New Results

The use of DNA barcoding for the identification of giant hogweeds in the European North-East of Russia

D.M. Shadrin, I.V. Dalke, I.G. Zakhozhiy, D.S. Shilnikov, M.N. Kozhin, I.F. Chadin
doi: https://doi.org/10.1101/2023.02.21.529251

Abstract

Qualitative differences in the nrDNA ITS and ETS sequences in the total sample were determined to allow identification of the following groups of samples: H. mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi × H. cf. mantegazzianum among other representatives of the Heracleum genus presented in the genetic databases. The study revealed the absence of correlation between a sample type in the group H. mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi × H. cf. mantegazzianum and qualitative transformations in the ITS and ETS sequences. The qualitative analysis of the rbcL, matK, rps16 intron, intergenic spacers psbA-trnH, trnQ-rps16, rps16-trnK and rpl32-trnL cpDNA sequences for the samples identified as Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi × H. cf. mantegazzianum demonstrated no correlation between them as well.

Introduction

Apiaceae Lindl. (Umbelliferae Juss.) is a large family of flowering plants comprised of compound species complexes with a high level of morphological variability (Kljuykov et al., 2018). The most popular classification for Umbelliferae at present is a system based on the anatomy and morphology of the fruit (Pimenov, Leonov, 1993; Pimenov, 2012; Valuyskikh et al., 2021). The classification system presented is not absolute and at times fails to accurately reflect relationships within the family (Downie et al., 2010). The genus Heracleum includes 53 taxa globally (Tkachenko, 2014). With regard to the Heracleum genus, most of the researchers’ attention is given to a group consisting of three species: Heracleum mantegazzianum Sommier & Levier, H. sosnowskyi Manden., H. persicum Desf. These three species are combined into a group of “large, tall or giant” hogweeds with a height of 3-5 meters (Mandenova, 1950, Nielsen et al., 2005; Jahodová et al., 2007a). Despite the fact that these species are referred to the section of the genus Heracleum – Pubescentia by a number of taxonomists, which includes some more species, allocating of the giant hogweed group is convenient for practical purposes. It is generally recognized that determination of the boundaries of the Pubescentia species is challenging (Mandenova, 1950; Jahodová et al., 2007a; Pimenov, Ostroumova, 2012; Ebel et al., 2018; Vladimirov et al., 2019).

Ornamental attributes and nutritional properties of these giant hogweed species caused their intentional introduction in a range of European countries. Following the second half of the 20th century, these species began to actively spread beyond the boundaries of cultivated crops. H. mantegazzianum Somm. et Lev. – Central and Western Europe (https://www.gbif.org/species/3034825), H. persicum Desf. ex Fisch., – European North (https://www.gbif.org/species/3628745) and H. sosnowskyi Manden – Eastern Europe (https://www.gbif.org/species/3642949. Currently, all the three species are considered invasive in a large area of the European continent, with the exception of the Caucasus, where H. mantegazzianum and H. sosnowskyi were originally described as endemic species (Nielsen et al., 2005; Ecology and management…, 2007; Zakhozhiy et al., 2021).

The Heracleum genus claims a complex evolutionary history, including hybridization, which resulted in complex variations in its species morphology, thus making identification of the species with traditional taxonomic methods more difficult (Pyšek et al., 2007; Sortland, 2010). Studies of the biology of the H. mantegazzianum and H. sosnowskyi species carried out within their invasive habitats in order to develop measures to control distribution of these species showed that they can be considered ecological twins for a number of individual development, structural and population dynamic features (Pyšek et al., 2007; Dalke et al., 2015). It should also be taken into account that their identification and documentation were not always carried out when introducing plants of this genus (Kudinov et al., 1980; Skupchenko, 1989; Pyšek et al., 2007). According to some authors (Pyšek et al., 2007), it would be extremely difficult for introducers to collect mericarps of solely one species in the common growth area for both H. mantegazzianum and H. sosnowskyi, since the key distinction feature of these two species – the degree of dissection of leaf lobes – is almost impossible to be used in the fruiting time. Some authors (Ebel et al., 2018) noted that “…the use of the name Heracleum sosnowskyi for wildings of the “giant hogweed” is rather tentative”. We believe that the statement about the conditionality of such naming can be equally applied to H. mantegazzianum.

