Functional Ecology Supporting Information Table S1 A list of the 24 species used to test for differences in pollen limitation and autofertility among native, invasive alien and non-invasive alien plant species in Germany. The table gives information on the taxonomic group (Family), the life cycle, the reported breeding system and pollination biology according to BiolFlor (www2.ufz.de/biolflor/index.jsp), the flower color, the plant group for each species; the native range according to GRIN (http://www.ars-grin.gov/cgi-bin/npgs/html/tax_search.pl) and the year of first record in Germany or in the Czech Republic, or the year of first cultivation (*) for the alien species. Family Species Life cycle Breeding system Pollination biology Flower color Group Native range First record Apiaceae Daucus carota Biennial Self-compatible Autonomous selfpollination, insect pollination White native - - Apiaceae Heracleum mantegazzianum Biennial Self-compatible Selfpollination, White invasive Caucasus 1862 White noninvasive Northern Africa, NA Western Asia, Caucasus, Middle Asia, East Europe, Southeastern Europe, Southwestern Europe insect pollination Apiaceae Pimpinella peregrina Biennial - - Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 1 Functional Ecology Family Species Life cycle Breeding system Pollination biology Flower color Group Native range First record Asteraceae Senecio inaequidens Perennial Self-compatible Selfpollination, Yellow invasive Southern Africa 1889 Purple native - - insect pollination Asteraceae Aster amellus Perennial Self-compatible Selfpollination, insect pollination Asteraceae Aster laevis Perennial - - Purple noninvasive Eastern Canada, Western Canada, Northeastern USA, North-Central USA, Southeastern USA 1851 Asteraceae Bidens connata Annual Self-compatible Selfpollination, Yellow noninvasive Northeastern USA, North-Central USA, Southeastern USA 1865 Yellow invasive Subarctic America, Eastern Canada, Western Canada, Northeastern USA, North-Central USA, Northwestern USA, Southeastern USA, 1891 insect pollination Asteraceae Bidens frondosa Annual Self-compatible Selfpollination Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 2 Functional Ecology Family Species Life cycle Breeding system Pollination biology Flower color Group Native range First record South-Central USA, Southwestern USA Asteraceae Bidens tripartita Annual Self-compatible Selfpollination, Yellow native - - insect pollination Asteraceae Solidago canadensis Perennial Self-incompatible Insect pollination Yellow invasive Eastern Canada, Western Canada, Northeastern USA, North-Central USA, Southeastern USA 1838 Asteraceae Solidago graminifolia Perennial Self-incompatible Insect pollination Yellow noninvasive Subarctic America, Eastern Canada, Western Canada, Northeastern USA, North-Central USA, Northwestern USA, Southeastern USA 1848 Asteraceae Solidago virgaurea Perennial Self-incompatible Insect pollination Yellow native - - Balsaminaceae Impatiens balfourii Perennial - Insect pollination Pink noninvasive Indian Subcontinent 1901* Balsaminaceae Impatiens glandulifera Annual Self-compatible Insect pollination Pink invasive Indian Subcontinent 1854 Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 3 Functional Ecology Family Species Life cycle Breeding system Pollination biology Flower color Group Native range First record Balsaminaceae Impatiens noli-tangere Annual Self-compatible Cleistogamy, Insect pollination Yellow native - - Brassicaceae Lepidium densiflorum Biennial Self-compatible Selfpollination, White noninvasive Subarctic America, Eastern Canada, Western Canada, Northeastern USA, North-Central USA, Northwestern USA, Southeastern USA, South-Central USA, Southwestern USA, Northern Mexico 1879 White native - - White invasive Subarctic America, Eastern Canada, Western Canada, Northeastern USA, North-Central USA, Northwestern USA, Southeastern USA, South-Central USA, Southwestern USA, 1786 insect pollination