Abstract
In vitro regeneration is a pre-requisite for developing transgenic plants through tissue culture-based genetic engineering approaches. Huge variations among different genotypes of the genus Brassica necessitate the identification of a set of regeneration conditions for a genotype, which can be reliably used in transformation experiments. In this study, we evaluated the morphogenesis potential of four commercial cultivars (Faisal canola, Punjab canola, Aari canola, Nifa Gold) and one model, Westar, from four different explants namely cotyledons, hypocotyls, petioles and roots on three different Brassica regeneration protocols, BRP-I, -II and -III. The regeneration efficiency was observed in the range of 6–73%, 4–79.3%, 0–50.6%, and 0–42.6% from cotyledons, petioles, hypocotyls and roots, respectively, whereas, the regeneration response in terms of average shoots per explant was found to be 0.76–10.9, 0.2–3.2, 0–3.4 and 0–2.7 from these explants. Of the commercial varieties tested, almost all varieties showed poorer regeneration than Westar except Aari canola. In comparison to Westar, its regeneration frequency from cotyledons was up to 7.5-fold higher on BRP-I, while it produced up to 21.9-fold more shoots per explant. Our data show that the explant has strong influence on the regeneration response, ranging from 24% to 92%. While the growth of commercial cultivars was least affected by the regeneration conditions provided, the effect on Westar was twice that of the commercial cultivars. After determining the optimal explant type and regeneration conditions, we also determined the minimum kanamycin concentration levels required to selectively inhibit the growth of untransformed cells for these cultivars. Regenerated shoots of Aari canola could be successfully grown to maturity within 16–18 weeks, with no altered phenotype noted and normal seed yields obtained. Therefore, the commercial variety, Aari canola, could be a good candidate for future genetic transformation studies.
Highlights
Brassica, from the family Brassicaceae, is an economically important genus
The explants would swell in the first week of incubation, and shoot primordia will start appearing in the second week
The present study was carried out to evaluate the regeneration potential of commercial cultivars of Brassica belonging to Brassica juncea and Brassica napus
Summary
From the family Brassicaceae, is an economically important genus. Brassica oilseed varieties producing oil low in anti-nutritive aliphatic glucosinolates and erucic acid as well as rich in unsaturated fatty acids are generally termed as ‘canola’. Was more often used for B. napus but some canola quality varieties of B. rapa and B. juncea are available [1,2,3,4]. Being rich in omega-6 and omega-3 fatty acids and low saturate fats, canola oil is Plants 2019, 8, 558; doi:10.3390/plants8120558 www.mdpi.com/journal/plants. Plants 2019, 8, 558 considered as a heart-healthy oil. Due to its high quantity of proteins, its meal for poultry and livestock is considered as good as soybean [5,6]
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