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In the present work, hydroxyapatite-polymer materials were developed. The preparation, as well as characterization of the ceramic-polymer composites based on polyvinylpyrrolidone, sodium alginate, and gelatin were described. The system was enriched with the addition of common sage extract (Salvia officinalis). The antioxidant potential of sage aqueous extract and total polyphenol content was determined. The antioxidant capacity and total phenolic content of extract were equal to 86.06 ± 0.49% and 16.21 ± 0.58 mg gallic acid equivalents per gram of dry weight, respectively. Incubation studies in selected biological liquids were carried out to determine the biomineralization capacity on the surface of the composites and to examine the kinetics of release of the active substances from within the material. As a result of the incubation, a gradual release of the extract over time from the polymer matrix was observed, moreover, the appearance of new apatite layers on the composite surface was recorded as early as after 14 days, which was also confirmed by energy-dispersive X-ray spectroscopy (EDS) microanalysis. The composites were analyzed with Fourier transform infrared spectroscopy (FTIR) spectroscopy, and the morphology was recorded by scanning electron microscope (SEM) imaging. The in vitro biological studies allowed their cytotoxic effect on the reference L929 fibroblasts to be excluded. Further analysis of the biomaterials showed that enrichment with polyphenols does not support the adhesion of L929 cells to the surface of the material. However, the addition of these natural components stimulates human monocytes that constitute the first step of tissue regeneration.

Topicality. According to WHO, more than 4 % of the population globe suffer from a variety of joint and spine disorders. In Ukraine, more than 34,500 patients with primary joint diseases are registered annually. Diseases of the musculoskeletal system have a chronic course that requires long-term treatment and puts increased demands on the effectiveness and safety of anti-inflammatory drugs. Therefore, the above justifies the feasibility and relevance of the search for new highly effective and safe domestic drugs.Aim. To make an experimental study of the pharmacological activity of a combined gel for the treatment of inflammatory joint disease if chronic inflammation – adjuvant arthritis in rats.Materials and methods. The subjects of the study were model samples of a combined gel with active pharmaceutical ingredients - white willow bark, common sage leaves dried extract and methyl salicylate. The anti-inflammatory properties of the new gel were studied using a model of adjuvant arthritis in rats (sensitization to the base of the tail by PAF at 0.1 ml/animal, and the introduction on the seventh day of a resolving dose by PAF 0.1 ml/animal under aponeurosis of the right hind limb subplantarly). Diclofenac sodium gel, 5 %, was used as the comparison drug. The test specimens were applied to the affected paw surface daily at a dose of 0.2 ml/animal. Indicators: volume of edema of animal paws (3, 7, 10, 14 and 21 days of experiment); leukocyte and FAN content in the blood and alkaline and acid phosphatase activity and serum CIK content (21 days).Results and discussion. According to the results of studies, against the background of chronic inflammation, the combined gel shows an efficiency comparing to the diclofenac sodium gel, 5 %, conceding to it only in its anti-exudative effect. However, a positive feature of the new gel, in contrast to the drug of comparison, is the ability to maintain the cellular level of non-specific animal immunity, which is significantly impaired under the condition of this pathology.Conclusions. The obtained experimental data substantiate the prospect of further pharmacological studies of the developed combination gel based on the active ingredients of white willow bark, sage medicinal leaves dried extract and methyl salicylate., Актуальность. По данным ВОЗ свыше 4 % населения земного шара болеют разными заболеваниями суставов и позвоночника. В Украине ежегодно регистрируется более 34500 пациентов с первичными заболеваниями суставов. Заболевания опорно-двигательного аппарата имеют хроническое течение, что требует длительного лечения и выдвигает повышенные требования к эффективности и безопасности противовоспалительных средств. Таким образом, вишеизложенное обосновывает целесообразность и ктуальность поиска новых высокоэффективных и безопасных отечественных препаратов.