Brassicaceae Lepidium graminifolium Perennial Self-compatible Selfpollination, insect pollination Brassicaceae Lepidium virginicum Biennial Self-compatible Selfpollination, insect pollination Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 4 Functional Ecology Family Species Life cycle Breeding system Pollination biology Flower color Group Native range First record Northern Mexico, Southern Mexico, Caribbean, Mesoamerica Geraniaceae Geranium macrorrhizum Perennial Self-compatible Insect pollination Pink noninvasive Middle Europe 1588* (Austria), Southeastern Europe, Southwestern Europe Geraniaceae Geranium pratense Perennial Self-compatible Insect pollination Purple native - Geraniaceae Geranium pyrenaicum Perennial Self-compatible Selfpollination, Purple invasive Northern Africa, 1800 Western Asia, Caucasus, Northern Europe, Middle Europe (not Germany), East Europe, Southeastern Europe, Southwestern Europe Yellow native - insect pollination Papaveraceae Chelidonium majus Perennial Self-compatible Selfpollination, - - insect pollination Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 5 Functional Ecology Family Species Papaveraceae Papaveraceae Life cycle Pollination biology Flower color Group Native range First record Eschscholzia californica Annual Self-incompatible Insect pollination Yellow noninvasive Northwestern USA, South-Central USA, Southwestern USA, Northern Mexico 1818* Papaver somniferum Self-incompatible Selfpollination, Purple invasive Macaronesia, <1500 Northern Africa, Western Asia, Southeastern Europe, Southwestern Europe Annual Breeding system insect pollination Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 6 Functional Ecology Table S2 Plant material used to test for differences in pollen limitation and autofertility among native, invasive alien, and noninvasive alien plant species in Germany. The table gives information on the taxonomic group (Triplet), the plant group, the number of grid cells in which the species have been recorded in Germany since 1980 according to Floramap, the sources of plant materials used in the experiment, the pot size, the first month of treatment and the year of treatment, the number of plants used in the three treatments (PS: pollen supplementation, OP: open pollination, PE: pollinator exclusion) for each species. The range of the number of treated flower units per plant is given in parentheses. Triplet Species Group Number of Source(s) of seeds grid cells or seedlings Pot size Month and Number of Number of Number of year of plants PS plants OP plants PE treatment Apiaceae Daucus carota native 2656 Hofberg Garten; 2.5 L June 2013 12(10) 15(10) 15(10) 4L July 2013 3(6-9) 3(6-9) - UFA Apiaceae Heracleum invasive 1928 mantegazzianum Apiaceae Forest or grassland near the Universtity Pimpinella non- 157 BG Konstanz 3L May 2013 11(10) 15(5-10) 7(10) peregrina invasive Asteraceae I Senecio inaequidens invasive 1332 BG Konstanz 4L June 2013 15(3-11) 15(1-14) 15(10-12) Asteraceae I Aster amellus native 313 Hofberg Garten 3L Aug 2013 15(4-10) 15(10) 15(10-15) Asteraceae I Aster laevis non- 37 Holland 4/98 3L July 2013 15(9-11) 15(10) 15(5-11) invasive Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 7 Functional Ecology Triplet Species Group Number of Source(s) of seeds grid cells or seedlings Pot size Month and Number of Number of Number of year of plants PS plants OP plants PE 21(4-10) 21(5-10) 18(5-10) treatment Asteraceae II Bidens connata non- 181 BG Konstanz 1.9 L invasive Aug 2012, Sep 2013 Asteraceae II Bidens frondosa invasive 1637 BG Konstanz; B&T 3L Sep 2012 15(8-13) 15(8-12) 15(8-16) Asteraceae II Bidens tripartita native 2252 UFA; B&T; BG 3L Sep 2012 15(3-12) 15(10-11) 14(4-11) Konstanz Asteraceae III Solidago canadensis invasive 2331 BG Göttingen; B&T 3L Sep 2013 2(11-28) 3(14-21) 3(9-22) Asteraceae III Solidago non- 19 B&T 2.5 L Aug 2013 15(3-17) 15(4-17) 15(5-10) graminifolia invasive Solidago virgaurea native 2162 Rieger-Hofmann; 3L