Empleo de los Sistemas de Inmersión Temporal (SIT) en el cultivo in vitro de plantas
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Resumen
El desarrollo de Sistemas de Inmersión Temporal (SIT) para el cultivo in vitro, a fines de la década de 1990, anunció nuevos métodos semiautomáticos de micropropagación de plantas con fines agrícolas y de conservación. Para 2005, se habían desarrollado y probado varios sistemas y sus beneficios fueron usados con una variedad de cultivos de importancia comercial como la caña de azúcar, el café, el banano y el ñame. El empleo SIT ha demostrado ser una variante más eficiente de la micropropagación vía organogénesis sin comprometer la estabilidad genética del material vegetal. Se ha revisado el progreso en el campo y la adopción de estos sistemas en años anteriores. Se informan mejoras en los protocolos de SIT desarrollados anteriormente para especies económicamente importantes, y esto continúa dando fe de sus beneficios en comparación con los protocolos que emplean medios de cultivo semisólidos o líquidos. Aunque recientemente se han desarrollado protocolos en SIT para otras especies de plantas, esta lista es limitada. Por otro lado, existe un interés cada vez mayor en esta tecnología para la producción de metabolitos secundarios y estudios fisiológicos.
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Cómo citar
Toledo Rodríguez, E. A., Bernal Villegas, A., & Gómez Kosky, R. (2021). Empleo de los Sistemas de Inmersión Temporal (SIT) en el cultivo in vitro de plantas. Agricultura Tropical, 7(2), 1–9. Recuperado a partir de https://agriculturatropical.edicionescervantes.com/index.php/inivit/article/view/193
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AFREEN, F. 2006. Temporary immersion bioreactor, In: Gupta, S. D. and Y. Ibaraki (eds). Plant Tissue Culture Engineering. Springer Netherlands, p. 187-201.
ALAMILLA-MAGAÑA, J.C.; J.H. CAAMAL-VELÁZQUEZ, M.A. CRIOLLO-CHAN; J.E. VERA-LÓPEZ; J.A. REYES-MONTERO. 2019. Biofactory and temporary immersion bioreactor: In vitro propagation of Anthurium andreanum L., and economic viability. Revista Agroproductividad, 12(10): 23-29.
ALVARD, D.; F. COTE and C. TEISSON.1993. Comparison of methods of
liquid médium culture for banana micropropagation. Effects of
temporary immersion of explants. Plant Cell, Tissue and Organ
Culture, 32: 55-60.
ARAGÓN, C.E. 2015. Physiological characteristics as analyzed by hormone profile, metabolic pathways and expression of specific induced genes of C3, C4 and CAM tropical crops propagated by Temporary Immersion Bioreactors (TIB). Tesis presentada en opción al grado científico de Doctor en Ciencias Agronómica. Instituto de Agronomía, ʺUniversidad Técnica de Lisboa". 142 p.
ARAGÓN, C.E.; M. ESCALONA; R. RODRÍGUEZ; M.J. CAÑAL; I. CAPOTE; D. Pina and J. GONZÁLEZ-OLMEDO. 2010. Effect of sucrose, light, and carbon dioxide on plantain micropropagation in temporary immersion bioreactors. In Vitro Cell Dev. Biol. Plant, 46:89-94.
ARENCIBIA, A.D.; C. VERGARA; K. QUIROZ; B. CARRASCO. 2013. An approach for micropropagation of blueberries (Vaccinium corymbosum L.) plants mediated by temporary immersion bioreactors (TIBs). Am. Journal Plant Sciencie, 4(5): 1022-1028.
ARENCIBIA, A.D.; A. BERNAL VILLEGAS; C.M. ZAYAS; E.R. CARMONA; C. CORDERO; G. GONZÁLEZ; R. GARCÍA and I. SANTANA. 2012. Hydrogen peroxide induced phenylpropanoids pathway eliciting a defensive response in plants micropropagated in Temporary Immersion Bioreactors (TIBs). Plant Science, 195: 71-79.
BELLO-BELLO, J.J.; S. SCHETTINO-SALOMÓN; J. ORTEGA-ESPINOZA. 2021. A temporary immersion system for mass micropropagation of pitahaya (Hylocereus undatus). 3 Biotech, 11(437): 2-8.
