Research Article |
Corresponding author: Sylvain G. Razafimandimbison ( sylvain.razafimandimbison@nrm.se ) Academic editor: Hanno Schaefer
© 2018 Elisette M. Rahelivololona, Eberhard Fischer, Steven B. Janssens, Sylvain G. Razafimandimbison.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Rahelivololona EM, Fischer E, Janssens SB, Razafimandimbison SG (2018) Phylogeny, infrageneric classification and species delimitation in the Malagasy Impatiens (Balsaminaceae). PhytoKeys 110: 51-67. https://doi.org/10.3897/phytokeys.110.28216
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The species-rich genus Impatiens (Balsaminaceae) is represented in Madagascar by no less than 260 species. We conducted molecular phylogenetic analyses of the Malagasy Impatiens based on nuclear and plastid data and 52 accessions (representing 33 species) to: 1) reassess the monophyly of the Malagasy Impatiens; 2) assess the monophyly of the sections Preimpatiens (Humblotianae and Vulgare groups) with spurs and Trimorphopetalum without spurs as delimited by Perrier de la Bâthie, as well as that of the subgenera Impatiens and Trimorphopetalum as defined by Fischer and Rahelivololona; 3) infer the current status of some morphologically variable species; and 4) test whether the species of Impatiens from the Marojejy National Park form a monophyletic group. The Malagasy Impatiens are further confirmed to be paraphyletic with respect of the Comorian I. auricoma. The present sectional and subgeneric classifications of the Malagasy Impatiens are partly supported, with strong support for the monophyly of the sect. or subgen. Trimorphopetalum. Section Preimpatiens was not supported as monophyletic and neither the Humblotianae group nor the Vulgare group is monophyletic. Impatiens elatostemmoides, I. “hammarbyoides”, I. inaperta, I. lyallii and I. manaharensis are either para- or polyphyletic and may represent morpho-species. The Impatiens species from the Marojejy National Park do not form a monophyletic group and therefore are suggested to be derived from numerous independent colonisation events from all over Madagascar followed by subsequent diversifications.
Balsaminaceae , infrageneric classification, Impatiens , Madagascar, monophyly, species delimitation, systematics, taxonomy
The genus Impatiens L. (Balsaminaceae) is a monophyletic and diverse genus with more than 1000 species (e.g.
Representatives of Impatiens subgen. Impatiens. A, B Impatiens bicaudata, Montagne d’Ambre C I. lyallii, Montagne d’Ambre D, E I. bisaccata, Montagne d’Ambre F I. max-huberi, Marojejy G I. nomenyae, Marojejy H, I I. masoalensis, Marojejy KI. cf. manaharensis, Marojejy L, O I. marojejyensis, Marojejy M I. susan-nathansoniae, Marojejy N I. hendrikii, Marojejy. Photos: E. Fischer.
Representatives of Impatiens subgen. Trimorphopetalum. A Impatiens lutzii, Montagne d’Ambre B I. galactica, Marojejy C, D I. “capuronii”, Marojejy E, F I. furcata, Marojejy G, K I. navicula, Marojejy H, L I. “humillima”, Marojejy I I. “hammarbyoides”, Marojejy M I. elatostemmoides, Marojejy NI. aff. elatostemmoides 3, Montagne d’Ambre. Photos: E. Fischer.
The category of species is widely accepted as the basic or working unit of biological classification (
The first molecular phylogenetic study of the family Balsaminaceae by
The Marojejy National Park is located in north-eastern Madagascar within the SAVA Region. With its tallest peak rising to 2137 m, the area is home to a diverse flora of upland species. The wide range of elevations and rugged topography of Marojejy create diverse habitats, which transition quickly with changes in altitude. There are four types of forests within the park: lowland rainforest below 800 m (Fig.
The main objective of this study was to reconstruct a new and larger phylogeny of the Malagasy Impatiens, with a particular emphasis on taxa collected from Marojejy, using two nuclear AP3/DEF homologues (ImpDEF1 and ImpDEF2) and the plastid atpB-rbcL spacer. The resulting phylogeny was subsequently used to (i) reassess the monophyly of the Malagasy Impatiens as stated by
The taxon sampling for this study was focused on the Malagasy representatives of the genus Impatiens. We expanded the previous dataset of
List of taxa studied, voucher information and accession numbers of the selected markers. ‘–’ refers to a missing sequence.
