Features of creating a collection of Crimean native grape varieties in vitro

В статье приводятся результаты работы по созданию коллекции крымских автохтонных сортов in vitro. Нынешнее состояние требует принятия экстренных мер по поддержанию ее жизнеспособности. Одним из возможных способов сохранения ценного генетического материала является создание вегетирующих коллекций растений in vitro . Процесс создания коллекции растений in vitro автохтонных сортов винограда состоял из отдельных этапов: этап «сбор исходного материала», этап «получение первичного экспланта», этап «стерилизация и введение в условия in vitro одно- двухглазковых эксплантов побегов», этап «индукция развития побега», этап «укоренение развившихся побегов», этап «минимальное размножение микрочеренкованием до объема 10-15 растений», этап «перевод растений из активного роста в режим глубокого покоя для сохранения». Получена асептическая культура 47 автохтонных крымских сортов винограда, что составляет 65,3% от общего числа собранных на Ампелографической коллекции Института «Магарач». Основные причины, лимитирующие сортовой спектр, были вызваны отсутствием пролиферации почки in vitro и гибелью растений на стадии индукции побега. Таким образом, вместе с образцами, которые уже поддерживались в культуре, вегетирующая коллекция крымских автохтонных сортов составляет 50 образцов. The article presents the results of work on the creating of a collection of Crimean native varieties in vitro . The current state of collection requires urgent measures to maintain its viability. One of the possible ways to preserve valuable genetic material is to create vegetating collections of plants in vitro . The process of creating a collection of native grape varieties in vitro consisted of different stages: the stage of “collecting the initial material”, the stage of “obtaining the primary explant”, the stage of “sterilizing and introducing one- or two-eye shoot explants into the in vitro conditions”, the stage of “induction of the shoot development”, the stage of “rooting of the developed shoots”, the stage of “minimal reproduction by microcutting to the volume of 10-15 plants”, the stage of "plants transfer from active growth to deep dormancy for conservation". The aseptic culture of 47 Crimean native grape varieties was obtained, which is 65.3% of the total number collected in the Ampelographic Collection of the Magarach Institute. Main reasons limiting the varietal spectrum were caused by the lack of bud proliferation in vitro and the loss of plants at the stage of shoot induction. Thus, together with samples already preserved in the culture, the vegetating collection of Crimean native varieties consists of 50 samples.

Chemical Compositions, Somatic Embryogenesis, and Somaclonal Variation in Cumin

This is the first report evaluating the relationship between the chemical compositions of cumin seeds (based on the analysis of the content of catalase, ascorbate peroxidase, proline, protein, terpenic compounds, alcohol/phenols, aldehydes, and epoxides) and the induction efficiency of somatic embryogenesis in two Iranian superior cumin landraces (Golestan and North Khorasan). Cotyledons isolated from Golestan landrace seeds cultivated on MS medium supplemented with 0.1 mg/L kinetin proved to be the best primary explant for the induction of somatic embryogenesis as well as the regeneration of the whole plantlet. Results indicated that different developmental stages of somatic embryos were simultaneously observed on a callus with embryogenic potential. The high content of catalase, ascorbate peroxidase, proline, and terpenic hydrocarbons and low content of alcoholic and phenolic compositions had a stimulatory effect on somatic embryogenesis. Band patterns of RAPD markers in regenerated plants were different from those of the mother plants. This may be related to somaclonal variations or pollination system of cumin. Generally, measurement of chemical compositions can be used as a marker for evaluating the occurrence of somatic embryogenesis in cumin. Also, somaclonal variations of regenerated plants can be applied by the plant breeders in breeding programs.

Subpopulation of nestin positive glial precursor cells occur in primary adult human brain cultures

Glial fibrillary acidic protein (GFAP) is an intermediate filament protein considered to be the best astroglial marker. However, the predominant cell population in adult human brain tissue cultures does not express GFAP; these cells have been termed “glia-like” cells. The basic question about histological origin of adult human brain cultures remains unanswered. Some authors showed that “glia-like” cells in adult human brain cultures might be of non-glial origin. We examined primary explant tissue cultures derived from 70 adult human brain biopsies. Within first 5–10 days approximately 5–10% of the small explants became attached. Outgrowing cells were mostly flat cells. These cells formed confluent layer over 3–6 weeks in culture. At confluence the cultures contained 2–5% of microglial cells, 0.1% GFAP-positive astrocytes, less than 0.01% oligodendrocytes and 95–98% GFAP-negative “glia-like” cells. This population of flat “glia-like” cells was positively stained for vimentin, fibronectin, and 20–30% of these cells stained for nestin. Our findings revealed that 1 mM dibutyryl-cAMP addition, in serum free conditions, induced a reversible stellation in 5-10% of the flat “glia-like” cells but did not induce the expression of GFAP or nestin in morphologically changed stellate cells. These results demonstrate that “glia-like” cells in primary adult human brain cultures constitute heterogeneous cell populations albeit with similar morphological features. Two distinct subpopulations have been shown: (i) the one immunostained for nestin; and (ii) the other reactive for dibutyryl-cAMP treatment.