Abstract
The objective of this study was to evaluate the effects of 6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA) combinations, basal media and beta-lactam antibiotics on in vitro organogenesis from mature stem segments of 'Pêra', 'Valência' and 'Bahia' sweet oranges and 'Cravo' rangpur lime. For induction of shoot regeneration, the segments of the four cultivars were placed on Murashige and Skoog (MS) medium containing the following BAP/NAA concentrations: 0.0/0.0; 0.25/0.0; 0.25/0.25; 0.5/0.0; 0.5/0.5; 1.0/0.0; 2.0/0.0; 2.0/0.25; 2.0/0.5; and 2.0/1.0 mg L-1. In order to test the influence of the culture media on shoot-bud induction, (MS), Murashige and Tucker (MT), and woody plant medium (WPM) formulations were evaluated, associated with the best combination of plant growth regulators obtained in the previous experiment. The influence of four beta-lactam antibiotics (timentin, cefotaxime sodium salt, meropenem trihydrate and augmentin) on shoot regeneration was determined. Better regeneration responses were achieved when internodal segments were cultured onto MS-based medium with 500 mg L-1 cefotaxime with the following BAP/NAA concentrations: 0.5 + 0.25 mg L-1 for 'Cravo', 1.0 + 0.25 mg L-1 for 'Valência' and 'Bahia', and 1.0 + 0.5 mg L-1 for 'Pêra'. Genotype, growth regulators, basal media and beta-lactam antibiotics affect the morphogenetic response in mature tissues of citrus.
Highlights
Several methods for genetic transformation of citrus have been described in the literature, but so far the most effective are those using Agrobacterium‐mediated transformation of juvenile materials, such as zygotic embryos, hypocotyls, epicotyls, and cotyledons (Fleming et al, 2000; Al‐Bahrany, 2002; Costa et al., 2002; Oliveira et al, 2009)
The objective of this study was to evaluate the effects of different 6‐benzylaminopurine (BAP) and α‐naphthaleneacetic acid (NAA) combinations, basal media and beta‐lactam antibiotics on in vitro shoot regeneration from mature stem segments of 'Pêra', 'Valência' and 'Bahia' sweet oranges and 'Cravo' rangpur lime
Plant regeneration via direct and indirect organogenesis was obtained on the surface of **Significant at 1% probability
Summary
Several methods for genetic transformation of citrus have been described in the literature, but so far the most effective are those using Agrobacterium‐mediated transformation of juvenile materials, such as zygotic embryos, hypocotyls, epicotyls, and cotyledons (Fleming et al, 2000; Al‐Bahrany, 2002; Costa et al., 2002; Oliveira et al, 2009) Plants regenerated from these explant sources have long juvenile stages before initial fruit production, and many years are necessary before evaluating the horticultural and commercial traits introduced into the transgenic plants. For successful genetic transformation of mature tissue, the first step is the establishment of an efficient plant regeneration system, since in vitro culture using mature tissue as explants is still far from routine (Almeida et al, 2003; Rodríguez et al, 2008) Reasons for this include the relatively low responsiveness of woody plants to exogenous growth regulators and the failure of standard surface sterilization techniques (Cervera et al, 2008). The addition of antibiotics to the culture medium may help to eliminate contamination by bacteria, which hinder the in vitro establishment of explants from mature tissues
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