Phylogenetic taxonomy of Artemisia L. species from Kazakhstan based on matK analyses

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INTRODUCTION

Artemisia of the family Asteraceae is a genus with great economic potential and importance. Species of this genus have been used in many aspects of life throughout the course of history (Lachenmeier, 2010; Petrovska, 2012).

They have been used as medicinal, food, and ornamental plants (Weatherset al., 2011). A number of species have a very important medicinal significance, especiallyArtemisia annua L. (Klayman, 1985) and Artemisia absinthium L.

(Lachenmeier, 2010). One of the most common examples is tarragon — A. dracunculus L. It is widespread in the wild across much of Eurasia and North America, and is culti- vated for culinary and medicinal purposes. Its essential oils are used as antibacterial agents; dry leaves are added as fla- vourings for meat and fish (Aglarovaet al., 2008; Obolskiy et al.,2011).

Of the family Asteraceae, the tribe Anthemideae, genusAr- temisiais one of the largest genera (Bremer and Humphries, 1993; Oberpreiler et al., 1995). It includes about 500 spe- cies, which are distributed in five subgenera (Vallès and McArthur, 2001). Species of this genus are widespread in the Northern Hemisphere, particularly in Eurasian temper- ate zone and North America, and in South and North Africa

(Bremer, 1994; Torrelet al., 1999). Due to the large amount of species in the genus, their classification is still complex and not fully completed. In earlier studies, the genus was subdivided into three subgenera (Poljakov,1961; Kornkven et al., 1998). Absinthium and Tridentataesubg. were con- sidered as the sections of ArtemisiaandSeriphidum subg., respectively. In more recent work already four subgenera were declared (Persson et al., 1974). Absinthium was proposed as a separate subgenus originated from theArtemisia subg. Based only on the capitula type and florets fertility, five major groups described as subgeneric or sectional rank (Absinthium,Artemisia,Dracunculus,Seriphidium, andTri- dentatae) are more or less constantly found in classic stud- ies confirmed by molecular data (Torrelet al., 1999). Pre- vious phylogenetic studies onArtemisiashowed monophyly of the genus and monophyly of the three main infrageneric groups (Dracunculus, Seriphidium, Tridentatae), whereas subgenera Absinthium and Artemisia were described as polyphyletic (Watson et al., 2002). Classical subgeneric separation based only on morphological traits was rear- ranged, because in some cases it was not supported by the traditional classifications (Sanz et al., 2008; Tkach et al., 2008). Moreover, processes such as hybridisation, introgres- sion, and polyploidisation are very common for these plants

PROCEEDINGS OF THE LATVIAN ACADEMY OF SCIENCES. Section B, Vol. 72 (2018), No. 1 (712), pp. 29–37.

DOI: 10.1515/prolas-2017-0068

PHYLOGENETIC TAXONOMY OF ARTEMISIA L. SPECIES FROM KAZAKHSTAN BASED ON MATK ANALYSES

Yerlan Turuspekov

^1,5

, Yuliya Genievskaya

¹

, Aida Baibulatova

¹

, Alibek Zatybekov

¹

,

Yuri Kotuhov

²

, Margarita Ishmuratova

³

, Akzhunis Imanbayeva

⁴

, and Saule Abugalieva

^1,5,#

1Institute of Plant Biology and Biotechnology, 45 Timiryazev Street, Almaty, KAZAKHSTAN 2Altai Botanical Garden, Ridder, KAZAKHSTAN

3Karaganda State University, Karaganda, KAZAKHSTAN

4Mangyshlak Experimental Botanical Garden, Aktau, KAZAKHSTAN

5Al-Farabi Kazakh National University, Biodiversity and Bioresources Department, Almaty, KAZAKHSTAN

#Corresponding author, absaule@yahoo.com

Communicated by Isaak Rashal

The genusArtemisiais one of the largest of the Asteraceae family. It is abundant and diverse, with complex taxonomic relations. In order to expand the knowledge about the classification of Kazakhstan species and compare it with classical studies, matK genes of nine local species in- cluding endemic were sequenced. The infrageneric rank of one of them (A. kotuchovii) had re- mained unknown. In this study, we analysed results of sequences using two methods — NJ and MP and compared them with a median-joining haplotype network. As a result, monophyletic origin of the genus and subgenus Dracunculus was confirmed. Closeness ofA. kotuchoviito other spe- cies of Dracunculus suggests its belonging to this subgenus. Generally, matK was shown as a useful barcode marker for the identification and investigation ofArtemisia genus.

