MOLECULAR PHYLOGENETICS AND EVOLUTION

Molecular Phylogenetics and Evolution 24 (2002) 274–301 www.academicpress.com

Phylogenetics of asterids based on 3 coding and 3 non-coding chloroplast DNA markers and the utility of non-coding DNA at higher taxonomic levels
Birgitta Bremer,a,e,* K are Bremer,a Nahid Heidari,a Per Erixon,a Richard G. Olmstead,b €,d and Edit Barkhordariana Arne A. Anderberg,c Mari K€ allersjo
Department of Systematic Botany, Evolutionary Biology Centre, Norbyv€ agen 18D, SE-752 36 Uppsala, Sweden b Department of Botany, University of Washington, P.O. Box 355325, Seattle, WA, USA c Department of Phanerogamic Botany, Swedish Museum of Natural History, P.O. Box 50007, SE-104 05 Stockholm, Sweden d Laboratory for Molecular Systematics, Swedish Museum of Natural History, P.O. Box 50007, SE-104 05 Stockholm, Sweden e The Bergius Foundation at the Royal Swedish Academy of Sciences, P.O. Box 50017, SE-104 05 Stockholm, Sweden Received 25 September 2001; received in revised form 4 February 2002
a

Abstract Asterids comprise 1/4–1/3 of all flowering plants and are classified in 10 orders and >100 families. The phylogeny of asterids is here explored with jackknife parsimony analysis of chloroplast DNA from 132 genera representing 103 families and all higher groups of asterids. Six different markers were used, three of the markers represent protein coding genes, rbcL, ndhF, and matK, and three other represent non-coding DNA; a region including trnL exons and the intron and intergenic spacers between trnT (UGU) to trnF (GAA); another region including trnV exons and intron, trnM and intergenic spacers between trnV (UAC) and atpE, and the rps16 intron. The three non-coding markers proved almost equally useful as the three coding genes in phylogenetic reconstruction at the high level of orders and families in asterids, and in relation to the number of aligned positions the non-coding markers were even more effective. Basal interrelationships among Cornales, Ericales, lamiids (new name replacing euasterids I), and campanulids (new name replacing euasterids II) are resolved with strong support. Family interrelationships are fully or almost fully resolved with medium to strong support in Cornales, Garryales, Gentianales, Solanales, Aquifoliales, Apiales, and Dipsacales. Within the three large orders Ericales, Lamiales, and Asterales, family interrelationships remain partly unclear. The analysis has contributed to reclassification of several families, e.g., Tetrameristaceae, Ebenaceae, Styracaceae, Montiniaceae, Orobanchaceae, and Scrophulariaceae (by inclusion of Pellicieraceae, Lissocarpaceae, Halesiaceae, Kaliphoraceae, Cyclocheilaceae, and Myoporaceae + Buddlejaceae, respectively), and to the placement of families that were unplaced in the APG-system, e.g., Sladeniaceae, Pentaphylacaceae, Plocospermataceae, Cardiopteridaceae, and Adoxaceae (in Ericales, Ericales, Lamiales, Aquifoliales, and Dipsacales, respectively), and Paracryphiaceae among campanulids. Several families of euasterids remain unclassified to order. Ó 2002 Elsevier Science (USA). All rights reserved.
Keywords: Asterids; Phylogeny; rbcL; ndhF; matK; trnT-F; trnV-atpE; rps16; coding DNA; non-coding DNA

1. Introduction The asterids constitute one of the major clades of the flowering plants. They represent an evolutionary successful group with over 80,000 species or 1/4–1/3 of all flowering plants. Four of the 10 largest plant families belong to this group, Asteraceae (c. 22,750 species),
*

Corresponding author. Fax: +46-8-612 90 05. E-mail address: birgitta.bremer@bergianska.se (B. Bremer).

