MOLECULAR GENETIC IDENTIFICATION OF SCOTS PINE AND SIBERIAN LARCH POPULATIONS IN PERM KRAI BASED ON POLYMORPHISM OF ISSR-PCR MARKERS
Ya. V. Sboeva1, Yu. S. Vasil’eva1, N. V. Chertov1, N. A. Pystogova1, S. V. Boronnikova1, R. N. Kalendar2, N. A. Martynenko1,3
1Perm State National Research University, Perm, Russian Federation
2National Center for Biotechnology, Nur-Sultan, Republic of Kazakhstan
3Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, Moscow, Russian Federation
Keywords: полиморфизм ДНК, генетическое разнообразие, ISSR-анализ, Larix sibirica Ledeb, Pinus sylvestris L, генетические формулы, штрих-коды, DNA polymorphism, genetic diversity, ISSR-analysis, genetic formulas, barcodes
Abstract
The use of DNA-fingerprinting of forest-forming woody plants is considered the most promising tool for genetic control of wood’s geographic origin, the formation of a reliable management system for harvesting and turnover of lumber. The purpose of this work was to search for identification markers, genotyping trees, and molecular genetic identification of previously not studied 2 populations of the Siberian larch Larix sibirica Ledeb. and 4 populations of Scots pine Pinus sylvestris L. of different regions of the Perm Krai. To DNA extraction, from each plant specimens wood were individually obtained and a modified method of extracting DNA from wood was used. In total, the analysis used the DNA of 114 Scots pine trees and 55 Siberian larch trees. For genetic testing, we used ISSR (Inter Simple Sequence Repeats)-method of DNA polymorphism analysis. Genetic identification was performed based on the original author’s method proposed by S. V. Boronnikova and I. V. Boboshina (2014). As a result of molecular genetic analysis, 74 ISSR markers were found and analyzed in populations of Scots pine, 85 ISSR markers were identified in populations of the Siberian larch, and the share of polymorphic loci in both species was high. As a result of molecular genetic identification, identification of species and polymorphic ISSR-markers and their combinations were found that characterize the belonging of trees to a species, as well as to a specific population. Molecular genetic formulas and barcodes for each individual population of two species have been compiled. The effectiveness, stability, and reproducibility of detected identification markers and their combinations have been proven through anonymous testing. The data obtained are the basis for determining the place of origin of wood, which will allow us to recommend measures to counter illegal logging and reduce the damage to the budget of logging regions of Russia, such as the Perm Krai.
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