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: http://herba.msu.ru/shipunov/plans/sh-pstr1.pdf
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Дата индексирования: Sun Apr 10 02:46:24 2016
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DACTYLORHIZA (ORCHIDACEAE) DIVERSITY FROM EUROPEAN RUSSIA: A PLASTID MICROSATELLITE STUDY A.B. Shipunov, M.W.Chase Jodrell Laboratory, Royal Botanic Gardens, Kew, Richmond, Surrey, TW9 3DS, UK INTRODUCTION Dactylorhiza Nevski is distributed across temperate and subtropical Eurasia and North Africa. The number of accepted species varies from 12 to 75 (Pedersen, 1998). There are around 12 Dactylorhiza species in European Russia and most of them have probably a West European origin. Investigations of allozyme markers and AFLP (Hedren, 2002) showed that part of Dactylorhiza is diploid (e.g., D. incarnata), another part is autopolyploid (D. maculata) and third has allotetaploid origin (D. traunsteineri and others). Microsatellites are short repeats in DNA that have high diversity on species level and thus can be useful for classification of closely related species and determining origin and geographical patterns. Orchid plastid microsatellites can also help to determine possible maternal parents of allopol yploids. Microsatellite study of Russian Dactylorhiza was never performed. The goal of our work is to screen plastid microsatellites and then to compare results with West European data. MATERIAL AND METHODS 107 samples were collected in different parts of European Russia and West Caucasus, from Krasnodar to Murmansk region--more than 3000 km from south to north. Then samples were dried in silica gel to prevent DNA degradation. DNA was extracted by common CTAB protocol (Doyle and Doyle, 1987). PCR was performed with set of primers to amplify four polymorphic loci in three different plastid DNA regions: trnS-trnG spacer, trnL intron and trnL-trnF spacer. RESULTS AND DISCUSSION We found ten combinations of fragment lengths, referred below as "haplotypes". In general, our data strongly coincide with analogous West European research. Among these haplotypes D. fuschii (A) haplotype is most frequent. We found also three specific haplotypes: RU1, RU2 and RU3. The first probably has independent origin from the typical for this region (North Karelia) D. fuchsii haplotype. RU2 is from specimens determined as D. flavescens and close to D. romana haplotypes from Mediterranean region. RU3 haplotype from D. euxina is close (but not identical) to two haplotypes reported from samples of this species in Turkey. thus Russian D. elodes can belong to D. maculata. Dactylorhiza cruenta now often accepted as form of D. incarnata (see, for example, Hedren et al., 2001), which coincides with our data because all our D. cruenta samples contain typical for D. incarnata haplotype E. Haplotype H (initially founded in Georgian material) was found in typical D. incarnata samples from Central Russia. This haplotype (and also Q and X) probably have wider distribution than was previously imagined. Interestingly also is the absence of haplotype C, reported for European D. traunsteineri and other allotetraploids. All our possible tetraploid specimens shared haplotypes either with D. fuschii A (for D. traunsteineri) or with D. incarnata E (for D. baltica). In addition, haplotypes N and X from Western Europe often belong to allotetraploids and thus another undescribed allotetraploid form could exist in Russia. We founded significant geographical pattern of haplotype distribution from south to north (Kruskal-Wallis 2 = 21.4529, p-value << 0.05). Distribution of haplotypes in Russia also has links to Western Europe flora (fig. 2). These observations support Averjanov (1990) point of view for main areas of Dactylorhiza distribution.