One of the main subjects of the AMIGA (Assessing and Monitoring the Impacts of Genetically modified plants on Agro-ecosystems) project of F7P was to establish protocols for risk assessment in GM crops providing reliable evaluation of their impact on the environment. We were able to develop an efficient bioassay where micropropagated plants of cisgenic potato lines resistant to late blight disease caused by Phytophthora infestans Mont de Bary, were used as feed for larvae of Colorado potato beetle as Non Target Organism. The studies performed under contained laboratory conditions show no impact of Phytophthora resistant potato lines on life cycle parameters and dynamics of larval growth of the Non Target Organism.

The omics disciplines applied in the context of nutrition and health have the potential to deliver biomarkers for health and comfort, reveal early indicators of disposition to disease, and discover bioactive, beneficial food components. These technologies are aimed at unraveling the overall expression of genes, proteins, and metabolites in a functionally relevant context, and provide insights into the molecular basis of various fundamental processes involved in growth and development of plants and their environment. Genomics is an entry point for looking at the other omics sciences. The information in the genes of an organism, its genotype, is largely responsible for the final physical makeup of the organism, referred to as the phenotype. The main purpose of the application of genomics is to gain a better understanding of the whole genome of plants. Agronomically important genes may be identified and targeted to produce more nutritious and safe food while at the same time preserving the environment. Proteomics is known as protein expression profiling, whereby the expression of proteins in an organism resulting from a stimulus is identified at a certain time. Proteomics can also be applied to CONteNtS

Germination of seeds of root parasites like broomrapes (Orobanchaceae) is tightly regulated by chemical products exuded from the roots of the host plant, known as germination stimulants (GSs). Changes in the levels of synthesis and emission of GS can allow the development of practical measures for control of the crops-harming parasitic species. However, the genes encoding enzymes responsible for GS biosynthesis are still unknown. We performed a large-scale screening of 62,000 Arabidopsis activation-tag mutants for alteration in susceptibility to Phelipanche ramosa and to identify lines with altered GS production among them. After five successive screenings we identified 36 lines with altered susceptibility to P. ramosa. Seven of them displayed altered levels of GS production. By using a combination of Southern blot and thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR), we pinpointed the location of activation-tag constructs in these lines. A combination of differential display and quantitative real-time PCR (qRT-PCR) allowed us to identify several affected genes. Two of them are directly involved in isoprenoid biosynthetic pathway in chloroplasts, and we believe that their activation led to increased levels of GS production. We believe that these genes are responsible for increased GS production in five of the Arabidopsis lines resistant to P. ramosa.

Somatic embryogenesis being one of the in vitro techniques that produce high regeneration rates, serves for high frequency propagation, gene transfer and germplasm storage.The aim of this work is to study the possibilities for in vitro long-term storage of grapevine genetic resources via repetitive somatic embryogenesis.Thirteen years after induction from leaves explants embryogenic cultures of Bulgarian grapevine variety "Velika" are still propagating via repetitive somatic embryogenesis in frame of clusters keeping their embryogenic and morphogenic competence.Plantlents regeneration from cotyledonary stage somatic embryos showed a normal phenotype compared with shoot-tip propagated controls.The genetic fidelity of regenerated plants was assessed by DNA analysis using Inter Simple Sequence Repeat (ISSR) markers.The amplified loci showed no differences between mother and regenerated plants, confirming the genetic authenticity of in vitro stored and propagated plants.

Extracts of the resurrection plant Haberlea rhodopensis have been tested for fungitoxic activity against four important plant pathogens. No effect was found against Alternaria alternata and Fusarium oxysporum. The growth of Botrytis cinerea was strongly inhibited, in particular by apolar fractions of the extracts. An interesting stimulating effect was found for the same fractions when applied to Phytophthora citricola. Further studies on the component(s) with fungitoxic (for B. cinerea) or affectors (for P. Citricola) potential are in progress.

Orobanchaceae (broomrapes) is a morphologically diverse family of predominantly herbaceous, parasitic plants. The majority of species are facultative or obligate root parasites that subsist on broad-leaf plants, thereby depleting them of nutrients, minerals and water. The taxonomy status of the family Orobanchaceae among other flowering plants is often subject of debate. They possess only a few morphological features suitable for taxonomy purposes and yet even they are quite changeable. The variability within the species is too high and hampers the attempts to create proper determination keys. During last two decades several molecular markers were used for reevaluate taxonomy, biodiversity and phylogenetic relationships within the family. Recent investigations supported by molecular taxonomy analyses have resulted in re-definition of Orobanchaceae family. According to this classification Orobanchaceae consists of 89 genera, containing 2061 species. On the Balkans the family Orobanchaceae is represented by 3 genera: Orobanche includes 25 species; Phelipanche comprises of 9 species and some putative hybrids; Diphelypaea occurs with single species, Diphelypaea boissieri, in Macedonia and Greece. Only a few recent studies based on modern methods took place during last decade. Their findings confirmed differences between Phelipanche and Orobanche genera, but raised new question about their internal structure. Several broomrape species parasitize important crops. They are widely spread in Bulgaria, Southern Europe, Russia, Middle East and Northern Africa. They cause losses in crop productivity estimated at hundreds of millions of dollars annually than affect the livelihoods of 100 million farmers. A wide variety of approaches have been explored to control broomrapes, but none have been found to be sufficiently effective and affordable. The new findings about their life cycle and the recent genomic project focused on sequences of Ph. aegyptiaca genome open new perspectives for management of the harmful broomrape species and for understanding of their biology and evolution as well.

Colorado potato beetle, Leptinotarsa decemlineata Say, is the most destructive insect pest of potatoes. When the population of beetles is high, plants can be completely defoliated and commercial potato production is nearly impossible without control of the beetle. The beetles have shown a tremendous ability to develop resistance against insecticides. Previously, a biotechnology approach to control Colarado potato beetle based on the use of the synthetic Bt gene was developed. In this article, a transformation procedure for three commercial Bulgarian potato cultivars was developed and potentially commercial transgenic lines have been selected based on field resistance to Colorado potato beetles and yield. Plants were transformed with the Bacillus thuringiensis (Bt) cry3A gene using Agrobacterium-mediated transformation. 110 plants from the three cultivars were regenerated and tested by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). The Cry3A protein accumulation varied across the transgenic lines, rating from very low to 71.5 μg/g fresh weight. 21 transgenic lines expressing the Cry3A protein at levels above 10 μg/g fresh weight were tested in two successive years in filed conditions at two different locations of the country. All transgenic lines compared with the controls, nontransgenic potatoes from the respective cultivar, were consistently protected from foliar damages from all developmental stages of the beetle. The comparison of all properties of the tested transgenic lines, including variety phenotypes and tuber yield, allowed the selection of the most promising 2–3 lines per cultivar. Selected lines produced tuber amounts 80–100% higher compared with the control, non-transgenic plants. Those lines were grown for mass propagation during the third year of field experiments. The presence of the transgene in these lines was confirmed with the use of primers specific to the transgene by polymerase chain reaction (PCR). Additionally, the results from the insect bioassay showed that these lines were highly resistant to insect feeding, leading to 100% of mortality of larval populations. In summary, we generated potentially commercial potato lines highly resistant to Colorado potato beetle using Bt technology that may have a profound impact on development of sustainable agriculture in Bulgaria. This is one of the several agriculture biotechnology products entirely developed and tested in Bulgaria.