Исследования коллектива кафедры общей фармацевтической и биомедицинской технологии в области биоинженерии растительных продуцентов биологически активных веществ

Article is devoted to the complex researches of various morphological groups of raw materials, a cellular biomass and a productive part of metaboloms of vegetative superproducers of a natural and biotechnological origin. All investigations were carried out according to the Federal special purpose program «Scientific and scientific and pedagogical shots of innovative Russia» for 2009-2013, action item № 1.2.1"Carrying out of scientific researches by scientific groups under the direction of doctors of sciences», Project: «Bioengineering as a basis of mobilization of adaptive potential of bioobjects - superproducers BAS» GK П555/05.08.2009 - НК-97П.Статья посвящена комплексным исследованиям различных морфологических групп сырья, клеточной биомассы и продуктивной части метаболома растительных суперпродуцентов природного и биотехнологического происхождения. Все исследования выполнялись в соответствии с Федеральной целевой программой «Научные и научно-педагогические кадры инновационной России» на 2009-2013 гг., в рамках реализации мероприятия № 1.2.1 «Проведение научных исследований научными группами под руководством докторов наук». Проект: «Биоинженерия как основа мобилизации адаптивного потенциала биообъектов - суперпродуцентов БАВ» ГК П555/05.08.2009 - НК-97П

Характеристика представителей рода Atropa в условиях интродукции

Four taxa of the genus Atropa L. introduced in the central regions of the Russian Federation: Atropa acuminata Royle, A. komarovii Blin. & Shalyt, A. belladonna L. f. lutea (Döll) G.Dietr. and A. belladonna L. were studied. The studied Atropa taxa were evaluated by the following criteria: phenological phases, seed and raw materials productivity, pest damaging, content of active compounds. After winter rest, on the second year of vegetation, all introduced taxa have passed all phenological phases, entered fruiting stage and formed seeds with high rates of germination energy and germination capacity. The studies of pollen size and fertility have shown variation in pollen fertility from 81.0 to 94.0%. The obtained information on the resistance to regional biotic and abiotic factors indicates high adaptive potential of the species. The introduced Atropa taxa, as compared to A. belladonna cv. Bagira, produce less raw materials and seeds but has higher content of bioactive compounds. Being protected, the studied introduced Atropa taxa of different geographical origin can be considered as potential source of alkaloids for pharmaceutical plant raw materials. According to the karyological studies of belladonna species their chromosome numbers were determined as 2n = 72 and 2n = 74. The low variability in karyotype indicators of the studied Atropa species proves the stability of their chromosome sets. REFERENCES 1. Maqbool F., Singh S., Kaloo Z.A., Jan M. 2014. Medicinal importance of Genus Atropa Royle – A review. – Int. J. Adv. Res. 2(2): 48–54. http://www.journalijar.com/uploads/637_IJAR-2611.pdf 2. Atlas arealov i resursov lekarstvennykh rasteniy SSSR. 1983. [Atlas of the distribution areas and resources of the medicinal plants of the USSR]. Moscow. 340 p. (In Russian) http://plant.geoman.ru/books/item/f00/s00/z0000004/ 3. Rhodes J.B., Abrams J.H., Manning R.T. 1978. Controlled clinical trial of sedative-anticholinergic drugs in patients with the irritable bowel syndrome. – J. Clin. Pharmacol. 18(7). P. 340–345.https://doi.org/10.1002/j.1552-4604.1978.tb01603.x 4. Chopra. R.N., Nayar S.L., Chopra I.C. 1986. Glossary of Indian Medicinal Plants (Including the Supplement). New Delhi. 330 p. 5. Tyler V.E., Brady L.R., Robbers J.E. 1988. Pharmacognosy. – 9th Edition. P hiladelphia. 537 p. 6. Srivastava V., Negi A.S., Ajayakumar P.V., Khan S.A., Banerjee S. 2012. Atropa belladonna hairy roots: Orchestration of concurrent oxidation and reduction reactions for biotransformation of carbonyl compounds. – Appl Biochem Biotechnol – 166(6): 1401–1408. https://doi.org/10.1007/s12010-011-9533-3 7. Waterman P.G. 1993. Alkaloids: general observations. In: Methods in plant biochemistry: Volume 8. Alkaloids and sulphur compounds. New York. P. 1–16. 8. Zhang L., Kai G.Y., Lu B.B., Zhang H.M., Tang K.X., Jiang J.H., Chen W.S. 2005. Metabolic engineering of tropane alkaloid biosynthesis in plants. – J. of Integrative Plant Biology. 47:136–143. 9. Ashtiana F. 2011. Tropane alkaloids of Atropa belladonna L. and Atropa acuminate Royle ex Miers plants. – J. Medicinal Plants Research. 5(29):6515–6522. https://doi.org/10.5897/JMPR2011.482 10. Guggisberg A., Hesse M. 1984. Chapter 3. Putrescine, spermidine, spermine and related polyamine alkaloids. – The Alkaloids: Chemistry and Pharmacology. 22: 85–188. https://doi.org/10.1016/S0099-9598(08)60178-9 11. Fitopreparapty VILAR (Spravochnik). 2009. [VILAR herbal preparations (Reference book)]. M. 255 p. (In Russian) 12. Chavan N. 2017. Coin image of action of Atropa belladonna in crude and in homoeopathic potentised form. – J. Dev. Drugs. 6(2): 175. https://doi.org/10.4172/2329-6631.1000175 13. Klimakhin G.I. 2000. IntroduktsioИзучены таксоны рода Atropa L., интродуцируемые в условиях Нечерноземной зоны РФ: Atropa acuminata Royle, A. komarovii Blin. & Shalyt, A. belladonna L. f. lutea (Döll) G.Dietr. в сравнении с сортом Багира (A. belladonna). Дана оценка изучаемых таксонов на ряд признаков: фенологические фазы, продуктивность сырья и семян, повреждаемость вредителями, содержание действующих веществ. Показано, что интродуценты рода Atropa уступают по продуктивности сырья и семян районированному сорту Багира, но характеризуются повышенным содержанием биологически активных соединений и могут рассматриваться в качестве потенциальных продуцентов алкалоидов для фармацевтического сырья. Проведены кариологические исследования таксанов Atropa и установлены их хромосомные числа (2n = 72; 2n = 74). Выявленная невысокая изменчивость кариотипических показателей свидетельствует о стабильности хромосомного набора кариотипов исследуемых таксонов