BERNAL VILLEGAS, A. 2019. Compuestos fenólicos de caña de azúcar (Saccharum spp.) obtenidos en Biorreactores de Inmersión Temporal con actividad protectora contra patógenos. Tesis presentada en opción al grado científico de Doctor en Ciencias Agrícola. Universidad Agraria de la Habana "Fructuoso Rodríguez". 95p.
BERNAL VILLEGAS, A. Z. OCCEGUERA; M. JIMÉNEZ; O. RIVERA; L. GARCÍA and M. DE FERIA. 2002. Use of the Temporary Immersion Systems for sugarcane vitroplants production. Biotecnología Vegetal, 2(4): 201-206.
BERTHOULY, M.; H. ETIENNE. 2005. Temporary immersion system: a new concept for use liquid medium in mass propagation. In: Hvoslef-Eide, A.K. and W. Preil (eds). Liquid culture systems for in vitro plant propagation. Springer, Dordrecht. 165-195.
DE FERIA, M.; E. JIMÉNEZ; M. CHÁVEZ. 2002. Empleo de sistemas de inmersión temporal para la multiplicación in vitro de brotes de Saccharum spp. var. IBP 89-112. Biotecnología Vegetal, 2(3): 143-147.
DELFINO, P.; R. RIVATA; P. BIMA. 2020. Sistema de Inmersión Temporal (SIT): alta eficiencia en la propagación in vitro del portainjerto híbrido Prunuspersica x P. amygdalus. Nexo Agropecuario, 8(1): 13-18.
DUCOS, J.P.; L. CHARLES; P. VINCENT. 2007. Bioreactors for coffee mass propagation by somatic embryogenesis. International Journal of Plant Developmental Biology, 1(1): 1-12.
ESCALONA, M.; J.C. LORENZO; B. GONZÁLEZ; M. DAQUINTA; Z. FUNDORA; C.G. BORROTO; D. ESPINOSA; E. ARIAS and M.E. ASPIOLEA. 1999. New system for in vitro propagation of pineapple (Ananas comosus L. Merr). Pineapple News, 5:5-7.
GEORGIEV, M. and J. WEBER. 2014. Bioreactors for plant cells: hardware configuration and internal environment optimization as tools for wider commercialization. Biotechnology Lett, 1-9.
GEORGIEV, V.; A. SCHUMANN; A. PAVLOV and T. BLEY. 2014.Temporary immersion systems in plant biotechnology. Engineering in Life Sciences, 14(6): 1-15.
GEORGIEV, V.; I. IVANOV; S. BERKOV and A. PAVLOV. 2013. Temporary immersion systems for Amaryllidaceae alkaloids biosynthesis by Pancratium maritimum L. shoot culture. J. Plant Biochem. Biotechnol, 17(1):33-44.
GEORGIEV, V.; M. ILIEVA; T BLEY and A. PAVLOV. 2008. Betalain production in plant in vitro systems. Acta Physiology Plant, 30: 581-593.
GÓMEZ- KOSKY, R.; M. DE FERIA; L. POSADA; T. GILLIARD; F.M. BERNAL; M.V. REYES; M.M. CHÁVEZ and E.M. QUIALA. 2002. Somatic embryogenesis of the banana hybrid cultivar ‘FHIA-18’ (AAAB) in liquid medium and scale-up in a bioreactor. Plant Cell, Tissue and Organ Culture, 68: 21-26.
GONZÁLES, R.; S. RÍOS; F. AVILÉS and M. SÁNCHES-OLATE. 2011. In vitro multiplication of Eucalyptus globulus by temporary immersion system. Bosque. (in spanish), 32:147-154.
HUANG, T.K. and K.A. McDONALD. 2012. Bioreactor systems for in vitro production of foreign proteins using plant cell cultures. Biotechnoly. Adv., 30: 398-409.
IVANOV, I.; V. GEORGIEV; S. BERKOV and A. PAVLOV. 2012. Alkaloid patterns in Leucojum aestivum shoot culture cultivated at temporary immersion conditions. Journal Plant Physiology, 169: 206-211.
JIMÉNEZ, E.; N. PÉREZ- ALONSO; M. DE FERIA; R. BARBÓN; A. CAPOTE; M. CHÁVEZ; E. QUIALA and J.C. PÉREZ. 1999. Improved Production of potato microtubers using a Temporary Immersion System. Plant Cell Tiss Org Cult., 59: 19 23.
