One of the objective of the QualiRedFruits project was to analyse the influence of biotisation with Glomus intraradices isolate LPA8 and Sinorhizobium meliloti, production site (Switzerland France, Slovenia, Bulgaria) and cultivar ('Tulameen', 'Polka') on antioxidant molecules (AOM) content and antioxidant activities in raspberry fruits. 'Polka' was significantly richer in anthocyanins and showed much higher ORAC values than 'Tulameen', whereas for total phenolic content and FRAP values no differences were observed between both cultivars. Growing location significantly influenced AOM content and antioxidant capacities of raspberry fruits for both cultivars with, in general, fruits from soil production systems showing the higher contents of health related compounds than production systems on substrate. However, the treatments with and without biotisation showed no clear differences on AOM production and antioxidant activities in fruits at each location due to high natural colonisation by indigenous mycorrhizal fungi. In conclusion, the most important factors influencing AOM content and antioxidant activities in raspberry fruits in this experiment were the plant genotype (cultivar) and to a lesser extent the production site.

Leaves of wild Vaccinium spp. berries are considered a valuable source of antioxidants, mainly belonging to polyphenols, specifically flavonoids provide significant protection against the development and progression of many chronic pathological conditions including cancer, diabetes, osteoporosis, cardio-vascular and neurodegenerative diseases. Targeted high-performance liquid chromatography-electro spray ionization-high resolution mass spectrometry operated in the tandem mode (LC-ESI/HRMS2) approach was applied to evaluate and compare the metabolite profiles of bilberry (Vaccinium myrtillus L.) and lingonberry (Vaccinium vitis-idaea L.) leaves originating from the Rhodope Mountain in Bulgaria during growing season. In total, 55 different phenolic compounds, 52 in bilberry and 48 in lingonberry were identified and quantified, including flavan-3-ols, proanthocyanidins, flavonols and their glycosides, iridoids, cinchonain isomers, various phenolic acid conjugates. The contents of different metabolites varied remarkably in bilberry leaves, whereas in lingonberry leaves the metabolites maintained quite constant levels during the growing season.

CHRISTOVA, P. and R. BATCHVAROVA, 2015. Pyramiding of four transgenes in one tobacco line by sexual crosses. Bulg. J. Agric. Sci., 21: 829–834 To combine four transgenes (Np, CP, ttr and ahas 3R) in one tobacco line a cascade of sexual crosses between four transgenic tobacco lines resistant to TSWV, PVY, P. syringae pv. tabaci and herbicide Glean®, respectively, was performed. The first step of the experiment where genes for resistance were combined in pairs was previously described (Docheva et al., 2002). Here we report the second step of pyramiding of four transgenes and combining the resistance to TSWV, PVY, P. syringae pv. tabaci and herbicide Glean® in one tobacco line (cross 2002). Complex resistance to all tree pathogens and the herbicide was analyzed in F1 and F2 generations and integration of all four transgenes in tobacco genome was proved by PCR analyses. Thirteen F2 tobacco lines with pyramided genes for resistance to TSWV, PVY, P. syringae pv. tabaci and herbicide Glean® were selected.

The aim of this work is to study the influence of different mycorrhizal fungi and their further treatment in the nursery on plant mineral uptake and dry matter accumulation. Additional biotizations were preliminary done in some of the experimental treatments. The analysis of dry matter accumulation was performed twice in vegetation period (in May and October). Each sample was analyzed also for plant mineral uptake at six basic macro and microelements (N, P, K, Ca, Mg, Fe). Generally, all additional mycorrhiza treatments have a positive effect on the dry matter accumulation. In Trichoderma experimental plots highest increase is observed in Trichoderma (BS)(TG) and Trichoderma (SL)(TG) treatments, which showed more than 55% dry matter accumulation rates. For comparison, Trichoderma non-treated increase rate is less than 10%. Similar results are obtained in Glomus and Glomus+Agrobacterium treatments where additional treated plots overrun non treated ones with 21.1% and 82.7% respectively. Concerning the mineral uptake, additional mycorrhiza application show advantages in all treatments.

The soil-born nematodes and especially some groups of them are susceptible to different biological and chemical factors occurring in the environment. That is why investigations related to influence of mycorrhizal fungi including Trichoderma harzianum and Glomus intraradices and bio-control bacteria such as Agrobacterium radiobacter on soil structure and nematode population dynamics present a specific interest. The main results of this study showed that inoculation of GF677/‘Royal Glory’ nursery plants with mycorrhizas and bio control bacteria lead to reduction of the number of ectoparasite virus-transmitting phytonematodes while maintaining the density of semi-parasite and free leaving ones. Therefore, mycorrhiza and bio-control bacteria treatments could be recommended as a useful tool for biological control in peach nurseries.