In the last decade, in order to identify systematically complex plants, alongside with traditional taxonomy methods DNA barcoding of plants has been widely used, which enables identification of organisms based on short fragments of the DNA sequence (Hebert et al., 2003; Kress, 2017). In order to barcode representatives of the plant kingdom the following is used: rbcL and matK genes, intergenic spacer trnH-psbA of chloroplast DNA (cpDNA), sequence ITS1-5.8s-ITS2 of the ribosomal nuclear DNA cluster (nrDNA) (Shneyer, Rodionov, 2018; Shekhovtsov et al., 2019; Shadrin, 2021). By now, the most detailed description of the molecular phylogenetic relationships, as well as the origin and migration of representatives of the Apiaceae family has been carried out using the ITS region of nrDNA (Downie et al., 2001, 2010; Banasiak et al., 2013). To identify a number of subtaxa of the Apiaceae family, an attempt was made to use the cpDNA trnH-psbA intergenic spacer region (Degtjareva et al., 2012), as well as a combination of ITS and trnH-psbA sequences (Liu et al., 2014; Valuyskikh, Shadrin, 2021). Another work declared high conservation of the intergenic spacer psbA-trnH to act as a limitation for the identification of representatives of the Heracleum genus and related taxa (Logacheva et al., 2007). An effective marker for phylogenetic studies of the tribe Tordylieae, which comprises the Heracleum genus, is the nrDNA ETS (external transcribed spacer) sequence, especially in combination with the ITS sequence (Logacheva et al., 2010). In addition to the markers listed above, for the molecular phylogenetic analysis of representatives of the Heracleum genus in the Chinese flora, noncoding cpDNA sequences were used: rps16 intron, trnQ-rps16, rps16-trnK and rpl32- trnL (Yu et al., 2011). The discrepancy between the phylogenies of the Heracleum genus based on the comparison of nrDNA and cpDNA sequences was also noted there, which, according to the authors, is an unprecedented case in the molecular phylogenetic system of the Apiaceae system (Yu et al., 2011). The Apiaceae is a large and taxonomically complex family, especially at the low systematic level, which requires new approaches for the analysis and search for taxonomic attributes.

The work aims at finding cpDNA and nrDNA markers that can make it possible to differentiate between H. sosnowskyi and H. mantegazzianum, which is essential to solve the taxonomic problems for this group of plants and assess the vector and rate of microevolutionary processes of these species under the invasion conditions.

Materials and methods

Sampling

The samples of the Heracleum genus representatives were collected in five Russian regions (Fig. 1.1, 1.2): the Kabardino-Balkarian Republic (5 samples), Karachay-Cherkess Republic (14 samples), Komi Republic (16 samples), Krasnodar Krai (11 samples), Murmansk Region (4 samples). The samples were taken from reproductive plants in the flowering or initial fruiting phase. Samples dedicated for genetic analysis were put into paper bags. Herbarium specimens were obtained from each sample: a small segment of the inflorescence and the middle leaf cut into several parts according to the size of the herbarium sheet. Each plant was photographed from several angles, the height and diameter at the base were measured.

Fig. 1.
Fig. 1.

Heracleum samples collection sites within invasive and natural habitats.

Fig. 2.
Fig. 2.

Boundaries of natural habitats for Heracleum mantegazzianum, H. sosnowsky, H. persicum and sample collection sites. Habitat boundaries are given according to Jahodová et al., 2007a.

Morphological analysis

Samples of Heracleum sibiricum L. For the molecular phylogenetic analysis were identified using the key published in «Flora of the Northeast of the European Part of the USSR» (Flora…, 1977). We analyzed the attributes differentiating H. mantegazzianum and H. sosnowskyi, based on the data of a number of authors who had an opportunity to work with both species (Table 1) (Mandenova, 1950; Tsvelev, 2000; Pimenov, Ostroumova, 2012; Vladimirov et al., 2019). In most cases, the quantitative characteristics given for both species are overlapping sets. An exception is the plant height (up to 150 cm) indicated for H. sosnowskyi, according to I.P. Mandenova’s monograph (Mandenova, 1950). On this basis, all the giant hogweed samples we collected could not be referred to H. sosnowskyi. The most striking difference between these two species, as noted by all the authors, is the shape of segments and leaf lobes. I.P. Mandenova also points out the difference in leaf shape between H. mantegazzianum and H. sosnowskyi, however, she mentions that the leaf shape in plants of this genus is very variable and cannot serve as a stable diagnostic property. Other authors (Tsvelev, 2000; Vladimirov et al., 2019) consider features associated with leaf shape to be sufficient to distinguish between these two species.