JOVA, M.; R. GÓMEZ-KOSKY; E.E. CUELLAR and A.E. CUELLAR. 2012. Efficiency of semi-automated culture systems on microtubers formation of yam (Discorea alata L.). Biotechnology. Agron. Soc. Environ., 16: 45-47.
LORENZO, J. C. 2000. Micropropagación de la caña de azúcar en Sistemas de Inmersión Temporal. Tesis para optar por el grado científico de Doctor en Ciencias Agrícolas. Centro de Bioplantas. UNICA. 100 p.
LUDWING-MULLER, J.; M. GEORGIEV and T. BLEY. 2008. Metabolite and hormonal status of hairy root cultures of Devil’s claw (Harpagophytum procumbens) in flasks and in a bubble column bioreactor. Process Biochemistry, 43(1): 15-23.
MARTÍNEZ RIVERO, A.; M.A. RAMÍREZ-MOSQUEDA; O. MOSQUEDA FRÓMETA; M. ESCALONA MORGADO, M. RIVAS PANECA; R. C. RODRÍGUEZ ESCRIBA; M. A. DAQUINTA GRADAILLE; J. J. BELLO-BELLO. 2020. Influence of Vitrofural® on sugarcane micropropagation using temporary immersion system. Plant Cell Tiss Organ Cult, 141: 447-453.
PALAZÓN, J.; A. MALLOL; R. EIBL; C. LETTENBAUER; R.M. CUSIDÓ and M.T. PIÑOL. 2003. Growth and ginsenoside production in hairy root cultures of Panax ginseng using a novel bioreactor. Planta Médica, 69(4): 344-349.
PAVLOV, A. and T. BLEY. 2008. Betalains biosynthesis by Beta vulgaris L. hairy root culture in a temporary immersion cultivation system. Process Biochem, 41: 848-852.
PÉREZ-ALONSO, N.; A. CAPOTE; A. GERTH and E. JIMÉNEZ. 2012. Increased cardenolides production by elicitation of Digitalis lanata shoots cultured in temporary immersion systems. Plant Cell Tiss. Organ Cult, 110: 153-162.
PÉREZ-ALONSO, N.; D. WILKEN; A. GERTH; A. JAHN; H.M. NITZSCHE; G. KERNS; G. CAPOTE-PÉREZ and E. JIMÉNEZ. 2009. Cardiotonic glycosides from biomass of Digitalis purpurea L. cultured in temporary immersion systems. Plant Cell. Tissue. Organ Cult, 99: 151-156.
RAMÍREZ-MOSQUEDA, M. A.; J.J. BELLO-BELLO.2021. SETIS™ bioreactor increases in vitro multiplication and shoot length in vanilla (Vanilla planifolia Jacks. Ex Andrews). Acta Physiol Plant, 43-52.
RITALA, A.; L. DONG; N. IMSENG and T. SEPPÄNEN-LAAKSO. 2014. Evaluation of tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) hairy roots for the production of geraniol, the first committed step in terpenoid indole alkaloid pathway. Journal Biotechnology, 176: 20-28.
SANTANA, I.; J. RODRÍGUEZ; O. NODARSE; A. BERNAL VILLEGAS; J.L. MONTE DE OCA; M. JIMÉNEZ; P. MACHADO; J.M. ROLO, A. PÉREZ y C.M. ZAYAS. 2014. Biofábrica de 5ta generación. En XXXVI Convención y EXPOATAM 2014 "Héctor M. Sáenz Couret", septiembre 2 al 5 de 2014. Revista ATAM, México. ISSN: 2007610: 10 p.
SCHUMANN, A.; S. BERKOV; D. CLAUS and A. GERTH. 2012. Production of galanthamine by Leucojum aestivum shoots grown in different bioreactor systems. Appl. Biochem. Biotechnol, 167: 1907-1920.
SENGAR, K.; R.S. SENGAR and G.S. KUMAR. 2011. The effect of in vitro environmental conditions on some sugarcane varieties for micropropagation. Afr. J. Biotechnol, 10(75): 17122-17126.
SEQUEIDA, A.; E. TAPIA; M. ORTEGA; P. ZAMORA; A. CASTRO; C. MONTES; G.E. ZÚÑIGA and H. PRIETO. 2012. Production of phenolic metabolites by Deschampsia antarctica shoots using UV-B treatments during cultivation in a photobioreactor. Electronic Journal of Biotechnology, 15(4): 10-18.
