ANNOTATION of thesis for the scientific degree of Doctor of Philosophy (Ph.D) Specialty «6D061300-Geobоtany»

Karasholakova Lyazzat Naushabayevna

“ Structure of populations, current state, rational use and preservation of rare endemic species Lonicera iliensis Pojark»

General description of the work. The thesis is dedicated to the study and assessment of the current state of populations of rare, endemic species Lonicera iliensis Pojark. from Ili-Balkhash region for rational use and solve of ex situ conservation problems. Relevance of the research topic. Protection and conservation of the gene pool of plants, especially of rare, endangered and endemic plants at this time are the actual problem. All plant species are vulnerable and an important part of the flora, the loss of each species can lead to the destruction of ecosystems and the depletion of biodiversity. For the safety of rare and endemic species there is need to examine and assess the current status of their populations. Lonicera iliensis Pojark. – rare and almost endemic species, with sharply decreasing range, which belongs to the family of Caprifoliaceae Juss., subsection of Caerulea Rehd.. Under the government regulation of the RK from 21.06.2007 № 521 was included in the list of objects of environmental protection that have particular ecological, scientific and cultural importance. The range of ili honeysuckle decrease every year, it was entered ih tne “Red Book” for the preservation as Ili-Balkhash region endemic. The relevance of this study is to examine and assess the current state of Lonicera iliensis Pojark. populations. According to the literature there are reports of high antimicrobial, anticarcinogenic and antioxidant activity of essential oils from the flowers of Japanese honeysuckle. The phytochemical studies of honeysuckle species conducted insufficiently until today. Therefore, the study of component composition and antimicrobial activity of Lonicera iliensis Pojark essential oils is relevant. In world practice, along with the traditional methods of genetic resources preservation are widely used methods for cryopreservation of germplasm accessions. Cryopreservation is a deep freezing and storage of cells, tissues and

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organs of plants in liquid nitrogen at very low temperature (-196ºC). Therefore it is important for the conservation of this rare, endemic species to use all the advanced technologies, including the creation in vitro aseptic plants and seed collection at different storage temperatures. Created collections of in vitro aseptic plants and seed can be used as material for reintroduction (for recovery) this species in nature, as well as for the exchange of genetic material between research organizations. The goals and tasks of the work. The study of population structure, assessment of the current state, the definition of phytochemical characteristics for the rational use and preservation of rare endemic species of Ile-Balkhash region Lonicera iliensis Pojark. The objectives of the work: 1. To study the number, age structure of coenopopulations and to determine the floristic composition of plant communities with the participation of Lonicera iliensis Pojark.; 2. To study of anatomical and morphological features of Lonicera iliensis Pojark. stem and leaf structure from different populations; 3. To obtain the essential oils of Lonicera iliensis Pojark., to determine of qualitative composition and quantitative content of essential oil and determine the antimicrobial activity of essential oils; 4. To develop ex situ conservation regulations and to create collections of in vitro aseptic plants and Ili honeysuckle seeds on different storage temperatures (+4°С; –18°С...–20°С; –196°С) 5. To develop recommendations of in situ and ex situ conservation of rare, endemic species Lonicera iliensis Pojark. Object of the study: three natural populations of Lonicera iliensis: the first population of Ili honeysuckle was found in the floodplain of river Ili near village in the , , the second population – in the floodplain of river Chilik near village , , Almaty region, the third population – in the floodplain of the Charyn River in the tract in Aktogai, Raiymbek district, Almaty region. Research methods. In the work used geobotanical, biotechnological and microbiological methods, as well methods of gas chromatography with flame ionization detector and mass spectrometry. Scientific novelty of the research. As a result of our research for the first time:

