RESEARCH ARTICLE: AGRICULTURAL BIOTECHNOLOGY AND FOOD REVISTA DE CIENCIAS AGRÍCOLAS Volumen 35(2):5-13 ISSN Impreso 0120-0135 e-ISSN 2256-2273 doi: http://dx.doi.org/10.22267/rcia.183502.86 In vitro arrow cane (Gynerium sagitatum Aubl.) multiplication in double phase medium Multiplicación in vitro Gynerium sagitatum Aubl.) en medio doble fase de caña flecha ( Claudia López D.1; Isidro E. Suárez2 1 Colombia, [email protected]. 2 Investigadora, Instituto de Biotecnología Aplicada para el Caribe (IBAC), Universidad de Córdoba, Montería, Colombia, [email protected]. Docente, Ph.D. Instituto de Biotecnología Aplicada para el Caribe (IBAC), Universidad de Córdoba, Montería, In vitro Gynerium sagitatum Aubl.) multiplication Revista de Ciencias Agrícolas Citar: Lopez, C. & Suarez, I. (2018). arrow cane ( in double phase médium. 35(2): 5-13 doi:http://dx.doi.org/10.22267/ rcia.183502.86 Received: august 25 2017. Accepted: september 24 2018. ABSTRACT Gynerium sagitatum Arrow cane ( Aubl.) is a Poaceae species used as fiber source to make traditional and valuable handmade craftsmanship by indigenous communities in Northern Colombia. Since no commercial crops are established fiber needs are taken from natural plant populations affecting ecosystem. A micropropagation protocol to clonally multiply large quantities of arrow cane plant material for planting commercial crops has been developed; however, micropropagated plants are costly compared to naturally extracted plant material. To reduce micropropagated plants costs, in the present research a double phase medium formulation along with continuous shoot culture with no periodic transfers to fresh medium was compared to semisolid medium system with subculture every four weeks with respect to multiplication rate and costs of micropropagated plants. The results showed that continuous culture of explants with double phase medium and no periodic transfers resulted in higher multiplication rates and larger shoots compared to shoots cultured using the conventional semisolid medium system and transfer to fresh medium every four weeks. Plants from both, semisolid and double phase culture system, fully adapted and recovered when transferred to ex vitro conditions. Keywords:The cost analysis In vitro showed that double phase cultured shoots are ≥20% lessex vitro expensive.. propagation, double phase, axillary meristems, BAP, UNIVERSIDAD DE NARIÑO Rev. Cienc. Agr. Julio - Diciembre 2018, 35(2): 5-13 6 López y Suárez - In vitro multiplication of cane arrow RESUMEN Gynerium sagitatum Aubl.) es una especie de la familia Poaceae utilizada como Caña flecha ( necesidadesfuente de fibra de parala industria elaborar son tradicionales tomadas de y plantacionesvaliosas artesanías naturales por afectando comunidades el ecosistema. indígenas Parade la proveerCosta Norte material Colombiana. vegetal paraDebido la siembraa que no de existen cultivos cultivos comerciales, comerciales, un protocolo la fibra para para las la multiplicación clonal masiva de plantas de caña flecha ha sido desarrollado; sin embargo, las plantas micropropagadas resultan costosas comparadas con los propágulos extraídos periódicasde las poblaciones a medio naturales.fresco comparado Con el fin con de el reducir sistema los convencional costos de las en plantas, medio semisólidoen el presente con transferenciasestudio se evaluó cada el cuatrouso de semanasmedio doble a medio fase fresco,y cultivo con continuo respecto de a brotesla tasa sinde multiplicacióntransferencias doble fase sin transferencias periódicas a medio fresco resultó en tasas de multiplicación más y costos de las plantas micropropagadas. Los resultados mostraron que el cultivo en medio altas y brotes de mayor longitud al compararlos con los brotes obtenidos en medio semisólido trasplantadosy transferencias a condiciones cada cuatro ex semanas. vitro Tanto los brotes cultivados en medio doble fase como los cultivados en medio semisólido se adaptaron y establecieron normalmente cuando fueron mediante transferencias mensuales .a El medio análisis fresco. de costos mostró que los brotes multiplicados en medio doble fase son ≥20% menos costosos y se recuperan ex vitro similar a los obtenidos Palabras clave: Propagación in vitro ex vitro. , doble fase, meristemos axilares, BAP, INTRODUCTION A Gynerium sagitatum Aubl.) is a Poaceae Arrow cane propagates by sexual and clonal meth and,- species native to West India and distributed ods; plants are dioceaus and flowers can be -1wind rrow cane ( pollinated, seeds index is around 1.7°C. million Flowering Kg usu when viable, seeds can germinate from three to sev- from México through Paraguay in the American en days after imbibition at 20-30 - continent. The plant is well adapted to inter tropical ally occurs at the