Transient expression of β-glucuronidase following biolistic delivery of foreign DNA into coffee tissues

Abstract

Some conditions related to the transient expression of β-glucuronidase in biolistically-treated Coffea spp. tissues were investigated, and subsequently used in a promoter study. Bombardments were performed on different types of tissue (leaves, somatic embryos and suspension cultures) of genotypes of C. arabica, C. canephora and Arabusta, using 4 different promoter sequences. Tobacco leaves were used as a comparison. In general, similar large variation and mean values of transient expression were observed between coffee and tobacco leaves. With regard to the coffee tissues effect, transient expression was best detectable and most frequently observed with bombarded leaves of microcuttings. Disturbing endogenous light blue staining was found with control treatments of somatic embryos. For the three coffee species tested, the most effective promoter was the EF1α-A1 promoter of Arabidopsis thaliana.

Coffee biotechnology

“…Furthermore, a system for genic transfer is required that is simple, cheap, and reproducible. In coffee, there are efficient regeneration systems through somatic embryogenesis from several tissues and the genetic transformation of coffee plants has been achieved successfully by several research groups (Barton et al, 1991;Spiral et al, 1993;Sugiyama et al, 1995;van Boxtel et al, 1995;Hatanaka et al, 1999;Spiral et al, 1999;Leroy et al, 2000;Fernández-Da Silva and Menéndez-Yuffá, 2003;Ogita et al, 2004).…”

“…The application of the biobalistic system with coffee tissues was described for the first time by van Boxtel et al (1995). The experiments were carried out on different tissues and revealed that suspension cultures and somatic embryos were less appropriate for transient expression studies of β-glucuronidase.…”

Intake of caffeine and other methylxanthines during pregnancy and risk for adverse effects in pregnant women and their foetuses

“…A few years later, plantlets were also produced from somatic embryos transformed via Agrobacterium tumefaciens (Leroy et al, 1997). Van Boxtel et al (1995) reported transient expression of the GUS marker gene following the delivery of foreign DNA into leaves of micro cuttings, somatic embryos, and suspension cultures of Coffea arabica, Coffea canephora, and Arabusta (a hybrid between the two species) by particle acceleration technology (with a "gene gun"). The author stressed the critical use of tissue in order to obtain success in the regeneration of plants from protoplats.…”

Transformation of coffee (Coffea Arabica L. cv. Catimor) with the cry1ac gene by biolistic, without the use of markers

“…Hatanaka et al (1999), Leroy et al (2000), Perthuis et al (2005), Ribas et al (2006), Alpizar et al (2006) and Kumar et al (2006) have all reported regeneration of such transformed tissues. The genes gus, bar and nptII have also been incorporated by electroporation; notable are the studies of van Boxtel et al (1995) who described the transitory expression of the gus gene, Barton et al (1991) who transformed coffee-plant embryos with the nptII gene, and Fernández-Da Silva and Menéndez-Yuffá (2003) who reported the regeneration of transformed tissues with positive results for the GUS and PCR reactions for gus and bar genes, while da Cunha et al (2004) achieved the transformation of C. arabica using biolistics with the nptII and gus genes. The insertion of the cry1ac gene into coffee plants has been accomplished by transformation with A. tumefaciens (Leroy et al 2000); however, the biolistic procedure has the advantage of simple construction vectors besides easier transformation protocols as complex plant-bacteria interactions are eliminated (Gray andFiner, 1993, cited by Ribas, et al 2006).…”