Целью работы стало экспериментальное изучение фармакологической активности комбинированного геля для терапии воспалительных заболеваний суставов на модели хронического воспаления – адъювантного артрита у крыс.Материалы и методы. Объектами исследования были модельные образцы геля с активными фармацевтическими ингредиентами – ивы белой коры и шалфея лекарственного листья экстрактами сухими, а также метилсалицилатом. Противовоспалительные свойства нового геля исследовали на модели адъювантного артрита у крыс самцов (сенсибилизация в основу хвоста по ПАФ по 0,1 мл/животное и введение на седьмые сутки разрешающей дозы ПАФ по 0,1 мл/животное под апоневроз правой задней концовки субплантарно). Как препарат сравнения использовали гель Диклофенак натрия, 5 %. Исследуемые тест-образцы наносили на пораженную поверхность лапы ежедневно в дозе 0,2 мл/животное. Показатели: объем отека лапы животного (3, 7, 10, 14 и 21 сутки испытаний); содержание лейкоцитов и ФАН в крови и активность основной и кислой фосфатаз, а также содержание ЦИК в сыворотке крови (21 сутки).Результаты и их обсуждение. По результатам исследований на фоне хронического воспаления на модели адъювантного артрита разработанный комбинированный гель проявляет эффективность, сравнимую с гелем Диклофенак натрия, 5 %, уступая ему только по антиэкссудативному действию, но обнаруживая также и иммуно-модулирующее действие. Также позитивным со стороны фармакологического действия исследуемого средства является сохранение клеточного звена неспецифического иммунитета животных, которое существенно нарушается при условии данной патологии.Выводы. Полученные экспериментальные данные обосновывают перспективность дальнейших фармакологических исследований разработанного комбинированного геля на основе ивы белой коры и шалфея лекарственного листьев экстрактов сухих, а также метилсалицилата., Актуальність. За даними ВООЗ понад 4 % населення земної кулі страждають на різні захворювання суглобів та хребта. В Україні щорічно реєструється більше 34500 пацієнтів із первинними захворюваннями суглобів. Захворювання опорно-рухового апарату мають хронічний перебіг, що потребує довготривалого лікування та створює підвищені вимоги до ефективності та безпечності протизапальних засобів. Отже, вищевикладене обґрунтовує доцільність та актуальність пошуку нових високоефективних та безпечних вітчизняних препаратів.Метою роботи стало експериментальне вивчення фармакологічної активності комбінованого гелю для терапії запальних захворювань суглобів на моделі хронічного запалення – ад’ювантного артриту у щурів.Матеріали та методи. Об’єктами дослідження були модельні зразки комбінованого гелю з активними фармацевтичними інгредієнтами – верби білої кори та шавлії лікарської листя екстрактами сухими та метилсаліцилатом. Протизапальні властивості нового гелю досліджували на моделі ад’ювантного артриту у щурів самців (сенсибілізація в основу хвоста по ПАФ по 0,1 мл/тварину та введення на сьому добу розрізнювальної дози ПАФ по 0,1 мл/тварину під апоневроз правої задньої кінцівки субплантарно). Як препарат порівняння використовували гель Диклофенак натрію, 5 %. Досліджувані тест-зразки наносили на уражену поверхню лапи щоденно у дозі 0,2 мл/тварину. Показники: об’єм набряку лапи тварин (3, 7, 10, 14 та 21 доба досліду); вміст лейкоцитів та ФАН у крові та активність лужної та кислої фосфатаз і вміст ЦІК у сироватці крові (21 доба).Результати та їх обговорення. За результатами досліджень на тлі хронічного запалення комбінований гель виявляє ефективність, порівняну з гелем Диклофенак натрію, 5 %, поступаючись йому лише за антиексудативною дією. Проте позитивною рисою нового гелю на відміну від препарату порівняння є здатність до збереження клітинної ланки неспецифічного імунітету тварин, який суттєво порушується за умови даної патології.Висновки. Отримані експериментальні дані обґрунтовують перспективність подальших фармакологічних досліджень розробленого комбінованого гелю на основі активних інгредієнтів верби білої кори та шавлії лікарської листя екстрактів сухих та метилсаліцилату.