STANLY, C.; A. BHAT; A.M.D. ALI; C. L. KENG and B.P. LIM. 2010. A comparative study of Curcuma zedoaria and Zingiber zerumbet plantlet production using different micropropagation systems. Afr. J. Biotechnol, 9: 4326-4333.
STEINGROEWER, J.; T. BLEY; V. GEORGIEV; I. IVANOV; C. STANLY; A. BHATT; A.M.D. ALI; C. L. KENG and B.P. LIM. 2013. Bioprocessing of differentiated plant in vitro systems. Eng. Life Sc,13: 26-38.
WATT, P. 2012. The status of temporary immersion system (TIS) technology for plant micropropagation. Africa. J. Biotechnol, 3(3): 14025-14035.
ALAMILLA-MAGAÑA, J.C.; J.H. CAAMAL-VELÁZQUEZ, M.A. CRIOLLO-CHAN; J.E. VERA-LÓPEZ; J.A. REYES-MONTERO. 2019. Biofactory and temporary immersion bioreactor: In vitro propagation of Anthurium andreanum L., and economic viability. Revista Agroproductividad, 12(10): 23-29.
ALVARD, D.; F. COTE and C. TEISSON.1993. Comparison of methods of
liquid médium culture for banana micropropagation. Effects of
temporary immersion of explants. Plant Cell, Tissue and Organ
Culture, 32: 55-60.
ARAGÓN, C.E. 2015. Physiological characteristics as analyzed by hormone profile, metabolic pathways and expression of specific induced genes of C3, C4 and CAM tropical crops propagated by Temporary Immersion Bioreactors (TIB). Tesis presentada en opción al grado científico de Doctor en Ciencias Agronómica. Instituto de Agronomía, ʺUniversidad Técnica de Lisboa". 142 p.
ARAGÓN, C.E.; M. ESCALONA; R. RODRÍGUEZ; M.J. CAÑAL; I. CAPOTE; D. Pina and J. GONZÁLEZ-OLMEDO. 2010. Effect of sucrose, light, and carbon dioxide on plantain micropropagation in temporary immersion bioreactors. In Vitro Cell Dev. Biol. Plant, 46:89-94.
ARENCIBIA, A.D.; C. VERGARA; K. QUIROZ; B. CARRASCO. 2013. An approach for micropropagation of blueberries (Vaccinium corymbosum L.) plants mediated by temporary immersion bioreactors (TIBs). Am. Journal Plant Sciencie, 4(5): 1022-1028.
ARENCIBIA, A.D.; A. BERNAL VILLEGAS; C.M. ZAYAS; E.R. CARMONA; C. CORDERO; G. GONZÁLEZ; R. GARCÍA and I. SANTANA. 2012. Hydrogen peroxide induced phenylpropanoids pathway eliciting a defensive response in plants micropropagated in Temporary Immersion Bioreactors (TIBs). Plant Science, 195: 71-79.
BELLO-BELLO, J.J.; S. SCHETTINO-SALOMÓN; J. ORTEGA-ESPINOZA. 2021. A temporary immersion system for mass micropropagation of pitahaya (Hylocereus undatus). 3 Biotech, 11(437): 2-8.
BERNAL VILLEGAS, A. 2019. Compuestos fenólicos de caña de azúcar (Saccharum spp.) obtenidos en Biorreactores de Inmersión Temporal con actividad protectora contra patógenos. Tesis presentada en opción al grado científico de Doctor en Ciencias Agrícola. Universidad Agraria de la Habana "Fructuoso Rodríguez". 95p.
BERNAL VILLEGAS, A. Z. OCCEGUERA; M. JIMÉNEZ; O. RIVERA; L. GARCÍA and M. DE FERIA. 2002. Use of the Temporary Immersion Systems for sugarcane vitroplants production. Biotecnología Vegetal, 2(4): 201-206.
BERTHOULY, M.; H. ETIENNE. 2005. Temporary immersion system: a new concept for use liquid medium in mass propagation. In: Hvoslef-Eide, A.K. and W. Preil (eds). Liquid culture systems for in vitro plant propagation. Springer, Dordrecht. 165-195.