Rubiaceae (c. 10,200 species), Lamiaceae (c. 6700 species), and Apocynaceae s.l. (c. 4800 species). They are often herbaceous plants with bisexual, insect-pollinated flowers, stamens in one circle, and sympetalous corollas. Plants with such corollas, known as Sympetalae, have been recognised as a natural group since the 18th century (Jussieu, 1789). Takhtajan (1964, 1969) renamed the group as subclass Asteridae, although he later (Takhtajan, 1987, 1997) restricted his Asteridae to the core of the order Asterales (sensu APG, 1998). Cronquist (1981)

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B. Bremer et al. / Molecular Phylogenetics and Evolution 24 (2002) 274–301

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maintained a more wide circumscription of the Asteridae, including the Asterales, Dipsacales, Gentianales, Lamiales, and Solanales as currently understood (APG, 1998). Dahlgren (1983), who stressed the importance of chemical characters for classification, placed Apiales ( ¼ Araliales) and Cornales close to Asterales and Dipsacales, respectively, and in his diagrams Ericales were surrounded by Cornales, Dipsacales, Gentianales, Lamiales, and Solanales. These placements were based on the occurrence of polyacetylenes and iridoids, which are common compounds in the Asteridae. With molecular data, particularly from the rbcL gene of the chloroplast genome, it became evident that the ‘‘core’’ Asteridae (Asterales, Dipsacales, Gentianales, Lamiales, and Solanales) are nested in a larger monophyletic group, including not only Cornales, Ericales, and Apiales but also Garryales and Aquifoliales (Chase et al., 1993; Downie and Palmer, 1992; Olmstead et al., 1992, 1993). Later analyses including more taxa and/or based on more genes, in particular ndhF, atpB, and 18S rDNA, have corroborated the first molecular analyses and generated more detailed knowledge of the group (Backlund and Bremer, 1997; Hempel et al., 1995; Gustafsson et al., 1996; Morton et al., 1996; Plunkett et al., 1996; Savolainen et al., 1994; Soltis and Soltis, 1997; Soltis et al., 1997). The results from these studies are considered in the classification by the Angiosperm Phylogeny Group (APG, 1998), which is the starting point for the present study. Subsequent analyses are also considered here and in the forthcoming revision of the APG-system (Albach et al., 2001a; Backlund et al., 2000; K arehed, 2001; Olmstead et al., 2000; Oxelman et al., 1999; APGII, in prep.). So far we know that all asterids form a strongly supported monophyletic group including 10 orders, viz. Cornales, Ericales, Garryales, Gentianales, Lamiales, Solanales, Aquifoliales, Apiales, Asterales, and Dipsacales. The last eight of these constitute the euasterids, which form two major subgroups, known as asterids I and II (Chase et al., 1993) or euasterids I (Garryales, Gentianales, Lamiales, and Solanales) and II (Apiales, Aquifoliales, Asterales, and Dipsacales) (APG, 1998). Since these names are awkward and easily confused, we here take the opportunity to rename euasterids I as lamiids and euasterids II as campanulids. More global analyses of the flowering plants (Soltis et al., 2000) have corroborated the monophyly of asterids and euasterids, and partly also the monophyly of lamiids, campanulids, and the ten APG-orders (APG, 1998, APGII, in prep.). Much has been learned from the published analyses, but many questions remain to be answered. Still there is no convincing support for the interrelationship among the three basal groups, i.e., Cornales, Ericales, and the euasterids. Different analyses, with low bootstrap or jackknife support values for the groupings, show contradictory results; rbcL/atpB/18S rDNA data (Soltis