Comparative cytogenetic study of the tetraploid Matricaria chamomilla L. and Matricaria inodora L.

A comparative cytogenetic study of the autotetraploid breed of Matricaria chamomilla L. (M. recutita L.) and Matricaria inodora L. was carried out by DAPI-banding, fluorescent hybridization in situ (FISH) with 26S and 5S rDNA probes, and analysis of meiosis. All chromosomes were identified in both karyotypes on the basis of DAPI-banding images and 26S and 5S rDNA distribution, and species-specific idiograms were composed for both M. chamomilla and M. indora taking into account the polymorphous variants of DAPI-banding images, showing the location of the 26S and 5S rDNA sites. © 2014 Pleiades Publishing, Inc

A comparative analysis of karyotypes of three species of Macleaya-producers of a complex of isoquinoline alkaloids

Using a set of methods (C-banding, DAPI-staining, fluorescence hybridization in situ (FISH) with probes of 26S and 5S rDNA, and analysis of meiosis), the first comparative cytogenetic study of three species of Macleaya, producers of complex isoquinoline alkaloids, cordate Macleaya cordata (Willd.) R. Br. (2n = 20), small-fruited Macleaya microcarpa (Maxim.) Fedde (2n = 20) and Macleaya kewensis Turrill (2n = 20), was first carried out. On the basis of morphometric analysis, formulas of karyotypes were made for each species. Species ideograms for M. cordata, M. microcarpa, and M. kewensis were constructed taking into account the polymorphic variants of the C-banding patterns and indicating the location of 26S and 5S rDNA sites. A comparative study revealed that the karyotypes of M. microcarpa and M. kewensis have more in common with each other than with M. cordata. Analysis of meiotic chromosomes suggests of genetic stability of Macleaya genomes. The results of chromosome analysis were used to confirm the close relationship of Macleaya and to clarify their phylogenetic relationships. © 2012 Pleiades Publishing, Ltd

A comparative analysis of karyotypes of three species of Macleaya-producers of a complex of isoquinoline alkaloids

Using a set of methods (C-banding, DAPI-staining, fluorescence hybridization in situ (FISH) with probes of 26S and 5S rDNA, and analysis of meiosis), the first comparative cytogenetic study of three species of Macleaya, producers of complex isoquinoline alkaloids, cordate Macleaya cordata (Willd.) R. Br. (2n = 20), small-fruited Macleaya microcarpa (Maxim.) Fedde (2n = 20) and Macleaya kewensis Turrill (2n = 20), was first carried out. On the basis of morphometric analysis, formulas of karyotypes were made for each species. Species ideograms for M. cordata, M. microcarpa, and M. kewensis were constructed taking into account the polymorphic variants of the C-banding patterns and indicating the location of 26S and 5S rDNA sites. A comparative study revealed that the karyotypes of M. microcarpa and M. kewensis have more in common with each other than with M. cordata. Analysis of meiotic chromosomes suggests of genetic stability of Macleaya genomes. The results of chromosome analysis were used to confirm the close relationship of Macleaya and to clarify their phylogenetic relationships. © 2012 Pleiades Publishing, Ltd

