The aim of the present study is to develop a system for micropropagation of bog blueberries. They are distributed from 1700 to 2500 m above sea level in the mountains Rila, Pirin, Western and Central Balkan Mountain, Vitosha, Osogovo, Western Sredna Gora and Western and Middle Rhodopes. An approach for the statistical optimization of micropropagation media under laboratory conditions has been applied for the first time. Sterile explants were introduced in vitro on WPM, enriched with 3 mg.l-1 2-iP and pH 4.2. To optimize proliferation, nine variants of WPM were tested with the addition of growth regulators (zeatin and 2-iP) in varying concentrations (1.5 - 3.5 mg.l-1), organic additives (casein hydrolisate and myo-inositol and pH values (4.3 - 4.8). The best result regarding the reproduction coefficient was found on M1 nutrient medium (3.77 pcs/explant), enriched with 3.5 mg.l-1 zeatin and 2-iP, 250 mg.l-1 casein hydrolisate, pH 4.8. The largest average length of shoots was reported on the modified medium M9 with 2.63 cm. The resulting micro shoots were rooted on WPM nutrient medium, halved macrosol content and different levels of IAA (0.5 - 1 mg.l-1), myo-inositol and pH. The highest percentage of rooting was found on R8 nutrient medium with 56.7%.

The production of Vaccinium sp. has been a growing worldwide interest after understending their importance for diet and human health. The aim of the work was direct regeneration, propagation and rooting of axillary buds isolated from five highbush blue-berry cultivars 'Bluecrop', 'Bluegold' 'Bluejay', 'Spartan' and 'Patriot' and adapt of suitable protocol, developed and applied for Vaccinium corymbosum L. cultivars in our laboratory. For micropropagation, a basal cultural medium WPM was used, supplemented with 3.0 mg/L zeatin and 2.0 mg/1 2-iP, pH 4.2. The highest proliferation capacity was reported for cv. 'Bluegold' after fourth passage of sub cultivation (4.4 shoots per explant) and the largest length of the shoots was reported for the cv. 'Bluejay' (3.9 cm). The rooting ability was established on the same basal medium (WPM) with half-reduced salt concentration enriched with 1.0 mg/L IAA, pH 4.2. The high rooting percentage (33.3%) and the highest average number of roots (3.9) was reported for cv. 'Spartan' and 'Patriot'. The results demonstrate that regeneration potential is genotyp dependent.

Georgieva, M., Kondakova, V. & Yancheva, S. (2020). A comparative study on raspberry cultivars in micropropa- gation. Bulg. J. Agric. Sci., 26 (3) , 527-532 The aim of the present study was to clarify the effectiveness of a simplified protocol for micropropagation of raspberry cultivars, such as Samodiva (control), Meeker, Willamette and the candidate cultivar of Magdalena (passed through DUS test) as prospective ones, which are suitable for cultivation in the mountain and hilly regions of Bulgaria. The effect of growth regulators BAP (0.5 mg l(-1) ) and IBA (0.01 mg l(-1) ) on proliferation capacity and shoot length over six passages was established. The highest multiplication potential 3.9 at the fifth passage and average length of the shoots 3.76 cm (fourth passage) were registered in the candidate cultivar of Magdalena. The best rhizogenic ability was recorded in Samodiva- 80.5%. These results show that genotype is the primary factor determining a high propagation and economically significant efficiency in this pro- cess. The application of simple medium for all studied genotypes is towards commercial propagation, comparing their growth characteristics and obtaining pre-base material for planting in a production nursery and further investigations.

