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2016, № 63

UDC: 575.174.015.3: 577.21:502.753::582.893
GSNTI: 34.15.23

Approbation of ISSR DNA markers for genotyping of sea holly (Eryngium maritimum L.)

The paper presents the results of a study aimed at finding informative ISSR DNA markers promising for analysis of genetic homogeneity of micropropagated plants of sea holly (Eryngium maritimum L.). This species has declining numbers and could fall into the category of endangered species. In this regard, urgent is the development of methodological aspects of in vitro clonal micropropagation of this species with subsequent reintroduction into natural habitats. Reintroduction must be strictly controlled at the point of the genetic stability of the regenerated plants, ie, genetic identity of the micropropageted plants to original plant, serve as a source of explants. One of the promising methods for the analysis of genetic stability is ISSR-genotyping. The work had the task of testing ISSR-markers in relation to this species and selection the most promising for further genotyping of in vitro plants. Testing 20 ISSR-markers was performed. Among ISSR-markers with detected amplification was identified from 2 (M1 and UBC 845 markers) to 13 fragments (UBC818 marker) in electrophoretic spectrum. For the markers UBC827, UBC817, 3A30, 3A35, 3A37 and M8 amplification has not been identified. In accordance with the results identified, the following markers with high information content, are promising to perform genotyping samples species Eryngium maritimum: UBC844, UBC843, UBC818, UBC811, ASSR02, 3A39. These markers can be effectively used for further control of plant genetic homogeneity obtained in in vitro culture. It was confirmed by a test of sample of regenerated plants for genetic homogeneity.
Keywords: Sea holly, micropropagation in vitro, genetic stability, genotyping, ISSR-markers
DOI: 10.21515/1999-1703-63-95-101

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Authors:

  1. Suprun Ivan Ivanovich, Ph.D., Head. laboratory of genetics and microbiology, North Caucasian Region Research Institute of Horticulture and Viticulture.
  2. Stepanov Ilya Vladimirovich, junior researcher laboratory of genetics and microbiology, North Caucasian Region Research Institute of Horticulture and Viticulture.
  3. Malyarovskaya Valentina Ivanovna, Ph.D. Head. laboratory of biotechnology, Russian Research Institute of Floriculture and Subtropical Crops.
  4. Kolomiets Tatyana Michailovna, k. s. h. n. (Senior Researcher), Leading Researcher, Russian Research Institute of Floriculture and Subtropical Crops.
  5. Samarina Lidia Sergeevna, Ph.D., Senior researcher, Russian Research Institute of Floriculture and Subtropical Crops.