Micropropagation protocol for the wild Brazilian greenberry (Rubus erythroclados)
- Paulo Mauricio Centenaro Bueno
, - Luiz Antonio Biasi,
- Mauro Brasil Dias Tofanelli
Abstract
This study presents the first micropropagation protocol for greenberry (Rubus erythroclados), a wild Brazilian species with edible green fruits. In the in vitro multiplication stage, three concentrations of benzyladenine (BA) were tested (0, 5 and 10 μM), combined with three concentrations of indolebutyric acid (IBA) (0, 3 and 6 μM) in two subsequent subcultures. In the rooting stage, in and ex vitro rooting were compared after pulse treatment of the microcutting for 10 seconds in IBA (0, 2.46, 4.92 and 7.38 mM). For the in vitro trial, the microcuttings were maintained in glass bottles with an MS medium under controlled conditions inside a growth room. For the ex vitro trial, the microcuttings were planted in styrofoam containers with vermiculite and maintained inside a greenhouse with an intermittent mist system. R. erythroclados multiplication was obtained with the addition of BA to the culture medium, while IBA reduced the shoot proliferation and increased mortality. The ex vitro rooting showed the best results, reaching 95.8% for rooted and acclimatizated plants without IBA. An efficient and simple protocol can be used for R. erythroclados micropropagation with 5 μM BA for in vitro shoot proliferation and ex vitro rooting of microcuttings with intermittent misting.
Keywords
Native species, tissue culture, rooting, microcutting, explant, growth regulators
Author Biography
Paulo Mauricio Centenaro Bueno
Agronomist, PhD in Plant Sciences.
Professor at IFPR, teaching plant phisiology, plant breeding, fruitculture and anual crops.
Luiz Antonio Biasi
Departamento de Fitotecnia e Fitossanitarismo
Setor de Ciências Agrárias
Mauro Brasil Dias Tofanelli
Departamento de Fitotecnia e Fitossanitarismo
Setor de Ciências Agrárias
References
- Augusto, C.S.S., L.A. Biasi, and C.A. Telles. 2006. Enraizamento e aclimatização de plantas micropropagadas de amoreira-preta cv. Brazos. Rev. Bras. Frutic. 28(3), 473-476. Doi: 10.1590/S0100-29452006000300029
- Campagnolo, M.A. and R. Pio. 2012. Enraizamento de estacas caulinares e radiculares de cultivares de amoreira-preta coletadas em diferentes épocas, armazenadas a frio e tratadas com IBA. Ciênc. Rural 42, 232-237. Doi: 10.1590/S0103-84782012000200008
- Clapa, D., A. Fira and N. Joshee. 2013. An efficient ex vitro rooting and acclimatization method for horticultural plants using float hydroculture. HortScience 48(9), 1159-1167.
- Cordeiro, J., C.V. Roderjan, and W.A. Rodrigues. 2011. Plantas lenhosas da floresta ombrófila mista do parque municipal das Araucárias – Guarapuava (PR). Ambiência 7, 441-460. Doi: 10.5777/ambiencia.2011.03.03
- Debnath, S.C. 2003. Micropropagation of small fruits. pp. 465-506. In: Jain, S.M. and K. Ishii (eds.). Micropropagation of woody trees and fruits. Kluwer Academic Publishers, Dordrecht, The Netherlands. Doi: 10.1007/978-94-010-0125-0_15
- Debnath, S.C. 2004. Clonal propagation of dwarf raspberry (Rubus pubescens Raf.) through in vitro axillary shoot proliferation. Plant Growth Reg. 43, 179-186. Doi: 10.1023/B:GROW.0000040110.53216.6a
- Del Castillo, A.R. and A.A. Zerda. 1990. Estudios preliminaries para la propagación clonal in vitro de mora (Rubus glaucus L.). Agron. Colomb. 7(1-2), 17-25.