View this table:
Table 1.

Attributes distinguishing H. sosnowskyi from H. mantegazzianum

For the morphological identification of the giant hogweed samples described in this work, we used the set of features given in the monograph by M.G. Pimenov and T.A. Ostroumova, 2012 (Pimenov, Ostroumova, 2012) (Table 1). These authors recognize two species – H. sosnowskyi and H. mantegazzianum, but for the latter one they consider H. wilhemsii Fissch. & Avé-Lall to be a preferable name, though they accept the use of H. mantegazzianum due to its being widely distributed. All the specimens collected in the Karachay-Cherkess Republic and Krasnodar Krai were associated by us with H. mantegazzianum based on morphological features, which is also consistent with the known boundaries of the natural habitats of the giant hogweed (Jahodová et al., 2007a) (Fig. 1). The identification was carried out mainly on the basis of the shape of the segments and leaf lobes of the samples. The use of the shape of the stylopodium, size of the stiles, and color of the anthers as attributes was challenged either due to the difficulties in correlating the verbal description of the morphology of these structures with the observed condition of the attributes, or because the observed condition of such attributes failed to correspond to any one described in the species diagnosis. In some cases, not only quantitative, but also qualitative properties turned out to be midpoint between the attributes typical for H. sosnowskyi and H. mantegazzianum. Each specimen of the giant hogweed was assigned to either H. sosnowskyi or H. mantegazzianum based on a count of the number of attributes that matched the description of the species. With an equal number of attributes in the sample assigned to each of these two species or with a predominance of intermediary characteristics, such samples were considered hybrids of H. sosnowskyi x H. mantegazzianum.

Molecular Analysis

DNA Extraction, Amplification, Sequencing

Total DNA was extracted from dried leaves using the DNA-Extran-3 kit (Sintol, Russia) according to the manufacturer’s manual. Polymerase chain reaction (PCR) of the fragments was carried out in 50 μl of a mixture containing 10 μl of ScreenMix (Eurogen, Russia), 10 μl of each primer (0.3 μM) (Evrogen, Russia), 18 μl of ddH2O (PanEko, Russia) and 2 μl of DNA nanomatrix (1 ÷ 100 ng). Primers for the amplification of DNA sections used in the analysis and the conditions for carrying out the polymerase chain reaction are presented in Table 2.

View this table:
Table 2.

Used primers and conditions for the polymerase chain reaction

The PCR products were separated in a 1.5% agarose gel based on tris-acetate-ethylene-diamine-tetraacetic acid (TAE) and further purified using the ColGen DNA purification system (Sintol, Russia) according to the manufacturer’s manual. The sequencing products were separated on a NANOFOR 05 DNA sequencer (Sintol, Russia). DNA isolation, PCR, and sequencing were performed using the equipment of the “Molecular Biology” Center for Collective Use, the Institute of Biology, Federal Research Center of the Komi Research Center, Ural Branch of the Russian Academy of Sciences, Syktyvkar.

Multiple alignment of nucleotide sequences was performed using the ClustalW algorithm in MegaX (Thompson et al., 1994; Kumar et al., 2018). Phylogenetic trees were built in MegaX using the maximum likelihood method (Maximum Likelihood, ML). When constructing molecular phylogenetic trees, we used the nucleotide sequences obtained by us, as well as data from other authors available in Genbank (NCBI, 2023) and BOLD Systems (BOLD Systems, 2023) databases. Azilia eryngioides (Pau) Hedge & Lamond was used as an outgroup in the reconstruction of phylogenetic trees. The analysis resulted in obtainment of characteristics for both individual sequences and combined data. The list of samples of Heracleum genus considered in this study, including their location, voucher information, and GenBank (NCBI) number, is presented in Table 3.

