Efficient, one step and cultivar independent shoot organogenesis of potato

Abstract

An efficient, one step and genotype independent protocol of shoot organogenesis was developed from leaf and internodal explants taken from microshoots of different cultivars of potato ( L.). Initially, microshoots were cultured on basal Murashige and Skoog medium additionally supplemented with 10 µM AgNO (MS1 medium) to achieve healthy shoot growth required to get the quality explants. Shoot organogenesis was induced from both types of explants (leaf and internodal) on MS1 medium variously supplemented with 6-benzyladenine (BA) and gibberellic acid (GA). Maximum explants were induced shoot organogenesis on MS1 medium supplemented with 10 µM BA and 15.0 µM GA from both the cultivars namely 'Kufri Chipsona 1' and 'Kufri Pukhraj'. Among the types of explants used, better response was observed from internodal segments as compared to leafs. This optimized medium combination was found to be equally effective for all the eight cultivars tested namely 'Kufri Pukhraj', 'Kufri Chipsona 1', 'Kufri Chipsona 2', 'Kufri Jyoti', 'Kufri Surya', 'Kufri Chandramukhi', 'Kufri Khyati' and 'Desiree'. The clonal uniformity of the regenerated shoots was confirmed using random amplified polymorphic DNA and inter-simple sequence repeats markers.

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

“…The microshoot cultures of Solanum tuberosum L. cultivar 'Kufri Chipsona 1' (CS-1) maintained at Thapar Institute of Engineering & Technology, Patiala, were subcultured using nodal explants at the regular interval of 21 days on MS1 medium (Kaur et al 2017;Murashige and Skoog (1962) medium supplemented with 10 µM silver nitrate (AgNO3), 74mM sucrose and 0.7% (w/v) agar as gelling agent) to obtain fully expanded leaves for experimentation. All the experiments were carried out in glass culture bottles (Kasablanka, Mumbai) containing 30ml regeneration medium (Kaur et al 2017; MS1 medium supplemented 10 µM BA and 15 µM GA3), here onwards referred to as MS2 medium. All tissue culture grade chemicals were purchased from HiMedia Laboratories, Mumbai, India.…”

“…Regenerated shoots (randomly selected) were multiplied on MS1 medium and total genomic DNA was isolated using method described by Doyle and Doyle (1990). PCR amplifications were carried out using five each of RAPD and ISSR primers using previously described amplification conditions (Kaur et al 2017). Amplified products were separated on ethidium bromide stained agarose gel 1.2% (w/v) and visualised under UV trans-illuminator (Gel Doc Mega; Biosystematica, USA).…”

“…In potato, an important rabi food crop, shoot organogenesis is extensively studied using different explants such as leaf, internode and tuber discs (Yasmin et al 2003, Elaleem et al 2009, Molla et al 2011, Rezende et al 2013, Ghosh et al 2014, Abdelaleem 2015, Campos et al 2016, Kaur et al 2017). Most of the reports are mainly focussed on Plant Growth regulators (PGRs), but, studies on the role of other factors such as gelling agents, light intensity, medium pH, silver nitrate etc.…”

“…This can induce or inhibit differentiation of plant cells into adventitious shoots (Gonzalez et al 1997). The use of ethylene biosynthesis or action inhibitors such as silver nitrate, silver thiosulphate, aminoethoxyvinylglycine have shown improvement in shoot organogenic efficiencies in many plants including potato (Naik and Chand 2003, Mookkan and Andy 2014, Kaur et al 2017).…”

The Morphogenic Potential of Siberian Potato Cultivars in Tissue Cultures

“…У сортов культурного вида картофеля S. tuberosum L. получены растения-регенеранты практически из всех органов: из черешков листа (Yee et al, 2001), листовых пластинок (Andersson et al, 2003;Banerjee et al, 2006), стеблевых эксплантов (Kaur et al, 2017), клубневых дисков (Vasquez, Clarence, 2002), протопластов (Ehsanpour, Jones, 2001;Craig et al, 2005).…”