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Open AccessArticle

Investigating the In Vitro Regeneration Potential of Commercial Cultivars of Brassica

Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad 38000, Pakistan
Nuclear Institute for Agriculture and Biology, Faisalabad 38000, Pakistan
Department of Crop Genetics, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK
Authors to whom correspondence should be addressed.
Plants 2019, 8(12), 558;
Received: 10 November 2019 / Revised: 24 November 2019 / Accepted: 26 November 2019 / Published: 29 November 2019
(This article belongs to the Special Issue Plant Tissue Culture)
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. View Full-Text
Keywords: in vitro regeneration; tissue culture; commercial cultivars; Brassica juncea; Brassica napus in vitro regeneration; tissue culture; commercial cultivars; Brassica juncea; Brassica napus
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MDPI and ACS Style

Farooq, N.; Nawaz, M.A.; Mukhtar, Z.; Ali, I.; Hundleby, P.; Ahmad, N. Investigating the In Vitro Regeneration Potential of Commercial Cultivars of Brassica. Plants 2019, 8, 558.

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