Genetic transformation of three sweet orange cultivars from explants of adult plants

The difficulty in adult tissue genetic transformation in woody species is still an obstacle to be overcome, including in most sweet orange cultivars of the Brazilian citrus industry. This work reports that, after in vitro culture adjustments, transgenic adventitious buds of 'Hamlin', 'Pra', and 'Valencia' sweet oranges (Citrus sinensis L. Osbeck) were recovered using adult material as explant source, in genetic transformation experiments via Agrobacterium tumefaciens. The transgenic buds were identified by the GUS histochemical analysis and confirmed by PCR analysis, which indicated the presence of an amplified fragment of 817 bp corresponding to the uidA gene sequence. The efficiencies of genetic transformation for 'Hamlin', 'Pra', and 'Valencia' sweet orange cultivars were 2.5, 1.4, and 3.7%, respectively. Media supplemented with auxins and cytokinins during co-culture, and media with high concentrations of cytokinins (3 mg L-1) during transgenic selection led to the transformation and, consequently, the regeneration of adequate number of adventitious buds for the three cultivars. The use of sonication during the explant disinfection was not effective to reduce endophytic contamination and reduced transformation efficiency.

Somatic embryogenesis from ovaries of sweet orange cv. Tobias

Callogenesis, somatic embryogenesis, and regeneration were obtained from tissues of unfertilized ovaries of sweet orange (Citrus sinensis Osbeck.) cv. Tobias. The influence of two modified basal media, woody plant medium (WPM) and N6 medium, to induce callus formation from pistils was determined. Overall, high frequencies of callogenesis were observed when either medium was used. However, initial culture of explants in WPM medium followed by transfer of callus to N6 medium resulted in higher frequency of callus induction (of 2.30 callus per explant that were larger than 0.5 cm in size), and of subsequent development of embryogenic callus (10%). A total of 125 somatic embryos were obtained. After 6 months of culture, 72% of somatic embryos germinated into plantlets. These plantlets were subsequently micrografted in vitro, and then acclimatized. Ploidy of these plants were determined using flow cytometry and TRAPS molecular markers were used to confirm their maternal origin.

Rootstocks for high fruit yield and quality of 'Tahiti' lime under rain-fed conditions

Despite considerable research conducted on 'Tahiti' lime [Citrus latifolia (Yu Tanaka) Tanaka] in several countries, few long-term studies have focused on rootstock effects on fruit production and quality under non-irrigated conditions. As for many other fruit crops, rootstock studies for 'Tahiti' lime are often based on the evaluation of several horticultural responses simultaneously, instead of considering multivariate statistical approaches which may provide with more comprehensive information. Consequently, a trial was installed to evaluate the horticultural performance of non-irrigated 'Tahiti' lime trees budded onto the following 12 rootstocks: 'HRS 801' and 'HRS 827' hybrids; 'Rubidoux', 'FCAV' and 'Flying Dragon' trifoliates; 'Sun Chu Sha Kat' and 'Sunki' mandarins; 'Cravo Limeira' and 'Cravo FCAV' 'Rangpur' limes; 'Carrizo' citrange, 'Swingle' citrumelo, and 'Orlando' tangelo. The trial was installed in 2001, in an 8 m x 5 m spacing with no supplementary irrigation. Measurements of yield, fruit quality oriented to different consuming markets, canopy volume and tree tolerance to drought, were performed. A multivariate cluster analysis identified both 'Rangpur' lime rootstocks as those inducing larger cumulative yield and higher percentage of fruits for the domestic market, with highest drought tolerance to the trees. Despite of their high susceptibility to drought stress under non-irrigated conditions, the 'Flying Dragon' and 'FCAV' trifoliate rootstocks performed outstandingly for 'Tahiti' lime, inducing higher yield efficiency, early bearing and larger percentage of high-quality fruits for foreign markets, with smaller trees more suitable for high-density plantings. (c) 2012 Elsevier B.V. All rights reserved.