DE FERIA, M.; E. JIMÉNEZ; M. CHÁVEZ. 2002. Empleo de sistemas de inmersión temporal para la multiplicación in vitro de brotes de Saccharum spp. var. IBP 89-112. Biotecnología Vegetal, 2(3): 143-147.
DELFINO, P.; R. RIVATA; P. BIMA. 2020. Sistema de Inmersión Temporal (SIT): alta eficiencia en la propagación in vitro del portainjerto híbrido Prunuspersica x P. amygdalus. Nexo Agropecuario, 8(1): 13-18.
DUCOS, J.P.; L. CHARLES; P. VINCENT. 2007. Bioreactors for coffee mass propagation by somatic embryogenesis. International Journal of Plant Developmental Biology, 1(1): 1-12.
ESCALONA, M.; J.C. LORENZO; B. GONZÁLEZ; M. DAQUINTA; Z. FUNDORA; C.G. BORROTO; D. ESPINOSA; E. ARIAS and M.E. ASPIOLEA. 1999. New system for in vitro propagation of pineapple (Ananas comosus L. Merr). Pineapple News, 5:5-7.
GEORGIEV, M. and J. WEBER. 2014. Bioreactors for plant cells: hardware configuration and internal environment optimization as tools for wider commercialization. Biotechnology Lett, 1-9.
GEORGIEV, V.; A. SCHUMANN; A. PAVLOV and T. BLEY. 2014.Temporary immersion systems in plant biotechnology. Engineering in Life Sciences, 14(6): 1-15.
GEORGIEV, V.; I. IVANOV; S. BERKOV and A. PAVLOV. 2013. Temporary immersion systems for Amaryllidaceae alkaloids biosynthesis by Pancratium maritimum L. shoot culture. J. Plant Biochem. Biotechnol, 17(1):33-44.
GEORGIEV, V.; M. ILIEVA; T BLEY and A. PAVLOV. 2008. Betalain production in plant in vitro systems. Acta Physiology Plant, 30: 581-593.
GÓMEZ- KOSKY, R.; M. DE FERIA; L. POSADA; T. GILLIARD; F.M. BERNAL; M.V. REYES; M.M. CHÁVEZ and E.M. QUIALA. 2002. Somatic embryogenesis of the banana hybrid cultivar ‘FHIA-18’ (AAAB) in liquid medium and scale-up in a bioreactor. Plant Cell, Tissue and Organ Culture, 68: 21-26.
GONZÁLES, R.; S. RÍOS; F. AVILÉS and M. SÁNCHES-OLATE. 2011. In vitro multiplication of Eucalyptus globulus by temporary immersion system. Bosque. (in spanish), 32:147-154.
HUANG, T.K. and K.A. McDONALD. 2012. Bioreactor systems for in vitro production of foreign proteins using plant cell cultures. Biotechnoly. Adv., 30: 398-409.
IVANOV, I.; V. GEORGIEV; S. BERKOV and A. PAVLOV. 2012. Alkaloid patterns in Leucojum aestivum shoot culture cultivated at temporary immersion conditions. Journal Plant Physiology, 169: 206-211.
JIMÉNEZ, E.; N. PÉREZ- ALONSO; M. DE FERIA; R. BARBÓN; A. CAPOTE; M. CHÁVEZ; E. QUIALA and J.C. PÉREZ. 1999. Improved Production of potato microtubers using a Temporary Immersion System. Plant Cell Tiss Org Cult., 59: 19 23.
JOVA, M.; R. GÓMEZ-KOSKY; E.E. CUELLAR and A.E. CUELLAR. 2012. Efficiency of semi-automated culture systems on microtubers formation of yam (Discorea alata L.). Biotechnology. Agron. Soc. Environ., 16: 45-47.
LORENZO, J. C. 2000. Micropropagación de la caña de azúcar en Sistemas de Inmersión Temporal. Tesis para optar por el grado científico de Doctor en Ciencias Agrícolas. Centro de Bioplantas. UNICA. 100 p.
LUDWING-MULLER, J.; M. GEORGIEV and T. BLEY. 2008. Metabolite and hormonal status of hairy root cultures of Devil’s claw (Harpagophytum procumbens) in flasks and in a bubble column bioreactor. Process Biochemistry, 43(1): 15-23.
