Reproductive potential, metabolic and genetic profile, in <i>in situ</i> and <i>ex situ</i> conditions, of medicinal plants species from the Bulgarian flora with resource deficit

Department of Botany and Agrometeorology. The main research fields in the department are Anatomy and Plant Morphology, Plant Systematic, Cytology and Histology, Biology of Seed, Mushrooming, Herbal medicine, Applied Dendrology and Phytocoenology. It has an experimental area for field experiments.

The object of research in this project are the following valuable medicinal plants, with conservation significance for the Bulgarian flora, under a special regime of use, according to an annual Order of the Minister of Environment and Water: Alkanna tinctoria , Helichrysum arenarium, Primula veris and Arctostaphilus uva ursi.

Alkanna tinctoria (L.) Tausch (Anchusa tinctoria) (Boraginaceae) is a species included in the Red Data Book of the Republic of Bulgaria (2015), in the Red List of Higher Plants (Petrova & Vladimirov 2009) with the category „endangered species“ and in Appendix 3 of the Biodiversity Act (BDA 2002) for species forbidden for collection from their natural habitats.

Primula veris L. (Medicinal primrose) (Primulaceae) is a protected species according to the BDA, included in Annex 4 of the BDA and in the List of Species of Medicinal Plants under special regime of conservation and use based on MPA (Art.10, parag. 1 , 2 and 3). Species with annual quantities of herbs eligible for collection for commercial purposes from their natural habitats, outside the territories of the national parks determined by the Ministry of Environment and Water (MEW). The above-ground parts and roots of the primrose are allowed to collect from the natural habitats, and the MEW sets annual quotas for their collection.

Arctostaphilus uva ursi. (Sweet Grapes) (Ericaceae) ) is a protected species according to the BDA, included in Annex 4 of the BDA and in the List of Species of Medicinal Plants under special regime of conservation and use based on MPA (Art.10, parag. 1, 2, and 3). The species is forbidden to be collected for commercial purposes from the natural habitats throughout the country. It is included in the list of medicinal plants of the Annexes to the Convention on International Trade with Endangered Species of Wild Fauna and Flora.

Helichrysum arenarium (L.) Moench (everlasting) (Asteraceae) is a protected species according to the BDA, included in Annex 4 of the BDA and in the List of Species of Medicinal Plants under special regime of conservation and use based on MPA (Art.10, parag. 1, 2, and 3). The species is forbidden to be collected for commercial purposes from the natural habitats throughout the country.

Project objectives

Strategic objectives:

Providing raw material from valuable medicinal plants with resource deficit, demanded on the national and international markets.

Preservation of the natural populations of medicinal plant species of conservation significance for the Bulgarian flora.

Increasing the scientific capacity of project participants, training of PhD students and students, enhancing the mobility of scientists for a fuller integration into the European scientific community.

Increasing the ecological consciousness of the civil society and popularizing the conservation measures of the studied species, respectively of the biodiversity in Bulgaria.

The strategic objectives correspond to the priorities of the research in Bulgaria and worldwide in the last decades, related to the preservation of biodiversity.

Operational objectives:

Evaluation and update of the state of the target species populations, their degree of threat from human activities and climate changes.

Study on the ecological and biological characteristics of the species in situ and ex situ under different environmental conditions (altitude, soil type, light, etc.).

Establishment of the reproductive potential of the target species in their native populations and the possibilities for their ex situ multiplication.

Exploration of the reproductive systems and evolutionary mechanisms of the studied species (hybridization, polyploidy, apomixis, etc.).

Establishment of the character of the populations in connection with the realization of the reproductive potential under specific environment conditions (including stress) and offering of recommendations for effective conservation of the studied species.

. Establishment of the influence of environmental factors on the metabolism by identifying changes in the spectrum of surface flavonoids.

Examination of the content and antiradical activity of BAS and metabolic determination of selected species, taking in consideration the changes in qualitative and quantitative composition under the influence of origin and environmental parameters in situ and ex situ.

Determination of the level of intrapollution and inter-population genetic diversity using modern methods and DNA markers in relation to the conservation of the genofond of the species.

Application of plant biotechnology methods for effective in vitro multiplication of the target plant species by experimental testing of different culture conditions, incl. induction of root cultures; Development of specific protocols for each of the studied species.