Vitalija Misevičiūtė. Master’s thesis “The technology of oral hydrogel and the quality research” thesis supervisor Assoc. Dr. Z. Kalvėnienė; Lithuanian University of Health Sciences, Medical Academy, Faculty of Pharmacy, Pharmaceutical Technology and Social Pharmacy Department. - Kaunas. The aim of this study: To create and construct a hydrophilic gel for the purpose of managing oral wounds and evaluate the gel's quality and stability. Research objectives: 1. To conduct a secondary research about the possible mouth wounds to be managed with hydrogels and the consistency, of the hydrogel, that is to be needed to achieve a positive outcome; 2. To produce hydrogels with lidocaine hydrochloride and sage extract; 3. To evaluate how polymer concentrations with sage extract are affecting the quality and stability of the hydrogel. Methods: Determination of pH value, viscosimetry, texture analysis, determination of antioxidative activity by DPPH*, lidocaine hydrochloride quantitative and stability tests. The results and conclusions: 1) Mouth hydrogels with lidocaine hydrochloride and sage extract are made using three different jellifying agents: Chitosan, Xanthan gum, and Acrylate / C10-30 alkyl acrylate crosspolymer (Ultrez 21); preservatives – EDTA, citral; taste enhancing saccharin; greasy and moisturizing plant glycerol; solvent – purified water. 2) pH analysis showed that Chitosan hydrogels are acidic (pH 3,59 – pH 3,65) and unstable when mixed with sage extract. While Xanthan gels have the pH value of 5,25 – 5,35. It is found, however, that once sage extract is added; it stays stable for up to one month. Ultrez 21 gels with a pH of (6,25 – 6,48) are the closest ones to our saliva environment. 3) Viscosimetry tests exhibited a strong correlation between jellifying agents and the dynamical viscosity formed by the same agents. It was found that Ultrez 21 polymer of a lower concentration creates a higher viscous hydrogel. The research of hydrogels consistency showed that there is statistically significant dependency of deformation force, consistency, cohesiveness, and viscosity index with the polymer and its concentration. 4) After one month, xanthan gum rubble was destroyed. Hencethe research was continued with Ultrez 21 gels only. DPPH* method showed that sage extract creates an antioxidant activity of 55, 05± 0, 4%. Furthermore, if it is inserted in 15% hydrogel concentration, the antioxidant activity increases significantly, regardless of the concentration of the polymer. 5) The amount of lidocaine hydrochloride in the hydrogels was determined by titration and the calculated release of the active substance was 95%. Polymer concentration did not affect the release of the active substance (p> 0.05). 6) Stability studies have shown that in the presence of darkness at 25 ± 2 ° C for 6 months, the pH value of both concentrations of Ultrez 21 remains good for the oral mucosa; the dynamic viscosity was statistically insignificant. Statistically significant changes in texture parameters - viscosity index and consistency occur after 3 months. Changes in hydrogel coughing remain unimportant for all 6 months of observation, as well as a deformative force of 1.0%. hydrogel (p