July 2013 6(4-10) 12(10) 8(9-11) Asteraceae III Hofberg Garten Balsaminaceae Impatiens balfourii non- 5 BG Bern 3L Aug 2012 15(1-11) 15(10) 15(1-10) 2299 Forest near the 3L Aug 2012 15(2-10) 15(3-16) 14(4-10) 3L Aug 2012 23(1-8) 23(1-10) 25(1-8) 1.9 L June 2013 15(1-15) 15(1-14) 14(10) invasive Balsaminaceae Impatiens invasive glandulifera Balsaminaceae Impatiens noli- Universtity native 2335 tangere Brassicaceae Forest near the Universtity Lepidium non- densiflorum invasive 57 Lab Collection oder Wayne Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 8 Functional Ecology Triplet Species Group Number of Source(s) of seeds grid cells or seedlings Pot size Month and Number of Number of Number of year of plants PS plants OP plants PE treatment Brassicaceae Lepidium native 64 graminifolium Brassicaceae Lepidium Botanical Garden 1.9 L Aug 2013 15(5-10) 15(7-10) 15(10) (BG) Konstanz invasive 633 BG Konstanz 3L May 2013 15(10-12) 15(10) 15(10) Geranium non- 132 Jelitto 2.5 L May 2014 4(5) 4(5) 3(5) macrorrhizum invasive Geranium pratense native 1737 Hofberg Garten; 2.5 L May 2014 11(5) 11(5) 10(5) virginicum Geraniaceae Geraniaceae Rieger-Hofmann Geraniaceae Geranium invasive 1829 B&T; UFA 2.5 L May 2014 10(5) 10(5) 10(5) pyrenaicum Papaveraceae Chelidonium majus native 2549 Rieger-Hofmann 3L May 2013 15(2-10) 15(5-10) 15(5-10) Papaveraceae Eschscholzia non- 143 Hofberg Garten; 1.9 L June 2013 13(1-10) 14(1-10) 11(3-10) californica invasive Papaver invasive 2.5 L July 2013 3(2-3) 4(3-4) 4(3) Papaveraceae somniferum Wyss 1277 B&T; Hofberg Garten Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 9 Functional Ecology Table S3 Results of two generalized linear mixed-effects models from which Impatiens nolitangere was dropped testing for the main and interactive effects of pollination treatment (pollen supplementation, open pollination and pollinator exclusion) and plant group (native, invasive alien and non-invasive alien) and the main effect of year of treatment on mean fruit set and seed production per fruit. Fruit set (n=1855) Seeds per fruit (n=6544) Fixed factors Df Chi2 p-value Df Chi2 p-value Pollination treatment 2 345.04 <0.0001 2 156.34 <0.0001 Plant group 2 0.052 0.974 2 2.973 0.226 Year 2 1.253 0.534 2 0.565 0.754 Pollination treatment x plant 4 2.504 0.644 4 28.007 <0.0001 group Random factors Std. dev. Std. dev. Taxonomic group 1.27 1.126 Species: Taxonomic group 5.158 1.077 Plant individual: (Species: 4.458 0.527 Observation* - 0.206 Std. dev. residuals 1.045 0.697 Taxonomic group) *To avoid overdispersion of the number of seeds per fruit, we included an observation-level random factor. Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 10 Functional Ecology Table S4 Results of four phylogenetically informed linear models from which Impatiens nolitangere was dropped testing for the effects of plant group (native, invasive alien and noninvasive alien) on pollen limitation and autofertility. The analyses were conducted using raw and adjusted values for the indices of pollen limitation and autofertility. Values of Grafen’s rho close to zero indicate a weak phylogenetic signal. Chi2 df P-value Grafen’s rho Index of pollen limitation 2.4588 2 0.2925 0.0463 Adjusted index of pollen limitation 1.038 2 0.5952 <0.0001 Index of autofertility 0.6318 2 0.7291 0.1136 Adjusted index of autofertility 0.7569 2 0.6849 0.1282 Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 11 Functional Ecology Table S5 A list of references used to resolve polytomies within families for the phylogeny of native, invasive, and non-invasive alien plant species for which the degrees of pollen limitation and autofertility were assessed. Family References Asteraceae Goertzen, L.R., Cannone, J.J., Gutell, R.R. & Jansen, R.K. (2003) ITS secondary structure derived from comparative analysis: implications for sequence alignment and phylogeny of the Asteraceae. Molecular Phylogenetics and Evolution, 29, 216-234. Panero, J.L. & Funk, V.A. (2008) The