View this table:
Table 3.

Origin, voucher information (or references), and GenBank accession numbers for plants used in the present study

Results and Discussion

The molecular phylogenetic analysis included 50 samples of plants of the Heracleum genus (Table 3). DNA sequences of markers were not obtained for all the samples included in the analysis, since originally the analysis for each marker was carried out on a selective group that included plants classified by us as H. sosnowskyi and H. mantegazzianum. If the specimens of the sample group did not differ or produces differences not correlating with species specificity, the marker was not used further. A total of 224 sequences were generated for nine markers (Table 4).

View this table:
Table 4.

Characteristics of sequences of Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H.cf. sosnowskyi × H.cf. mantegazzianum.

Based on the analysis of the published sequences of the rbcL, matK genes and cpDNA psbA-trnH intergenic spacer of the plants belonging to the Heracleum genus (NCBI, 2023; BOLD Systems, 2023), it was assumed that the studied sequences are highly conserved and cannot be used for DNA barcoding of hogweed plants. Sequence analysis of the rbcL and matK genes of 27 samples collected by us (Table 3) jointly with other data for other species of the Heracleum genus (NCBI, 2023; BOLD Systems, 2023), showed a very low level of divergence both within the analyzed group of giant hogweeds and for the entire series of Heracleum taxa. The degree of pairwise genetic differences between the compared samples was from 0 to 1.1% and from 0 to 1.5% for the rbcL and matK genes, respectively. Among the plant samples assigned by us to H. sosnowskyi and H. mantegazzianum, the degree of pairwise genetic differences was 0%. Thus, it was confirmed that these markers are not applicable for identification of plants of the Heracleum genus.

The analysis of 41 sequences of the psbA-trnH intergenic spacer revealed variations in patterns TTGTTCTATCA and TGATAGAACAA at positions from 60 to 76 bp, which were repeated twice in the analyzed sequences (Fig. 3). Previously, discrete variations in the psbA-trnH sequence were published for plants of the Heracleum genus (Logacheva et al., 2007).

Fig. 3.
Fig. 3.

The variable sites from an alignment of trnH-psbA cpDNA sequences.

The variations described above occur both within our giant hogweed sample series and among other species included in the analysis. The analysis of variants showed no correlation between this type of variability and species. The presence of variations was explained by a combination of two independent evolutionary events (inversion and duplication), assuming that they occurred several times independently of each other (Logacheva et al., 2007). Following the psbA-trnH intergenic spacer sequence alignment, the studied sample series showed another polymorphic site – at position 30 (G/T). The revealed polymorphism, like the previous ones, did not correlate with the species. Pairwise genetic variability in the sample analyzed was 5.3%.

Next, 10 plant samples were studied to assess the variability of the sequences of rps16 intron and three non-coding cpDNA intergenic spacers – trnQ-rps16, rps16-trnK and rpl32-trnL (Table 3.1). Previously, these markers were used to study molecular phylogenetic relationships in the genus Heracleum (Yu et al., 2011) and to identify H. mantegazzianum (Identification…, 2019). Sequence alignment showed that rps16 intron and intergene spacer trnQ-rps16 have a less pronounced level of variability than intergenic spacers rps16-trnK and rpl32-trnL (Table 4). The polymorphism of the rps16 intron fragments and the trnQ-rps16 intergenic spacer is mainly explained by single nucleotide substitutions. The variability of the rps16-trnK intergenic spacer was determined by single nucleotide insertions and deletions. In the hogweed samples under study, the rpl32-trnL intergenic spacer, which included three indelia 12, 10, and 6 bp long, with a total alignment length of 978 bp attracted special attention. For other angiosperms, specific indeliums were described in the sequence of the intergenic spacer rpl32-trnL (Shaw et al., 2007; Miller et al., 2009).

Nevertheless, the revealed variability for the rps16 intron sequences and cpDNA intergenic spacers trnQ-rps16, rps16-trnK and rpl32-trnL of the sample series under study did not allow to allocate the samples based on their reference to H. mantegazzianum, H. cf. sosnowskyi or H. cf. mantegazzianum (Fig. 4).

