Hostname: page-component-586b7cd67f-rdxmf Total loading time: 0 Render date: 2024-11-25T04:01:47.069Z Has data issue: false hasContentIssue false

Molecular markers to assess genetic diversity of Gentiana lutea L. from the Ukrainian Carpathians

Published online by Cambridge University Press:  12 December 2014

Maryana Z. Mosula
Affiliation:
Volodymyr Hnatiuk Ternopil National Pedagogical University, M. Kryvonis street 2, Ternopil46027, Ukraine
Igor O. Andreev*
Affiliation:
Institute of Molecular Biology and Genetics NAS of Ukraine, Akad. Zabolotnogo street 150, Kyiv03680, Ukraine
Olena M. Bublyk
Affiliation:
Institute of Molecular Biology and Genetics NAS of Ukraine, Akad. Zabolotnogo street 150, Kyiv03680, Ukraine
Vitaliy M. Mel'nyk
Affiliation:
Institute of Molecular Biology and Genetics NAS of Ukraine, Akad. Zabolotnogo street 150, Kyiv03680, Ukraine
Iryna I. Konvalyuk
Affiliation:
Institute of Molecular Biology and Genetics NAS of Ukraine, Akad. Zabolotnogo street 150, Kyiv03680, Ukraine
Nadiya M. Drobyk
Affiliation:
Volodymyr Hnatiuk Ternopil National Pedagogical University, M. Kryvonis street 2, Ternopil46027, Ukraine
Viktor A. Kunakh
Affiliation:
Institute of Molecular Biology and Genetics NAS of Ukraine, Akad. Zabolotnogo street 150, Kyiv03680, Ukraine
*
*Corresponding author. E-mail: [email protected]

Abstract

The aim of the study was to develop the system of polymerase chain reaction (PCR)-based markers for the assessment of genetic diversity and population genetic studies of Gentiana lutea L. as well as to determine the utility of two indices of marker informativeness. The informativeness was determined for 40 PCR primers of different types (random amplified polymorphic DNA, inter simple sequence repeat, inter-retrotransposon amplified polymorphism, resistance gene analog polymorphism and conserved DNA-derived polymorphism markers) by evaluating discriminating power (DL) and resolving power (Rp) in a sample of 30 plants from two populations. Analysis of correlation between the index value and the number of differentiated pairs of genotypes in the given sample revealed that DL is more efficient than Rp; therefore, we selected primers based on the DL value. In total, 12 primers with the largest values of DL were chosen. Analysis of genetic relationship among 86 plants from six populations showed that the number of bands produced by the three of selected primers was sufficient to give average bootstrap support across six key nodes in the dendrogram higher than 85%, while using six of the primers resulted in average bootstrap value exceeding 99%. Thus, a minimal set of three to six selected primers are sufficient for a quick assessment of genetic diversity of G. lutea populations, depending on the sample size and degree of differentiation between populations, while the rest of the primers with DL values above 0.8 may be used for ecogenetic surveys. Preliminary results obtained with selected primers indicate the moderate level of genetic variation within the species and significant differentiation among individual populations.

