ISSR-ANALYSIS OF POLYMORPHISM OF NATURAL AND INTRODUCED SAPOSHNIKOVIA DIVARICATA (APIACEAE) POPULATIONS
Tatyana E. Skvortsova, Tatyana V. Elisafenko, Irina N. Kuban, Viktorya N. Yakk, Olga V. Dorogina
Central Siberian Botanical Garden, SB RAS, Novosibirsk, Russia
Keywords: Apiaceae, Saposhnikovia divaricata, natural population, introduction, ISSR-PCR, genetic diversity, intrapopulation variability, interpopulation variability, Republic of Buryatia
Abstract
Saposhnikovia divaricata (Turcz.) Schischk. is endemic to Southeast Asia. Biologically active substances have been found in the roots of plants. Consequently, a massive and unregulated withdrawal of S. divaricata from plant communities has been noted, which may lead to a disruption in the balance of genes in cenopopulations of this species. The assessment of the genetic variability of natural populations and the development of S. divaricata cultivation technology is relevant. The optimal ISSR-PCR protocol for S. divaricata has been developed in this study: the most informative primers were selected, conditions for their specific amplification were selected, the polymorphism of ISSR markers in the natural population and plants of the 3-rd generation introducers were analyzed. We studied 16 samples from the natural population of Siberia (Republic of Buryatia) and 15 samples of introduced plants grown at the exposition “Rare and endangered species of plants of Siberia” of the Central Siberian Botanical Garden SB RAS (Novosibirsk). Total DNA was isolated by a modified CTAB method. Of the tested 17 ISSR primers, 4 informative ones for S. divaricata were selected, for which a comparative analysis of genetic variability was carried out. As a result, genetic distances, the level of polymorphism, intra-and inter-population variability were determined. We identified 114 PCR fragments ranging in length from 220 to 3000 bp, 108 of which were polymorphic (94.7 %). Using various statistical programs (TREECON 1.3b, GenALEx 6.51), it was found that the natural population of S. divaricata is characterized by high genetic variability: the Shannon indexis 0.48 ± 0.02, intrapopulation variability is 89 %. We have established that ISSR-PCR is a method for fast and accurate analysis of morphologically close populations of S. divaricata for the detection of genetic changes. For rapid screening, it is sufficient to use one of the four markers (HB12, HB14, 17899A, 17899B), which have shown high efficiency. As a result of the study, it was found that the genetic diversity of S. divaricata did not change under the conditions of introduction during the change of generations compared with the original natural population.
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