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Article
Polymorphism of alien Erigeron canadensis L. (Asteraceae) along the Trans-Siberian Railway
Maria A. Galkina1* , Viktoria N. Zelenkova2 , AndreyYu. Kurskoy2 , Mikhail Yu. Tretyakov2 , Valeriy K. Tokhtar2 , Yulia K. Vinogradova1
1 N.V. Tsitsin Main Botanical Garden, Russian Academy of Sciences, ul. Botanicheskaya 4, Moscow, 127276 Russia
2 Belgorod State University, ul. Pobedy 85, Belgorod, 308015 Russia *mawa.galkina@gmail.com
Received: 22.02.2022 Abstract. Transport corridors serve as one of the main vectors of
Revised: 04.05.2022 plant invasion over long distances. The Trans-Siberian Railway,
Accepted: 11.05.2022 connecting two parts of the world with a different set of native
Published online: 19.08.2022 species, is a unique research object for analyzing the distribution
of alien plants on a global scale. The invasive species of North DOI: 10.23859/estr-220222 American origin, Erigeron canadensis L., found throughout the
UDC 581.9:575.2:58.02 Trans-Siberian Railway has been set as a model object. This
species grows directly on the railway track and on the adjacent Translated by D.M. Martynova slopes, therefore, its spreading is likely along the transport corri-
dor, but not repeatedly from settlements located nearby the railway. All plants have been divided into three haplotypes in accordance to the structure of chloroplast DNA sites (rpl32-trnL and trnL-trnF). The first two haplotypes are represented in the samples from European Russia, the third one includes all samples from the Urals, Western Siberia, the Far East, and all the rest of material collected in European part of Russia. These data confirm our hypothesis about the leading role of the Trans-Siberian Railway in the distribution of E. canadensis in Russia from west to east. However, the isolated haplotypes indicate a low degree of polymorphism of the studied genome regions of E. canaden-sis. Therefore, its successful invasion is mainly associated with modification variability.
Keywords: Canadian horseweed, railway flora, invasive species, populations, ITS1-2, rpl32-trnL, trnL-trnF, modification variability
To cite this article. Galkina, M.A. et al., 2022. Polymorphism of alien Erigeron canadensis L. (Asteraceae) along the Trans-Siberian Railway. Ecosystem Transformation 5 (3), 14-20. https://doi.org/10.23859/estr-220222
Introduction
Erigeron species, or horseweed (section Conyza), have naturalized in Europe; E. bilbaoanus (J. Remy) Cabrera, E. blakei Cabrera, E. bonariensis L., E. canadensis L., and E. sumatrensis Retz.; E. trilobus (Decne) Boiss. is also reported as ephemerophyte (Vinogradova, 2012). E. bonariensis, E. canadensis, and E. sumatrensis are three most common species, they are currently recorded in Russia as well (Galkina and Vinogradova, 2011; Vinogradova, 2012).
Canadian horseweed, E. canadensis, is an annual or winter biennial plant of North American origin. It is characterized by seed reproduction, and even small individuals of E. canadensis can produce about 2000 seeds per growing season. This explains active invasion of the species to the new territories, so it has been included in the top-100 most dangerous invasive species in Russia in 2018 (Dgebuadze et al., 2018: Vinogradova et al., 2018). According to DAISIE (Handbook..., 2009) and GT IBM A databases, the species is included in the top ten most aggressive invasive species in Europe. The identification of such plant taxa and the study of their adaptive capabilities is an urgent task of modern botany. Invasive species are the second largest threat to natural biodiversity after habitat destruction (Bellard et al., 2016; Olmstead, 2006). In phytocenoses, where first invasive species have penetrated, the other invaders are easily introduced (Hess et al., 2019).
All species of plants on the Earth, both in their native area and the secondary distribution range, are subject to microevolutionary processes. The polymorphism of the populations of the species in any part of the range indicates a high degree of genome variability, which may be associated with the influence of both abiotic and biotic environmental factors. In turn, high degree of genome variability indicates high plasticity and a wide adaptive potential, which may be associated with a wide range of responses and modification variability, or with a high degree of polymorphism. The intensity of these processes, as well as the initiating reasons, may be completely different. The influence of phytophagous animals is reported as one of such reasons for other representatives of the Asteraceae family, such as Hieracium x robustum Fr. and phytophagous gall-forming Aulacidea hieracii (L., 1758) (Kritskaya et al., 2019). The other reason is the transition to apomixis, as in some representatives of the genera Chondrilla and Taraxacum (Kashin et al., 2019; Van Dijk et al., 2020).
