In vitro plant regeneration of Albizia lebbeck (L.) from seed explants

Abstract

Objectives: An efficient and reproducible regeneration protocol for rapid multiplication of Albizia lebbeck (L.) was developed by using intact seed explants.Methods: Murashige and Skoog's (MS) medium supplemented with different hormones (BA, Kn, GA3 and TDZ) was used for the induction of multiple shoots from the seed explants. Ex-vitro rooting was performed by using pulse treatment method in auxins (IBA and NAA) and the complete plantlets were transferred to the field.Results: High frequency direct shoot induction was found in aseptic seed cultures of A. lebbeck on Murashige and Skoog medium supplemented with 5.0 µM TDZ (Thiadiazuron). Seeds were germinated after 7 days of culture and induced maximum 8 shoots from the region adjacent to the apex of the primary shoot of the seedling upto 25 days of incubation. Proliferating shoot cultures with increased shoot length was established by sub-culture of excised sprouting epicotyls on MS medium supplied with reduced concentrations of TDZ. Maximum shoot regeneration frequency (76 %) with highest number of shoots (21) and shoot length (5.1 cm) per sprouting epicotyl was observed in the MS medium supplemented with 0.5 µM TDZ after 8 weeks of culture. Different concentrations of Indole-3-butyric acid (IBA) and α-naphthalene acetic acid (NAA) were tested to determine the optimal conditions for ex-vitro rooting of the microshoots. The best treatment for maximum ex-vitro root induction frequency (81 %) was accomplished with IBA (250 µM) pulse treatment given to the basal end of the microshoots for 30 min followed by their transfer in plastic cups containing soilrite and eventually established in normal garden soil + soilrite (1:1) with 78 % survival rate. In addition, histological study was undertaken to gain a better understanding of the regenerated shoots from the epicotyl region.Conclusion: The findings will be fruitful in getting a time saving and cost effective protocol for the in vitro propagation of Albizia lebbeck.Keywords: Woody legume; TDZ; multiplication; epicotyl segment; shoot stumps.Abbreviations used: GA3 (Gibberellic acid); BA (6-Benzyladenine); Kn (Kinetin); TDZ (Thidiazuron); MS (Murashige and Skoog Medium); IBA (Indole-3-butyric acid); NAA (α-Naphthalene acetic acid).

Physiological and biochemical parameters influencing ex vitro establishment of the in vitro regenerants of Albizia lebbeck (L.) Benth.: an important soil reclaiming plantation tree

“…In vitro regenerated shootlets of A. lebbeck rooted by ex vitro rooting method gave the highest number of root regeneration response as compared to in vitro rooting (Perveen et al 2011;Perveen et al 2013b) and is less time consuming because of rooting and acclimatization took place simultaneously. The importance of ex vitro rhizogenesis over in vitro rooting have been reported by many researchers (Rogers and Smith 1992;Nas and Read 2004).…”

“…The in vitro regenerated shootlets were rooted successfully by using previously standardized ex vitro rooting (Perveen et al 2013b). The primary hardened plants on soilrite for 4 weeks (Fig.…”

“…For rooting 4-5 cm shootlets were rooted and acclimatized in soilrite by utilizing Perveen et al (2013b) method. After 4 weeks of soilrite acclimatization, plants were transferred to pots containing a mixture of garden soil [loam to clayey ] ?…”

“…Unsystematic and illicit erosion, low natural rejuvenation, constricted habitat specificity and insufficient conventional methods resulted in the rigorous depletion of A. lebbeck population (Perveen et al 2011). Therefore, keeping in views the outputs of the tissue culture techniques in woody trees, a number of efforts have been made for the in vitro propagation of A. lebbeck by using hypocotyl, root, cotyledon, leaflet, stem, petiole and seed explants (Gharyal and Maheshwari 1983;Mamun et al 2004;Gharyal and Maheshwari 1990;Perveen et al 2011Perveen et al , 2012Perveen et al , 2013b. For clonal plantlets production in vitro propagation through axillary shoots proliferation is a useful technique, while adventitious bud induction is a promising tool in order to obtained high frequency of plantlets at commercial level in a shorter period of time.…”

Cobalt induced augmentation of in vitro morphogenic potential in Erythrina variegata L.: a multipurpose tree legume

“…Implying, nutrients alone were not able to break the dormancy of the quiescent cells. Cytokinin, especially BA has been reported to overcome dormancy in many species including Acacia ehrenbergiana , Albizia lebbeck (Perveen et al 2011), Euphorbia cotinifolia (Perveen et al 2013) and Withania somnifera (Kulkarni et al 2000). These observations corresponds with reports that cytokinins can make quiescent cell competent to divide by activating various cell cycle check points (Francis and Sorrell 2001), thus breaking the dormancy.…”