TRIFONOVA, Z., I. TSVETKOV, N. BOGATZEVSKA and R. BATCHVAROVA, 2014. Efficiency of Pseudomonas spp. for biocontrol of the potato cyst nematode Globodera rostochiensis (Woll.). Bulg. J. Agric. Sci., 20: 666-669 The potential of Pseudomonas spp. as biocontrol agent against potato cyst nematode Globodera rostochiensis was tested in pot experiment under glasshouse conditions. The best control of nematode population was provided by the bacterial strains Pseudomonas putida 3 (2) and Pseudomonas aurantiacea 13 (2). The plant growing parameters in the treatments enhanced as compared with the inoculated with Globodera rostochiensis. The two bacterial strains significantly reduced nematode popula tions by 40.7% - 42.2 % compared to the control.

Red raspberry (Rubus idaeus) fruit quality at different harvest stages is a function of the fruit’s metabolite content, which results from physiological changes during fruit growth and ripening. In order to investigate raspberry fruit development, targeted GC-MS metabolic profiling analysis was conducted. Principal component analysis (PCA) was employed to explore the polar metabolite profiles and represent a strong separation between fruit samples at four developmental stages. Each stage of fruit development had a characteristic metabolic profile. Significant changes in metabolite levels were found in fruits turning red in comparison with fruits over-ripening. The levels of free amino acids decreased gradually before the red ripening stage, but increased in the over-ripening stage. In addition, fruit development is characterized by rapidly increasing sugar levels and decreasing organic acids.

We compared the effect of biotisation techniques on growth and mineral uptake in nursery-grown micropropagated Myrobolan 29C plants, against conventional techniques. Biotisation techniques consisted of inoculation of mycorrhizas (Glomus intraradices) and bioagents (Trichoderma harzianum), moreover, the soil borne pathogens were controlled through antagonist microorganisms. Nutrients (N, P, K) were supplied based on plant demand for biotised plants whilst they were empirically supplied for the conventionally managed block. Biotised plants showed a double growth (total dry matter) compared to control plants (34 g/plant against 18 g/plant) despite a significant reduction of the amounts of nutrients supplied. A fertigation plan is proposed to reduce undue mineral elements application by matching the plant demand for nutrients.

The resistance of common bean to Colletotrichum lindemuthianum is controlled by a number of race-specific genes. To combine various genes for resistance to anthracnose alternating crossing was performed. The resulting recombinant inbred lines (RILs) are based on the cross DG 2-36 [Red Hawk/TO//AB136/3/Red Hawk/TO//TU/4/DG 98-53]. Breeding for resistance to C. lindemuthianum using several physiological races of anthracnose identified in Bulgaria was performed from F-1 to F-5 generation. Seven RILs of F, progeny, possessing more than one specific gene for resistance were selected by RAPD and SCAR markers linked to Co-1 and Co-4 genes. The line DG 2-36-58-3 was determined as the most promising by quality complex of growth habit type, vegetation period, type of seeds, yield and presence of Co-1 and Co-4 genes that confer resistance to 74 out of 78 worldwide recognized anthracnose races.Several common bean varieties with useful breeding traits were investigated as a source of Co-2 specific gene. Cultivar 'Drezden' and the new Bulgarian variety 'Beslet' carrying of two genes for resistance in the Co-2 locus were determined as promising sources of the gene for the future breeding programs to obtain durable resistance to C. lindemuthianum.

An independent and competent evaluation of AgroBioInstitute (ABI) functioning in the area of plant biotechnology was implemented in the frame of project “Biotechnology Approaches in ABI at the Service of Crop Breeding” (PlantBioServ), funded by EC. It was achieved by evaluation of research capacity building, training and education capability; coordination and network potential and human capacity of ABI in the fields of plant biodiversity, plant abiotic and biotic stress, and molecular genetics of plants. Three evaluators, nominated by EC, worked in close and permanent contact and thus gave a complete and fully valuable evaluation of ABI. The staff of ABI contributed actively to the coordination and effectiveness of the support mechanisms and accomplishment of the associated work plan. The main results of the project were development of a SWOT analysis of the AgroBioInstitute, an Action Plan to improve the ABI's RTD capacity with a Plan for implementation and a Strategic Framework Programme (2010–2013) which defines ABI's development goals and objectives and thematic areas in which the ABI will focus its work. The information about the project was disseminated at national level at Open Information Day (OID) for scientists and farmers and at international level on the IXth meeting of International Consultative Council (ICC) organized by ABI. Meetings with journalists during the conducting the project and during the OID and ICC were organized. Short communications and popular papers in English and Bulgarian were published and a bilingual webpage of the project was created. Target groups for whom the results of the project could be relevant are national and international plant research organizations, seed companies, farmers, PhD and Postgraduate students, biotech specialists.

