Establishment of efficient regeneration protocol for three rapeseed cultivars

Abstract

Economically, rapeseed is one of the most important crops in the world. Over the past decades, rapeseed research has been focused on improving biotechnological methods to facilitate breeding. The effectiveness of these methods depends on efficient tissue culture techniques. The aim of the present study was to establish an efficient protocol for regeneration of rapeseed. It was conducted in three stages. The first stage was to determine the effect of different concentrations (10%, 20%, 30% and 40%) of sodium hypochlorite solutions on seedling growth parameters. In the second stage, the effects of different growth media (Murashige and Skoog, MS, and Gamborg) and plant growth regulators (6-benzylaminopurine, 1-naphthaleneacetic acid and thidiazuron) at different concentrations on the regeneration capacity of stem explants of three rapeseed cultivars were investigated. In the last stage, the shoots of ‘Spok’ were cultured for three weeks on MS medium containing 1.5, 3 and 4.5 mg L−1 of indol butyric acid for rooting. The best results in germination, seedling growth and root length were obtained by using 10% disinfectant concentration for 25 minutes. The highest values for shoot regeneration were obtained from the stem explant cultured on MS medium containing 3 mg L−1 BAP and 0.2 mg L−1 NAA. It was found that MS containing 1.5 mg L−1 IBA was the most efficient medium for root formation.

Investigating the In Vitro Regeneration Potential of Commercial Cultivars of Brassica

“…Zhang and Bhalla [34] reported huge variations (0-96.7%) in seven Australian cultivars of Brassica napus. Similar genotype-dependent variations among different Brassica species have been reported in several studies [23,33,[35][36][37][38][39][40]43,[45][46][47]49].…”

“…Regeneration in Brassica has been reported from several tissues and organs including leaves, stem sections, petioles, roots, hypocotyls, cotyledons, immature zygotic embryos, protoplasts and cell suspension cultures [16,27,[33][34][35][36][37][38][39][40][41]. Out of these, cotyledons, hypocotyls and roots have been frequently used for genetic transformation [2,16,19,37,42,43].…”

“…The regeneration of these cultivars, including that of Aari canola, could be further improved upon by optimizing the plant regeneration conditions. It has been observed that varying the media components affects the shoot formation efficiencies significantly [33,[35][36][37]42,43,[45][46][47]. The fact that huge variations have been observed among different explants, a generalized recommendation cannot be made.…”

“…The explants were subjected to regeneration under three different regeneration protocols termed as Brassica Regeneration Protocols, BRP-I [23], BRP-II [53] and BRP-III [19]. A number of protocols for the in vitro regenerations of different Brassica genotypes have been reported in the literature, which are highly specialized for a certain genotype and are often complicated [1,3,16,28,29,[32][33][34][35][36][37]40,[42][43][44][45][46]48,[54][55][56][57][58][59][60]. Further, different explant types have been shown to respond differently to different hormonal combinations [44], which is further complicated by the fact that regeneration in Brassica is highly variable among different genotypes [42].…”