Abstract
We established an advanced protocol for in vitro propagation of Aloe vera via comparison of basal media, sucrose contents, growth hormone combinations, and additional supplementation with various polyamines. The maximal number and growth of shoots after 5 weeks was obtained using MS media including 30 g L−1 sucrose supplemented with 1.0 mg L−1 BA and 0.1 mg L−1 NAA. To improve shoot production, various concentrations of putrescine, spermidine, and spermine were added under optimal growth hormone conditions (MS media supplemented with 30 g L−1 sucrose, 1.0 mg L−1 BA, and 0.1 mg L−1 NAA). Maximal shoot number and growth after 5 weeks were achieved with supplementation of 50 mg L−1 spermidine. Regenerated plants were successfully acclimatized in soil with 100% efficiency. Cytogenetic inspection revealed that the regenerated plants maintained intact chromosomes identical to those of plants grown in field conditions. This protocol provides a valuable alternative for mass production of elite Aloe vera.
Similar content being viewed by others
Abbreviations
- BA:
-
6-benzylaminopurine
- B5 media:
-
Gamborg B5 medium
- MS media:
-
Murashige and Skoog media
- NAA:
-
1-naphthalene acetic acid
- SH media:
-
Schenk and Hildebrandt media
References
Abadi D, Kaviani B. 2010. In vitro proliferation of an important medicinal plant Aloe-A method for rapid production. Aust. J. Crop Sci. 4: 216–222
Adesoye A, Okooboh G, Akande S, Balogun M, Odu B. 2012. Effect of phytohormones and genotype on meristem and shoot tip culture of Telfairia occidentalis Hook F. J. Appl. Biosci. 49: 3415–3424
Ahlawat KS, Khatkar BS. 2011. Processing, food applications and safety of Aloe vera products: a review. J. Food Sci. Technol. 48: 525–533
Ahmad N, Guo B, Fazal H, Abbasi BH, Liu C-Z, Mahmood T, Shinwari ZK. 2011. Feasible plant regeneration in black pepper from petiole explants. J. Med. Plants Res. 5: 4590–4595
Baque MA, Shin Y-K, Elshmari T, Lee E, Paek K. 2011. Effect of light quality, sucrose and coconut water concentration on the microporpagation of Calanthe hybrids (‘Bukduseong’x‘Hyesung’and ‘Chunkwang’x‘Hyesung’). Aust. J. Crop Sci. 5: 1247–1254
Brandham P. 1971. The chromosomes of the Liliaceae: II: Polyploidy and karyotype variation in the Aloineae. Kew Bull. 25: 381–399
Brandham P, Johnson M. 1977. Population cytology of structural and numerical chromosome variants in the Aloineae (Liliaceae). Plant Syst. Evol. 128: 105–122
Chen T, Xu Y, Wang J, Wang Z, Yang J, Zhang J. 2013. Polyamines and ethylene interact in rice grains in response to soil drying during grain filling. J. Exp. Bot. 64: 2523–2538
Cheruvathur MK, Abraham J, Mani B, Dennis Thomas T. 2010. Adventitious shoot induction from cultured internodal explants of Malaxis acuminata D. Don, a valuable terrestrial medicinal orchid. Plant Cell Tiss. Org. 101: 163–170
Choi S, Chung M. 2003. A review on the relationship between Aloe vera components and their biologic effects. Semin. Integr. Med. 1: 53–62
Cvikrová M, Binarová P, Cenklová V, Eder J, Machácková I. 1999. Reinitiation of cell division and polyamine and aromatic monoamine levels in alfalfa explants during the induction of somatic embryogenesis. Physiol. Plant. 105: 330–336
Davidonis G. 1995. Changes in polyamine distribution during cotton fiber and seed development. J. Plant Physiol. 145: 108–112
Gamborg O, Miller R, Ojima K. 1968. Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50: 151–158
Grace O, Simmonds M, Smith G, Van Wyk A. 2008. Therapeutic uses of Aloe L.(Asphodelaceae) in southern Africa. J. Ethnopharmacol. 119: 604–614
Hashemabadi D, Kaviani B. 2008. Rapid micro-propagation of Aloe vera L. via shoot multiplication. Afr. J. Biotechnol. 7: 1899–1902
Kakkar R, Nagar P, Ahuja P, Rai V. 2000. Polyamines and plant morphogenesis. Biol. Plant. 43: 1–11
Liao Z, Chen M, Tan F, Sun X, Tang K. 2004. Microprogagation of endangered Chinese aloe. Plant Cell Tiss. Org. 76: 83–86
Meyer H, Staden J. 1991. Rapid in vitro propagation of Aloe barbadensis Mill. Plant Cell Tiss. Org. 26: 167–171
Murashige T, Skoog F. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 15: 473–497
