Induction of somatic embryogenesis and analysis of genetic fidelity of in vitro-derived plantlets of Bambusa nutans Wall., using AFLP markers

Abstract

Bambusa nutans Wall., is an evergreen, perennial, and multipurpose bamboo having strong culms, which are largely used for construction, scaffolding, craft purposes, pulp, and paper industry. Multiple shoots from nodal segments (3-4 cm) of young branches of mature culms were established in Murashige and Skoog (1962) (MS) medium supplemented with various concentrations of 6-benzylaminopurine (BAP) (1.0-6.0 mg l -1 ) or in combination with a-naphthaleneacetic acid (NAA) (0.5-1.0 mg l -1 ) or kinetin (Kn) (1.0-2.0 mg l -1 ). February-March and December were found to be the best seasons for culture establishments. Maximum shoots were achieved on MS medium fortified with BAP (2.0 mg l -1 ). Embryogenic callus (slightly greenish compact, globular, and slow growing) was initiated from the base of severed sprouted buds in 2-3 subsequent subcultures on MS medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) (5.0 mg l -1 ) under dark incubations. Maturation and germination of well-organized somatic embryos was achieved on MS medium containing BAP and 2,4-D (1.0 mg l -1 each) with 20.0 mg l -1 ascorbic acid. Fullstrength MS medium supplemented with 2% glucose favored further development of proliferated somatic embryos into plantlets. Genetic variations of fieldestablished B. nutans plants regenerated through tissue cultures were assessed by amplified fragment length polymorphism (AFLP) analysis using 6 primer combinations. Four hundred and seven scorable fragments were amplified, of which 402 (98.8%) have recorded conservation at various morphogenetic stages leading to plantlets regeneration, therefore, revealed a high level of genetic stability.

The Effect of Gelling agent, medium pH and silver nitrate on adventitious shoot regeneration inSolanum tuberosum

“…Since shoot organogenesis was achieved through intermediate callus phase, therefore, clonal fidelity using regenerated shoots was tested using RAPD and ISSR markers. The utility of molecular markers in studying clonal fidelity of in vitro propagated plants is well documented (Khatab and El-Banna 2011, Kumar et al 2010, Rani and Raina 2000, Nag et al 2012, Mehta et al 2011). All the scored markers (both by RAPD and ISSR) were found to be monomorphic indicating the genetic uniformity of regenerated plants ( Figure 1C-D).…”

Induction of somatic embryogenesis and evaluation of genetic stability in regenerated plants of Magnolia dealbata

“…The values of genetic instability in this study (≤ 10 %) correspond to absent to low levels of variation reported for SE (0 -10 %) as in Cassia occidentalis, Ipomoea batatas, Picea abies, Distylium chinense, Quercus robur, Psidium guajava, Olea maderensis, O. europea, etc. (Fourré et al 1997, Lopes et al 2006, Valladares et al 2006, Triqui et al 2008, Yang et al 2008, Mehta et al 2011, Rai et al 2012, Bobadilla et al 2013, Li et al 2014, Carra et al 2016, Morais-Lino et al 2016, Naz et al 2016, Vinoth and Ravindhran 2016. It is possible that the explants selected may be less susceptible to genetic instability (DeVerno et al 1999) and, as a consequence, the results differ from those reported by others using the same morphogenetic route with intermediate levels of variation (17 -31 %) (Müller et al 1990, López et al 2004, Wilhelm et al 2005, Burg et al 2007, Prado et al 2010, Dey et al 2015.…”

“…SE in M. dealbata may reduce the genetic uniformity of the regenerants through the callus phase presented both during primary and secondary SE (Sharma et al 2007, Rai et al 2012. The culture environment might also present stresses leading to a loss of programmed cellular control (Mehta et al 2011). In Pinus pinaster and Quercus robur, prolonged propagation reduced genetic stability (Endemann et al 2001, Marum et al 2009.…”

Biotic Contamination and Possible Ways of Sterilization: A Review with Reference to Bamboo Micropropagation.

“…1% Bavistin is reported as most widely used concentration as surface sterilants during in vitro propagation of bamboo. Though there are exceptions also, Bavistin reported @ 0.1% in Bambusa nutans Wall 68 ; 0.5% in B.nutans 80 ; 0.2% in D. strictus 81 . Besides Bavistin, other fungicides that are reported till date as surface sterilants for in vitro propagation of bamboo are Benomyl and Mancozeb.…”

“…Explant collected at rainy season is having higher bud breaking coupled with high chances of contamination. More than 60% endogenous contamination during rainy season reported in B. nutans 68 . Collection of nodal explants (Arundinaria callosa Munro) during rainy season showed 30% more contaminations than that of after monsoon season 69 .…”