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- carried out a comprehensive study of the 3 populations, 9 coenopopulations of rare endemic species Lonicera iliensis Pojark. based on the number, age spectrum; - conducted the analysis of the biological diversity of plant communities with the participation of Lonicera iliensis; - investigated biometrics of morphological and anatomical structure of the aerial vegetative organs; - detailed component composition of essential oils extracted from various organs on the different vegetation phases of three natural populations of Ili honeysuckle were identified. A total of 47 constituents making up 67.4–89.4% of the total essential oils volume were identified. Differences in the essential oils constituent composition were found between populations and samples; - the antimicrobial activity of Lonicera iliensis essential oils obtained from different organs on the different vegetation phases was determine. The study of the antimicrobial activity of essential oils was performed with relative to strains of gram-positive bacteria: Bacillus cereus, Bacillus subtilis, Staphylococcus aureus 10 and strain gram-negative Escherihia coli 113; - regulations of medium-term and long-term seed storage of Ili honeysuckle at different temperatures (+ 4°C and –18°C...–20°C) and cryopreservation (– 196°C) have been developed. The seed collection of Ili honeysuckle from three natural populations at different storage conditions was created. The composition of the medium for introduction to in vitro and micropropagation of Ili honeysuckle were optimized, also was created in vitro collection of aseptic plants from three natural populations. Scientific and practical relevance of dissertation. The scientific significance of the results of the study is that the obtained new scientific information about features of coenopopulations rare endemic species Lonicera iliensis, which will allow to objectively assess the state of the populations studied species and to predict the prospects of its existence and further development. The new information on the component composition of essential oils of Lonicera iliensis depending on the part of the plant, the growing phase and the study population were obtained. The obtained results can be used for the reintroduction of the purpose of preservation of unique populations, as well as the organization of work for the ex situ conservation (genebanks seed, in vitro culture). The act of implement the research results into practice was obtained.

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Published methodic recommendations "Essential oils of honeysuckle", in which outlined the basic principles of the work on obtaining essential oils from different parts of the plant and determine the qualitative composition and quantitative content of components, as well as on identifing the antimicrobial activity of essential oils. The results obtained during the execution of the work are used in the teaching of theoretical and special courses to undergraduates in the Department of biodiversity and bioresources of the Al-Farabi Kazakh National University. Main provisions for the defense: 1. The results of the analysis of Lonicera iliensis 3 population, 9 coenopopulations structure based on the number and age composition. 2. Evaluation of the biological diversity of plant communities with the participation of Lonicera iliensis. 3. Features of the anatomical and morphological structure of ground vegetative organs of Lonicera iliensis. 4. The results of the analysis of the component composition of essential oils extracted from various organs of plants of three natural Ili honeysuckle populations. 5. Essential oils samples of Lonicera iliensis with high antimicrobial activity. 6. Regulations of medium-term and long-term preservation of Ili honeysuckle seeds at different temperatures (+4°С, –18°С...–20ºС and –196°С). The personal contribution of the author. The author of the work, have participated in the selection of the concept and research objective, determination of the work goal, research setting targets, also in the design and performance of all experiments, collection and analysis of received data and writing of the dissertation. The connection with the plan of the basic scientific works. The dissertation was performed within the projects «The evaluation of the genetic state of populations of endemic, rare and threatened plant species from Ile- Balkhash region with important ecological, scientific and cultural value by morpho-anatomical and molecular genetic analysis methods» (state registration number 0110РК00419) for 2011-2013 and «Development of production biotechnology for biologically active preparations of Kazakhstan species of honeysuckle» (№ state registration number 0112РК00867), which was carried out in the framework of scientific and technical program 0.0585 ”Biotechnology for creation of medicinal preparations, biological active supplements and specialized

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diet products and other materials on base of cultivated, introduced and wild plant species of Kazakhstan’s flora” for 2012-2014. Testing of the work: The research results and main dissertation provisions were reported and provided on the international scientific conferences: - International Scientific Conference dedicated to the 90th anniversary of the birth of G.B.Kedrov (16-21 September, 2013, Moscow); - III International Conference dedicated to the 70th anniversary of the National Academy of Sciences of BPI, year of ecological culture and environmental protection (CIS) and the 150th anniversary of the birth of VI Vernadsky (September 17-21, 2013, Bishkek); - 10th International Symposium on the Chemistry of Natural Compounds (21- 23 November, 2013, Tashkent-Bukhara); - International Conference "The systematic and floristic study of northern Eurasia" (dedicated to the 85th anniversary (of the birth) of Prof. AG Yelenevskii.) (2013, Moscow); - Plant Biology and Biotechnology International Conference (28-30 May, 2014, Almaty); - Abstracts of 17th European Congress on Biotechnology (15-18 May, 2014, Lecce). Publications. Dissertation main provisions, results, findings and conclusion are set forth in 13 printed works, among them 1 article in international journal, which are included into the database of Thomson Reuters; 4 articles in republican scientific journals, recommended by the Committee on control in education and science sphere of the Ministry of Education and Science of RK; 4 articles and 3 abstracts in the materials of the conferences of republican and international level, 1 guidelines. The structure of the dissertation. Dissertation is stated on 152 pages of computer text and consists of normative references, symbols and abbreviations, introduction, literature review, materials and methods, results and discussion, conclusions, list of references from 224 titles, contains 24 pictures, 6 tables and appendix 7.

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