Approaches for gene targeting and targeted gene expression in plants.

Transgenic science and technology are fundamental to state-of-the-art plant molecular genetics and crop improvement. The new generation of technology endeavors to introduce genes 'stably' into 'site-specific' locations and in 'single copy' without the integration of extraneous vector 'backbone' sequences or selectable markers and with a 'predictable and consistent' expression. Several similar strategies and technologies, which can push the development of 'smart' genetically modified plants with desirable attributes, as well as enhance their consumer acceptability, are discussed in this review.

Vehicles and ways for efficient nuclear transformation in plants.

Transgenic science and technology are fundamental to the state-of-art plant molecular genetics and crop improvement. The new generation of technology endeavors to introduce genes 'stably' into 'site-specific' locations and in 'single copy' without the integration of extraneous vector 'backbone' sequences or 'selectable markers'. Numerous plant transformation technologies have developed with the aim of achieving these objectives. Here we discuss some of these technologies, which can push the development of 'better transgenic plants with desirable characters only'.

Pre- and post-agroinfection strategies for efficient leaf disk transformation and regeneration of transgenic strawberry plants.

Following previously described Agrobacterium tumefaciens-mediated transformation procedures for Fragaria x ananassa Duch. 'Chandler', we undertook several experiments to establish the importance of some parameters affecting transformation. The most important factor that increased the percent recovery of transformants was the introduction of a pre-selection phase, in-between co-cultivation and selection, in which leaf disks were cultured on pre-selection regeneration medium containing validamycin A, timentin, and cefotaxime. The average percentage of leaf disks forming shoots on selection medium containing cefotaxime (250 mg l(-1)) + timentin (250 mg l(-1)) was 5.4% and about three shoots per regenerating leaf disk. Maximum transformation percentage, based on polymerase chain reaction, was 31.25%. Transgene integration and copy number were assessed by Southern hybridization confirming single copy as well as multiple copies of transgene integration in shoots as well as roots separately. This confirmed the non-chimeric nature of these transgenic plants. The system is very promising for the regeneration of genetically transformed cells and obtaining transgenic strawberry plants at high efficiency.

Overexpression of the HMG-CoA reductase gene leads to enhanced artemisinin biosynthesis in transgenic Artemisia annua plants.

An effective and affordable treatment against malaria is still a challenge for medicine. Most contemporary drugs either are too expensive to produce or are not effective against resistant strains of the malaria parasite Plasmodium falciparum. The plant Artemisia annua L. is the source of artemisinin, an effective drug against malaria for which no resistant strains of the bacterium have been reported. However, the artemisinin content of A. annua is very low, which makes its production expensive. Here we report the use of transgenic technology to increase the artemisinin content of A. annua. We report the production of transgenic plants of A. annua into which we transferred 3-hydroxy-3-methylglutaryl CoA reductase (HMGR) gene from Catharanthus roseus (L.) G. Don using Agrobacterium-mediated gene transfer technology. Transgene integration and copy number were assessed by PCR and Southern hybridization, which confirmed the stable integration of multiple copies of the transgene in 7 different transgenic lines of A. annua. The leaf tissue of three of the A. annua transgenic lines possessed significantly higher HMGR activity compared with wild-type controls, and this activity was associated exclusively with microsomal membranes. The artemisinin content of the shoots of one of the transgenic lines depicted an increase of 22.5 % artemisinin content compared with wild-type control A. annua plants.