MARTÍNEZ RIVERO, A.; M.A. RAMÍREZ-MOSQUEDA; O. MOSQUEDA FRÓMETA; M. ESCALONA MORGADO, M. RIVAS PANECA; R. C. RODRÍGUEZ ESCRIBA; M. A. DAQUINTA GRADAILLE; J. J. BELLO-BELLO. 2020. Influence of Vitrofural® on sugarcane micropropagation using temporary immersion system. Plant Cell Tiss Organ Cult, 141: 447-453.
PALAZÓN, J.; A. MALLOL; R. EIBL; C. LETTENBAUER; R.M. CUSIDÓ and M.T. PIÑOL. 2003. Growth and ginsenoside production in hairy root cultures of Panax ginseng using a novel bioreactor. Planta Médica, 69(4): 344-349.
PAVLOV, A. and T. BLEY. 2008. Betalains biosynthesis by Beta vulgaris L. hairy root culture in a temporary immersion cultivation system. Process Biochem, 41: 848-852.
PÉREZ-ALONSO, N.; A. CAPOTE; A. GERTH and E. JIMÉNEZ. 2012. Increased cardenolides production by elicitation of Digitalis lanata shoots cultured in temporary immersion systems. Plant Cell Tiss. Organ Cult, 110: 153-162.
PÉREZ-ALONSO, N.; D. WILKEN; A. GERTH; A. JAHN; H.M. NITZSCHE; G. KERNS; G. CAPOTE-PÉREZ and E. JIMÉNEZ. 2009. Cardiotonic glycosides from biomass of Digitalis purpurea L. cultured in temporary immersion systems. Plant Cell. Tissue. Organ Cult, 99: 151-156.
RAMÍREZ-MOSQUEDA, M. A.; J.J. BELLO-BELLO.2021. SETIS™ bioreactor increases in vitro multiplication and shoot length in vanilla (Vanilla planifolia Jacks. Ex Andrews). Acta Physiol Plant, 43-52.
RITALA, A.; L. DONG; N. IMSENG and T. SEPPÄNEN-LAAKSO. 2014. Evaluation of tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) hairy roots for the production of geraniol, the first committed step in terpenoid indole alkaloid pathway. Journal Biotechnology, 176: 20-28.
SANTANA, I.; J. RODRÍGUEZ; O. NODARSE; A. BERNAL VILLEGAS; J.L. MONTE DE OCA; M. JIMÉNEZ; P. MACHADO; J.M. ROLO, A. PÉREZ y C.M. ZAYAS. 2014. Biofábrica de 5ta generación. En XXXVI Convención y EXPOATAM 2014 "Héctor M. Sáenz Couret", septiembre 2 al 5 de 2014. Revista ATAM, México. ISSN: 2007610: 10 p.
SCHUMANN, A.; S. BERKOV; D. CLAUS and A. GERTH. 2012. Production of galanthamine by Leucojum aestivum shoots grown in different bioreactor systems. Appl. Biochem. Biotechnol, 167: 1907-1920.
SENGAR, K.; R.S. SENGAR and G.S. KUMAR. 2011. The effect of in vitro environmental conditions on some sugarcane varieties for micropropagation. Afr. J. Biotechnol, 10(75): 17122-17126.
SEQUEIDA, A.; E. TAPIA; M. ORTEGA; P. ZAMORA; A. CASTRO; C. MONTES; G.E. ZÚÑIGA and H. PRIETO. 2012. Production of phenolic metabolites by Deschampsia antarctica shoots using UV-B treatments during cultivation in a photobioreactor. Electronic Journal of Biotechnology, 15(4): 10-18.
STANLY, C.; A. BHAT; A.M.D. ALI; C. L. KENG and B.P. LIM. 2010. A comparative study of Curcuma zedoaria and Zingiber zerumbet plantlet production using different micropropagation systems. Afr. J. Biotechnol, 9: 4326-4333.
STEINGROEWER, J.; T. BLEY; V. GEORGIEV; I. IVANOV; C. STANLY; A. BHATT; A.M.D. ALI; C. L. KENG and B.P. LIM. 2013. Bioprocessing of differentiated plant in vitro systems. Eng. Life Sc,13: 26-38.
WATT, P. 2012. The status of temporary immersion system (TIS) technology for plant micropropagation. Africa. J. Biotechnol, 3(3): 14025-14035.