et al., 2000) place Cornales as sister to Ericales while ndhF data alone (Olmstead et al., 2000) or ndhF together with rbcL/atpB/18S rDNA data (Albach et al., 2001a) show Cornales as sister to the rest of the asterids. Several studies indicate that lamiids and campanulids are sister taxa, although both groups have low to only medium support. Lamiids have jackknife or bootstrap values of 53/66% (Olmstead et al., 2000), 56% (Soltis et al., 2000), or 40% (Albach et al., 2001a). Campanulids have 68% (Olmstead et al., 2000), 88% (Soltis et al., 2000), or below 33% (Albach et al., 2001a). Despite all these studies based on many taxa and both three and four genes, the relationships among the orders within lamiids and campanulids, respectively, are in most parts unclear. The same applies to most family interrelationships within the orders. As noted above, most molecular studies of higherlevel (orders and families) phylogenetic interrelationships in asterids, and in flowering plants in general, are based on coding chloroplast DNA. In particular, the chloroplast genes rbcL, ndhF, and atpB have been used, but also nuclear 18S rDNA has been used. Non-coding chloroplast DNA have hitherto been utilised almost entirely for phylogenetic analyses at lower levels, and is generally taken to be phylogenetically uninformative at €hle et al., 1994; Gielly and Taberlet, higher levels (e.g., Bo 1994; Kelchner, 2000; Soltis and Soltis, 1998), since homoplasy from repeated mutations in saturated positions is assumed to swamp the phylogenetic signal. There are, however, analyses indicating that this latter assumption is erroneous, at least for silent mutations in third positions of coding chloroplast DNA (e.g., € et al., 1998; Sennblad and Bremer, 2000). K€ allersjo Kelchner (2000) discussed the potential difficulties in using non-coding DNA, since it is highly structurally constrained and not randomly evolving. He gave several evolutionary mechanisms for non-coding sequence evolution (slipped-strand mispairing, stem-loop secondary structure, minute inversions, nucleotide substitutions, intramolecular recombination) which will influence the sequences and can cause problems with alignment. Kelchner argued that all matrices should be inspected prior to phylogenetic analyses and that the different mechanisms should be considered in the alignment. As in this paper, non-coding DNA in chloroplasts is generally meant to include the non-coding single copy regions in the chloroplast DNA molecule. However, it should be noticed that from the conserved inverted repeat regions of the chloroplast DNA, it has been demonstrated that the very slowly evolving non-coding introns are informative for the basal angiosperms (Graham and Olmstead, 2000; Graham et al., 2000). Here we explore the phylogenetic utility of non-coding DNA (from the large single copy region) at the family/ order level of asterid flowering plants, a level where this non-coding DNA generally is assumed to be useless.

we selected some further interesting taxa. rbcL. have earlier been described as separate families or they have been transferred from other families. Materials and methods 2. Proboscidea. ndhF. 2000). Amplified products were cleaned with Qiaquick PCR purification kit (Qiagen). (2000). If easily available we chose a species from the type genus of the family. ndhF. Desfontainia from Columelliaceae (Savolainen et al.. and matK. Both these genera assume basal positions in the core eudicots in general. Pterostyrax of Styracaceae (Soltis et al.. Hempel et al. Paeonia of the Paeoniaceae and Vitis of the Vitaceae. Ten new rbcL sequences. on a MegaBACE 1000 capillary machine (Amersham Pharmacia Biotech). 1998). In the final analyses.. All sequenced markers for this genus are considerably different from those of the other taxa. Cardiopteris. In all data sets one or a few sequences were 2. 2001b. Lissocarpa.g. These genera. we included seven more genera since the number of families and their interrelationships within the order is very unclear. from the list of families with uncertain position in APG (1998).. Carlemanniaceae and Sphenostemonaceae.276 B. 128 new trnL sequences. (2) a region including trnV exons and intron.. 1996) and Schima of . 1995. Three represent coding genes.1. trnV-atpE. Savolainen et al. Sanango. Soltis et al. Theaceae (Morton et al. Antirrhinum. Three others represent non-coding DNA: (1) a region including trnL exons and the intron and intergenic spacers between trnT (UGU) to trnF (GAA). Peltanthera. Quintinia of Escalloniaceae (Gustafsson et al. three genera of Icacinaceae (K arehed. based on both coding and non-coding DNA. Paracryphia. Because the monophyly of the asterids already has been convincingly demonstrated (Soltis et al. we included in total 132 genera. was excluded due to difficulties in alignment.g. Sequencing We used six different DNA sequence regions from the chloroplast genome. Taxon sampling The sampling strategy was to include one member of each of the 106 asterid families from the APG-system (APG. 2.. and (3) the rps16 intron. The present study aims at presenting such a phylogeny and a refined classification of the asterids. The genus Hydrostachys (Hydrostachyaceae in APG. and Sladenia. 2. families. 2001. Appendix A) due to failure to amplify the targeted region. 2000) indicate that Hydrostachys is nested within or close to the family Hydrangeaceae and we therefore decided to exclude it from our analyses. here abbreviated trnL. Bremer et al. Androya. rps16) is phylogenetically more or less informative and useful than the commonly used coding DNA (e.. viz... In addition to the representatives of the APG-families. 31 new ndhF sequences. We also included Maesa of the family Maesaceae. where asterids constitute one of the major clades (APG. 129 new matK sequences. Montiniaceae. 2000). All PCR and sequencing primers are listed in Appendix B. a pseudogene from Orobanche. Published analyses (Albach et al. Dipentodon. The particular aims are to present: (1) supported phylogenetic interrelationships among the orders.3.. will increase support and resolution for the phylogeny of the asterids. and 125 new rps16 sequences were produced for this study.. trnT-F. From Lamiales. 1998) caused many problems. From one bigeneric family. All new sequences are listed in Appendix A. matK) at this higher taxonomic level and if such non-coding DNA. Most of the sequencing (or 538 of the in all 547 new sequences) was done in the Evolutionary Biology Centre labs in Uppsala according to the following procedure. 2000). Protocols followed that provided by the manufacturer. 2000a). Data matrices Six separate matrices were produced for the six markers. and (3) to test if noncoding DNA (e. (2) as far as possible re-circumscribe the asterid orders to include families presently unclassified to order. Kaliphora. described by Anderberg et al. rbcL. 1998). with our data. either with BigDyeTM terminator cycle sequencing kit (Applied Biosystems) and analysed on an ABI 377 (Applied Biosystems) or with DYEnamicTM ET termination cycle sequencing premix kit (Amersham Pharmacia Biotech). We obtained DNA representing 104 families and we failed to get material from two. here abbreviated rps16. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 The asterids are a biologically very diverse and speciesrich group and their evolutionary success could be explained or traced with a resolved and well supported phylogeny as a basis for further research. One rbcL sequence. Globularia. we also sequenced for the first time the genus Grevea. trnM and intergenic spacers between trnV (UAC) and atpE. In five other cases. Sequencing reactions were performed using two different protocols. 2000a.2. Olmstead et al. the phylogenetic position of Hydrostachys could not be established with any degree of certainty. there have been indications that families are non-monophyletic and hence we have included additional genera. Hence. Furthermore. They were difficult to align and the analyses resulted in phylogenies with very long branches for Hydrostachys. and Selago.. A limited number of taxa were not possible to sequence for some of the markers (cf. and informal groups. PCR reactions were performed using Taq polymerase. here abbreviated trnV. 124 new trnV sequences. we have only selected two non-asterid outgroup taxa. in combination with coding DNA. Pentaphylax. we included seven taxa. 1997).