Birçok ülkede ekonomik olarak yetiştirilen adaçayı (Salvia officinalis L.)'nın üretimi aşamasında zararlı böceklerin neden olduğu önemli kayıplar gibi bir takım zorluklarla karşılaşılmaktadır. Bu çalışmada ilk olarak S. officinalis üzerindeki zararlı böcekler ve doğal düşmanları saptanmıştır. Çalışmanın ikinci aşamasında ise adaçayının önemli zararlılarından nane yaprakbiti Eucarazzia elegans (Ferrari) (Hemiptera: Aphididae)'ın zararı, ekonomik etkisi ve yaprakbiti popülasyonunu etkileyen çevresel faktörler üzerinde durulmuştur. Çalışmada zararlıların teşvik ettiği bir sentetik bitki uçucu yağlarının olan (HIPVs) metil salisilat (MeSA)'ın adaçayı alanlarındaki doğal düşmanlar üzerindeki çekici etkisi ve nane yaprakbiti, E. elegans popülasyon gelişimi üzerine etkisi araştırılmıştır. Çalışmada son olarak nane yaprakbiti E. elegans biyolojik savaşında kullanılması muhtemel Coccinella septempunctata Linnaeus ve Adalia bipunctata (Linnaeus) (Coleoptera: Coccinellidae) predatörlerinin tür içi ve türler arası etkileşimleri ortaya konulmuştur. Çalışma boyunca S. officinalis üzerinde 11 ayrı takıma ait 31 familyadan 51 böcek türü toplanmıştır. Bu türlerden 19'u zararlı, 3'u polinatör ve 29 tür doğal düşman olarak sınıflandırılmıştır. Zararlı böceklerden Micantulina stigmatipennis (Mulsant & Rey), Eupteryx gyaurdagicus Dlabola (Hemiptera: Cicadellidae), Aphis passeriniana (Del Guercio) ve Eucarazzia elegans (Ferrari) (Hemiptera: Aphididae), Dysmiscoccus angustifrons (Hall) (Hemiptera: Pseudococcidae), Chrysolina americana (Linnaeus) (Coleoptera: Chrysomelidae) ve Thrips tabaci Lindeman (Thysanoptera: Thripidae) adaçayı üzerinde en yoğun olarak görülen anahtar zararlı türler olarak saptanmıştır. Bu çalışma ile A. passeriniana'nın Türkiye'de bulunduğu ve E. elegans'ın ise İzmir'de bulunduğu ilk kez ortaya konulmuştur. Ayrıca M. stigmatipennis türünün ilk kez S. officinalis üzerinde beslendiği ve konukçusu olduğu saptanmıştır. Eucarazzia elegans'ın S. officinalis üzerindeki zarar oranını ve ekonomik etkisini belirlemek üzere sürvey ve laboratuvar çalışmaları gerçekleştirilmiştir. Çalışma sonuçlarına göre E. elegans'ın, erken ilkbahar döneminde %9.5 zarar oranı ile bir önceki yıldan kalan yaşlı yapraklar üzerinde beslendiği belirlenmiş ve daha sonra yaz başında %1.1 zarar oranı ile genç yapraklara, çiçek tomurcuklarına ve çiçeklere göç ettiği saptanmıştır. Yaprakbitinin mevsimsel dağılımının nem ve yağışa göre sıcaklıktan daha fazla etkilendiği görülmüştür. Yaprakbiti zararı dolayısıyla uçucu yağ miktarı üzerinde bir farklılık oluşturmamasına rağmen, uçucu yağ bileşenlerinden bazıları kontrol grubuna göre büyük ölçüde değişime uğramıştır. Toplam yaş ve kuru yaprak ağırlık artışı iki kat popülasyon seviyesinde sırasıyla kontrol ve normal yaprakbiti popülasyon seviyesinde 1/3 ve 1/2 oranında azalmıştır. Çalışmalarda E. elegans'ın adaçayında 8-10 haftadan fazla beslenmesi sonrasında %50'den fazla ürün kaybına neden olduğu saptanmıştır. En yüksek maliyet yarar oranı altı kez insektisit uygulanmış parsellerde gerçekleşmiştir. Zararlıların teşvik ettiği bitki uçucu yağlarından (HIPVs) olan ve yapay olarak elde edilmiş olan MeSA ile yapılan denemelerde, Syrphidae familyasına bağlı türler (Diptera), Rhagonycha fulva (Scopoli) (Coleoptera: Cantharidae) ve C. septempuctata (Coleoptera: Coccinellidae) türler yoğun olarak öne çıkmıştır. Yarı açık tarla denemelerinde, MeSA uygulanan ve dolayısıyla doğal düşmanların cezbedildiği adaçayı bitkilerinde E. elegans popülasyonları önemli biçimde azalmıştır. Laboratuvar koşullarında gerçekleştirilen denemelerde nane yaprakbiti E. elegans sayısı ve popülasyon artış oranının MeSA uygulanmış ve uygulanmamış bitkilerde farklı olmadığı ortaya konulmuştur. Bu durum, MeSA uygulamasının zararlı nane yaprakbiti türünün mücadelesinde doğal düşmanların rolünü vurgulamıştır. Yaprakbiti E. elegans'ın mücadelesinde etkili iki genel predatör, C. septempunctata (Cs) ve A. bipunctata (Ab) ile yapılan çalışmalarda; kültür kavanozu denemelerinde aynı tür (Cs + Cs) ve farklı tür (Cs + Ab) kombinasyonlarında antagonistik etki görülürken, aynı tür Ab + Ab kombinasyonunda belirgin sinerjistik etki görülmüştür. Bununla birlikte petri denemelerinde, Ab + Ab aynı tür kombinasyonu belirgin olarak, ilave etki şeklinde, fakat diğer predatör türün benzer kombinasyonunda (Cs + Cs) ve farklı tür kombinasyonunda (Cs + Ab) sinerjistik bir etki görülmüştür. Bununla birlikte petri alanında, aynı tür kombinasyonu Ab + Ab toplamsal etki göstermiş, fakat aynı tür kombinasyonu Cs + Cs ve farklı tür kombinasyonu Ab + Cs uygulamalarında sinerjistik etki ortaya çıkmıştır. Bu sonuca göre söz konusu predatörlerin av tüketim alışkanlıkları, farklı tür / aynı tür (F = 410.86; df = 4; P < 0.000;) kombinasyonlarında ve iki bağımsız faktörden (tür × deneme alanı) etkilenirken (F = 58.87; df = 4; P < 0.000) tek başına deneme alanının özelliklerinden etkilenmemiştir (F = 0.817; df = 1; P = 0.369). Buna göre söz konusu Coccinellidae türlerinin avı tercihinin öncelikle yaşam alanı yapısı karmaşıklığı yerine predatör çeşitliliği ile ilgili olduğu belirlenmiştir., Common sage (Salvia officinalis L.) have been economically cultivated in many countries, but the production is still facing a number of challenges including significant production losses due to insect pests. This research studied: 1) insect pests of S. officinalis and their natural enemies, 2) incidence and economic impact of a key insect pest, Eucarazzia elegans (Ferrari) (Hemiptera: Aphididae) on S. officinalis and evaluations of the environmental factors influenced the aphid population, 3) the attractiveness of methyl salicylate (MeSA) lures, a synthetic herbivore-induced plant volatiles (HIPVs), to the natural enemies in the sage fields and its impact on the mint aphid, E. elegans growth population, and 4) the interaction of two generalist predators, Coccinella septempunctata Linnaeus and Adalia bipunctata (Linnaeus) (Coleoptera: Coccinellidae) to control the mint aphid, E. elegans. Throughout the study, 11 insect orders spread into 31 families and 51 species consist of 19 natural enemies, 3 pollinators and 29 insect pests. Micantulina stigmatipennis (Mulsant & Rey), Eupteryx gyaurdagicus Dlabola (Hemiptera: Cicadellidae), Aphis passeriniana (Del Guercio), Eucarazzia elegans (Ferrari) (Hemiptera: Aphididae), Dysmiscoccus angustifrons (Hall) (Hemiptera: Pseudococcidae), Chrysolina americana (Linnaeus) (Coleoptera: Chrysomelidae) and Thrips tabaci Lindeman (Thysanoptera: Thripidae) were determined as the key insect pests. A. passeriniana was a new species recorded in Turkey whilst E. elegans were known as the new species recorded in Izmir. M. stigmatipennis has been found to feed on S. officinalis and report as a host plant for the first time. A survey and laboratory experiments on incidence rates and economic impact of E. elegans on S. officinalis revealed that E. elegans infested old leaves with an incidence rate of 9.5% in the early spring and then migrated to young leaves and blossom at the beginning of the summer with the incidence rate of 1.1%. The seasonal distribution of the aphid was affected more by temperature than humidity and rainfall. Although the aphid infestation had no differences on the quantity of the essential oil, some of the essential oil components were markedly different from the control. The total plant weights (fresh and dry) with the normal and double infestation rate were decreased about a third and more than a half compared to the control, respectively. Aphids feeding on common sage beyond 8-10 weeks caused more than 50% yield loss. The highest cost-benefit ratio was achieved in plots where six insecticide applications were made. The experiment with MeSA lures as an artificial herbivore-induced plant volitiles (HIPVs) recorded several numbers of natural enemies of which Syrphidae (Diptera), Rhagonycha fulva (Scopoli) (Coleoptera: Cantharidae), and C. septempuctata (Coleoptera: Coccinellidae) were abundantly caught on traps adjacent to the MeSA odors. In semi-field experiments, E. elegans populations were significantly