybrids via organogenesis. Plant Cell Tiss Org Cult, v. 25, p. 61-67, 1991.
KAUL, K. Factors influencing in vitro micropropagation of Pinus strobus L. Biol Plant, v. 32, n. 4, p. 266-272, 1990.
LAMBARDI, M.; SHARMA, K. K.; THORPE, T. A. Optimization of in vitro bud induction and plantlet formation from mature embryos of Aleppo pine (Pinus halepensis Mill.). In vitro Cell Dev Biol Plant, v. 29, p. 189-199, 1993.
LITVAY, J. D et al. Conifer suspension culture medium development using analytical data from developing seeds. Institute Paper Chemistry, Appleton (U. S.), 1981.
MEHRA-PALTA, A.; SMELTZER, R. H.; MOTT, R. L. Hormonal control of induced organogenesis – Experiments with excised plant parts of loblolly pine. Tappi J, v. 61, n. 1, p. 37-40, 1978.
MONTALBÁN, I. A. et al. Testing novel cytokinins for improved in vitro adventitious shoots formation and subsequent ex vitro performance in Pinus radiate. Forestry, v. 84, n. 4, p. 363-373, 2011.
NANDWANI, D.; KUMARIA, S.; TANDON, P. Micropropagation of Pinus kesiya Royle ex Gord (Khasi pine). Eur J Hort Sci, v. 66, p. 68-71, 2001.
OLIVEIRA, L. F. et al. Micropropagation of Pinus taeda L. from juvenile material. Tree For Sci Biotech, v. 6, n. 1, p. 96-101, 2012.
PULLMANN, G.S.; JOHNSON, S. Somatic embryogenesis in loblolly pine (Pinus taeda L.): improving culture initiation rates. Ann For Sci, v. 59, p. 663-668, 2002.
SEN, S. et al. Abscisic acid: a role in shoot enhancement from loblolly pine (Pinus taeda L.) cotyledon explants. Plant Cell Rep, v. 8, p. 191-194, 1989.
SILVEIRA, V. et al. Effect of plant growth regulators on the cellular growth and levels of intracellular protein, starch and polyamines in embryogenic suspension cultures of Pinus taeda. Plant Cell Tiss Org Cult, v. 76, n. 1, p. 53-60, 2004.
STOJICIC, D. et al. Micropropagation of Pinus peuce. Biologia Plantarum, v. 56, n. 2, p. 362-364, 2012.
TANG, W.; GUO, Z. In vitro propagation of loblolly pine via direct somatic organogenesis from mature cotyledons and hypocotyls. Plant Growth Regul, v. 33, p. 25-31, 2001.
TANG, W.; OUYANG, F. Plant regeneration via organogenesis from six families of loblolly pine. Plant Cell Tiss Org Cult, v. 58, n. 3, p. 223-226, 1999.
TANG, W.; WHETTEN, R.; SEDROFF, R. Genotypic control of high-frequency adventitious shoot regeneration via somatic organogenesis in loblolly pine. Plant Sci, v. 16, p. 167-272, 1998.
TEASDALE, R. D.; DAWSON, P. A.; WOOLHOUSE, H. W. Mineral nutrient requirements of a Loblolly Pine (Pinus taeda) cell suspension culture – Evaluation of a medium formulated from seed composition data. Plant Physiol, v. 82, p. 942-945, 1986.
WEBB, D. T.; FLINN, B. S.; GEORGIS, W. Micropropagation of eastern white pine (Pinus strobus L.). Can J For Res, v. 18, p. 1570-1580, 1988.
ZEL, J. Micropropagation of Pinus sylvestris. In: Ahuja MR (Ed.) Micropropagation of woody plants. Kluwer: Academic Publishers, Dordrecht, p. 347-365, 1993.