Taxa | Voucher information | nrITS | atpB–rbcL | ImpDEF1 | ImpDEF2 |
---|---|---|---|---|---|
Impatiens andringitrensis H.Perrier | Bot. Gard. Bonn 36655 (BONN), Madagascar | – | MH157104 | MH157123 | – |
Impatiens auricoma Baill. 1 | Bot. Gard. Bonn 34154 (BONN), Comores | – | DQ147815 | EF133562 | EF133615 |
Impatiens auricoma Baill. 2 | E. Fischer 1270 (Bot. Gard. Zürich, E.Fischer s.n.) (BONN), Comores | MH881113 | MH881068 | – | – |
Impatiens bicaudata H.Perrier 1 | E. Fischer 1340 (Bot. Gard. Bonn 36586) (BONN), Madagascar | MH881114 | MH881069 | – | MH881160 |
Impatiens bicaudata H.Perrier 2 | E. Fischer 1437 (BONN), Madagascar | MH881115 | MH881070 | MH881199 | MH881161 |
Impatiens bicaudata H.Perrier 3 | Bot. Gard. Zürich, E.Fischer s.n. (BONN), Madagascar | – | – | MH881198 | – |
Impatiens bisaccata H.Perrier 1 | E. Fischer 1271 (Bot. Gard. Bonn 36496) (BONN), Madagascar | MH157152 | MH881071 | – | – |
Impatiens bisaccata H.Perrier 2 | E. Fischer 1435 (BONN), Madagascar | MH881117 | MH881072 | MH881200 | – |
Impatiens “capuronii” Humb. ex Eb.Fisch. & Raheliv. ined. | E. Fischer 1432 (Bot. Gard. Bonn 36427) (BONN), Madagascar | MH157171 | MH157106 | MH157127 | MH157135 |
Impatiens catati H.Perrier 1 | E. Fischer 1347 (Bot. Gard. Bonn 35920) (BONN), Madagascar | MH881120 | MH881074 | – | – |
Impatiens catati H.Perrier 2 | E. Fischer 1278 (Bot. Gard. Bonn 28424) (BONN), Madagascar | MH157142 | FJ826634 | FJ826686 | – |
Impatiens cecili N.E.Br. | Knox 4353 (LV), Zimbabwe | – | FJ826635 | FJ826687 | FJ826741 |
Impatiens cf. manaharensis Baill.2 | E. Fischer 1427 (BONN), Madagascar | MH881123 | MH881078 | MH881204 | MH881166 |
Impatiens cf. manaharensis Baill. 3 | E. Fischer 1348 (Bot. Gard. Bonn 36384)(BONN), Madagascar | MH881139 | – | – | MH881182 |
Impatiens elatostemmoides H.Perrier 1 | E. Fischer 1284 (Bot. Gard. Bonn 26821) (BONN), Madagascar | MH157156 | MF567403 | – | MF567460 |
Impatiens elatostemmoides H.Perrier 2 | E. Fischer 1420 (BONN), Madagascar | MH881124 | MH881080 | MH881205 | – |
Impatiens aff. elatostemmoides H.Perrier 3 | E. Fischer 1439 (BONN), Madagascar | MH881110 | MH881065 | MH881194 | MH881156 |
Impatiens elatostemmoides H.Perrier 4 | E. Fischer 1429 (BONN), Madagascar | MH881121 | MH881076 | – | – |
Impatiens elianae S.Abrahamczyk & Eb.Fisch | E. Fischer 1326 (Bot. Gard. Bonn 36144) (BONN), Madagascar | MH157157 | MF567404 | – | MF567461 |
Impatiens eriosperma H.Perrier | E.Fischer 1342 (Bot. Gard. Bonn 35921) (BONN), Madagascar | MH157158 | MF567414 | – | MF567466 |
Impatiens furcata H.Perrier | E. Fischer 1441 (BONN), Madagascar | MH881127 | MH881083 | MH881206 | MH881170 |
Impatiens galactica Eb.Fisch., Raheliv. & S.Abrahamczyk 1 | E. Fischer 1319 (Bot. Gard. Bonn 36393) (BONN), Madagascar | MH881153 | MH881107 | – | – |
Impatiens galactica Eb.Fisch., Raheliv. & S.Abrahamczyk 2 | E. Fischer 1426 (BONN), Madagascar | MH881128 | MH881108 | MH881225 | MH881192 |
Impatiens „hammarbyoides“ Eb.Fisch. & Raheliv. 1 ined. | E. Fischer 1430 (BONN), Madagascar | MH157165 | MF567417 | MF567445 | MF567469 |
Impatiens “hammarbyoides” Eb.Fisch. & Raheliv.2 ined. | E. Fischer 1447 (Bot. Gard. Bonn 37437) (BONN), Madagascar | MH157144 | MH157099 | MH157121 | – |
