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БИОЛОГИЧЕСКИЕ НАУКИ
y^K 577.151.64:581.143.6
O.A.Seldimirova*, I.R.Galin**
*PhD in Biology, Leader Scientific Researcher **PhD in Biology, Junior Scientific Researcher Ufa Institute of Biology RAS Yu.A. Yanbaev Doctor of Biology, Professor Vice-Chancellor for Academic Work Bashkir State University, Ufa
ISOZYME ANALYSIS FOR EVALUATION OF VARIABILITY OF ANDROGENIC REGENERANTS OF SPRING WHEAT OBTAINED THROUGH POLYEMBRYOIDOGENESIS IN VITRO
Abstract
Isozyme markers and polyacrylamide gel electrophoresis have been used to reveal isozyme loci of some enzyme systems of androgenic regenerants obtained through polyembryoidogenesis in wheat anther culture in vitro. A complete lack of variability in electrophoretic spectra of regenerants was revealed.
Keywords
Androgenesis in vitro, anther culture, polyembryoids, isozymes, wheat
Androgenesis in vitro, based on the method of anther culture, is a promising biotechnological approach widely used in the commercial breeding of spring wheat [1]. Biotechnologically optimal way to obtain haploids in anther culture in vitro is embryoidogenesis - the formation of embryo-like structure - embryoid, which immediately develops into regenerants [2]. A special case of embryoidogenesis in vitro is the formation of embryoids with multiple shoot apical meristem - polyembryoids [6].
On the one hand polyembryoids formations must provide genetic uniformity of regenerants and therefore to be one of the ways to optimize biotechnology of wheat valuable genotypes cloning [2, 5]. On the other hand, the entering of plants in culture in vitro is the strong stress factor which may cause a variety of genetic changes [4]. In this connection, the evaluation of genetic homogeneity of regenerants obtained through polyembryoidogenesis is very important.
Materials and methods
The objects of the study were regenerants of spring wheat cultivar Zhnitsa. They were obtained through polyembryoidogenesis in anther culture in vitro [3]. The some isozymes of enzyme systems, described earlier as promising for the detection of genetic changes in wheat regenerants obtained in culture in vitro [7], were used as a molecular markers. The method of polyacrylamide disk electrophoresis have been used The enzymes were separated in vertical plates of polyacrylamide gel and histochemically detected as described in [8].
Results and discussions
During the studies we have found a complete similarity of the of enzyme systems spectra in androgenic regenerants. On the figure 1 the electrophoretic spectra for glutamate dehydrogenase and formate dehydrogenase (fig. 1a), alanine aminopeptidase (fig. 1b) and aspartate aminotransferase (fig. 1c) are presented. The figure shows that the electrophoregrams do not reveal differences in component compositions of regenerants, although sometimes we identified differences in the intensity of bands.
It is known that the culture in vitro of plant organs and tissues is associated with an effect on explants such factors as mechanical damage of tissues, physical factors, chemical effects of exogenous plant hormones, minerals, and other additives. These factors may cause a powerful stress and therefore mutagens. The frequency of de novo
Figure 1 - Electrophoretic spectra of some enzymes of regenerants, obtained through polyembryoidogenesis in
anther culture in vitro of spring wheat Zhnitsa: a - glutamate dehydrogenase (GDH, 1.4.1.2) and formate dehydrogenase (FDH, 1.2.1.2), b - alanine aminopeptidase (AAP, 3.4.11.2) and c - aspartate aminotransferase
(AAT, 2.6.1.1).
events significantly exceeds the background values and can reach 10-2 [4]. Therefore the estimation of genetic homogeneity of regenerants is very important.
Isozymes are useful tool for this purpose. They allow estimating the frequency of mutation-like events, including in culture in vitro. Also they have a monogenic genetic control, in contrast to some types of DNA markers. Their resolution is high enough.
