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Using the nuclear LEAFY gene to reconstruct phylogenetic relationships among invasive knotweed (Reynoutria, Polygonaceae) populations

Published online by Cambridge University Press:  23 April 2021

Nicholas P. Tippery*
Associate Professor, Department of Biological Sciences, University of Wisconsin–Whitewater, Whitewater, WI, USA
Alyssa L. Olson
Undergraduate Student, Department of Biological Sciences, University of Wisconsin–Whitewater, Whitewater, WI, USA
Jenni L. Wendtlandt
Undergraduate Student, Department of Biological Sciences, University of Wisconsin–Whitewater, Whitewater, WI, USA
Author for correspondence: Nicholas P. Tippery, Department of Biological Sciences, University of Wisconsin–Whitewater, Whitewater, WI53190. (Email:


Knotweed species in the genus Reynoutria are native to eastern Asia but have become noxious weeds in Europe and North America. In the United States, invasive populations of Japanese knotweed (Reynoutria japonica Houtt.), giant knotweed [Reynoutria sachalinensis (F. Schmidt) Nakai], and their interspecific hybrid known as Bohemian knotweed (R. × bohemica Chrtek & Chrtková) continue to expand their ranges. Although these plants are among the most invasive terrestrial species, there are relatively few molecular tools for identifying the parental species, the F1 hybrid, or subsequent hybrids or introgressed individuals. We studied Reynoutria populations in Wisconsin, a state where all three taxa grow, to determine whether molecular data would be useful for distinguishing species and identifying hybrids. We obtained DNA sequence data from the plastid matK gene and the nuclear LEAFY gene and compared these to previously published sequences. Data from the uniparentally inherited matK region included haplotypes attributable to R. japonica and R. sachalinensis. Nuclear data indicated that R. sachalinensis plants are most similar to native plants in Japan, with two Wisconsin accessions exhibiting a monomorphic genotype for the LEAFY gene. Three Wisconsin accessions of R. japonica were each characterized by having three distinct kinds of LEAFY sequence. Most plants in our study were found to possess two or three phylogenetically distinct copies of the LEAFY gene, with the copies being most closely related to R. japonica and R. sachalinensis, respectively, and these were inferred to be interspecific hybrids. Altogether, five kinds of interspecific hybrids were identified, reflecting various combinations of LEAFY sequence types from the parental species. The widespread existence of hybrid plants in Wisconsin, many of which are morphologically identifiable as R. japonica, indicates a cryptic genetic diversity that should be examined more broadly in North America using molecular tools.

Research Article
© The Author(s), 2021. Published by Cambridge University Press on behalf of the Weed Science Society of America

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Associate Editor: Marie Jasieniuk, University of California, Davis


Bailey, JP (1994) Reproductive biology and fertility of Fallopia japonica (Japanese knotweed) and its hybrids in the British Isles. Pages 141159 in de Waal, LC, Child, LE, Wade, PM, Brock, JH, eds. Ecology and Management of Invasive Riverside Plants. Chichester, UK: Wiley Google Scholar
Bailey, JP (2003) Japanese knotweed s.l. at home and abroad. Pages 183196 in Child, L, Brock, JH, Prach, K, Pysek, P, Wade, PM, Williamson, M, eds. Plant Invasions: Ecological Threats and Management Solutions. Leiden, Netherlands: Backhuys Google Scholar
Bailey, JP (2013) The Japanese knotweed invasion viewed as a vast unintentional hybridisation experiment. Heredity 110:105110 CrossRefGoogle ScholarPubMed
Bailey, JP, Bímová, K, Mandák, B (2007) The potential role of polyploidy and hybridisation in the further evolution of the highly invasive Fallopia taxa in Europe. Ecol Res 22:920928 CrossRefGoogle Scholar