Fig. 4.
Fig. 4.

Molecular-phylogenetic trees constructed based on sequence comparison of 10 plant samples of giant hogweed. Trees were constructed using the maximum likelihood method. The specimen GenBank accession numbers in parentheses follows the names. a) - rps16 intron, b) - trnQ-rps16, c) - rps16-trnK, d) - rpl32-trnL.

The nrDNA ITS sequence analysis revealed resolution in the topology of the phylogenetic tree for most species of the genus Heracleum (Fig. 5). The pairwise genetic distance between different species ranged from 0 to 2.1%. Meanwhile, for the group of samples including, according to our data, Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi×H. cf. mantegazzianum a very low level of variability in the ITS sequence may be noted. The pairwise genetic distance ranged from 0 to 0.3%. A value of 0.3% was associated with a hybrid peak (R) in the 5.8s region in three samples of H. cf. sosnowskyi collected in the North Caucasus (NCBI: OQ222183, OQ222184, OQ222185). The use of ITS sequences for some representatives of the Heracleum genus is known not to lead to their resolution on the topology of the molecular phylogenetic tree in every instance (Logacheva et al., 2010). Based on the variability of ITS sequences, other authors suggested the origin and possible migration routes of 29 species of the Heracleum genus in the Chinese flora (Yu et al., 2011).

Fig. 5.
Fig. 5.

Optimal phylogenetic tree from Maximum Likelihood analyses for the ITS nrDNA Heracleum data. The specimen GenBank accession numbers in parentheses follows the names. Samples obtained by the authors are highlighted in bold.

The samples assigned by us to Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi×H. cf. mantegazzianum with a high bootstrap value (99) grouped into a separate clade differing from the rest of the species of the genus under (Fig. 5). After overall alignment of the sequences of different species of the Heracleum genus at position 125 for ITS1, a unique substitution (A/G) typical for just the mentioned plant samples was found. The sequences we obtained for H. sphondylium subsp. sibiricum were found to be identical to the Genbank data for this species (CCЫЛКA), as shown in the molecular phylogenetic tree (Fig. 5). It can be assumed that the ITS sequence has sufficient polymorphism to enable identification of some species of the Heracleum genus, but cannot be used to distinguish between the samples of Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi×H. cf. mantegazzianum in the European part of Russia.

Thus, using the unique substitution we identified in the nrDNA ITS sequence, we identified a group of samples assigned to Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi×H. cf. mantegazzianum from among other representatives of the Heracleum genus, with their ITS sequences presented in genetic databases.

Another genetic marker that was used as a potential DNA barcode to distinguish between the analyzed samples was the nrDNA ETS sequence. A revision of genetic databases (NCBI; BOLD Systems) showed that data on ETS are not available for H. sosnowskyi, and for H. mantegazzianum are presented with regard to one sample.

For 46 samples of plants classified as Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi×H.cf mantegazzianum ETS sequences were obtained. A total of 81 ETS sequences for 32 species of the genus Heracleum were included in the analysis (Fig. 6). The ratio of variable sites to the total alignment length within the analyzed group (Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi×H. cf. mantegazzianum) was 4.2 % (Table 4).

Fig. 6.
Fig. 6.

Optimal phylogenetic tree from Maximum Likelihood analyses for the ETS nrDNA Heracleum data. The specimen GenBank accession numbers in parentheses follows the names. Samples obtained by the authors are highlighted in bold.

Based on the comparison of H. sosnowskyi plant sequences with the only available in genetic databases H. mantegazzianum sample, we found the use of the ETS sequence for species distinction upon involving more H. mantegazzianum samples as promising (Shadrin et al., 2021). However, the data obtained as a result of follow-up work, analysis of the literature and data on the morphological features of this species group suggest that not all plants previously traditionally assigned to H. sosnowskyi based on their confinement to Russia’s European northeast can be attributed specifically to H. sosnowskyi.

As a result, the samples that we previously identified as H. sosnowskyi (Shadrin et al., 2021) were divided into four groups after reidentification using a set of features (Table 1): Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi×H. cf. mantegazzianum.