Comparative cytogenetic study of the forms of Macleaya cordata (Willd.) R. Br. From different localities

A comparative cytogenetic study of two introduced forms of Makleaya cordata (Willd.) R. Br. = syn. Bocconia cordata Willd. grown in different ecological and geographical regions (Moscow and Donetsk areas) was carried out. In the study, a complex of methods utilizing various chromosomal markers, i.e., C- and DAPI-banding technique, fluorescence in situ hybridization (FISH) with probes of 26S and 5S rDNA, as well as estimation of the total area of C-positive regions (C-HCH) in prophase nucleoli and meiosis analysis, was used. In the karyotypes (2n = 20), each chromosome was identified on the basis of C-banding and FISH patterns and the chromosome ideograms were built. Pericentrometric and telomeric C-positive bands in chromosomes of the Moscow form karyotype were found to be smaller and intercalary bands, larger than the corresponding bands in the M. cordata form grown in Donetsk. It was found that the content of C-HCH in prophase nucleoli in the form of M. cordata grown in Donetsk was higher than in the form grown in Moscow. In both forms sites of 26S rDNA and 5s rDNA were localized on satellite chromosome 1 and on chromosome 4 respectively but the signals were more intensive in the plant form grown in Donetsk. The results of this study enable selecting M. cordata forms for use in pharmacology and recommending them for cultivation in various ecological and geographical regions. © 2012 Pleiades Publishing, Ltd

Comparative cytogenetic study of the tetraploid Matricaria chamomilla L. and Matricaria inodora L.

A comparative cytogenetic study of the autotetraploid breed of Matricaria chamomilla L. (M. recutita L.) and Matricaria inodora L. was carried out by DAPI-banding, fluorescent hybridization in situ (FISH) with 26S and 5S rDNA probes, and analysis of meiosis. All chromosomes were identified in both karyotypes on the basis of DAPI-banding images and 26S and 5S rDNA distribution, and species-specific idiograms were composed for both M. chamomilla and M. indora taking into account the polymorphous variants of DAPI-banding images, showing the location of the 26S and 5S rDNA sites. © 2014 Pleiades Publishing, Inc

Molecular cytogenetic characterization, leaf anatomy and ultrastructure of the medicinal plant Potentilla alba L.

Potentilla alba L. is a valuable medicinal plant widely used in folk and traditional medicine and particularly promising in complex treatment of thyroid pathology. Natural resources of this species are insufficient due to ever-growing use in contemporary medicine. Comprehensive investigations of different P. alba populations are essential for the successful extension of P. alba plantings. Aiming for a better understanding of karyotype structure, chromosome behaviour in meiosis and developing new diagnostic characters, we performed molecular cytogenetic characterization and leaf structure and ultrastructure analyses of two introduced P. alba samples originating from different habitats. Based on chromosome morphology, distribution of 45S/5S rDNA and DAPI-banding patterns, all chromosomes in the karyotypes were identified and the P. alba chromosomal idiogram was constructed. Our findings confirmed P. alba karyotype stability and also revealed several diagnostic characters of this species: the features of cells of upper and lower leaf epidermis, the presence of calcium oxalate druses and three types of leaf indumentum, essential for evaluation of genetic diversity in different populations, validation of raw materials and further selection progress. The meiotic abnormalities were detected probably related to low pollen activity and indicated the advantages of vegetative propagation in the development of a P. alba plantation system. © 2018, Springer Science+Business Media B.V., part of Springer Nature

Comparative cytogenetic study of the forms of Macleaya cordata (Willd.) R. Br. From different localities

A comparative cytogenetic study of two introduced forms of Makleaya cordata (Willd.) R. Br. = syn. Bocconia cordata Willd. grown in different ecological and geographical regions (Moscow and Donetsk areas) was carried out. In the study, a complex of methods utilizing various chromosomal markers, i.e., C- and DAPI-banding technique, fluorescence in situ hybridization (FISH) with probes of 26S and 5S rDNA, as well as estimation of the total area of C-positive regions (C-HCH) in prophase nucleoli and meiosis analysis, was used. In the karyotypes (2n = 20), each chromosome was identified on the basis of C-banding and FISH patterns and the chromosome ideograms were built. Pericentrometric and telomeric C-positive bands in chromosomes of the Moscow form karyotype were found to be smaller and intercalary bands, larger than the corresponding bands in the M. cordata form grown in Donetsk. It was found that the content of C-HCH in prophase nucleoli in the form of M. cordata grown in Donetsk was higher than in the form grown in Moscow. In both forms sites of 26S rDNA and 5s rDNA were localized on satellite chromosome 1 and on chromosome 4 respectively but the signals were more intensive in the plant form grown in Donetsk. The results of this study enable selecting M. cordata forms for use in pharmacology and recommending them for cultivation in various ecological and geographical regions. © 2012 Pleiades Publishing, Ltd