In vitro Cultivation of Hancornia speciosa Gomes : The Physical Constitution of the Culture Medium, Sucrose Concentrations and Growth Conditions

Abstract

The effect of constitution of the culture medium, different concentrations of sucrose, and different growth conditions on the germination of seeds and growth of explants of ʺmangabaʺ (Hancornia speciosa Gomes) were evaluated in two in vitro assays. In assay one, the physical constitution of a medium (solid and liquid) and different concentrations of sucrose (0, 15, 30, 45, 60, 75, and 90 g/l) were tested, and seed germination was found to be variable. In assay two evaluation of the effect of different growth conditions (with and without agitation) on the in vitro growth of explants. Present results showed that ʺmangabaʺ had the highest percentage of germination and potentiality when inoculated in a liquid medium with (15 g/l) or without sucrose. The medium without agitation resulted in a better growth of explants the average length number of buds.

Clonal Fidelity and Genetic Diversity of Micropropagated Hancornia speciosa Gomes (Apocynaceae) as Evaluated by Molecular Markers

“…The seeds were manually washed under running water to remove the tegument and submersed for 10 min in a container with running water with three drops of Tween (80%). Then, they were immersed in a bowl with 70% (v/v) alcohol for 1 min and in a solution of sodium hypochlorite (20%) for 20 min, followed by a triple wash with autoclaved distilled water in a laminar flow, as described by [38]. For in vitro establishment, the seeds were germinated in glass tubes (25 × 150 mm) containing 20 mL on half-strength Murashige and Skoog salts ( 12 MS [42]) supplemented with vitamins [43], 30 g L −1 sucrose (Sigma ® ) (St. Louis, MO, USA) and 3.5 g L −1 agar (Sigma ® ) (St. Louis, MO, USA).…”

“…The pH of media was adjusted to 5.7 ± 0.03 prior to autoclaving. Culture tubes were maintained under a photoperiod of 16 h at 25 ± 3 • C, a relative humidity of 45%-46%, and an active photosynthetic radiation of 45-55 mol m −2 s −1 [38]. Once the in vitro establishment was completed (about 60 days), these plants were used as explants donors for further procedures (Figure 1).…”

“…Total genomic DNA was extracted from leaves of donor plants and their respective shoots (derived from nodal segments) or calluses (derived from leaves of donor plants) collected at each performed subculture, following standard procedures [38,44]. The concentration and quality of the extracted DNA was assessed via determination of A260/A280 absorbance ratio using a spectrophotometer (NanoDrop™, Technologies; Thermo Fisher Scientific, Inc., Wilmington, DE, USA).…”

“…Despite efforts to develop a reliable protocol for in vitro establishment and shoot multiplication for H. speciosa [12,20,[38][39][40], there is no research regarding the SV at the molecular level focused on this species. Accessing the genetic stability of plantlets recovered by tissue culture techniques at early stage through molecular markers is recommended since morphological variations in plants raised through tissue culture can be detected usually at later stages [41].…”

Coletânea bibliográfica acadêmica sobre a mangabeira (Hancornia speciosa Gomes)

“…Dos 13 estudos agrupados na área reprodução vegetal, 12 se referem especificamente à reprodução in vitro da mangabeira, tanto de maneira sexuada, isto é, por meio de sementes (Pereira-Netto 1996;Machado et al 2004;Lédo et al 2007;Reis 2011;Cabral et al 2013), quanto assexuada, por meio de segmentos nodais Lédo et al 2011;Jesus Sá et al 2012), segmentos caulinares (Soares et al 2007), microestacas (Jesus Sá et al 2011) e calos (Fráguas et al 2009). Todos os autores justificam seus estudos no fato de a mangabeira obter baixas taxas de germinação em viveiros, o que interfere no aproveitamento da espécie em larga escala, quer para usos econômicos ou para conservação florestal.…”