- Deng, R. and D.J. Donnelly. 1993. In vitro hardening of red raspberry through CO2 enrichment and relative humidity reduction on sugar-free medium. Can. J. Plant Sci. 73(4), 1105-1113. Doi: 10.4141/cjps93-149
- Erig, A.C., A. De Rossi, and G.R.L. Fortes. 2002. 6-benzilaminopurina e ácido indolbutírico na multiplicação in vitro da amoreira-preta (Rubus idaeus), cv. Tupy. Ciênc. Rural 32(5), 765-770. Doi: 10.1590/S0103-84782002000500005
- Jin, W., Y. Gu, and S.Z. Zhen. 1992. In vitro propagation of Rubus species. Sci. Hortic. 49(3-4), 335-340. Doi. 10.1016/0304-4238(92)90169-D
- Lazic, T. and D. Ruzic. 2007. Organogenesis in vitro from the leaf of blackberry cv. Cacanska Bestrna. Genetika 39, 69-78. Doi: 10.2298/GENSR0701069L
- Leitzke, L.N., C.R. Damiani, and M.W. Schuch. 2009. Meio de cultura, concentração de AIB e tempo de cultivo no enraizamento in vitro de amoreira-preta e framboeseira. Rev. Bras. Frutic. 31(2), 582-587. Doi: 10.1590/S0100-29452009000200037
- Leitzke, L.N., C.R. Damiani, and M.W. Schuch. 2010. Influência do meio de cultura, tipo e concentração de citocininas na multiplicação in vitro de amoreira-preta e framboeseira. Ciênc Agrotec. 34(2), 352-360. Doi: 10.1590/S1413-70542010000200012
- Maia, A.J. and R.V. Botelho. 2008. Reguladores vegetais no enraizamento de estacas lenhosas da amoreira-preta cv. Xavante. Semina: Ciênc. Agrár. 29, 323-330. Doi: 10.5433/1679-0359.2008v29n2p323
- Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant 15, 473-479. Doi: 10.1111/j.1399-3054.1962.tb08052.x
- Oliveira, A.F., N.N.J. Chalfun, A.A. Alvarenga, J. Vieira Neto, R. Pio, and D.L. Oliveira. 2009. Estaquia de oliveira em diferentes épocas, substratos e doses de AIB diluído em NaOH e álcool. Ciênc. Agrotec. 33(1), 79-85. Doi: 10.1590/S1413-70542009000100011
- Oliveira, R.P. and A.F.P. Nino. 2009. Potencial de multiplicação in vitro de cultivares de framboeseira. Rev. Bras. Frutic. 31(1), 280-284. Doi: 10.1590/S0100-29452009000100040
- Oliveira, R.P., A.F.P. Nino, and L.V. Ferreira. 2008. Potencial de multiplicação in vitro de cultivares de amoreira-preta. Rev. Bras. Frutic. 30(3), 585-589. Doi: 10.1590/S0100-29452008000300004
- Pasa, M.S., G.L. Carvalho, M.W. Schuch, J.D. Schmitz, M.M. Torchelsen, G.K. Nickel, L.R. Sommer, T.S. Lima, and S.S. Camargo. 2012. Qualidade de luz e fitorreguladores na multiplicação e enraizamento in vitro da amoreira-preta ‘Xavante’. Ciênc. Rural 42(8), 1392-1396. Doi: 10.1590/S0103-84782012000800010
- Pelizza, T.R., J. Muniz, P. Camargo, A.A. Kretzschmar, and L. Rufato. 2013. Enraizamento ex vitro e aclimatização de plântulas micropropagadas de amoreira-preta 'Xavante'. Rev. Bras. Frutic. 35(1), 329-332. Doi: 10.1590/S0100-29452013000100039
- Poothong, S. and B.M. Reed. 2015. Increased CaCl2, MgSO4, and KH2PO4 improve the growth of micropropagated red raspberries. In Vitro Cell Dev. Biol-Plant 51, 648-658. Doi: 10.1007/s11627-015-9720-y
- Sigarroa-Rieche, A.K. and C.L. García-Delgado. 2011. Establecimiento y multiplicación in vitro de mora de castilla (Rubus glaucus Benth) variedade sin espinas, mediante ápices meristemáticos. Acta Agron. 60(4), 347-354.
- Taiz, L. and E. Zeiger. 2013. Fisiologia vegetal. 5a ed. Artmed, Porto Alegre, Brazil.
- Villa, F., M. Pasqual, F.A. Assis, L.A.S. Pio, and G.A. Assis. 2008. Crescimento in vitro de amoreira-preta: efeito de reguladores de crescimento e da cultivar. Ciênc. Agrotec. 32(6), 1754-1759. Doi: 10.1590/S1413-70542008000600012
- Welander, M. 1985. In vitro culture of raspberry (Rubus idaeus) for mass propagation. J. Hortic. Sci. 60(4), 493-499. Doi: 10.1080/14620316.1985.11515656
- Wu, J., S.A. Miller, H.K. Hall, and P.A. Mooney. 2009. Factors affecting the efficiency of micropropagation from lateral buds and shoot tips of Rubus. Plant Cell Tiss. Org. 99, 17-25. Doi: 10.1007/s11240-009-9571-5