stem terminal in 18-20 months old zone conditions with a better growth rate in wet low stem; however, under Humid Caribbean conditions sexual propagation does not occur because seeds lands, organic soils and altitude up to 1600m above are unviable. Clonal propagation is the current way sea level (GRIN, 2013). Cultivation and processing Zenú Indian group, most of for plant multiplication and dissemination; the new of arrow cane have been the main income source shoots emerge from underground rhizomes that for communities of the expand radially up to 20m from the main stem. The them dedicated to make craftsmanship products newly grown stems eventually mature and flower from the plant´s central nerve becoming the most becoming a new source for basal growing shoots. famous handmade Colombian products (DANE, This growth and propagation habit is not only very 2005). Recent studies evidenced the potential for efficient for colonization new territorieset al but also landfill phytoremediation using arrow cane alone an effectiveet way al to preserve wetlands and shores or in association with other plant species (Madera- from erosion and degradation (Kalliola ., 1992; Parra, 2015a, 2015b). Araméndiz ., 2005). UNIVERSIDAD DE NARIÑO Rev. Cienc. Agr. Julio - Diciembre 2018, 35(2): 5-13 López y Suárez - In vitro multiplication of cane arrow 7 A rate and costs variables. rrow cane plant extraction from wild populations micropropagation with respect to multiplication to make craftsmanship (hats, rings, shoes, etc.), MATERIALS AND METHODS homes, musical instruments and ornamental products happens at a very high rate since Plant material. Plant material consisted of in vitro commercially crops are not available. It is estimated that more than 50% of arrow cane becomingnatural populations a cultural, economicin the Colombian and environmental Northern established arrow cane plants cultivar “Criolla” Coast have been eliminated in the past 10 years cultivated for 12 months with monthly subcultures. The culture medium was MS (Murashige-1 and issue. Several studies have been conducted aimed Skoog, 1962)® supplied with (in mg® L ) myo inositol (100), sucrose (30.000), thiamine HCl (0.4) and at developing an efficientet propagation al methodet foral., natural restoration and commercial crops for fiber Phytagel (3.000) (Sigma Co .). The explants production (Araméndiz ., 2005, Suarez consisted of clusters with three 3-4 cm long shoots obtained from established plants after four weeks 2013). of subculture. The cultures were sstored). at 25°C with 12 h photoperiod provided-2 -2by cool white Micropropagation is a clonal propagation technique periods of time, reduced space and under aseptic fluorescent tubes (40-50 µmol m that allows massive plant multiplication in short Shoot multiplication. et al. 3 Clusters with three stems conditions (Sinhg , 2013; Waikhom and were established3 into 750cm polycarbonate flasks Louis, 2014). Micropropagated plants are cultured containing3 double phase medium consisting of from explants, established in closed containers with nutrient media and hormone supply that 100cm MS semisolid bottom medium added with result in high levels of genetic and phenotypic 30cm liquid MS medium on top of the semisolid uniformity. Usually, micropropagation requires phase; four explants were established in each container and the liquid phase was re-plenished fully equipped labs, relatively high amount ofet to avoid contamination. For conventional every two weeks inside of a laminar flow hood al.,reagents, well trained people and many labor micropropagationhours which results strategiesin high plant for costs arrow (Shinde cane 3 MS semisolid medium in order2016). to Several produce studies massive related plant to material developing for containedsemisolid insystem, 125cm 3similarly processed explants were cultivated in 30cm et al. polycarbonate flasks; a single Riveraplanting et commercialal cropset have al. been reported explant was established in each container and total (Pastrana and Suarez, 2009, Suarez , 2009; explants were transferred to fresh medium of the planting material., 2009; from Suarez natural populations., 2017); however, Double same formulation every four weeks. micropropagated plants results in higher costs than to reduce costs of Vitis vinifera et al., The medium formulation for both treatments was phase Ananasmedium comosus system have been implementedet al., MS (Murashige-1 and Skoog, 1962) supplied with Whitania somnifera (Couseloet al (in mg L ) mio inositol (100), sucrose® (30.000), 2006), (Scherwinski-Pereira thiamine HCl (0.4) and BAP (0.5); the semisolid 2012) and (Singh ., 2016) phase was added with Phytagel (3.000) (Sigma® micropropagated plants by means of reduced Co.). All flasks were covered