Therefore, considering the populations of the same species that are sufficiently distant from each other makes it possible to assess the ecological flexibility of the species and to trace the degree of genome variability. Railroads and ground roads
play a dual role: on the one hand, they allow plants to spread freely over long distances, on the other, they serve as foci and donor sites, from which unintentionally introduced alien species spread to nearby phytocenoses (Christen and Matlack, 2006; Galkina et al., 2021; Vinogradova et al., 2020; Wagner et al., 2021). The Trans-Siberian Railway connects Europe and Asia, which have a different set of native species, but the invasive species E. canadensis is noted nowadays along its entire length. Since railway conditions are characterized by a high degree of stability, the studied populations of this species are affected by exclusively abiotic factors associated with climate. E. canadensis most often grows on the railway track and adjacent slopes, but sometimes it is also found in the right-of-way. In our opinion, the growth of this species directly on the railway track indicates that it spreads directly along the Trans-Siberian Railway, secondary dispersal to nearby settlements takes place from the railroad. This hypothesis has been tested in the present study by molecular genetic analysis.
The study aims to identify genetic differences between individuals of Erigeron canadensis growing in different parts of the Trans-Siberian Railway.
Materials and methods
DNA was isolated from the leaves of herbarium specimens E. canadensis collected along the railroad tracks in the European part of Russia (Yaroslavl, Kostroma, and Vladimir oblasts), in the Urals (Sverdlovsk Oblast), in Western Siberia (Tyumen Oblast) and in the Far East (Khabarovsk Krai and Primorsky Krai) (Table 1; Fig. 1). DNA was extracted using the DNA-Extran-3 kit (CJSC Sintol, Russia). Polymerase chain reaction (PCR) was carried out in an amplifier BioRad T-100 (USA). Primers nnc18s10 (forward) and c26A (reverse) were used for the nuclear ribo-somal internal transcribed spacer 1-2 (ITS1-2) at an annealing temperature of 58 °C. Primers rpl32F (forward) and trnL UAG (reverse) were used for chloro-plast highly variable non-coding intergenic rpl32-trnL spacer at an annealing temperature of 57 °C; for the second chloroplast intergenic spacer trnL-trnF, these were primers C and F at temperatures from 0.3 to 65 °C according to the J. Shaw method (Shaw et al., 2007). The PCR product for sequencing was purified in a mixture of ammonium acetate and ethanol. DNA nucleotide sequences were read an automatic sequencer (Sintol, Russia). Further processing of nucleotide sequences was carried out in BioEdit programs. v. 7.0.5.3. (Hall, 1999) and TCS 1.21 (Clement et al., 2000). The obtained data were downloaded to the GenBank (NCBI) database.
Results and discussion
In all the studied individuals, the nuclear region of ITS1-2 was similar, but slight differences were noted in the chloroplast regions. Nucleotide substitutions and deletions were found in intergenic spacers rpl32-trnL and trnL-trnF (Fig. 2A). The Siberian populations differ only in deletion in the rpl32-trnL region. In the Far East, only E. canadensis population from Dal-nerechensk was characterized by polymorphism. Individuals collected in European Russia has polymorphism as well: for example, the ET1 specimen from the Vladimir Oblast was distinguished by the structure of the trnL-trnF region (Fig. 2B). However, these differences in the structure of the chloroplast regions in some samples were not associated with their belonging to a certain region, and sometimes to a specific population. Thus, there are differences in samples ET6a and ET6b belonging to the same population, collected in the right-of-way in the city of Galich, Kostroma region, as well as in samples ET2a and ET2b from a population growing near the Ros-tov-Yaroslavsky train station in Rostov town between the tracks (Fig. 2). This means that E. canadensis has undergone microevolutionary changes in the secondary range, at least in the Russian part of the range.
After processing the data on the structure of both chloroplast DNA regions in the TCS 1.21 program, all individuals were divided into three haplotypes
(Fig. 3). The first haplotype included an ET1 sample from the Vladimir Oblast (Bogolyubovo railway station), the second one, a sample collected in the Yaroslavl Oblast (ET2a, Rostov-Yaroslavsky station), the third one, the remaining 23 samples from the European part of Russia, the Urals, Western Siberia, and the Far East. This haplotype network testifies that, on the one hand, the population polymorphism of E. canadensis in European Russia is higher than in Siberia and the Far East, on the other hand, most of the identified intrapopulation and interpopulation differences are not significant. Higher polymorphism of the European population is presumably explained by the fact that this species has appeared in the European part of Russia much earlier than in the Siberian and Far Eastern parts of the secondary range. Therefore, microevolutionary changes in this population are logically more pronounced.