The development of explants of 12 cultivars and breeding samples of essential oil rose (obtained with the participation of Rosa damascena Mill., R. gallica L., R. alba L.) during long-term micropropagation was investigated. At the second multiplication stage 5 subcultures were carried out. Increase of the studied morphometric parameters of explants to 3-4 subcultures was established. In some genotypes, the maximum multiplication index was in the third subculture (сultivars ‘Raduga’, ‘Zolushka’, ‘Lada’, ‘Krymskaya Krasnaya’, ‘Lany’, ‘Vesna’ and samples N37-24, М215), while in others (сultivar ‘Kazanlykskaya’ and samples N138, G2168, N37-2) – in the fourth. The largest increase of the multiplication index in 3-4 subcultures compared to the first (3.0-4.8 times) was found in the cultivars ‘Lada’, ‘Lany’, ‘Raduga’, ‘Krymskaya Krasnaya’ and sample N37-24. In the fifth subculture the multiplication index decreased. However, in most cultivars and breeding samples it was higher than in the first subculture. The best morphogenetic potential was noted for сultivar ‘Raduga’ and samples G2168, N37-2, in which the multiplication index reached 14.2, 14.4 and 11.8, respectively. The minimal ability to propagation in vitro was in samples М215 and N138 – their multiplication index did not exceed 1.14.0.

Woodland berries are among the most important berry species worldwide. They are a rich source of a wide variety of bioactive substances. They are demanded by the food and pharmacy processing industries, due to their delicious taste and high bioactivity value as well. In recent decades, in vitro micropropagation has become the standard for commercial propagation of certain plant species. However, the economic assessment of this technology shows that it is labor-intensive and the price of the plants obtained is high, thus directing the scientists to automate the in vitro propagation applying different bioreactor systems, which are characterized by constant environmental conditions of cultivation and high propagation rates, which influence directly on the reduction of the cost of propagated plants. This chapter provides information about traditional techniques for in vitro propagation of berry plants discussing the problems that appeared using this technology and summarizes also recent achievement in the development of bioreactor design and operation modes for the in vitro propagation of berry plants. The chapter presents a deep overview of phytochemical profiles and bioactivity of berry plants, because we believe that bioreactor technology is very prospective not only for micropropagation of plants but also for producing target metabolites that are responsible for the bioactivity of berry fruits as well as for the bioactivity of the extracts of different berry plant organs.

Plant tissue cultures have been widely used in both fundamental and applied types of research on various biological species, and the scientific interest to transfer that technology in industrial scale has been rapidly growing. The use of in vitro technology for commercial propagation of different plant species and the production of bioactive compounds from them has become profitable industry worldwide. In the past decades, the progress in plant tissue culture technology was directed towards the introduction of the liquid medium for cultivation under submerged conditions in different bioreactor types, and automation of the entire process. Some applications of modified bioreactor systems and their importance for the advancement of plant biotechnology in the fields of agriculture, medicine, and pharmacy are discussed in this review.

The application of plant tissue cultures in fundamental and applied studies on various biological species has been rapidly growing. The use of in vitro technology for commercial propagation of plant species and for the production of bioactive components from them has become profitable industry worldwide. Various regeneration systems (protoplast cultures and somatic embryogenesis) and their importance for the advance of strategically significant priorities in the development of biotechnological science in agriculture, medicine, and pharmacy are treated in the present chapter. We believe that in the future development of the in vitro technology the major priorities could be conservation of plant genetic resources; restoring the balance between research studies related to genetic transformation of plants with the aim of providing sufficient, quality and safety foods for the world population, on the one hand, and the studies aimed at determining the risk of growing and consuming them, on the other; creating transgenic plants maintaining a constant level of induced protein; and, last but not least, the use of plant resources possessing valuable biologically active substances.

The Climate Change is one of the major problems in the entire world. The limiting influence of harmful factors as climate and other environmental changes, it will be crucial to breed new varieties for improved resistance to abiotic and biotic stresses. Climate Change can be tackled through plant breeding and better use of Plant Genetic Resources. Generally, plant breeding is the art and science of genetically improving plants for the benefit of humankind. It is important to mention potential contribution of organically managed systems to climate change mitigation is identified in the careful management of nutrients and, hence, the reduction of N2O emissions from soils. The application of biotechnological approaches, developing and implementation of new “omics” technologies and infrastructure construction on a high scientific level will help in building a strategy to overcome the damage of Climate Change. Key words: Climate Change, plant breeding, organic agriculture, plant biotechnology, “OMICS” technologies.