Key words: Artemisia, Artemisia kotuchovii, DNA barcoding, haplotype network.

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and this makes understanding of their relations at the molecular level even more difficult (Winward and McArthur, 1995).

Central Asia is a centre of the genus Artemisia origin and one of the most important centres of its diversification (McArthur and Plummer, 1978; Wang, 2004). According to previous studies, most of the Asian species belong to the Seriphidium subg. (Poljakov, 1961; Tkachet al., 2008). In Kazakhstan, there are around 80 species mostly growing in steppes and deserts (Pavlov et al., 1966). In general, en- demic and rare species of the genus Artemisia in Ka- zakhstan are poorly studied and their place in subgenera classification is not well described. Only a few works were dedicated to their biochemistry and economical importance (Goryaevet al., 1962; Nikitinaet al., 1964). The main aim of our study was to reveal the complex relations of subgenera in the genusArtemisiagrowing in Kazakhstan and compare results with traditional classifications based on both morphological traits (Poljakov, 1961) and molecular data (Watsonet al.,2002; Sanzet al., 2008; Garciaet al., 2011).

Another important objective was to determine the place of one unranked local species —A. kotuchovii— in the subgenera classification. Additionally, we tested two different methods for phylogenetic taxonomy (joining and maximum parsimony) and compared them to haplotype networking analysis.

This work represents a new direction in the study of Ka- zakhstan native flora. In the past, only a few reports were related to the description of the genetic variation of local flora (Adamset al., 1998; Turuspekovet al., 2002). There- fore, the study is an expansion of a research oriented to- wards description of endemic, rare, and economically important species of the country and part of cooperative nation-wide project (Turuspekov and Abugalieva, 2015) for

genotyping of plant accessions using DNA barcoding. The project combined efforts of local botanists and geneticists from biotechnology research organisations, botanical gar- dens, state nature parks, and reserves. In the last 20 years, DNA barcoding has shown itself as a powerful and efficient tool for sample identification and phylogeny of new and poorly studied species (Hebert et al., 2003; Hebert et al., 2005; Kress et al., 2017).

MATERIALS AND METHODS

Materials sampling.Nine populations ofArtemisiaspecies were collected from different places of central, southeastern, eastern, and western regions of Kazakhstan (Table 1, Fig. 1). For the reconstruction of intragenus topology sequences of twenty oneArtemisiataxa were taken from Gen- Bank (https://www.ncbi.nlm.nih.gov/genbank/).

DNA extraction, amplification and sequencing. Three plants from each population were chosen for the genetic analysis. Total genomic DNA was extracted from dry leaf material according to the modified Dellaporta DNA extraction protocol (Dellaporta et al., 1983). Individual DNA samples were analysed separately. PCR fragments were am- plified for thematuraseK gene of the chloroplast genome (matK) (Naeem et al., 2014).

All PCR reactions were carried out in 16 µl volumes in a Veriti Thermo cycler (Applied Biosystems, Foster City, CA, USA). One PCR reaction contained 4 mM of each dNTP, 6.4 mM of primer mix, 1.6 U of Taq DNA po- lymerase and 80 ng of total genomic DNA. Protocols for PCR reactions were taken from Jun et al. (2012). Primers chosen for PCR included matK-F

(5’-CCTATCCATCTGGAAATCTTAG-3’) and matK-R (5’-GTTCTAGCACAAGAAAGTCG-3’) with annealing

T a b l e 1 LIST OFARTEMISIASPECIES COLLECTED IN KAZAKHSTAN AND THEIR GENBANK ACCESSIONS NUMBERS

Region Species No. of collected plants GenBank accession numbermatK

Central KZ

(Karkaraly, Bol’shoe lake)

A. radicansKupr. 20 plants MG282056

Eastern KZ

(Valley of Kurchum River)

A. gmeliniiWeb. ex Stechm. 27 plants MG282059

Eastern KZ

(Southern Altai-Tarbagatai spine)

A. kotuchoviiKupr. 13 plants MG282057

Eastern KZ (Kurchum River)

A. sublessingiana(Kell.) Krasch. ex Poljak. 20 plants MG282053

Southeastern KZ

(Almaty State Nature Reserve)

A. santolinifoliaTurcz. ex Besser. 20 plants MG282055

Southeastern KZ

(Karatau State Nature Reserve)

A. scopaeformis*Ledeb. 20 plants MG282054

Southeastern KZ

(Altyn Emel National Park)

A. terrae-albaeKrasch. 20 plants MG282052

Southeastern KZ

(Karatau State Nature Reserve)

A. transiliensis* Poljak. 21 plants MG282051

Western KZ (West Karatau)

A. gurganicaWilld. 20 plants MG282058

* indicates endemic species for the Kazakhstan territory (Pavlovet al., 1966).

