Abstract
Seedling production of canjerana (Cabralea canjerana (Vell.) Martius) has been limited by difficulty in germination, caused by recalcitrant behavior of their seeds. The objective of this study was to develop an efficient protocol for conservation and production of canjerana micropropagated plantlets. Seeds of canjerana were disinfected with 0, 2.5, 5.0, 7.5 and 10% of NaOCl solution, at immersion times of 10, 20 and 30 min to produce aseptic seedlings, which were cultivated on Murashige and Skoog (MS) and Woody Plant Medium (WPM) media. Nodal segments were cultivated in WPM medium supplemented with 0 and 2.5 µM of 6-benzylamine purine (BAP), kinetin (KIN) and thidiazuron (TDZ) or with 0, 1, 3, 6, 9 and 12 µM of BAP. Micro-cuttings were cultivated in MS and WPM media with 0 and 5.0 µM of indole-3-butyric acid (IBA) and naphthaleneacetic acid (NAA). The rooted micro-cuttings were acclimatized in a humid chamber in a greenhouse. The highest percentage of aseptic seedlings was produced with a solution of 7.5% of NaOCl and immersion times of 20 and 30 min. Adding BAP, KIN and TDZ or increasing concentrations of BAP to the WPM medium did not increase shoot number and length. Neither the base medium nor the auxin (IBA and NAA) had a significant effect on the survival of micro-cuttings after 60 days of cultivation, but the addition of 5.0 µM of NAA did increase the percentage of rooting and survival during the acclimatization. Both nodal segments and micro-stumps of canjerana have showed a low rate of multiplication. The addition of 5.0 µM of NAA to the basal medium increased the percentage of in vitro rooting and the percentage of survival during acclimatization of canjerana plantlets. Key words: In vitro propagation, in vitro multiplication, in vitro rooting, acclimatization, microclonal hedge.
Highlights
To complement conventional tree breeding programs, three specific biotechnological techniques have been successfully used, namely, tissue culture, for in vitro propagation and conservation; molecular markers, to assess the genetic diversity within and among populations; and encapsulation and cryopreservation, for long-term conservation (Varshney and Anis, 2014)
Seeds treated with 7.5% of NaOCl for 30 min had more than 80% of disinfection and more than 90% of these seeds germinated (Figure 2A and B)
The highest percentage of disinfection was found when seeds were treated with 7.5% of NaOCl for 20 or 30 min (Figure 2A), which was associated with the highest vigor, quantified by the lowest mean germination time (Figure 2D), the highest index of germination rate (IGR) (Figure 2E) and the highest percentage of normal seedlings (Figure 2C)
Summary
To complement conventional tree breeding programs, three specific biotechnological techniques have been successfully used, namely, tissue culture, for in vitro propagation and conservation; molecular markers, to assess the genetic diversity within and among populations; and encapsulation and cryopreservation, for long-term conservation (Varshney and Anis, 2014). Among these techniques, tissue culture is an appealing approach for conservation, because it aids in the propagation of species with limited number of explants, Afr. J. The fruit is used in popular medicine due to his biochemical properties against Trypanosoma cruzi, which causes chagasic disease (Fournet et al, 1996)
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