Accessions from different Helianthus species, cultivated sunflower forms and interspecific hybrid forms and lines were included in this investigation. It was established that broomrape resistance was transferred from 11 perennial Helianthus species to interspecific hybrids developed on the basis of new sunflower lines. Some of the new lines possessed other desirable agronomical characters, which could be successfully transferred to new sunflower hybrid cultivars. Differences were observed in the origin of broomrapes found in different locations of northern Bulgaria. Diverse origin of some of the obtained sunflower forms was confirmed using the RAPD analysis.

ABTRACT Over the past 10 years, researchers have become increasingly interested in polyphenols. The main reason for this interest is the recognition of the antioxidant properties of polyphenols, their great abundance in our diet, and their probable role in the prevention of various diseases associated with oxidative stress, such as cancer, cardiovascular and neurodegenerative diseases (51). Furthermore, polyphenols, which constitute the active substances found in many plants, modulate the activity of a wide range of enzymes and cell receptors (1). In this way, in addition to having antioxidant properties, polyphenols have several other specific biological actions that are poorly understood. Not all polyphenols are absorbed with equal efficiency. Knowledge of the bioavailability and metabolism of the various polyphenols is necessary in order to evaluate their biological activity within target tissues. The types and distribution of polyphenols in foods and the bioavailability of polyphenols are the topics of our knowledge. Several thousand molecules having a polyphenol structure have been identified in higher plants, and several hundred are found in edible plants. These molecules are secondary metabolites of plants and are generally involved in defense against ultraviolet radiation or aggression by pathogens. These compounds may be classified into different groups as a function of the number of phenols that they contain. This review aims to describe evidences for the effects of polyphenol consumption on health, and to specify which out of the hundreds of existing polyphenols are likely to provide the greatest protection in the context of preventive nutrition. If these objectives are to be attained, it is first essential to determine the nature and distribution of these compounds in fruits. Such knowledge will allow evaluation of polyphenol intake and enable epidemiologic analysis that will in turn provide an understanding of the relation between the intake of these substances and the risk of development of several diseases.

Joint Genomic Center is a new initiative for the efficient establishment of public/private partnership. In December 2006 the Sofia University “St. Kliment Ohridski” and the Agricultural Academy agreed to create jointly a Genomic Center on the principles of the Limited liability Company. It is the first initiative of this kind that joins the scientific efforts of such institutions with the aim to meet mostly the needs of the private sector.

The resistance to fungal pathogens is one of the most important qualities of the tomato varieties. The molecular biology methods and genetic transformation bring a new opportunity to improve the existing cultivars through supplementation of genes responsible for antifungal proteins production. The plant defensin Rs-AFP2 isolated from radish is a small (5kDa) cistatin-rich protein which showed a broadspectrum antifungal activity in vitro. In this study a gene encoding Rs-AFP2 sequence was successfully introduced in tomato plants via Agrobacterium mediated transformation. The response of tomato plants constitutively expressing Rs-afp2 to infection with Fusarium oxysporum f. sp. lycopersici and Botrytis cinerea was evaluated via detached leaves bioassay, A part of the transformed tomato plants showed significantly higher levels of resistance compared to the control plants. Transgenic plant T14 had the highest level of expression of Rs-afp2 in the Northern blot hybridization and it also showed the highest level resistance to both Pathogens. The size of the colonized tissue in T14 was reduced with 91 % under the infection of F.oxysporum f sp. lycopersici and the lesions caused by B. cinerea were 68 % smaller compared with the control plants. Our results showed that the constitutive expression of a plant defensin gene in tomato plants can increase the resistance to more than one fungal pathogen, thus to improve their agro-ecological behavior

KOSTOV, K., R. BATCHVAROVA and S. SLAVOV, 2007. Application of chemical mutagenesis to increase the resistance of tomato to Orobanche ramosa L. Bulg. J. Agric. Sci., 13: 505-513 In this study chemical mutagenesis of tomato seeds with Ethylmethanesulfonate was used to obtain lines with increased resistance to Orobanche ramosa L. The M2 progeny of the mutant tomato plants was screened for the response to broomrape attack in large scale experiment conducted in greenhouse. Sixteen non-infested by the parasite tomato plants were selected. Their offspring was a subject of another screening for broomrape resistance using an artificial polyethylene bags system. As a result of the experiments six lines with significantly increased level of resistance to Orobanche ramosa L., were selected.