Bailey, JP, Bímová, K, Mandák, B (2009) Asexual spread versus sexual reproduction and evolution in Japanese knotweed s.l. sets the stage for the “Battle of the Clones.” Biol Invasions 11:11891203 CrossRefGoogle Scholar
Bailey, JP, Child, LE, Conolly, AP (1996) A survey of the distribution of Fallopia × bohemica (Chrtek & Chrtková) J. Bailey (Polygonaceae) in the British Isles. Watsonia 21:187198 Google Scholar
Bailey, JP, Wisskirchen, R (2006) The distribution and origins of Fallopia × bohemica (Polygonaceae) in Europe. Nord J Bot 24:173199 CrossRefGoogle Scholar
Barney, JN (2006) North American history of two invasive plant species: phytogeographic distribution, dispersal vectors, and multiple introductions. Biol Invasions 8:703717 CrossRefGoogle Scholar
Barney, JN, Tharayil, N, DiTommaso, A, Bhowmik, PC (2006) The biology of invasive alien plants in Canada. 5. Polygonum cuspidatum Sieb. & Zucc.[= Fallopia japonica (Houtt.) Ronse Decr.]. Can J Plant Sci 86:887906 CrossRefGoogle Scholar
Beerling, DJ, Bailey, JP, Conolly, AP (1994) Fallopia japonica (Houtt.) Ronse Decraene. J Ecol 82:959979 CrossRefGoogle Scholar
Benoit, LK, Les, DH, King, UM, Na, HR, Chen, L, Tippery, NP (2019) Extensive interlineage hybridization in the predominantly clonal Hydrilla verticillata . Am J Bot 106:16221637 CrossRefGoogle ScholarPubMed
Bímová, K, Mandák, B, Kašparová, I (2004) How does Reynoutria invasion fit the various theories of invasibility? J Veg Sci 15:495504 CrossRefGoogle Scholar
Brock, JH, Child, LE, de Waal, LC, Wade, M (1995) The invasive nature of Fallopia japonica is enhanced by vegetative regeneration from stem tissues. Pages 131139 in Pyšek, P, Prach, K, Rejmánek, M, Wade, M, eds. Plant Invasion—General Aspects and Special Problems. Amsterdam, Netherlands: SPB Academic Google Scholar
Buggs, RJA, Doust, AN, Tate, JA, Koh, J, Soltis, K, Feltus, FA, Paterson, AH, Soltis, PS, Soltis, DE (2009) Gene loss and silencing in Tragopogon miscellus (Asteraceae): comparison of natural and synthetic allotetraploids. Heredity 103:7381 CrossRefGoogle ScholarPubMed
Bzdega, K, Janiak, A, Książczyk, T, Lewandowska, A, Gancarek, M, Sliwinska, E, Tokarska-Guzik, B (2016) A survey of genetic variation and genome evolution within the invasive Fallopia complex. PLoS ONE 11:e0161854 CrossRefGoogle ScholarPubMed
Chernomor, O, von Haeseler, A, Minh, BQ (2016) Terrace aware data structure for phylogenomic inference from supermatrices. Syst Biol 65:9971008 CrossRefGoogle ScholarPubMed
Clements, DR, Larsen, T, Grenz, J (2016) Knotweed management strategies in North America with the advent of widespread hybrid Bohemian knotweed, regional differences, and the potential for biocontrol via the psyllid Aphalara itadori Shinji. Invasive Plant Sci Manag 9:6070 CrossRefGoogle Scholar
Crawford, KM, Whitney, KD (2010) Population genetic diversity influences colonization success. Mol Ecol 19:12531263 CrossRefGoogle ScholarPubMed
Decraene, LPR, Akeroyd, JR (1988) Generic limits in Polygonum and related genera (Polygonaceae) on the basis of floral characters. Bot J Linn Soc 98:321371 CrossRefGoogle Scholar
Doyle, JJ, Doyle, JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:1115 Google Scholar
Doyle, JJ, Flagel, LE, Paterson, AH, Rapp, RA, Soltis, DE, Soltis, PS, Wendel, JF (2008) Evolutionary genetics of genome merger and doubling in plants. Annu Rev Genet 42:443461 CrossRefGoogle ScholarPubMed
[EDDMapS] Early Detection & Distribution Mapping System (2020) Home page. Accessed: December 28, 2020Google Scholar
Forman, J (2003) Of Knotweed and Other Weeds: Invasive Plants at Home and Abroad. Ph.D dissertation, Boston: University of MassachusettsGoogle Scholar
Forman, J, Kesseli, RV (2003) Sexual reproduction in the invasive species Fallopia japonica (Polygonaceae). Am J Bot 90:586592 CrossRefGoogle Scholar