. respectively.4. 750 autapomorphic (singletons). 20 in trnV. trnV. namely. The coding matrix comprises 5717 aligned positions of which 1878 are constant. To obtain the most comprehensive data set and the most well supported phylogeny for the asterids we merged all data into a combined matrix and analysis. but less than 50% support in the non-coding data.. 3. Six of the eight possible nodes representing interrelationships among the 10 orders are supported (>50%) by the jackknife analysis (cf.0 (beta version 4. and 2941 parsimony-informative. 1996) with 1000 replicates with 5 RANDOM stepwise additions of sequences. 2.0b8. 36 nodes show contradictions between the three analyses. for short called the coding matrix or analysis. The strict consensus tree from the combined analysis with jackknife values for the nodes is shown in Figs. In two other cases.e. for short called the non-coding matrix or analysis. 2A–C. 14 in trnL. A few rbcL sequences from EMBL/GenBank were omitted. Support values for the nodes were obtained by jackknife analysis (Farris et al. their presence/ absence was coded with a question mark. In these cases. three in ndhF. see Section 4). There are no great differences in the data from the coding versus the non-coding analyses. Solanales are supported with 90% and the other 10 orders with 100% jackknife support. were coded as additional presence/ absence characters. and matK) into one matrix. One of the trees is shown with branch lengths in Figs. 1). one non-coding marker each is missing in our data (in Eucommia trnV and in Hydrolea rps16).e. and 15 in the matK gene. Of the 130 possible nodes (the number of taxa minus two). and 16 in the rps16 sequence. Seven of the ingroup taxa represented . none in rbcL. In the combined analysis and in the coding analysis there is high support for Sphenoclea and Hydrolea as sister taxa. Phylogenetic analyses Each data matrix was analysed using PAUP* 4. Results Table 1 includes number of parsimony-informative characters. i. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 277 missing. 1A– C. and the number of nodes with P 95% jackknife support. and 1989 parsimonyinformative. Only informative characters were analysed. the different analyses support different phylogenies between Apocynaceae/Gelsemiaceae/Gentianaceae/Loganiaceae. tree lengths. If a taxon appeared in different jackknifesupported positions in the different trees. The 1989 parsimony-informative characters include 50 indel characters. i. In some taxa where alignment left doubts about the homology of indels. and rps16). and other data from the three analyses. The number of nodes with P 67% jackknife sup- port. it was taken as an indication that the sequences could be erroneous and such taxa were re-sequenced. Each separate matrix was parsimony-jackknifed (see below) to get a preliminary phylogenetic tree from each DNA marker. in a few cases also from a new DNA preparation. and we did the same for the non-coding markers (trnL.B. The 2941 parsimony-informative characters include 18 indel characters. ndhF. We did not follow Kelchner’s (2000) prealignment procedure but used a standard pragmatic alignment. Fig. Swofford. The coding genes were aligned manually by using the reading frames of the corresponding amino acid sequences. Bremer et al. particularly poly-N-sequences (streches of the same nucleotide) of different length (probably due to slipped-strand mispairing) could not be aligned. Total jackknife support in relation to the number of aligned characters was calculated as the quotient between these two numbers. In two cases the support is high in two of the analyses for a particular node not occurring in the third analysis. 898 autapomorphic (singletons). and 37% of the characters deleted in each replicate. Most of these cases concern clades with low to medium support (<95%). consistency and retention indices. the non-coding analysis. All except 17 of the 129 ingroup taxa are placed in well supported clades representing orders of the APG (1998). respectively. There are some differences in the topology of the trees from the three different analyses. 1994) followed by manual corrections. the coding analysis. and the combined analysis. After the preliminary analyses. To investigate the phylogenetic utility of coding and non-coding DNA for the taxonomic level of this study we merged the data from the coding genes (rbcL. because they turned out to be clearly erroneous following the results of our preliminary analyses.. and between Buddleja/Scrophularia/Selago. a high or strong support. The non-coding DNA sequences were first aligned by Clustal W (Thomson et al. 1998) and parsimony analyses with a heuristic search strategy with 100 replicates of RANDOM stepwise additions of sequences and TBR branch swapping. Most of these differences are within clades of few taxa and are not in conflict with family or order classification. Presumably homologous indel events (gaps). number of equally parsimonious trees. All jackknife values P 50% in the strict consensus trees were summarised as a measure of total jackknife support for the whole tree. total jackknife support. 90% and 99%. were also calculated. In the noncoding markers some regions. There is 100% support for Eucommia in Garryales in the combined analysis and the coding analysis but less than 50% support in the non-coding analysis. MULTREES off. a medium to high support. and were excluded from further phylogenetic analyses. three data sets were constructed.. The non-coding matrix comprises 4197 aligned positions of which 1458 are constant. and only one tree saved at each replicate.

