Keywords
plant growth regulators
tdz
zeatine
iba
acclimatization
cultivo de tejidos
reguladores de crecimiento
tdz
aib
zeatina
aclimatización
How to Cite
Abstract
Sharpblue, Woodard, Bonita and Delite cultivars were propagated from leaf explants. Leaf sections from in vitro singlenode explants were compared with those from plants in the field. The explants were placed in WPM medium supplemented with 1.5 mg.l-1 TDZ to promote sprouting, then subcultured to fresh WPM medium supplemented with 0.5 or 1.0 mg.l-1 zeatin to enhance adventitious shoot growth. Shoot clump length and number of shoots >2mm and >1cm in each clump were determined. Shoots 8 to 10 cm were harvested at 8 weeks and subcultured in WPM medium supplemented with 0, 1, 2, 4 or 8 mg.l-1 IBA for rooting. After 10 weeks, shoots were transplanted to greenhouse for acclimatization. The leaf segments cultured in WPM medium with TDZ formed adventitious shoot clumps in all varieties, regardless of leaf source. A significant effect of cultivar, source of explant and interaction cultivar x leaf source was found for shoots >2 mm. Clump size varied significantly among cultivars, leaf source, and interaction cultivars x source x zeatin concentration, when transfering to a medium with 2 concentrations of the growth regulator, but no differences due to cytokinin concentration were found. Number of shoots >2mm and length of the longest shoot varied significantly with cultivar and the interaction cultivar x source of explant in WPM + zeatin medium. Woodard explants from in vitro leaf sections produced the highest number of shoots (44.70±13.34) and Delite the lowest (21.50±4.99). Shoots subcultured in WPM with different IBA concentrations had les than 10% rooting in all treatments. However, when transplanted to greenhouse 100% rooting was obtained, regardless of in vitro treatment with IBA or leaf explant source. All the microshoots of the 4 cultivars were successfully greenhouse acclimatized and plantlets later established in the field.References
BAÑADOS M. 2006. Blueberry Production in South America. Acta Hort. 715:165-172.
BRISSETTE L., TREMBLAY L., LORD D. 1990. Micropropagation of Lowbush Blueberry from Mature Field-grown Plants. HortScience 25(3):349- 351.
CAO X., HAMMERSCHLAG F. 2000. Improved Shoot Organogenesis from Leaf Explants of Highbush Blueberry. HortScience 35(5):945–947.
CASTRO D., ÁLVAREZ J. 2013. Micropropagación clonal de tres genotipos mortiño, Vaccinium meridionale sw., por proliferación de yemas axilares. Actual Biol. 35(99):135-144.
CRISTONI A., MAGISTRETTI M. 1987. Antiulcer and healing activity of Vaccinium myrtillus anthocyanosides. Farmaco 42(2):29-43.
DEBNATH S. 2005. A Two-step Procedure for Adventitious Shoot Regeneration from in vitro-derived Lingonberry Leaves: Shoot Induction with TDZ and Shoot Elongation Using Zeatin. HortScience 40(1):189-192.
DEBNATH S. 2007. Strategies to propagate Vaccinium nuclear stocks for the Canadian berry industry. Agriculture. Canadian Journal of Plant Science 87(4):911-922.
DEBNATH S. 2009. A two-step procedure for adventitious shoot regeneration on excised leaves of lowbush blueberry. In Vitro Cell.Dev.Biol.-Plant. 45:122–128.
DEBNATH S. 2011. Adventitious shoot regeneration in a bioreactor system and EST-PCR based clonal fidelity in lowbush blueberry (Vaccinium angustifolium Ait.). Scientia Horticulturae 128:124-130.
DEBNATH S., McRAE K. 2001. In vitro culture of lingonberry (Vaccinium vitis-idaea L.): the influence of cytokinins and media types on propagation. Small Fruits Review 1:3-19.
DEBNATH S., VYAS P., GOYALI J., IGAMBERDIEV A. 2012. Morphological and molecular analyses in micropropagated berry plants acclimatized under ex vitro condition. Can. J. Plant Sci. 92:1065-1073.
FINN C., OLMSTEAD J., HANCOCK J., BRAZELTON D. 2014. Welcome to the Party! Blueberry Breeding Mixes Private and Public with Traditional and Molecular to Create a Vibrant New Cocktail. Acta Hort. 1017:51-61.
FISCHER D.L., VIGNOLO G., ALDRIGHI M., FACHINELLO J., ANTUNES L. 2012. Rooting of Blueberry Hardwood Cuttings as Affected by Wood Type. Acta Hort. 926:273-278.
GAJDOŠOVÁ A., OSTROLUCKÁ M., LIBIAKOVÁ G. 2006. Characterization of in vitro obtained clones of selected Vaccinium sp., and cultivars using RAPD analysis. Institute of Plant Genetics and Biotechnology. Slovak Academy of Sciences. 22 p.
GONZÁLEZ M., LÓPEZ M., VALDES A., ROJAS R. 2000. Micropropagation of three berry fruit species using nodal segments from field grown plants. Ann. Appl. Biol. 137:073-078.
HINE A., ABDELNOUR A. 2013. Establecimiento in vitro de arándano (Vaccinium corymbosum L). Tecnología en Marcha 26(4):64-71.
HINRICHSEN P., CASTRO M., RAVEST G., ROJAS G., MÉNDEZ M., BASSIL N., MUÑOZ C. 2009. Minimal Microsatellite Marker Panel for
Fingerprinting Blueberry Cultivars. Acta Hort. 810:173-180.
HOWELL A. 2009. Update on Health Benefits of Cranberry and Blueberry. Acta Hort. 810:779-784.