Freeman, CC, Hinds, HR (2005) Fallopia. Pages 541–546 in Flora of North America Editorial Committee, ed. Flora of North America North of Mexico. Volume 5, Magnoliophyta: Caryophyllidae. Part 2. New York: Oxford University PressGoogle Scholar
Fung, C, González-Moreno, P, Pratt, C, Oliver, TH, Bourchier, RS, González-Suárez, M (2020) Effect of humidity and temperature on performance of Aphalara itadori as a biocontrol for Japanese knotweed. Biol Control 146:104269 CrossRefGoogle Scholar
Galasso, G, Banfi, E, De Mattia, F, Grassi, F, Sgorbati, S, Labra, M (2009) Molecular phylogeny of Polygonum L. s.l. (Polygonoideae, Polygonaceae), focusing on European taxa: preliminary results and systematic considerations based on rbcL plastidial sequence data. Atti Soc Ital Sci Nat Mus Civ Stor Nat Milano 150:113148 Google Scholar
Gammon, MA, Baack, E, Orth, JF, Kesseli, R (2010) Viability, growth, and fertility of knotweed cytotypes in North America. Invasive Plant Sci Manag 3:208218 CrossRefGoogle Scholar
Gammon, MA, Grimsby, JL, Tsirelson, D, Kesseli, R (2007) Molecular and morphological evidence reveals introgression in swarms of the invasive taxa Fallopia japonica, F. sachalinensis, and F. × bohemica (Polygonaceae) in the United States. Am J Bot 94:948956 CrossRefGoogle Scholar
Gammon, MA, Kesseli, R (2010) Haplotypes of Fallopia introduced into the US. Biol Invasions 12:421427 CrossRefGoogle Scholar
Gaskin, JF, Schwarzländer, M, Grevstad, FS, Haverhals, MA, Bourchier, RS, Miller, TW (2014) Extreme differences in population structure and genetic diversity for three invasive congeners: knotweeds in western North America. Biol Invasions 16:21272136 CrossRefGoogle Scholar
Grant, V (1981) Plant Speciation. 2nd ed. New York: Columbia University Press CrossRefGoogle Scholar
Griekspoor, A, Groothuis, T (2005) 4Peaks. Version 1.7. Google Scholar
Grimsby, JL, Kesseli, R (2010) Genetic composition of invasive Japanese knotweed s.l. in the United States. Biol Invasions 12:19431946 CrossRefGoogle Scholar
Grimsby, JL, Tsirelson, D, Gammon, MA, Kesseli, R (2007) Genetic diversity and clonal vs. sexual reproduction in Fallopia spp. (Polygonaceae). Am J Bot 94:957964 CrossRefGoogle Scholar
Hollingsworth, ML, Bailey, JP (2000) Evidence for massive clonal growth in the invasive weed Fallopia japonica (Japanese knotweed). Bot J Linn Soc 133:463472 CrossRefGoogle Scholar
Hollingsworth, ML, Bailey, JP, Hollingsworth, PM, Ferris, C (1999) Chloroplast DNA variation and hybridization between invasive populations of Japanese knotweed and giant knotweed (Fallopia, Polygonaceae). Bot J Linn Soc 129:139154 CrossRefGoogle Scholar
Hollingsworth, ML, Hollingsworth, PM, Jenkins, GI, Bailey, JP, Ferris, C (1998) The use of molecular markers to study patterns of genotypic diversity in some invasive alien Fallopia spp. (Polygonaceae). Mol Ecol 7:16811691 CrossRefGoogle Scholar
Kalyaanamoorthy, S, Minh, BQ, Wong, TKF, von Haeseler, A, Jermiin, LS (2017) ModelFinder: fast model selection for accurate phylogenetic estimates. Nat Methods 14:587589 CrossRefGoogle ScholarPubMed
Kim, JY, Park, CW (2000) Morphological and chromosomal variation in Fallopia section Reynoutria (Polygonaceae) in Korea. Brittonia 52:3448 CrossRefGoogle Scholar
Lewis, PO (2001) A likelihood approach to estimating phylogeny from discrete morphological character data. Syst Biol 50:913925 CrossRefGoogle ScholarPubMed
Li, A, Park, CW (2003) Reynoutria . Page 319 in Wu, CY, Raven, PH, Hong, DY, eds. Flora of China. Volume 5. St Louis, MO: Missouri Botanical Garden Press Google Scholar
Li, C, Ohadi, S, Mesgaran, MB (2020) Asymmetry in fitness-related traits of later-generation hybrids between two invasive species. Am J Bot 108:5162 CrossRefGoogle ScholarPubMed
Maddison, WP, Maddison, DR (2019) Mesquite: a modular system for evolutionary analysis. Version 3.61. Google Scholar