Impatiens hendrikii Eb.Fisch. &Raheliv. 1 | E. Fischer 1445 (BONN), Madagascar | MH881130 | MH881086 | MH881209 | MH881173 |
Impatiens hendrikii Eb.Fisch. &Raheliv. 2 | E. Fischer 1440 (BONN), Madagascar | MH881129 | MH881085 | MH881208 | MH881172 |
Impatiens “humillima” Humb. Eb.F isch. & Raheliv. ined. | E. Fischer 1431 (BONN), Madagascar |
MH881131
|
MH881087 | MH881210 | MH881174 |
Impatiens hydrogetonoides Launert | Dessein 719 (BR), Zambia | – | FJ826648 | FJ826699 | FJ826755 |
Impatiens inaperta (H-Perr.) H.Perrier 1 | E. Fischer 1346 (Bot. Gard. Bonn 27467) (BONN), Madagascar | – | MH157109 | – | – |
Impatiens inaperta (H-Perr.) H.Perrier 2 | E. Fischer 1357 (BONN), Madagascar | MH881132 | MH881089 | – | – |
Impatiens inaperta (H-Perr.) H.Perrier 3 | E. Fischer 1448 (BONN), Madagascar | MH881133 | MH881090 | MH881213 | MH881177 |
Impatiens laurentii Eb.Fisch. & Raheliv. | E. Fischer 1293 (Bot. Gard. Bonn 36132) (BONN), Madagascar | MH157159 | – | MH157120 | – |
Impatiens lutzii Eb.Fisch. & Raheliv. 1 | E. Fischer 1318 (Bot. Gard. Bonn 36381) (BONN), Madagascar | MH881135 | MH881092 | – | MH881179 |
Impatiens lutzii Eb.Fisch. & Raheliv. 2 | E. Fischer 1438 (BONN), Madagascar | MH881136 | MH881093 | MH881214 | MH881180 |
Impatiens lyallii H.Perrier 1 | E. Fischer 1294 (Bot. Gard. Bonn 152a) (BONN), Madagascar | MH157169 | MF567420 | MF567448 | MF567471 |
Impatiens lyallii H.Perrier 2 | E. Fischer 1341 (Bot. Gard. Bonn 152b)(BONN), Madagascar | MH881138 | MH881094 | – | – |
Impatiens manaharensis Baill. 1 | E. Fischer 1434 (Bot. Gard. Bonn 36384) (BONN), Madagascar | MH881139 | MH881077 | MH881203 | MH881182 |
Impatiens mandrakae Eb.Fisch. & Raheliv. | E. Fischer 1345 (Bot. Gard. Bonn 26822) (BONN), Madagascar | MH157166 | MF567421 | – | MF567472 |
Impatiens marojejyensis Humbert & H.Perrier | E. Fischer 1444 (BONN), Madagascar | MH881141 | MH881096 | MH881215 | MH881184 |
Impatiens masoalensis H.Perrier 1 | E. Fischer 1443 (BONN), Madagascar | MH881143 | – | MH881216 | – |
Impatiens masoalensis H.Perrier 2 | E. Fischer 1424 (Bot. Gard. Bonn 36386) (BONN), Madagascar | MH157161 | MF567422 | MF567449 | MF567473 |
Impatiens masoalensis H.Perrier 3 | E. Fischer 1424 (BONN), Madagascar | MH881144 | – | MH881217 |
MH881186 |
Impatiens max-huberi Eb.Fisch. &Raheliv. | E. Fischer 1421 (Bot. Gard. Bonn 36428) (BONN), Madagascar | MH157147 | MH157110 | MH157116 | MH157137 |
Impatiens navicula Eb.Fisch. & Raheliv. 1 | E. Fischer 1422 (BONN), Madagascar | MH881147 | MH881101 | MH881220 | MH881189 |
Impatiens navicula Eb.Fisch. & Raheliv. 2 | E. Fischer 1446 (BONN), Madagascar | MH881146 | MH881100 | MH881219 | MH881188 |
Impatiens nomenyae Eb.Fisch. & Raheliv. | E. Fischer 1425 (BONN), Madagascar | MH881148 | MH881102 | MH881221 | – |
Impatiens renae Eb.Fisch. & Raheliv. | E. Fischer 1442 (BONN), Madagascar | MH881149 | MH881103 | MH881222 | – |
Impatiens rutenbergii O.Hoffm | E. Fischer 1310 (Bot. Gard. Bonn 37463) (BONN), Madagascar | MH881150 | MH881104 | – | MH881190 |
Impatiens scripta H.Perrier | E. Fischer 1423 (BONN), Madagascar | MH881151 | MH881105 | MH881223 | MH881191 |
Impatiens aff. lyallii | E. Fischer 1428 (BONN), Madagascar | MH881152 | MH881106 | MH881224 | – |
Impatiens susan-nathansoniae Eb.Fisch. & Raheliv. | E. Fischer 1433 (BONN), Madagascar | MH881155 | MH881109 | MH881226 | MH881193 |