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temperature 50 °C and expected sizes of amplicons 784 bp (obtained from Asterales according to GenBank data).

Whole volume of each PCR product was checked by elec- trophoresis in 1.5% agarose gel at 80 V voltage for 40 min.

Single bands with expected sizes formatK were visualised, cut out from gel and purified using the ULTRAPrep® Aga- rose Gel Extraction Mini Prep Kit (AHN Biotechnologie GmbH, Nordhausen, Germany) according to the protocol provided by the company. Purified DNA amplicons were used for the sequence reactions with forward and reverse primers separately. All reactions were performed with the BigDye Terminator Cycle Sequencing technology (Applied Biosystems, Foster City, CA, USA) according to protocols of the company.

Alignment and phylogenetic analyses. Generated matK sequences of the samples were imported in MEGA 6 (Ta- muraet al., 2013) software. Results were evaluated by different methods used for phylogenetic reconstructions — neighbour-joining (Bhattacharyya, Mukherjee, 2017) and maximum parsimony (Bryant et al., 2017).

The final alignment was imported into DNASP v5.10 (Librado and Rozas, 2009) and converted into Roehlfile for- mat for the operations in the Network software (version 4.6;

http://fluxus-engineering.com). In addition, the nucleotide sequences formatK of local species were aligned with sequences ofArtemisiaspecies from the NCBI reference data-

base. The genetic structure was assessed through median- joining haplotype networks (Bandeltet al., 1999) using the Network software. Post-processing calculation was done without the MP criterion (e= 0).

RESULTS

DNA sequencing. DNA sequences of 784 bp of the matu- raseK gene (matK) were obtained from nine local Artem- isiaspecies and aligned in MEGA 6.06 together with available Artemisia references from the GenBank. Tanacetum parthenium L., Achillea ptarmica L., and Anthemis cotula L. from the same tribe Anthemideae (Asteraceae) were chosen as the outgroups. In total, 9 sites with gaps and 17 polymorphic sites were detected for the studied Artemisia species. Nine of those sites were singleton variable sites and other eight were parsimony informative sites (Fig. 2). In this study we used two sets of data: 1) Kazakhstan species with specimens from Genbank; 2) Kazakhstan species only.

There were no differences among DNA sequences in analysed three individual plants within nine studied species from Kazakhstan. The sequences ofmatK of the nine species were deposited to the NCBI database (Table 1).

Phylogenetic and haplotype network analyses of local species and GenBank specimens. The first tree was recon- structed using the NJ method (bootstrap 1000) for 33 species, including outgroups (Fig. 3). All species of the genus

Fig. 1.Collected sites ofArtemisiaspecies.

One number denotes sampling point for the population of one species: 1 –A. radicansKupr.; 2 –A. gmeliniiWeb. ex Stechm.; 3 –A. kotuchoviiKupr.; 4 – A. sublessingiana (Kell.) Krasch. ex Poljak.; 5 – A. santolinifoliaTurcz. ex Besser.; 6 – A. scopaeformis*Ledeb.; 7 – A. terrae-albaeKrasch.; 8 – A. transiliensis* Poljak.; 9 –A. gurganicaWilld. * Indicates endemic plants for Kazakhstan territory (Pavlovet al., 1966).

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Artemisia formed one large clade separately from outgroups. This Artemisia clade was further subdivided into

four subclades. The first subclade included two local species (A. terrae-albae and A. gurganica) belonging to the

Fig. 2.Polymorphic sites ofArtemisiaspecies detected inmatK region. * Indicates specimens take from GenBank with their accession numbers. Endemic species for Kazakhstan are indicated in bold.

Fig. 3. Neighbour-joining phylogenetic tree results from the analysis ofmatK sequences of nine local, twenty-one GenBankArtemisia species and three outgroup taxa. The subgenera classifications are given according to Poljakov, 1961. The lengths of branches are based on maximum composite likelihood and numbers at nodes shows a probability bootstrap. * denotes GenBank species with reference numbers from the NCBI database. Black arrows indicate endemic species.