Our results suggest that the selected method (polyembryoidogenesis) of regenerants obtaining in wheat anthers culture in vitro is biotechologically optimal. Our data are a new argument for the broader application of the discussed method as a method of clonal selection aimed at mass production of economically valuable genotypes of spring wheat - the main bread cereal [2]. References:
1. From microspore to variety / Batygina T.B., Kruglova N.N., Gorbunova V.Yu., Titova G.E., and Seldimirova O A., Moscow: Nauka, 2010. 174 p.
2. Kruglova N.N. Optimization of biotechnology of wheat plant obtaining in in vitro culture // Izvestia Ufimskogo Nauchnogo Tsentra RAN. 2012. № 3. P. 57-61.
_МЕЖДУНАРОДНЫЙ НАУЧНЫЙ ЖУРНАЛ «ИННОВАЦИОННАЯ НАУКА» №2/2016 ISSN 2410-6070_
3. Kruglova N.N., Batygina T.B. Guidelines on the use of the anther morphogenetic potential in biotechnological studies of spring wheat. Ufa: IB USC RAS, 2002. 39 p.
4. Kunakh V.A. Plant genome variation in the course of in vitro dedifferentiation and callus formation // Rus. J. Plant Physiol. 1999. V. 46. № 6. P. 808-817.
5. Seldimirova O.A., Kruglova N.N. Formation of polyembryoids in in vitro culture as a stage of wheat cloning biotechnology // Izvestia Ufimskogo Nauchnogo Tsentra RAN. 2014. № 1. P. 22-26.
6. Seldimirova O.A., Titova G.E. Kruglova N.N. Androclinic «siamese embryos» of wheat in vitro // Izvestia Ufimskogo Nauchnogo Tsentra RAN. 2015. № 4(1). P. 137-142.
7. Seldimirova O.A., Yanbaev Yu.A., Zaitsev D.Yu. Isoenzyme markers at researches of changeableness of spring soft wheat breed districted in Bashkortostan // Vestnik of the Orenburg State University. 2009. №6 (100). P. 337-339.
8. Yanbaev Yu. A., Bairamgulov N.R., Redkina N.N., Mullagulov R.Yu. Differentiation among populations of the Rhodiola iremelica Boriss. (Grassulaceae) in the Southern Urals // Rus. J. Genet. 2007. V. 43. № 11. P. 1314-1318.
© Сельдимирова О.А., Галин И.Р., Янбаев Ю.А., 2016
УКД 574.24
М.И. Скрипник, Н.П. Корогод
Студент; к.б.н., доцент Естественно- математический факультет Павлодарский Государственный Педагогический Институт
г. Павлодар, Республтка Казахстан
ОПРЕДЕЛЕНИЕ УРОВНЯ НАКОПЛЕНИЯ ЗОЛОТА И СЕРЕБРА В ВОЛОСАХ ДЕТЕЙ ПАВЛОДАРСКОЙ ОБЛАСТИ
Аннотация
Проведена оценка содержания золота и серебра (Ag, Аи) методом нейтронно-активационного анализа в пробах волосах детей Павлдарской области. Установлено, что наибольшее содержание серебра содержится в волосах жителей Аксуского района, а золота у жителей г.Павлодара.
Ключевые слова Высокотоксичные элементы, волосы, золото, серебро.
Избыточное накопление токсичных химических элементов могут быть причиной снижения уровня функциональных резервов организма человека [1, с. 838-845].
Правомерность и эффективность использования волос в анализе эколого-токсикологических корреляций доказана результатами нескольких международных координированных программ, таких как патент на изобретение № 2298212 «Способ определения участков загрязнения ураном окружающей среды» Рихванова Л.П., Язикова Е.Г., Барановской Н.В., Янкович Е.П и т.д.
Токсичность каждого элемента определяется общим его содержанием в различных химических состояниях [2]. Несмотря на то, что токсическая и летальная дозы золота для человека не определены, некоторые его органические соединения токсичны. Накапливаясь в почках, печени, селезёнке и гипоталамусе приводят к органическим заболеваниям, а также дерматитам, стоматитам, тромбоцитопении. Избыточные дозы серебра могут откладываться в тканях, что внешне выражается серой окраской слизистых оболочек и кожи [3].