2000) place Cornales as sister to Ericales while ndhF data alone (Olmstead et al. i. 1998). and it seems unnecessary to maintain two separate families for only three genera. Marcgraviaceae are sister to the rest. One genus. in prep.. and that the latter two are sister taxa. 1998) but recent analysis indicates that Curtisia is more closely related to Grubbiaceae and it is thus re-instated as a family in APGII (in prep. family interrelationships have hitherto been largely unknown or uncertain. 1998) and six are nested within the asterids. and Balsaminaceae and Tetrameristaceae are sister groups. Earlier studies have not resolved convincingly the relationships among the basal branches.274 0. is more closely related to the outgroup taxa and apparently does not belong in the asterids. Bremer et al. including unusual glandular pits on the inner surface of the sepals.99 1. shares many morphological similarities with Pelliciera (Cronquist. 1998.503 0. 2002).4 47. Ericales and euasterids.511 TJ 9831 8550 8009 TJ/na 0. still with partly unresolved interrelationships as Total jackknife support index Total jackknife support Retention index Consistency index Tree length Number of trees Percent informative of aligned Indels Table 1 Data from matrices. and Tetramerista in a single family Tetrameristaceae (also in APGII. K€ allersjo 1998) but the strongly supported monophyly of the rest of the order is new (and also found in Anderberg et al. as in this study..501 0.e.. which have also been found in analysis of chloroplast and mitochondrial genes in combination (Anderberg et al.. From the Cornales we have included four families and of these Cornaceae together with Grubbiaceae are the sister group to Hydrangeaceae and Loasaceae. Pelliciera was formerly in a family of its own (APG. 1981). Number of nodes with support P 95% nn-67 91 79 78 nn-95 64 61 41 4.. and trees Parsimonyinformative characters Aligned characters 6 Markers 3 Coding 3 Non-coding na 9914 5717 4197 nc 4930 2941 1989 ni 68 18 50 nc/na 49. but Pentamerista. Discussion We here show that Cornales are sister to the other asterids. identified in several earlier analyses (e. the sister genus of Tetramerista.91 .. The balsaminoid group is totally resolved and the relationships among its three families are strongly supported.67 1.329 RI 0. Table 2 summarizes classification of the asterids and the changes introduced. 1993. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 Number of nodes with support P 67% families of uncertain position (APG. unclassified in APG (1998). 2001a). a resolution hitherto not demonstrated with any degree of support. Xiang et al. Ericales comprise many families but except for the balsaminoid and the primuloid groups discussed below. The rbcL/atpB/18S rDNA data (Soltis et al. 2002). Hence we merge Pelliciera.7 51.).4 np 24 24 7452 steps 42203 26146 15954 CI 0. 2000) or ndhF together with rbcL/atpB/18S rDNA data place Cornales as sister to the rest of the asterids (Albach et al. and the results have been contradicting..278 B. We did not sample Curtisia in our study since it has been included in Cornaceae (APG.. Dipentodon of Dipentodontaceae. At the base Ericales are split in two strongly supported clades. The other basal clade of the Ericales comprises most of the families. the support values have been low.).g. The smaller balsaminoid group has been € et al. Here we identify a number of jackknife-supported family groups. analyses.. Pentamerista (not included in the analyses).294 0.