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subgenusSeriphidum. The next clade combined three species (A. gmelinii, A. santolinifolia, and A. roxburghiana) from the subg.Artemisia. The third subclade corresponded to the subg.Dracunculusand included all six species of this subgenus from GenBank. The local endemic speciesA. ko- tuchoviiwas also placed in this subclade. The last subclade was heterogeneous and represented four subgenera — Ar- temisia, Absinthium, Seriphidum, andTridentatae.

The optimal MP tree was chosen from ten replicates (bootstrap 1000) for the same 33 species (Fig. 4) that had been studied using the NJ method. The generated tree showed different topology when it was compared to the NJ tree. In the MP tree all Artemisia species formed one major clade with two subclades apart from the outgroups. The first subclade was subgenusDracunculus, which included A. kotu- chovii. The second subclade was heterogeneous and in- cluded all other subgenera except Dracunculus.

The matK dataset, which combined both the GenBank and local accessions, was used for the network association analysis. The Network incorporated 33 species with 16 haplotypes clustering in six major haplotype lineages, which corresponded to five Artemisia subgenera and the outgroup (Table 2). Mean haplotype diversity for the set was rela- tively high (Hd = 0.841), but nucleotide diversity was rather low (p = 0.0029, k = 2.211).

In order to compare topology of phylogenetic trees with the haplotype network, the studied nucleotide sequences combined in 16 haplotypes, including the outgroup, were analysed using Network 4.6. The obtained diagram shows con- sensus network with six groups of haplotypes corresponded to five subgenera and the outgroup cluster (Fig. 5). The largest haplotype H_6 in the centre of the diagram included 11 species from subg. Artemisia, Absinthium, and Seri- phidum. The second largest haplotype was H_3, which con- tained five species from the subgenusDracunculus, closely connected toA.kotuchoviiin H_12.The haplotype H_9 was represented byA. tridentateaoriginated from the subg.Ar- temisia group, not supporting the theory of Seriphidum as the ancestor of the Tridentataesubg.

Phylogenetic and haplotype network analyses of local species.The second dataset was restricted to only nine Ka- zakhstan species and three outgroup taxa. This restricted dataset was also used for phylogenetic reconstruction by NJ and MP methods. Two trees showed very similar profiles for this dataset (Fig. 6 A, B). In both cases, subg.Artemisia species A. santolinifolia andA. gmeliniiformed a separate subclade.A. kotuchoviiwas also placed apart from the oth- ers.

Local species sequences formed five haplotypes and were also used for median-joining network reconstruction (Table

Fig. 4.Maximum parsimony phylogenetic tree reconstructed from the analysis of matK sequences of nine local, twenty-one GenBank Artemisia species and three outgroup taxa. The subgenera classifications are given according to Poljakov, 1961. The lengths of branches are based on maximum composite likelihood and numbers at nodes shows a probability bootstrap. * denotes GenBank species with reference numbers from the NCBI database. Black arrows indicate endemic species.

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3). Haplotype diversity for this set of data is Hd = 0.806, nucleotide diversity is p = 0.024, k = 1.889. As a result, they were combined into two major groups corresponding toArtemisiaandSeriphidumsubgenera (Fig. 6 C). The largest haplotype H_1 includes four species, three of them be- longed to Seriphidum and one’s subgenus (A. radicans) is Artemisia. A. kotuchovii H_4 was placed separately from them, just like on phylogenetic trees.

DISCUSSION

Artemisiais one of the most complex genera and it is represented by the large number of species, diverse morphological types, ploidy and complicated genetic relationships

(Winward and McArthur, 1995). Because of this, the clarifi- cation of the genus’s taxonomy using classical botanical tools and morphological characteristics has many difficul- ties (Torrel et al., 1999). Therefore, usage of molecular markers is a valuable and promising addition to the traditional morphology-based classification. In this study, DNA barcoding approach based on the usage ofmatK marker was applied for the assessment of the Artemisia taxonomy of nine species collected in Kazakhstan. The analysis of the matK nucleotide sequences suggested that the marker pro- vides sufficient information for differentiation of studied taxa. Generated NJ and MP trees of the studied taxa allowed to determine a single clade suggesting monophyletic origin of the genus, which was proposed earlier in the classical ap- proaches (Torrel et al.,1999; Watson et al., 2002).

Previous molecular taxonomy studies with ITS using three main infrageneric groups of the genus (Dracunculus,Seri- phidium, and Tridentatae) suggested that they have mono- phyletic origin, while the two remaining subgenera Absin- thiumandArtemisiaappeared to be polyphyletic (Torrelet al., 1999; Sanz et al., 2008). Since the number of samples used in this study was limited, both NJ and MP phylogenetic trees only partially confirmed previously suggested taxonomic classification. For example, it is clearly visible, that theDracunculussubclade is very distinct in both trees.