Type
Research Article
Copyright
Copyright © NIAB 2014 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Botstein, D, White, RL, Skolnick, M and Davis, RW (1980) Construction of a genetic linkage map in man using restriction fragment length polymorphisms. American Journal of Human Genetics 32: 314331.Google Scholar
Caroli, S, Santoni, S and Ronfort, J (2011) AMaCAID: a useful tool for accurate marker choice for accession identification and discrimination. Molecular Ecology Resources 11: 733738.CrossRefGoogle ScholarPubMed
Collard, BCY and Mackill, DJ (2009) Conserved DNA-derived polymorphism (CDDP): a simple and novel method for generating DNA markers in plants. Plant Molecular Biology Reporter 27: 558562.Google Scholar
Dong, P, Wei, Y-M, Chen, G-Y, Li, W, Nevo, E and Zheng, Y-L (2009) Resistance gene analog polymorphisms (RGAPs) in wild emmer wheat (Triticum dicoccoides) and their ecological associations. Genetic Resources and Crop Evolution 56: 121136.Google Scholar
Fujii, H, Ogata, T, Shimada, T, Endo, T, Iketani, H, Shimizu, T, Yamamoto, T and Omura, M (2013) Minimal marker: an algorithm and computer program for the identification of minimal sets of discriminating DNA markers for efficient variety identification. Journal of Bioinformatics and Computational Biology 11: 1250022 doi: 10.1142/S0219720012500229 .Google Scholar
Jensen, SR and Schripsema, J (2002) Chemotaxonomy and pharmacology of Gentianaceae. In: Struwe, L and Albert, VA (eds) Gentianaceae: Systematics and Natural History. Cambridge: Cambridge University Press, pp. 573631.Google Scholar
Konvalyuk, II, Mel'nyk, VM, Drobyk, NM, Kravets, NB, Twardovska, MO and Kunakh, VA (2011) RAPD- and ISSR-analysis of genetic variation in Gentiana pneumonanthe L. tissue and organ culture. Visnik Ukrains'kogo Tovaristva Genetikiv i Selekcioneriv 9: 231.Google Scholar
Mayorova, OY, Grytsak, LR, Terehova, GI, Mel'nyk, VM, Andreev, IO and Drobyk, NM (2013) Gentiana lutea L. (Gentianaceae) in the flora of Ukrainian Carpathians: characteristics and current status of populations. Ukrajins'kyj Botanicnyj Zurnal 70: 780787.Google Scholar
Mel'nyk, VM, Spiridonova, KV, Andreev, IO, Strashniuk, NM and Kunakh, VA (2004) Variability of nuclear 18S-25S rDNA of Gentiana lutea L. in nature and in in vitro tissue culture. Cytology and Genetics 38: 1419.Google Scholar
Mosula, MZ, Konvalyuk, II, Mel'nyk, VM, Andreev, IO, Bublyk, OM, Drobyk, NM and Kunakh, VA (2013) Genetic diversity in Gentiana lutea L. populations from Svydivets ridge of the Ukrainian Carpathians. Visnik Ukrains'kogo Tovaristva Genetikiv i Selekcioneriv 11: 250259.Google Scholar
Mosula, MZ, Konvalyuk, II, Mel'nyk, VM, Bublyk, OM, Andreev, IO, Drobyk, NM and Kunakh, VA (2014) Analysis of genetic diversity in Gentiana lutea L. populations by method of marking between retrotransposon's sequences (IRAP-PCR). Fiziologiia Rastenii i Genetika 46: 4555.Google Scholar
Peakall, R and Smouse, PE (2006) GENALEX 6: genetic analysis in Excel. Population genetic software for teaching and research. Molecular Ecology Notes 6: 288295.Google Scholar
Peakall, R and Smouse, PE (2012) GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research – an update. Bioinformatics 28: 25372539.Google Scholar
Powell, W, Morgante, M, Andre, C, Hanfey, M, Vogel, J, Tingey, S and Rafalski, A (1996) The comparison of RFLP, RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis. Molecular Breeding 2: 225228.CrossRefGoogle Scholar
Prevost, A and Wilkinson, MJ (1999) A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theoretical and Applied Genetics 98: 107112.Google Scholar
Rogers, SO and Bendich, AJ (1985) Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues. Plant Molecular Biology 5: 6976.Google Scholar
Saini, M, Singh, S, Hussain, Z and Sikka, VK (2010) RAPD analysis in mungbean [Vigna radiata (L.) Wilczek.] II: a comparison of efficiency parameters of RAPD primers. Indian Journal of Biotechnology 9: 276282.Google Scholar
Schluter, PM and Harris, SA (2006) Analysis of multilocus fingerprinting data sets containing missing data. Molecular Ecology Notes 6: 569572.Google Scholar
Strashniuk, NM, Les'kova, OM, Zahrychuk, HYa, Mel'nyk, VM and Kunakh, VA (2006) Biologically active substances of Gentiana L. genus. 1. Biosynthesis and physiological influence. Phytotherapy 1: 3141.Google Scholar
Tessier, C, David, J, This, P, Boursiquot, JM and Charrier, A (1999) Optimization of the choice of molecular markers for varietal identification in Vitis vinifera L. Theoretical and Applied Genetics 98: 171177.Google Scholar
Twardovska, MO, Strashniuk, NM, Mel'nyk, VM, Konvalyuk, II and Kunakh, VA (2009) RAPD-analysis of the genome polymorphism for some Gentiana L. species from the Ukrainian flora. Reports of the National Academy of Sciences of Ukraine 5: 200204.Google Scholar
Yap, IV and Nelson, RJ (1996) WinBoot: a program for performing bootstrap analysis of binary data to determine the confidence of UPGMA-based dendrograms. IRRI Discussion Paper Series, No. 14. Manila, Philippines: International Rice Research Institute.Google Scholar
Supplementary material: File

Mosula Supplementary Material

Table S1

Download Mosula Supplementary Material(File)
File 154.6 KB