Regeneration from leaf explants and protoplasts of Brassica oleracea var. botrytis (cauliflower)

Chikkala, Veera R.N.; Nugent, Gregory D.; Dix, Philip J.; Stevenson, Trevor W.

doi:10.1016/j.scienta.2008.07.036

Cited by 21 publications

(10 citation statements)

References 30 publications

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“…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].…”

Section: Discussionsupporting

confidence: 84%

“…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].…”

Section: Discussionmentioning

confidence: 99%

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Investigating the In Vitro Regeneration Potential of Commercial Cultivars of Brassica

Farooq

Nawaz

Mukhtar

et al. 2019

Plants

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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.

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Section: Discussionsupporting

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Investigating the In Vitro Regeneration Potential of Commercial Cultivars of Brassica

Farooq

Nawaz

Mukhtar

et al. 2019

Plants

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“…Contrarily, GA 3 could not produce the highest shoot length response in some plant species (Abbasi et al 2010a, b). Chikkala et al (2009) concluded from their study that mature leaf explants produced fewer number of shoots than younger leaves of rapid-cycling B. oleracea and broccoli, but the number of shoots and regeneration frequency was higher in cauliflower. Kuginuki et al (1997) studied varietal differences in embryogenic and regenerative ability in the microspore culture of Chinese cabbage (B. rapa spp.…”

Section: Resultsmentioning

confidence: 99%

Efficient regeneration and antioxidative enzyme activities in Brassica rapa var. turnip

Abbasi

Khan

Guo

et al. 2010

Plant Cell Tiss Organ Cult

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The regeneration potential and antioxidative enzyme activities of economically important Brassica rapa var. turnip were evaluated. Calli were induced from leaf explants of seed-derived plantlets on Murashige and Skoog (MS) medium incorporated with different concentrations of various plant growth regulators (PGRs). The highest leaf explant response (83%) was recorded for 2.0 mg l -1 benzyladenine (BA) and 1.0 mg l -1 a-naphthaleneacetic acid (NAA). Subsequent subculturing of callus after 3 weeks of culture, on medium with similar compositions of PGRs, induced shoot organogenesis. The highest shoot induction response (83%) was recorded for 5.0 mg l -1 BA after 5 weeks of transfer. However, 7.8 shoots/explant were recorded for 2.0 mg l -1 BA. The transferring of shoots to elongation medium resulted in 5.1-cm-long shoots on 10 mg l -1 of gibberellic acid (GA 3 ). Rooted plantlets were obtained on MS medium containing different concentrations of indole butyric acid (IBA). The determination of activities of antioxidative enzymes (superoxide dismutase [SOD], ascorbate peroxidase [APX], catalase [CAT], glutathione peroxidase [GPX], and peroxidase [POD]) revealed involvement of these enzymes in callus formation and differentiation. All of the activities were interlinked with each other and played significant roles in the scavenging of toxic free radicals. This study will help in the advancement of a regeneration protocol for B. rapa var. turnip and the understanding of the functions of antioxidative enzymes in plant differentiation.

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“…To the best of our knowledge, there are still no reports on a stable genetic modification of cauliflower via the biolistic approach; only transient expression studies were reported by Brown and Wang (2004). Instead, this variety was modified by both direct DNA uptake (electroporation, PEG-mediated modification) and via the Rhizobium radiobacter and Rhizobium rhizogenes -mediated transformation (Chikkala et al 2008; Kumar Ray et al 2012; Radchuk et al 2002; Theriappan and Gupta 2014). However, despite those successful approaches, cauliflower is considered to be quite recalcitrant to genetic engineering and displays cultivar-dependent response to such modifications.…”

Section: Introductionmentioning

confidence: 99%

High efficiency transformation of Brassica oleracea var. botrytis plants by Rhizobium rhizogenes

et al. 2018

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Brassica oleracea var. botrytis, a very popular crop grown for its edible inflorescence, is bred only as a mutated annual cultivar and does not naturally occur in environment. Since cauliflower is still described as the most troublesome of all the B. oleracea vegetables regarding transformation processes, it is fully justified to focus on the improvement of tools for its genetic modifications. Here, we present a successful protocol for genetic transformation of cauliflower employing the process of agroinfection. The primary analysis of in vitro response of five cultivars allowed us to have chosen Pionier as the most promising cultivar; in consequence the Pionier was transformed via Rhizobium-mediated techniques in order to evaluate both, R. radiobacter (EHA 105, LBA 4404) and R. rhizogenes (ATCC 18534, A4) species. However, the latter system turned out to be more effective and, the A4 strain, in particular (72% transformation efficiency, 55% confirmed by GUS assay). That shows a promising technical advance especially when compared to the results of previous literature reports (e.g. 8.7% reported efficiency using R. rhizogenes). The transgenic cauliflower was obtained from hairy roots via organogenic callus induction. The potential transformants were analysed at the genomic and proteomic levels and their transgenic character was fully confirmed.Electronic supplementary materialThe online version of this article (10.1186/s13568-018-0656-6) contains supplementary material, which is available to authorized users.

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Regeneration from leaf explants and protoplasts of Brassica oleracea var. botrytis (cauliflower)

Cited by 21 publications

References 30 publications

Investigating the In Vitro Regeneration Potential of Commercial Cultivars of Brassica

Investigating the In Vitro Regeneration Potential of Commercial Cultivars of Brassica

Efficient regeneration and antioxidative enzyme activities in Brassica rapa var. turnip

High efficiency transformation of Brassica oleracea var. botrytis plants by Rhizobium rhizogenes

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