A. terrae-albae andA. gurganicafrom subgenus Seriphid- ium formed the separate subclade distantly apart from the other species. Another outcome from the analysis of the phylogenetic trees was the taxonomy of less studied species

—A. kotuchovii.The topology of both NJ and MP trees indicated that this species clearly belongs to the subgenus Dracunculus.

The haplotype networking diagram (Fig. 5) was rather more informative in comparison with the NJ and MP trees. First, the network analysis showed that haplotype H_6 was com- prised from 11 species of subgeneraArtemisia, Absinthium, andSeriphidium.Second, the H_6 was directly descendent

Fig. 5.Median-joining haplotype network. No MP criterion (e= 0). Red small dots are median vectors presumed unsampled or missing intermediates. Yellow dots denote haplotypes; size is proportional to their frequen- cies. Number of perpendicular dashes on branches is equal to the number of mutations between two neighbouring dots. Colours denote the major groups based on subgeneric division. The subgenera classifications are given according to Poljakov, 1961.

T a b l e 2 LIST OF HAPLOTYPES FORMED FROM THE ANALYSIS OFMATK GENE SEQUENCES OF LOCALARTEMISIA SPECIES, GENBANK SPECIMENS AND THE OUTGROUP

Haplotype Number of species

Species H_1 1 KJ372399.1Artemisia roxburghiana*

H_2 1 KF648716.1Artemisia vulgaris*

H_3 5 HM989797.1Artemisia scoparia*

JN894047.1Artemisia campestris*

JQ173388.1Artemisia capillaris*

KC474124.1Artemisia borealissubsp.

richardsoniana*

KF530805.1Artemisia japonica*

H_4 3 JQ173387.1Artemisia annua*

JQ173390.1Artemisia sacrorum*

KC474133.1Artemisia tilesii*

H_5 1 KC474129.1Artemisia hyperborea* (Artemisia furcata)

H_6 11 FN668458.1Artemisia arctisibirica*

GQ434109.1Artemisia gmelinii*

HM989726.1Artemisia argyi*

HM989729.1Artemisia lactiflora*

JQ173389.1Artemisia igniaria*

JQ173391.1Artemisia sieversiana*

JQ412200.1Artemisia afra*

Artemisia radicans Artemisia scopaeformis Artemisia sublessingiana Artemisia transiliensis

H_7 1 JN894044.1Artemisia absinthium*

H_8 1 HQ593182.1Artemisia dracunculus*

H_9 1 AF456776.1Artemisia tridentata*

H_10 2 Artemisia gurganica(Artemisia fragranssubsp.

gurganicaKrasch.) Artemisia terrae-albae H_11 1 Artemisia santolinifolia H_12 1 Artemisia kotuchovii H_13 1 Artemisia gmelinii

H_14 1 JN895338.1Tanacetum parthenium*

H_15 1 JN895745.1Achillea ptarmica*

H_16 1 JN895749.1Anthemis cotula*

* indicates specimens taken from the GenBank with their accession numbers. Endemic species for Kazakhstan are indicated in bold.

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from H_10, consisting of two species of subg.Seriphidium (Table 2). The presence ofA. scopaeformis (Seriphidium)in H_6 makes sense because it logically connects these two haplotypes. Third, the haplotype H_5 consisted of only one species (A. hyperborea) in this study, which descended from H_6. Fourth, the haplotype H_9 descended from H_5, which suggests that the subgenusTridentataeoriginated directly from species of subgenus Artemisia.

It is interesting that the network suggests three subgroups within the subgenusArtemisia.As the topology of the network suggests that H_13 (A. gmelinii) is a predecessor of H_1 (A. roxburghiana)and H_11 (A. santolinifolia),it re- sembles the outcome from the MP tree (Fig. 4). The other outcome from the networking analysis is that haplotype H_7 (A. absinthium, Absintium) is directly connected to haplotype H_6, which is a reasonable connection because A. sieversiana (Absintium) is a part of this most frequent

haplotype (H_6) in the study. Unlike in Absinthium, the haplotypes of Dracunculus (H_3 and H_8) are not connected to H_6 directly, but through intermediates mv1 and mv2, respectively (Fig. 5). As the mv2 connectsA. dracun- culuswithA. kotuchovii,this confirms that this species belongs to the subg. Dracunculus.