Mandák, B, Pyšek, P, Lysák, M, Suda, J, Krahulcová, A, Bímová, K (2003) Variation in DNA-ploidy levels of Reynoutria taxa in the Czech Republic. Ann Bot 92:265272 CrossRefGoogle ScholarPubMed
Mitchell, N, Owens, GL, Hovick, SM, Rieseberg, LH, Whitney, KD (2019) Hybridization speeds adaptive evolution in an eight-year field experiment. Sci Reports 9:112 Google Scholar
Müller, K (2005) SeqState—primer design and sequence statistics for phylogenetic DNA data sets. Appl Bioinf 4:6569 Google Scholar
Murrell, C, Gerber, E, Krebs, C, Parepa, M, Schaffner, U, Bossdorf, O (2011) Invasive knotweed affects native plants through allelopathy. Am J Bot 98:3843 CrossRefGoogle ScholarPubMed
Nguyen, LT, Schmidt, HA, von Haeseler, A, Minh, BQ (2015) IQ-TREE: a fast and effective stochastic algorithm for estimating maximum likelihood phylogenies. Mol Biol Evol 32:268274 CrossRefGoogle ScholarPubMed
Parepa, M, Fischer, M, Krebs, C, Bossdorf, O (2014) Hybridization increases invasive knotweed success. Evol Appl 7:413420 CrossRefGoogle ScholarPubMed
Parepa, M, Schaffner, U, Bossdorf, O (2012) Sources and modes of action of invasive knotweed allelopathy: the effects of leaf litter and trained soil on the germination and growth of native plants. NeoBiota 13:1530 CrossRefGoogle Scholar
Park, CW, Bhandari, GS, Won, H, Park, JH, Park, DS (2018) Polyploidy and introgression in invasive giant knotweed (Fallopia sachalinensis) during the colonization of remote volcanic islands. Sci Rep 8:16021 CrossRefGoogle ScholarPubMed
Pyšek, P, Brock, JH, Bímová, K, Mandák, B, Jarošík, V, Koukolíková, I, Pergl, J, Štěpánek, J (2003) Vegetative regeneration in invasive Reynoutria (Polygonaceae) taxa: the determinant of invasibility at the genotype level. Am J Bot 90:14871495 CrossRefGoogle Scholar
Ronquist, F, Teslenko, M, van der Mark, P, Ayres, DL, Darling, A, Höhna, S, Larget, B, Liu, L, Suchard, MA, Huelsenbeck, JP (2012) MrBayes 3.2: Efficient Bayesian phylogenetic inference and model selection across a large model space. Syst Biol 61:539542 CrossRefGoogle Scholar
Sanchez, A, Schuster, TM, Kron, KA (2009) A large-scale phylogeny of Polygonaceae based on molecular data. Int J Plant Sci 170:10441055 CrossRefGoogle Scholar
Schuster, TM, Wilson, KL, Kron, KA (2011) Phylogenetic relationships of Muehlenbeckia, Fallopia, and Reynoutria (Polygonaceae) investigated with chloroplast and nuclear sequence data. Int J Plant Sci 172:10531066 CrossRefGoogle Scholar
Simmons, MP, Ochoterena, H (2000) Gaps as characters in sequence-based phylogenetic analyses. Syst Biol 49:369381 CrossRefGoogle ScholarPubMed
Small, RL, Cronn, RC, Wendel, JF (2004) Use of nuclear genes for phylogeny reconstruction in plants. Aust Syst Bot 17:145170 CrossRefGoogle Scholar
Steward, AN (1930) The Polygoneae of eastern Asia. Contrib Gray Herbarium Harvard U 88:1129 Google Scholar
Suda, J, Trávníček, P, Mandák, B, Berchová-Bímová, K (2010) Genome size as a marker for identifying the invasive alien taxa in Fallopia section Reynoutria . Preslia 82:97106 Google Scholar
Tippery, NP, Pesch, JD, Murphy, BJ, Bautzmann, RL (2020) Genetic diversity of native and introduced Phragmites (common reed) in Wisconsin. Genetica 148:165172 CrossRefGoogle Scholar
Vrchotová, N, Šerá, B (2008) Allelopathic properties of knotweed rhizome extracts. Plant Soil Environ 54:301303 CrossRefGoogle Scholar
Yu, WG, Fa, SJ, Xu, CM, Zhu, LT, Hou, YT, Lin, FY, Li, FZ (2008) Systematic position of Reynoutria and Polygonum sibiricum inferred from sequences of chloroplast trnL-F and matK . J Syst Evol 46:676681 Google Scholar
Yan, P, Pang, QH, Jiao, XW, Zhao, A, Shen, YJ, Zhao, SJ (2008) Genetic variation and identification of cultivated Fallopia multiflora and its wild relative by using chloroplast matK and 18S rRNA gene sequences. Planta Med 74:15041509 CrossRefGoogle Scholar
Zika, PF, Jacobson, AL (2003) An overlooked hybrid Japanese knotweed (Polygonum cuspidatum × sachalinense; Polygonaceae) in North America. Rhodora 105:143152 Google Scholar