Total genomic DNA was isolated from silica-dried leaf material using a modified CTAB protocol (
Contiguous sequences were assembled using Geneious v7.0.6 (Biomatters, New Zealand). Automatic alignment of the datasets was carried out with MAFFT (
The aligned atpB-rbcL and ImpDEF1/ImpDEF2 matrices contained 924 bp and 812 bp, respectively. The phylogenetic tree, based on the combined data, is shown in Figure
The present analyses confirm the paraphyly of the Malagasy Impatiens with respect to the Comorian I. auricoma. This is consistent with
In addition, results of this molecular phylogenetic study further highlight the difficulties that the Impatiens taxonomists have faced when dealing with the species delimitation of the Malagasy Impatiens (e.g.
Finally, the Impatiens taxa from the Marojejy National Park do not form a monophyletic group, as they are spread across the tree (Fig.
The Malagasy Impatiens are paraphyletic with respect of the Comorian I. auricoma. The present subgeneric and sectional classifications of the Malagasy Impatiens are partly supported, with strong support for the monophyly of subgen. Trimorphopetalum. Neither the Humblotianae group nor the Vulgare group forms a monophyletic group. Impatiens elatostemmoides, I. “hammarbyoides”, I. inaperta and I. manaharensis are either para- or polyphyletic and may represent morpho-species. The Impatiens species from Marojejy do not form a natural group. A further study based on a much larger molecular data set and sampling from the entire geographic ranges of Impatiens in Madagascar is needed to produce a well-resolved phylogeny. This will hopefully allow for a retest of the monophyly of sect. Preimpatiens sensu
The first author (EMR) thanks the colleagues from Institute for Integrated Sciences, Department of Biology, University of Koblenz-Landau for training and funding, the Laboratory of Plant Systematics, Institute of Botany and Microbiology, Leuven for technical and financial support of the molecular laboratory work, P. Küpfer, Y.-M.Yuan, S. Wohlhauser, M. Callmander and P. Druart (all formerly University of Neuchâtel) for introducing her to molecular phylogenetics, technical training on molecular biology and financial support at the beginning of her studies; the DGF (Direction Générale des Forêts) and the MNP (Madagascar National Parks) in Madagascar for issuing collecting and exportation permits to EMR and EF; the Parc Botanique et Zoologique de Tsimbazaza (PBZT), Antananarivo, Madagascar, for allowing her to conduct the research on Impatiens; and the two anonymous reviewers and the Associate Editor (Hanno Schaefer) for their comments on an early version of the paper. The second author (EF) is indebted to the Missouri Botanical Garden and the Parc Botanique et Zoologique de Tsimbazaza for arranging our research and collecting permists. The Akademie der Wissenschaften und Literatur Mainz kindly sponsored the field trip of EF. Special thanks go to our colleagues Damien Ertz (Meise), Bernard Goffinet (Connecticut), Roger Lala Andriamiarisoa (Antananarivo) and Emmanuël Sérusiaux (Liège) who accompanied us in the field. Without the skills of Roger Lala Andriamiarisoa, the fieldwork would have been impossible.