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.

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

Wild berry species are known to exhibit a wide range of pharmacological activities. They have long been traditionally applied for their antiseptic, antimicrobial, cardioprotective and antioxidant properties. The aim of the present study is to reveal the potential for selective antiviral activity of total methanol extracts, as well as that of the anthocyanins and the non-anthocyanins from the following wild berries picked in Bulgaria: strawberry ( Fragaria vesca L.) and raspberry ( Rubus idaeus L. ) of the Rosaceae plant family, and bilberry ( Vaccinium myrtillis L.) and lingonberry ( Vaccinium vitis-idaea L) of the Ericaceae. The antiviral effect has been tested against viruses that are important human pathogens and for which chemotherapy and/or chemoprophylaxis is indicated, namely poliovirus type 1 (PV-1) and coxsackievirus B1 (CV-B1) from the Picornaviridae virus family, human respiratory syncytial virus A2 (HRSV-A2) from the Paramyxoviridae and influenza virus A/H3N2 of Orthomyxoviridae . Wild berry fruits are freeze-dried and ground, then total methanol extracts are prepared. Further the extracts are fractioned by solid phase extraction and the non-anthocyanin and anthocyanin fractions are eluted. The in vitro antiviral effect is examined by the virus cytopathic effect (CPE) inhibition test. The results reveal that the total extracts of all tested berry fruits inhibit the replication of CV-B1 and influenza A virus. CV-B1 is inhibited to the highest degree by both bilberry and strawberry, as well as by lingonberry total extracts, and influenza A by bilberry and strawberry extracts. Anthocyanin fractions of all wild berries strongly inhibit the replication of influenza virus A/H3N2. Given the obtained results it is concluded that wild berry species are a valuable resource of antiviral substances and the present study should serve as a basis for further detailed research on the matter.

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.

Wild berry species are known to exhibit a wide range of pharmacological activities. They have long been traditionally applied for their antiseptic, antimicrobial, cardioprotective and antioxidant properties. The aim of the present study is to reveal the potential for selective antiviral activity of total methanol extracts, as well as that of the anthocyanins and the non-anthocyanins from the following wild berries picked in Bulgaria: strawberry (Fragaria vesca L.) and raspberry (Rubus idaeus L.) of the Rosaceae plant family, and bilberry (Vaccinium myrtillis L.) and lingonberry (Vaccinium vitis-idaea L) of the Ericaceae. The antiviral effect has been tested against viruses that are important human pathogens and for which chemotherapy and/or chemoprophylaxis is indicated, namely poliovirus type 1 (PV-1) and coxsackievirus B1 (CV-B1) from the Picornaviridae virus family, human respiratory syncytial virus A2 (HRSV-A2) from the Paramyxoviridae and influenza virus A/H3N2 of Orthomyxoviridae. Wild berry fruits are freeze-dried and ground, then total methanol extracts are prepared. Further the extracts are fractioned by solid phase extraction and the non-anthocyanin and anthocyanin fractions are eluted. The in vitro antiviral effect is examined by the virus cytopathic effect (CPE) inhibition test. The results reveal that the total extracts of all tested berry fruits inhibit the replication of CV-B1 and influenza A virus. CV-B1 is inhibited to the highest degree by both bilberry and strawberry, as well as by lingonberry total extracts, and influenza A by bilberry and strawberry extracts. Anthocyanin fractions of all wild berries strongly inhibit the replication of influenza virus A/H3N2. Given the obtained results it is concluded that wild berry species are a valuable resource of antiviral substances and the present study should serve as a basis for further detailed research on the matter.