Cornales. 1996. Johnson et al. there are a number of well supported groups of families. Bremer et al. 1980. 1981) includes Maesaceae.g. 1987). Theophrastaceae. Actinidiaceae. Instead Ternstroemia forms a clade together with two genera of . Another group of families supported here is the ericoid group which contains six families with only weakly supported and uncertain interrelationships. In one of the first molecular analyses including Fouqueriaceae (Downie and Palmer.. as well as by Anderberg et al.). namely. The genus Lissocarpa was before molecular investigations placed close to Ebenaceae (Cronquist. and Ericaceae (Fig. e. 2000). Soltis et al. 1992) they were found to be sister taxon to Polemoniaceae. (C) Campanulids. (B) Lamiids. The enigmatic Fouquieriaceae are here supported (88%) as sister to Polemoniaceae.. 1981. The primuloid group of families (Primulales of Cronquist. 2000a). The position of Fouqueriaceae was much debated before molecular data was available.. or misplaced in Rutaceae (Savolainen et al.B. but in that study no other Ericales were included. Earlier classifications included Ternstroemia in Theaceae but this placement is not supported here. nor in other molecular investigations (Anderberg et al. It is now included in Ebenaceae (APGII. Savolainen et al. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 279 Fig. Myrsina€ ceae. (2002). 2000. 2000).. K€ allersjo et al. a position confirmed by this study. 1. Roridulaceae. 2000b. Strict consensus tree from the combined analysis of all 6 markers (coding and non-coding) with jackknife values for the nodes. 1A). The genus has recently been unplaced. However. Later studies (Johnson et al. 1983) or Violaceae (Cronquist.. Genera in bold have a new family placement and families in bold a new position compared to APG (1998). Cyrillaceae. Sarraceniaceae. indicated by the polytomy in Fig.. in prep. Takhtajan. 1999) including also Ericales taxa showed the same relationship to Polemoniaceae but with very low support (<50%). Clethraceae. 1A. close to Ericaceae (Dahlgren.. (A) Outgroups. and Ericales... 1981). as Lissocarpaceae with uncertain position by APG (1998). and Primulaceae (Anderberg et al. 2002.