In general, the analyses based on matK indicated that A.

terrae-albae andA. gurganica from the subg.Seriphidium are predecessors of all other taxa within the genusArtem- isia.Therefore, the conclusions based on the usage of plas- tid genome marker is not completely congruent with out- comes based on nuclear genome markers ITS (Torrelset al., 1999), as their parsimony analysis of 31 species resulted in a multifurcate type of the tree. Nevertheless, the authors indicated that the first clade of the tree was formed primarily from species of subg. Seriphidium. Later Watson with co- authors (Watsonet al., 2002) analysed a larger number of

Fig. 6.Phylogenetic trees and the haplotype network based on thematKsequences of nine local Artemisia species and three outgroup taxa.A.Neighbour-joining phylogenetic tree.

B.Maximum parsimony phylogenetic tree.C.

Median-joining haplotype network. The subgenera classifications are given according to Poljakov, 1961. The lengths of branches are based on Maximum Composite Likelihood and numbers at nodes shows a probability bootstrap. * denotes GenBank species. Black arrows indicate endemic species. Red small dots are median vectors presumed unsampled or missing intermediates. Yellow dots denote haplotypes; size is proportional to their fre- quencies. Number of perpendicular dashes on branches is equal to the number of mutations between two neighbouring dots. Colours denote the major groups based on subgeneric division.

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the genus Artemisia taxa consisting of 57 species of sub- tribe Artemisiinae from Old and New Worlds and pointed out on separation of subg.Dracunculus from all remaining Artemisia species. Therefore, one of the main conclusions in that study was the recognition of two subgenera within theArtemisia— subg.Dracunculusand an expanded subg.

Artemisia(Watsonet al., 2002).Phylogenetic trees and haplotype network generated using matK in this study hinted that the Dracunculus, although it is genetically a distinct subclade, descended from subg.Artemisia.

CONCLUSIONS

The application of phylogeny and haplotype network analyses indicated that some of the species of subgenusSeriphid- ium can be predecessors of the genus Artemisia. Specifi- cally, A. terrae-albae and A. gurganica of this subgenus were closest to outgroup species used in this study. The haplotype network analysis was more informative in comparison to generated NJ and MP trees, as it is suggested a hypothetical evolutionary pathway within the genus. The network showed that the most frequent haplotype H_6 was common for three subgeneraArtemisia, Absinthium,andSe- riphidium. The species of subgenera Tridentatae derived from the species ofArtemisia,whereas species of subgenera Dracunculus were distantly apart from the remaining species of the genus but via intermediate median vectors asso- ciated with the major haplotype H_6 of the genusArtemisia.

Also, it was shown that the earlier unstudied speciesA. ko- tuchovii is a part of the subgenusDracunculus.

ACKNOWLEDGEMENTS

The authors would like to acknowledge the funding from the Ministry of Education and Sciences of the Republic of Kazakhstan for national programme N0237.

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T a b l e 3 LIST OF HAPLOTYPES FORMED FROM THE ANALYSIS OFMATK GENE SEQUENCES OF LOCAL ARTEMISIA SPECIES AND THE OUTGROUP TAXA

Haplotype Number of species

Species

H_1 4 Artemisia radicans

Artemisia scopaeformis Artemisia sublessingiana Artemisia transiliensis

H_2 2 Artemisia gurganica(Artemisia fragranssubsp.

gurganicaKrasch.) Artemisia terrae-albae H_3 1 Artemisia santolinifolia H_4 1 Artemisia kotuchovii

H_5 1 Artemisia gmelinii

H_6 1 JN895338.1Tanacetum parthenium*

H_7 1 JN895745.1Achillea ptarmica*

H_8 1 JN895749.1Anthemis cotula*

* indicates specimens taken from the GenBank with their accession numbers. Endemic species for Kazakhstan are indicated in bold.

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Received 16 October 2017

Accepted in the final form 8 November 2017

KAZAHSTÂNASARTEMISIA L. SUGU FILOÌENÇTISKÂ TAKSONOMIJA, PAMATOJOTIES UZMATK ANALÎZI

Tika sekvencçti matK gçni deviòâm Kazahstânas Artemisia sugâm, t.sk. endçmiskâm. Tika apstiprinâta monofiletiskâ apakðìints Dracunculusizcelðanâs.A. kotuchoviituvums citâmDracunculussugâm norâda uz ðîs sugas piederîbu minçtai apakðìintij.matK gçnu var veiksmîgi izmantot kâ barkoda maríieriArtemisiaìints izpçtç.