reduced on treated common sage plants when it exposed to the natural enemies. When experiments were set in laboratory condition, the number and population growth rates of aphid did not differ between treated and untreated implicating no effects of MeSA on aphid population rate and emphasizing the role of natural enemies for controlling the mint aphid, E. elegans population. The study with two generalist adult predators, C. septempunctata (Cs) and A. bipunctata (Ab) (Coleoptera: Coccinellidae) to control aphid E. elegans resulted that in the culture jar the conspecific species Cs + Cs and heterospecific species Cs + Ab combinations showed antagonistic effects, but the synergistic effect for conspecific Ab+Ab. In the petri dish, however, the conspecific Ab + Ab combination exerted an additive effect, but a synergistic effect for conspecific Cs + Cs and heterospecific Cs + Ab. These prey consumption patterns were influenced by the combination hetero-/conspesific (F = 410.86; P < 0.000; df = 4) and the interaction among the two independent factors (species × arena)(F = 58.87; P < 0.000; df = 4) but not by type of arena alone (F = 0.817; P = 0.369; df = 1). Here, the prey preference in these ladybirds has primarily associated with a diversity of ladybird rather than habitat structure complexity.

V letih 2012–2014 smo v in vitro razmerah s tehniko tkivnih kultur razmnoževali žajbelj (Salvia officinalis L.). V poskus smo vključili izsečke S. officinalis kot izhodiščni material za gojenje v in vitro razmerah ter jih dali na indukcijsko gojišče. Rastlinski material smo z uporabo petih različnih metod sterilizacije predhodno površinsko sterilizirali. Najbolj uspešna je bila sterilizacija pri 15 minutnem obravnavanju s 50% etanolom in DICO. Druga naloga raziskave je bila najti čim bolj primerno gojišče za uspešno razmnoževanje žajblja. Pripravili smo tri različna gojišča G1, G2 in G3, ki so se med seboj razlikovala po vsebnosti rastnih regulatorjev. Največ poganjkov smo namnožili na gojišču G2, ki je vsebovalo BAP v koncentraciji 1 mg/l in IAA v koncentraciji 0,5 mg/l. Tretji del raziskave pa je bil povezan s koreninjenjem. Uporabili smo dve različni gojišči, K1 in K2. Število ukoreninjenih rastlin pri K1 je predstavljalo 82 %, pri K2 pa 79 % vseh rastlin. Na obeh gojiščih smo dobili dobre rezultate. V zaključnem delu raziskave smo rastline aklimatizirali v mini rastlinjakih. Po enem mesecu opazovanja smo prišli do zaključka, da so bile rastline uspešno aklimatizirane. Between 2012 and 2014 we applied the in vitro tissue culture technique to reproduce common sage (Salvia officinalis L.). The experiment involved ordinary cuts/fragments of S. officinalis as starting material for the in vitro cultivation, which was placed on the induction culture medium. The surface of the involved plant material was pre-sterilized using five different approaches. The most successful sterilization involved 50% ethanol and DICA (sodium dichloroisocyanurat) and lasted 15 min. The second task of the experiment was to find the most appropriate medium for sage reproduction. We prepared three different culture media: G1, G2 and G3, each with a different content of growth regulators. Most of the shoots were multiplied in the culture medium G2 that contained BAP (benylamino purine) in concentration of 0.5 mg/l. The aim of the third task was determine the best medium for rooting. We used two different culture media: K1 and K2. The number of rooted plants in the K1 culture medium represents 82% and in K2 79% of all plants. We got good results on both K1 and K2. The last task was the acclimatisation of plantlets in a miniature greenhouse. After a month of observations, we came to the conclusion that the majority of plants were successfully acclimatised.