JAAKOLA L., TOLVANEN A., LAINE K., HOHTOLA A. 2001. Effect of N6-isopentenyladenine concentration on growth initiation in vitro and rooting of bilberry and lingonberry microshoots. Plant Cell Tiss. Org. Cult. 66:73-77.
JIANG Y., HONG Y., DE-QIAO Z., SHAN-AN H., CHUANYONG W. 2009. Influences of Media and Cytokinins on Shoot Proliferation of ‘Brightwell’ and ‘Choice’ Blueberries In Vitro. Acta Hort. 810:581-586.
KALT W., FORNEY C. MARTIN A., PRIOR R. 1999.vAntioxidant capacity, vitamin C, phenolics, and anthocyanins after fresh storage of small fruits. J. Agr. Food Chem. 47(11):4638–4644.
KAMEI H., KOJIMA T., HASEGAWA M., KOIDE, T., UMEDA, T., YUKAWA T., TERABE K.1995. Suppression of tumor cell growth by anthocyanins in vitro. Cancer Invest. 13:590-594.
LI Y., MA H., ZHANG Z., WU L. 2006. Effect of Cytokinins on In Vitro Leaf Regeneration of Blueberry. Acta Hort. 715:417-419.
LILA M., KELLOGG J., GRACE M., YOUSEF G., KRAFT T., ROGERS R. 2014. Stressed for Success: How the Berry’s Wild Origins Result in Multifaceted Health Protections. Acta Hort. 1017:23-43.
LITWIN´CZUK W., SZCZERBA G., WRONA D. 2005. Field performance of highbush blueberries (Vaccinium corymbosum L.) cv. ‘Herbert’ propagated by cuttings and tissue culture. Scientia Horticulturae 106:162-169.
LIU C., CALLOW P., ROWLAND L., HANCOCK J., SONG G. 2010. Adventitious shoot regeneration from leaf explants of southern highbush blueberry cultivars. Plant Cell Tiss Organ Cult 103:137-144.
LLOYD G., McCOWN B. 1981. Commercially feasible micropropagation of Mountain Laurel, Kalmia latifolia, by use of shoot tip culture. Combined Proceedings - Internaticlonal Plant Propagator’s Society 30:421-427.
MADRIZ J. 1999. Vaccinium: Especies silvestres neotropicales, perspectivas para la domesticación de nuevos frutales arbustivos, en los bosques montanos del neotrópico. Memorias IX Congreso Nacional Agronómico. p. 2.
MARINO S., WILLIAMSON J., OLMSTEAD J. 2014. Vegetative Growth of Three Southern Highbush Blueberry Cultivars Obtained from micropropagation and Softwood Cuttings in Two Florida Locations. HortScience 49(5):556-561.
MEINERS J., SCHWAB M., SZANKOWSKI I. 2007. Efficient in vitro regeneration systems for Vaccinium species. Plant Cell Tiss Organ Cult 89:169-176.
MORRISON S., SMAGULA G., LITTEN W. 2000. Morphology, growth, and rhizome development of Vaccinium angustifolium Ait. seedlings, rooted softwood cuttings, and micropropagated plantlets. HortScience 4:550-570.
MURASHIGE T., SKOOG F. 1962. A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15(3):473-497.
OSTROLUCKÁ M., LIBIAKOVÁ G., ONDRUŠKOVÁ E., GAJDOŠOVÁ A. 2004. In vitro propagation of Vaccinium species. Acta Universitatis Latviensis, Biology 676:207-212.
PEREIRA M. 2009. Reversion to juvenility: the use of epicormics in the micropropagation of mature wild shrubs of Vaccinium cylindraceum Smith (Ericacaeae). Arquipélago. Life and Marine Sciences 26:63-68.
PRODORUTTI D., PERTOT I., GIONGO L., GESSLE C. 2007. Highbush Blueberry: Cultivation, Protection, Breeding and Biotechnology. The European Journal of Plant Science and Biotechnology 1(1):44-56.
RACHE L., PACHECO J. 2010. Propagación in vitro de plantas adultas de Vaccinium meridionale (Ericaceae). Acta Botanica Brasilica 24(4):1086-1095.
REED B., ABDELNOUR A. 1991. The Use of Zeatin to Initiate in Vitro Cultures of Vaccinium Species and Cultivars. HortScience 26(10):1320-1322.
SEDLAK J., PAPRSTEIN F. 2009. In Vitro Multiplication of Highbush Blueberry (Vaccinium corymbosum L.) Cultivars. Acta Hort. 810:575-580.
SONG G., HANCOCK J. 2011. Basic Botany of Vaccinium, pp. 197-221. In: C. Kole (ed.). Wild Crop Relatives: Genomic and Breeding Resources, Temperate Fruits, Springer-Verlag Berlin Heidelberg.
TETSUMURA T., MATSUMOTO Y., SATO M., HONSHO C., YAMASHITA K., KOMATSU H., UGIMOTO Y., KUNITAKE H. 2008. Evaluation of basal media for micropropagation of four highbush blueberry cultivars. Scientia Horticulturae 119:72-74.
TREVISAN R., FRANZON R., NETO R., DA SILVA R., DIAS G., CORRÊA A. 2008. Enraizamento de estacas herbáceas de mirtilo: Influência da lesão na base e do ácido indolbutírico. Ciênc. Agrotec. 32:402-406.
Van KOOTEN O., BROUNS F. 2014. X International Symposium on Vaccinium and Other Superfruits. Maastricht, The Netherlands. Acta Horticulturae 1017.
WANG X., LI H., LI W., WU W., HU S. 2012. In vitro Direct Plant Regeneration from Leaves of Blackberry Cultivar ‘Boysenberry’. Acta Hort. 926:279-285.
ZHIDONG Z., LIU H., WU L., LI Y. 2006. Technical System of Blueberry Micropropagation in China. Acta Hort. 715:421-425