Application of modern methods for plant material studies of Gentiana kurroo (Royle) somatic embryogenesis
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Nowadays somatic embryogenesis is being investigated with special attention paid to the identification of genes directly involved in triggering of cell competence and development of ontogenic stages. In order to trace successive cell divisions that lead to the formation of the globular structures, modern scanning and electron microscopy methods are applied. Plantlets, which come from somatic embryogenesis process, should be exactly the same as mother plants but in vitro culture conditions may induce many disturbances, which could be lasting (hereditary) or only transitory. These changes are usually called somaclonal variation and could be observed on different levels of the plant organization. To investigate this kind of variations both of the genetic and epigenetic types, specially designed molecular systems are needed. Here, we describe induction of somatic embryos from several explants of G. kurroo. In order to evaluate the particular ontogenic stages of somatic embryos and variability of regenerated plants, following methods were applied: light microscopy, scanning and transmission electron microscopy, 2D protein electrophoresis, flow cytometry of DNA content in the cell nucleus, cytogenetic analysis of chromosome number and molecular analysis with the use of AFLP.
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Agnieszka Fiuk, Ogrod Botaniczny ? Centrum Zachowania Roznorodnosci Biologicznej, Polska Akademia Nauk, 02-973 Warszawa, Poland