Nine new microsatellite markers (SSR) were isolated from Salvia officinalis L. A total of 125 alleles, with 8 to 21 alleles per locus, were detected in a natural population from the east Adriatic coast. The observed heterozygosity, expected heterozygosity, and polymorphic information content ranged from 0.46 to 0.83, 0.73 to 0.93 and 0.70 to 0.92, respectively. New microsatellite markers, as well as previously published markers, were tested for cross-amplification in Salvia brachyodon Vandas, a narrow endemic species known to be present in only two localities on the Balkan Peninsula. Out of 30 microsatellite markers tested on the natural S. brachyodon population, 15 were successfully amplified. To obtain evidence of recent bottleneck events in the populations of both species, observed genetic diversity (HE) was compared to the expected genetic diversity at mutation-drift equilibrium (HEQ) and calculated from the observed number of alleles using a two-phased mutation model (TPM). Recent bottleneck events were detected only in the S. brachyodon population. This result suggests the need to reconsider the current threat category of this endemic species.

Salvia officinalis (Lamiaceae), common or Dalmatian sage, is a Mediterranean aromatic and medicinal plant used in medicine since ancient times. Knowledge on current genetic patterns and genealogical history of its natural populations is required for both breeding efforts and species conservation. We used sequences of two chloroplast intergenic spacers, 3'rps16-5'trnK and rp132-trnL, from 83 individuals from eight natural populations to distinguish between anthropogenic vs natural origin of four disjunct inland populations found outside of the main Adriatic range of the species. We found seven haplotypes, high total gene diversity (H-r = 0.695) and genetic differentiation (G(ST) = 0.682), as well as a phylogeographic structure with two lineages, a sub-structured inland-Adriatic lineage (IAL, comprising inland and Adriatic sub-lineages) and a purely Adriatic lineage (PAL). All four inland and disjunct populations, which comprised the inland sub-lineage of IAL, were almost fixed for a distinct haplotype genealogically closely related to the ancestral haplotype and displayed other features of relict populations. Along with previous biogeographic data and other lines of evidence, assumptions on their anthropogenic origin were rejected. At present, a less diverse IAL (Hd = 0.426, pi = 0.00106) and a more diverse PAL (Hd = 0.403, pi = 0.00257), whose divergence was dated to the Pliocene (3.267 Mya), do not exhibit signs of recent demographic expansions and overlap on the SE Adriatic coast, a region delineated as the main glacial refugium of S. officinalis. Conservation measures accounting for the historical distinctiveness of populations and focusing on currently the most threatened populations are recommended.

Functional coordination between leaf and stem hydraulics has been proposed as a key trait of drought-resistant plants. A balanced water transport efficiency and safety of different plant organs might be of particular importance for plant survival in the Mediterranean climate. We monitored seasonal changes of leaf and stem water relations of Salvia officinalis L. in order to highlight strategies adopted by this species to survive in harsh environmental conditions. During summer drought, the water potential dropped below the turgor loss point thus reducing water loss by transpiration, whereas the photosynthetic efficiency remained relatively high. Leaves lost their water transport efficiency earlier than stems, although in both plant organs P50 (water potential inducing 50% loss of hydraulic conductivity) indicated surprisingly high vulnerability when compared with other drought-tolerant species. The fast recovery of leaf turgor upon restoration of soil water availability suggests that the reduction of leaf hydraulic conductance is not only a consequence of vein embolism, but cell shrinkage and consequent increase of resistance in the extra-xylem pathway may play an important role. We conclude that the drought tolerance of S. officinalis arises at least partly as a consequence of vulnerability segmentation.