E. canadensis dissipated along the Trans-Siberian Railway eastwards. The Asian populations of this invasive species have also been influenced by microevolutionary processes, but these changes are currently insignificant. Since all Far Eastern and Siberian collections belong to the same haplotype together with most European samples, our hypothesis about the railway as the main vector of E. canadensis dispersal to remote areas of Siberia and the Far East is confirmed.
Fig. 1. Sampling sites of Erigeron canadensis at the Trans-Siberian Railway.
Table 1. Samples of Erigeron canadensis used for molecular genetic analysis.
Region Sample no. ITS1-2 No. in GenBank rpl32—trnL trnL-trnF No. of herbarium sample Sampling site Year of sampling
ET1 OL853467 OL913131 OM731595 MHA0166597 Vladimir Oblast, Bogolyubovo railway station
ET2a ET2b OL853468 OL853469 OL913132 OL913133 OM731596 OM731597 MHA0166599 Yaroslavl Oblast, Rostov town, near the Rostov-Yaroslavsky railway station
European part of Russia ET 3 OL853470 OL913134 OM731598 MHA0166598 Yaroslavl Oblast, Rostov town
ET4 ET5a ET5b OL853471 OL853472 OL853473 OL913135 OL913136 OL913137 OM731599 OM731600 OM731601 MHA0166603 MHA0166602 MHA0166601 Kostroma Oblast, Kostroma city, Malyshkovo railway station Sendega railway station 2020
ET6a ET6b OL853474 OL853475 OL913138 OL913139 OM731602 OM731603 MHA0166600 MHA0166604 Kostroma Oblast, Galich town
Ural ET16a ET16b OL853487 OL853488 OL913151 OL913152 OM731617 OM731618 MHA0412853 MHA0412855 Sverdlovsk Oblast, Pervouralsk town 2021
ET17 OL853489 OL913153 OM731619 MHA0412854 Sverdlovsk Oblast, Ekaterinburg
Western Siberia ET13 - OM731592 OM731614 - Tyumen Oblast, Tyumen city, Voynovka railway station
ET14a ET14b — OM731593 OM731594 OM731615 OM731616 — Tyumen Oblast, Tyumen city, Utyashevo railway station 2021
ET7a ET7b ET8 ET9a OL853476 OL853478 OL853478 OL853479 OL913140 OL913142 OL913142 OL913143 OM731604 OM731605 OM731606 OM731607 MHA0333516 MHA0333514 MHA0333510 Khabarovsk Krai, Khabarovsk city, Khabarovsk-1 railway station Khabarovsk Krai, Khabarovsk city, Locomotive Depot railway station
Far East ET9b ET9c OL853480 OL853481 OL913144 OL913145 OM731608 OM731609 MHA0333509 MHA0333508 Primorsky Krai, Dalnerechensk town 2021
ET10 OL853482 OL913146 OM73161C) - Primorsky Krai, Vladivostok city, main railway station
ET11a ET11b OL853483 OL853484 OL913147 OL913148 OM731611 OM731612 MHA0333512 MHA0333511 Primorsky Krai, Ussuriysk city, Sakhzavod railway station
ET12 OL853485 OL913149 OM731613 MHA0333513 Primorsky Krai, Ussuriysk city
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Conclusions
The isolated haplotypes indicate a low degree of polymorphism of the studied genome regions of Erigeron canadensis. Successful invasion of this species is most likely due to modification variability. The differences in haplotypes in various populations indicate the spreading of the species eastwards along the Trans-Siberian Railway. The sites of formation of populations resistant to abiotic environmental factors act as invasion foci.
Funding
The work was performed within the framework of the State Task for Main Botanical Garden of Russian Academy of Sciences, MBG RAS (no. 1220426001413) and agreement no. EP/29-10-21-4 between MBG RAS and Komarov Botanical Institute of Russian Academy of Sciences. The study was supported by Russian Foundation for Basic Research (grant no. 19-54-26010) and the Ministry of Education and Science of Russian Federation (agreement no. 07515-2021-1056 and grant no. 075-15-2021-678 for the Centre for Collective Use "Herbarium of MBG RAS").
ORCID
M.A. Galkina 0000-0002-3707-1473 V.N. Zelenkova 0000-0002-5191-7359 A.Yu. Kurskoy 0000-0002-8400-0694 M.Yu. Tretyakov 0000-0001-6789-8060 V.K. Tokhtar 0000-0002-7417-4893 Yu.K. Vinogradova 0000-0003-3353-1230
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