In addition to the data already obtained (Shadrin et al., 2021), ETS sequences for 24 samples of H. mantegazzianum collected from natural populations of the North Caucasus were studied. The qualitative analysis of the ETS sequences of H. mantegazzianum from the North Caucasus revealed that they are more similar to plants growing in the European northeast of Russia than with the H. mantegazzianum sample presented in the genetic database (NCBI: FJ807509) (Fig. 6). The average pairwise genetic distance within the sample group under analysis ranged from 0.0 to 0.5%. The average pairwise genetic distance between other compared species ranged from 3.0 to 22.7%. A qualitative analysis of sequence variability showed no correlation between the identified changes and the species affiliation of the H. cf. sosnowskyi and H. mantegazzianum samples.

As a result of the analysis, we identified four substitutions at positions 34 (T/C), 186 (A/C, A/G), 259 (C/A) and 357 (A/G) attributable only to 46 samples of the plants classified by us as Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi×H. cf. mantegazzianum, including for a sample from the GenBank database (NCBI: FJ807509). The revealed variability can potentially be used as a marker for distinguishing H. sosnowskyi and H. mantegazzianum from other representatives of the Heracleum genus.

The analysis of the alignment of the combined sequences (ITS+ETS = 984 bp) showed the maintained general topology of the phylogenies constructed based on the comparison of the ITS and ETS sequences each separately.

The names H. mantegazzianum and H. sosnowskyi are likely to be used in one or another part of the vast invasive range of giant hogweed mainly following the established tradition. Obviously, the farms involved in the introduction of a new fodder crop did not carry out any special identification of these plants. Differentiation of two monocarpic species of giant hogweed is not an easy task even for professional botanists, especially since species of the Heraleum genus, even belonging to different sections, are capable of crossing (Stewart, Grace, 1984; Jahodová et al., 2007b). The conventionality of the name H. sosnowsky can explain the phenomenon of the existence of a clear boundary between the invasive ranges of H. mantegazzianum and H. sosnowskyi: in the former USSR, the giant invasive species of hogweed is called H. sosnowskyi, in Western Europe – H. mantegazzianum, in Eastern Europe both species are found. At first glance, from a practical point of view, the subtle taxonomic differences between the two species seem not to matter. However, misidentification of invasive species is scientifically unacceptable and can have serious implications in practice. The name H. sosnowskyi was included in official and unofficial methodological guidelines for the eradication of unwanted thickets of this species, in legal acts on regional and municipal levels in Russia, Belarus, Latvia, Lithuania and Estonia.

Conclusion

The comparative analysis of the following sequences: rbcL, matK, rps16 intron, cpDNA intergenic spacers psbA-trnH, trnQ-rps16, rps16-trnK and rpl32-trnL for the samples defined by us as Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi × H. cf. mantegazzianum did not reveal any qualitative differences that could be correlated with the samples. The comparative analysis of the nrDNA ITS and ETS sequences revealed qualitative differences in them, which make it possible to identify the groups of samples: Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi × H. cf. mantegazzianum among other representatives of the Heracleum genus, which are referred to in genetic databases at the moment. The comparison of ITS and ETS sequences for the samples of Heracleum mantegazzianum, H. cf. sosnowskyi, H. cf. mantegazzianum and H. cf. sosnowskyi × H. cf. mantegazzianum showed no correlation between their polymorphism and the type of samples in the given sample series. The data obtained through the study, as well as information on morphological features that make it possible to distinguish between H. mantegazzianum and H. sosnowskyi indicate that it is possible that the names H. mantegazzianum and H. sosnowskyi are used in one or another part of the vast invasive range of giant hogweed mainly following the established tradition.

Funding

The reported study was funded by RFBR and NSFB, project number No 20-54-18002.

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Posted February 22, 2023.
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The use of DNA barcoding for the identification of giant hogweeds in the European North-East of Russia
D.M. Shadrin, I.V. Dalke, I.G. Zakhozhiy, D.S. Shilnikov, M.N. Kozhin, I.F. Chadin
bioRxiv 2023.02.21.529251; doi: https://doi.org/10.1101/2023.02.21.529251
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