Nakhooda M, Watt MP, Mycock D. 2012. The properties and interaction of auxins and cytokinins influence rooting of shoot cultures of Eucalyptus. Afr. J. Biotechnol. 11: 16568–16578
Novero A, Jamiri F. 2012. Plant regeneration through direct shoot formation from Sago palm (Metroxylon sagu Rottb.) leaf explants. Asian J. Biotechnol. 4: 92–99
Park E-H, Bae H, Park WT, Kim YB, Chae SC, Park SU. 2012. Improved shoot organogenesis of gloxinia (Sinningia speciosa) using silver nitrate and putrescine treatment. Plant Omics 5: 6–9
Paul A, Mitter K, Raychaudhuri SS. 2009. Effect of polyamines on in vitro somatic embryogenesis in Momordica charantia L. Plant Cell Tiss. Org. 97: 303–311
Petri C, Alburquerque N, Perez-Tornero O, Burgos L. 2005. Auxin pulses and a synergistic interaction between polyamines and ethylene inhibitors improve adventitious regeneration from apricot leaves and Agrobacteriummediated transformation of leaf tissues. Plant Cell Tiss. Org. 82: 105–111
Pua E, Sim G, Chi G, Kong L. 1996. Synergistic effect of ethylene inhibitors and putrescine on shoot regeneration from hypocotyl explants of Chinese radish (Raphanus sativus L. var. longipinnatus Bailey) in vitro. Plant Cell Reps. 15: 685–690
Rathore MS, Chikara J, Mastan SG, Rahman H, Anand K, Shekhawat N. 2011. Assessment of genetic stability and instability of tissue culture-propagated plantlets of Aloe vera L. by RAPD and ISSR markers. Appl. Biochem. Biotech. 165: 1356–1365
Rocha LFdS, de Sousa PA. 2013. Relationship between gibberellins and carbohydrates in vegetable products. Afr. J. Plant Sci. 7: 445–447
Schenk R, Hildebrandt A. 1972. Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Can. J. Bot. 50: 199–204
Shim S-W, Hahn E-J, Paek K-Y. 2003. In vitro and ex vitro growth of grapevine rootstock’ 5BB’ as influenced by number of air exchanges and the presence or absence of sucrose in culture media. Plant Cell Tiss. Org. 75: 57–62
Shohael A, Chakrabarty D, Ali M, Yu K, Hahn E, Lee H, Paek K. 2006. Enhancement of eleutherosides production in embryogenic cultures of Eleutherococcus sessiliflorus in response to sucrose-induced osmotic stress. Process. Biochem. 41: 512–518
Singh B, Sood N. 2009. Significance of explant preparation and sizing in Aloe vera L.-A highly efficient method for in vitro multiple shoot induction. Sci. Hortic. 122: 146–151
Tang W, Newton RJ. 2005. Polyamines promote root elongation and growth by increasing root cell division in regenerated Virginia pine (Pinus virginiana Mill.) plantlets. Plant Cell Rep. 24: 581–589
Tu R, Zheng B. 2012. Highly efficient in vitro adventitious shoot regeneration of Adenosma glutinosum (Linn.) Druce using leaf explants. Afr. J. Biotechnol. 11: 7542–7548
Velcheva M, Faltin Z, Vardi A, Eshdat Y, Perl A. 2005. Regeneration of Aloe arborescens via somatic organogenesis from young inflorescences. Plant Cell Tiss. Org. 83: 293–301
Velcheva M, Faltin Z, Vardi A, Hanania U, Eshdat Y, Dgani O, Sahar N, Perl A. 2010. Aloe vera transformation: the role of Amberlite XAD-4 resin and antioxidants during selection and regeneration. In Vitro Cell Dev-Pl. 46: 477–484
Vig B. 1968. Spontaneous Chromosome Abnormalities in Roots and Pollen Mother Cells in Aloe vera L. Bull. Torrey. Bot. Club. 95: 254–261
Viu A, Viu M, Tavares A, Vianello F, Lima G. 2009. Endogenous and exogenous polyamines in the organogenesis in Curcuma longa L. Sci. Hortic. 121: 501–504
Wallace H, Fraser A, Hughes A. 2003. A perspective of polyamine metabolism. Biochem. J. 376: 1–14
Du WP, Shi DX, Xu LY, Yu GR, Wang ML. 2004. A preliminary study on the induction and propagation of adventitious buds for Aloe vera L. China J. Arg. Sci. 17: 224–227
Yoon Y-J, Murthy HN, Hahn EJ, Paek KY. 2007. Biomass production of Anoectochilus formosanus hayata in a bioreactor system. J. Plant Biol. 50: 573–576
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lee, Y.S., Park, H.M., Park, S.U. et al. Optimal protocol for mass propagation of Aloe vera . J. Crop Sci. Biotechnol. 16, 285–290 (2013). https://doi.org/10.1007/s12892-013-0099-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12892-013-0099-1