Nine new microsatellite markers (SSR) were isolated from Salvia officinalis L. A total of 125 alleles, with 8 to 21 alleles per locus, were detected in a natural population from the east Adriatic coast. The observed heterozygosity, expected heterozygosity, and polymorphic information content ranged from 0.46 to 0.83, 0.73 to 0.93 and 0.70 to 0.92, respectively. New microsatellite markers, as well as previously published markers, were tested for cross-amplification in Salvia brachyodon Vandas, a narrow endemic species known to be present in only two localities on the Balkan Peninsula. Out of 30 microsatellite markers tested on the natural S. brachyodon population, 15 were successfully amplified. To obtain evidence of recent bottleneck events in the populations of both species, observed genetic diversity (H$_E$) was compared to the expected genetic diversity at mutation-drift equilibrium (H$_{EQ}$) and calculated from the observed number of alleles using a two-phased mutation model (TPM). Recent bottleneck events were detected only in the S. brachyodon population. This result suggests the need to reconsider the current threat category of this endemic species.

Cilj ove disertacije bio je razvijanje i primjena mikrosatelitnih biljega na populacijama ljekovite kadulje (Salvia officinalis L.). Isto tako nastojalo se primijeniti razvijene biljege na drugim vrstama roda Salvia L. te vrsti Rosmarinus officinalis L.. Razvijeno je ukupno 29 biljega, a primjenom osam najinformativnijih provedena je populacijsko-genetička analiza 45 populacija ljekovite kadulje s Balkanskog poluotoka. Srodstveni odnosi između većine populacija bili su korelirani s njihovom geografskom udaljenošću. Izuzetak je pet populacija za koje se pretpostavlja da nisu prirodne nego potječu iz uzgoja. Prema alelnom bogatstvu središte genetičke raznolikosti ljekovite kadulje nalazi se u Srednjoj i Južnoj Dalmaciji. Primjena mikrosatelitnih biljega ljekovite kadulje na kratkozupčastoj kadulji (S. brachyodon Vandas), otkrila je prolaz populacije s poluotoka Pelješca kroz genetsko usko grlo. Dobiveni rezultat naglašava potrebu preispitivanja statusa ugroženosti ove endemične vrste. Mikrosatelitna analiza ljekovite i grčke kadulje na otoku Visu (S. fruticosa Mill.) potvrdila je postojanje hibridne svojte S x auriculata Mill.. The goal of this dissertation was development and application of microsatellite markers on common sage populations, as well as their application on other sage species and Rosmarinus officinalis L. Overall, 29 markers were developed, and by using eight of highest informativity, population genetic analysis of 45 common sage populations from Balkan peninsula was performed. Relationships between most of populations were corelated with their geographical distribution, with exeption of 5 populations which are believed to originate from cultivation. According to allelic richness, center of common sage genetic diversity is located in Central and Southern Dalmatia. Apllication of microsatellite markers from common sage on Salvia brachyodon Vandas revealed a recent bottleneck event in population from Pelješac peninsula, thus stressing the need for redefining of conservation status of this endemic species. Microsatellite analysis of common and greek sage (S. fruticosa Mill.) from Island of Vis confirmed presence of lineage of hybrid origin, S x auriculata Mill..

 Premise of the study: Microsatellite primers were identifi ed to study the genetic diversity, population genetic structure, and relationships of common sage and closely related species.  Methods and Results: Nine novel polymorphic trinucleotide microsatellite loci were identifi ed for Salvia offi cinalis . A total of 127 alleles were observed. The observed and expected heterozygosity values ranged from 0.375 to 0.880 and from 0.624 to 0.931, respectively. Nine new and 11 previously published microsatellite primers were tested for cross-amplifi cation. The species with the most successful cross-amplifi cation was S. fruticosa .  Conclusions: The nine new microsatellite markers will be useful in genetic studies of wild and cultivated populations of common sage and are potentially useful in genetic studies of closely related species.

Eleven polymorphic microsatellite markers were developed from common sage (Salvia officinalis L). The variability of specific microsatellite regions was assessed in wild population from the eastern Adriadic coast. All eleven primer pairs showed single-locus amplification with a total of 142 alleles and observed heterozygosity ranged from 0.300 to 0.833.