Tomato (Solanum lycopersicum L.) in the service of biotechnology

Originating in the Andes, the tomato (Solanum lycopersicum L.) was imported to Europe in the 16th century. At present, it is an important crop plant cultivated all over the world, and its production and consumption continue to increase. This popular vegetable is known as a major source of important nutrients including lycopene, bcarotene, flavonoids and vitamin C as well as hydroxycinnamic acid derivatives. Since the discovery that lycopene has anti-oxidative, anti-cancer properties, interest in tomatoes has grown rapidly. The development of genetic engineering tools and plant biotechnology has opened great opportunities for engineering tomato plants. This review presents examples of successful tissue culture and genetically modified tomatoes which resistance to a range of environmental stresses improved, along with fruit quality. Additionally, a successful molecular farming model was established

Expression of recombinant staphylokinase, a fibrin-specific plasminogen activator of bacterial origin, in potato (Solanum tuberosum L.) plants

One of the most dynamically developing sectors of green biotechnology is molecular farming using transgenic plants as natural bioreactors for the large scale production of recombinant proteins with biopharmaceutical and therapeutic values. Such properties are characteristic of certain proteins of bacterial origin, including staphylokinase. For many years, work has been carried out on the use of this protein in thrombolytic therapy. In this study, transgenic Solanum tuberosum plants expressing a CaMV::sak-mgpf-gusA gene fusion, were obtained. AGL1 A. tumefaciens strain was used in the process of transformation. The presence of the staphylokinase gene was confirmed by PCR in 22.5% of the investigated plants. The expression of the fusion transgene was detected using the β-glucuronidase activity assay in 32 putative transgenic plants. Furthermore, on the basis of the GUS histochemical reaction, the transgene expression pattern had a strong, constitutive character in seven of the transformants. The polyacrylamide gel electrophoresis of a protein extract from the SAK/PCR-positive plants, revealed the presence of a119 kDa protein that corresponds to that of the fusion protein SAK-mGFP-GUSA. Western blot analysis, using an antibody against staphylokinase, showed the presence of the staphylokinase domain in the 119 kDa protein in six analyzed transformants. However, the enzymatic test revealed amidolytic activity characteristic of staphylokinase in the protein extract of only one plant. This is the first report on a Solanum tuberosum plant producing a recombinant staphylokinase protein, a plasminogen activator of bacterial origin

Green Way of Biomedicine – How to Force Plants to Produce New Important Proteins

Rozdział książki Transgenic Plants - Advances and Limitations Edited by Yelda Özden Çiftç

Compendium on Food Crop Plants as a Platform for Pharmaceutical Protein Production

Tremendous advances in crop biotechnology related to the availability of molecular tools and methods developed for transformation and regeneration of specific plant species have been observed. As a consequence, the interest in plant molecular farming aimed at producing the desired therapeutic proteins has significantly increased. Since the middle of the 1980s, recombinant pharmaceuticals have transformed the treatment of many serious diseases and nowadays are used in all branches of medicine. The available systems of the synthesis include wild-type or modified mammalian cells, plants or plant cell cultures, insects, yeast, fungi, or bacteria. Undeniable benefits such as well-characterised breeding conditions, safety, and relatively low costs of production make plants an attractive yet competitive platform for biopharmaceutical production. Some of the vegetable plants that have edible tubers, fruits, leaves, or seeds may be desirable as inexpensive bioreactors because these organs can provide edible vaccines and thus omit the purification step of the final product. Some crucial facts in the development of plant-made pharmaceuticals are presented here in brief. Although crop systems do not require more strictly dedicated optimization of methodologies at any stages of the of biopharmaceutical production process, here we recall the complete framework of such a project, along with theoretical background. Thus, a brief review of the advantages and disadvantages of different systems, the principles for the selection of cis elements for the expression cassettes, and available methods of plant transformation, through to the protein recovery and purification stage, are all presented here. We also outline the achievements in the production of biopharmaceuticals in economically important crop plants and provide examples of their clinical trials and commercialization

Cytokinin Signaling and De Novo Shoot Organogenesis

The ability to restore or replace injured tissues can be undoubtedly named among the most spectacular achievements of plant organisms. One of such regeneration pathways is organogenesis, the formation of individual organs from nonmeristematic tissue sections. The process can be triggered in vitro by incubation on medium supplemented with phytohormones. Cytokinins are a class of phytohormones demonstrating pleiotropic effects and a powerful network of molecular interactions. The present study reviews existing knowledge on the possible sequence of molecular and genetic events behind de novo shoot organogenesis initiated by cytokinins. Overall, the review aims to collect reactions encompassed by cytokinin primary responses, starting from phytohormone perception by the dedicated receptors, to transcriptional reprogramming of cell fate by the last module of multistep-phosphorelays. It also includes a brief reminder of other control mechanisms, such as epigenetic reprogramming

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

Abstract 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