value of sampling anomalous taxa in phylogenetic studies: major clades of the Asteraceae revealed. Molecular Phylogenetics and Evolution, 47, 757-782. Brassicaceae Bailey, C.D., Koch, M.A., Mayer, M., Mummenhoff, K., O'Kane, S.L., Warwick, S.I., Windham, M.D. & Al-Shehbaz, I.A. (2006) Toward a global phylogeny of the Brassicaceae. Molecular Biology and Evolution, 23, 2142-2160. Geraniaceae Fiz, O., Vargas, P., Alarcón, M., Aedo, C., García, J.L. & Aldasoro, J.J. (2008) Phylogeny and historical biogeography of Geraniaceae in relation to climate changes and pollination ecology. Systematic Botany, 33, 326342. Papaveraceae Hoot, S., Kadereit, J., Blattner, F., Jork, K., Schwarzbach, A. & Crane, P. (1997) Data congruence and phylogeny of the Papaveraceae sl based on four data sets: atpB and rbcL sequences, trnK restriction sites, and morphological characters. Systematic Botany, 22, 575-590. Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 12 Functional Ecology Table S6 Indices of pollen limitation (PL) and autofertility (AF) of 24 native, invasive alien and non-invasive alien plant species in Germany. Triplet Species Group PL1* PL2* PL3* AF Apiaceae Daucus carota native -0.187 -0.229 -0.206 0.225 Apiaceae Heracleum mantegazzianum invasive -0.549 -1.216 -0.796 - Apiaceae Pimpinella peregrina non-invasive 0.135 0.119 0.127 0.208 Asteraceae I Senecio inaequidens invasive 0.05 0.05 0.05 0.02 Asteraceae I Aster amellus native 0.001 0.001 0.001 0.594 Asteraceae I Aster laevis non-invasive -0.021 -0.022 -0.022 0.056 Asteraceae II Bidens connata non-invasive 0.173 0.147 0.159 0.88 Asteraceae II Bidens frondosa invasive 0.13 0.115 0.123 0.673 Asteraceae II Bidens tripartita native 0.573 0.364 0.453 0.924 Asteraceae III Solidago canadensis invasive 0.296 0.228 0.259 0.156 Asteraceae III Solidago graminifolia non-invasive 0.147 0.128 0.137 0.060 Asteraceae III Solidago virgaurea native 0.047 0.045 0.046 0.751 Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 13 Functional Ecology Triplet Species Group PL1* PL2* PL3* AF Balsaminaceae Impatiens balfourii non-invasive -0.061 -0.065 -0.063 0.338 Balsaminaceae Impatiens glandulifera invasive -0.086 -0.094 -0.090 0.023 Balsaminaceae Impatiens nolitangere native -0.080 -0.087 -0.083 0.213 Brassicaceae Lepidium densiflorum non-invasive 0.772 0.436 0.572 0.844 Brassicaceae Lepidium graminifolium native 0.047 0.045 0.046 0.397 Brassicaceae Lepidium virginicum invasive 0.275 0.216 0.243 0.808 Geraniaceae Geranium macrorrhizum non-invasive 0.079 0.073 0.076 0.119 Geraniaceae Geranium pratense native 0.281 0.219 0.248 0.070 Geraniaceae Geranium pyrenaicum invasive 0.042 0.040 0.041 0.201 Papaveraceae Chelidonium majus native 0.516 0.340 0.416 0.394 Papaveraceae Eschscholzia californica non-invasive -0.118 -0.134 -0.126 0.407 Papaveraceae Papaver somniferum invasive -0.354 -0.549 *Indices of pollen limitation were calculated following different formulae. -0.437 1.215 PL1 = PL2 = Seeds per flower [pollen supplementation]− Seeds per flower [open pollination] Seeds per flower [open pollination] (Eckert et al. 2010). Seeds per flower [pollen supplementation]− Seeds per flower [open pollination] PL3 = ln Seeds per flower [pollen supplementation] Seeds per flower [pollen supplementation] Seeds per flower [open pollination] (Larson and Barrett 2000). (Vamosi et al. 2006; Harmon-Threatt et al. 2009). Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 14 Functional Ecology Table S7 Results of two phylogenetically informed linear models from which the Apiaceae, Balsaminaceae and Papaveraceae triplets were dropped testing for the effects of plant group (native, invasive alien and non-invasive alien) on pollen limitation and autofertility. The analyses were conducted using raw values for the indices of pollen limitation and autofertility. Values of Grafen’s rho close to zero indicate a weak phylogenetic signal. Chi2 df P-value Grafen’s rho Index of pollen limitation 0.276 2 0.871 <0.0001 Index of autofertility 1.594 2 0.4507 0.38 Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 15 Functional Ecology Fig S1 Fruit set and seed production (mean ±1 SE) in the three treatments (pollen supplementation, open pollination, pollinator exclusion) on eight confamilial triplets of native (black species names), invasive alien (red species names) and non-invasive alien (blue species names) plant species. Pollen supplementation Open pollination Pollinator exclusion 6 1.0 Seed production 0.4 3 2 0.2 1 0.0 0 D. carota H. mantegazzianum P. peregrina D. carota 1.0 Seed production 0.6 0.4 0.2 0.0 A. amellus H. mantegazzianum P. peregrina S. inaequidens A. laevis 80 0.8 Fruit set 4 ASTERACEAE I Fruit set 0.6 APIACEAE 5 0.8 60 40 20 0 S. inaequidens A. laevis A. amellus Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 16 Functional Ecology Pollen supplementation Open pollination Pollinator exclusion 40 Seed production 0.6 0.4 0.2 0.0 20 10 0 B. frondosa B. tripartita B. connata 1.0 25 0.8 20 Seed production Fruit set B. tripartita 30 0.6 0.4 0.2 0.0 S. virgaurea B. frondosa B. connata S. canadensis S. graminifolia ASTERACEAE III Fruit set 0.8 ASTERACEAE II 1.0 15 10 5 0 S. canadensis S. graminifolia S. virgaurea Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 17 Functional Ecology Pollen supplementation Open pollination Pollinator exclusion 8 Seed production 0.6 0.4 0.2 0.0 I. noli-tangere I. glandulifera 2 I. noli-tangere I. balfourii I. glandulifera I. balfourii L. virginicum L. densiflorum 2.0 Seed production 0.8 Fruit set 4 0 1.0 0.6 0.4 0.2 0.0 L. graminifolium 6 BRASSICACEAE Fruit set 0.8 BALSAMINACEAE 1.0 1.5 1.0 0.5 0.0 L. virginicum L. densiflorum L. graminifolium Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 18 Functional Ecology Pollen supplementation Open pollination 6 0.8 5 0.6 0.4 0.2 4 3 2 1 0.0 G. pratense GERANIACEAE 1.0 Seed production Fruit set Pollinator exclusion 0 G. pyrenaicum G. pratense G. macrorrhizum G. pyrenaicum G. macrorrhizum P. somniferum* E. californica 1.0 Seed production Fruit set 0.6 0.4 0.2 0.0 C. majus PAPAVERACEAE 40 0.8 30 20 10 0 P. somniferum E. californica C. majus *Seed production for Papaver somniferum was divided by 100. Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 19 Functional Ecology Fig. S2 Phylogenetic tree of the 24 species used to test for differences in pollen limitation and autofertility among native (black), invasive alien (red), and non-invasive alien (blue) plant species in Germany. Eschscholzia californica Papaver somniferum Chelidonium majus Geranium pyrenaicum Geranium pratense Geranium macrorrhizum Lepidium virginicum Lepidium graminifolium Lepidium densiflorum Impatiens noli-tangere Impatiens glandulifera Impatiens balfourii Solidago virgaurea Solidago graminifolia Solidago canadensis Aster laevis Aster amellus Senecio inaequidens Bidens tripartita Bidens frondosa Bidens connata Heracleum mantegazzianum Pimpinella peregrina Daucus carota Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 20 Functional Ecology Fig. S3 Correlation between autofertility and pollen limitation indices. To avoid mathematical non-independence of the indices, we randomly allocated half of the treated plants per species in the pollen supplementation treatment to the autofertility index and the other half to the pollen limitation index (Morris, W.F., Traw, M.B. & Bergelson, J. 2006 On testing for a tradeoff between constitutive and induced resistance. Oikos, 112, 102-110). Non-adjusted indices are shown in black and adjusted ones (values larger than one set to one for autofertility indices; negative values set to zero for pollen limitation indices) in red (n=23). 0.8 0.6 Pollen limitation index 0.4 0.2 0.0 -0.2 -0.4 -0.6 0.0 0.2 0.4 0.6 0.8 1.0 1.2 Autofertility index Non-invasive naturalized alien plants were not more pollen limited than invasive aliens and natives in a common garden Mialy Razanajatovo and Mark van Kleunen 21