Studies on the possibilities of applying hydroponic systems for reproduction of the target species and for stimulation of their root growth.

Establishment of appropriate conditions for ex vitro adaptation and acclimatization of plants propagated by the methods of plant biotechnology.

. Establishment and maintenance of ex situ collections for storing up of valuable genetic material necessary for future activities related with the protection of the target species and their sustainable use as medicinal plants.

Promotion of the results of the project among the scientific community and civil society to implement the strategic objectives of the project.

Scientific team

The scientific team consists of scientists with proven experience and competence in the subject of the project.

Work Package 1. In situ and ex situ studies and collection of initial material: WP 1 will be developed jointly by scientists from both partnering scientific organizations that are experts in plant diversity, monitoring and resource evaluation of conservatively significant, rare, medicinal and aromatic plants and creation of ex situ collections of them: Assoc. Prof. Dr. Antonina Vitkova has experience in the management of projects in this subject, incl. funded by the NSF. Assist. prof Stoyan Stoyanov is an expert in the monitoring of rare and protected plants of the Bulgarian flora, he has gained sufficient experience participating in projects to EMEPA and Life Plus Program. Dr. Ivanka Semerdzhieva as a lecturer in Agricultural University Plovdiv has excellent command of methods of analysis, resource and evaluation of medicinal plants. Prof. Valcho Zhelyazkov working in Oregon University USA is an expert on the ecology of medicinal plants and their natural products. He will work in the implementation of project tasks providing a several months each year in Bulgaria.

Work Package 2. Metabolic Analysis and Antioxidant Activity: The scientists included in WP 2 work in the field of phytochemical analysis of medicinal plants extracts. Prof. Strahil Berkov has experience in project management and is expert in alkaloid chemistry, analytical methods (GC-MS, HPLC-UV / MS, NMR) and their application in analyzes of BAS and in metabolomics, and Associate Prof Milena Nikolova - in the study of the activity of BAS /surface flavonoids, antiradical activity/, determination of content, composition and dynamics of accumulation of BAS in protected and economically valuable medicinal plants.

Work Package 3 Embryological studies: Assoc. Prof. Elina Iankova-Tsvetkova, Assoc Prof. Petka Yurukova-Grancharova, and the young scientist Rosalia Nikolova are representatives of the only scientific unit in Bulgaria, working in the field of evolutionary, ecological and experimental embryology of wild plants of Bulgarian flora, with a focus of research on the reproductive biology and reproductive potential of rare, threatened and medicinal plant species.

Work Package 4. Genetic Studies and Flow Cytometry: Assist. prof. Vladimir Vladimirov has experience in the field of flow cytometry and has specialized these techniques in the Institute of Botany, Vienna, Austria. Assoc. Prof. Peter Zhelev has experience in population-genetic and phylogenetic studies in plants and in the development and application of different genetic markers. A main part of his research has been conducted jointly with foreign scientists in the framework of international projects. The scientific interests and experience of Assist prof. Ina Aneva are in the field of protection of medicinal plants and their genetic resources.

Work Package 5 Plant Biotechnology and hydroponic cultures: WP 5 consists of young scientists and PhD students leded by Assoc. Prof. Marina Stanilova who founded the Biotechnology laboratory for medicinal plants within her guided international project NATO SfP 974,453-Bioproduction (2001-2006) and developed the department of plant biotechnology in IBER-BAS. Dr. M. Stanilova has experience in the guidence of graduates, PhD students, as well as of big projects, incl. such financed by the NSF, connected with the in vitro propagation of medicinal, threatened, rare and endemic plants. PhD students Asya Kozhuharova and Vladimir Ilinkin have very good results in the clonal propagation, and the young researcher Boryanka Traykova – in the ex vitro adaptation, maintenance of ex situ collections and hydroponic cultivation.

PROJECT КП-06-Н26-6

Reproductive potential, metabolic and genetic profile, in in situ and ex situ conditions, of medicinal plants species from the Bulgarian flora with resource deficit - scientific base for their cultivation

The strategic objectives correspond to the priorities of the research in Bulgaria and worldwide in the last decades, related to the preservation of biodiversity.