2000.. Shi) showing the same Ericales relationship as our sequence. Tang and S. in Cardiopteridaceae of the campanulids. in prep. The relationship between Sladenia. without support.Q. Diapensiaceae. The former was. and Styracaceae. Sladenia and Pentaphylax. although the family is here with low support close to Symplocaceae. and Ternstroemia was also found by Anderberg et al. whereas Pentaphylax appeared in a totally different position.280 B.H.). The exact position of Theaceae within Ericales is still unclear. placed in Ternstroemiaceae. Bremer et al. 1. (2002). namely. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 Fig. APGII. (continued ) uncertain position and listed as unplaced families by APG (1998). (2000a) investigated both Sladenia and Pentaphylax. The sequence that they used may be erroneous. Pentaphylax. the latter including Halesia of the former Halesiaceae (Soltis et al. There is a close relationship between Diapensiaceae and . Savolainen et al. since there is another sequence of Pentaphylax in GenBank (AF320785 submitted by S.

Olmstead et al. The sister group relationship between Ericales and the euasterids is here highly supported (100%) and so is also that between the two branches of the euasterids.. Soltis et al. Information from the six markers used here is not enough to resolve basal relationships among the lamiids. . 1981) or in Garryales (APG. In other studies (K arehed. but such a relationship is not supported here. Soltis et al. The basal relationships of the lamiids are still partly obscure. 2000) Icacinaceae have been suggested to be included in Garryales. the published jackknife or bootstrap values have not exceeded 66% (lamiids. The latter order is strongly supported (100%) with two families only. The family Icacinaceae has in recent studies been demonstrated to be at least biphyletic (Savolainen et al.B. Garryaceae (in- cluding Aucubaceae following APGII. lamiids and campanulids. 1. 2000). 1998).. 2000) and 88% (campanulids.. Another unplaced taxon at the base of the lamiids is Oncotheca (Oncothecaceae). 2001. 2000a. respectively. in prep. The monophyly of each group has been more or less accepted however hitherto without strong support. which has been classified earlier in Theales (Cronquist. The problems involve taxa of the Icacinaceae and the Garryales. In our limited sample of genera only Pyrenacantha and Icacina are supported (100%) as a group while the relationships to Casssinopsis and Apodytes are uncertain. respectively. 2002).b. The support for these two groups together is 100% and each group is supported as monophyletic by jackknife values of 100% and 99%. Bremer et al.. but relationships of Sapotaceae and Lecythidaceae are poorly supported and their positions are still unclear. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 281 Fig. 2000) with one part related to the campanulids and with a core of genera around Icacina positioned at the base of the lamiids.) and Eucommiaceae.. Soltis et al.. (continued ) Styracaceae (also found in Anderberg et al.

Olmstead et al. Above these unresolved basal branches in the tree there is a strongly supported (100%) and taxon-rich group of lamiids. and Ericales. 2000). Bremer et al.. Vahliaceae. Bremer et al.. however. In the 4-genes analysis of rbcL/atpB/ndhF/18S rDNA (Albach et al. but it disappears with the addition of 18S rDNA data (Soltis et al. but their exact sister group relationships remain an open question. both in terms of species number and number of families. 2000.. each strongly supported as monophyletic. Considering the other four families our combined analysis show supported relationships with Apocynaceae and Gentianaceae as sister groups. The Rubiaceae. 2000). 1993. are both here and in earlier molecular and morphological investigations shown to be the sister group to the rest of the order (Backlund et al. 2000. Ever since the first molecular cladistic analyses of a comprehensive asterid set of taxa (Chase et al.. 2. Olmstead et al. and . 2000b) there is weak bootstrap support (66%) for a sister group relationship between Gentianales and Lamiales and between Solanales and Boraginaceae (60%). relationships shown also in trees from ndhF analysis (Olmstead et al. 1993) it has been clear that these taxa are closely related. One of the 24 trees from the combined analysis of all 6 markers (coding and non-coding) drawn proportional to branch lengths.282 B. (C) Campanulids. Soltis et al. (B) Lamiids.... 2000). the second largest asterid family. Here belong the three orders Gentianales. Oxelman and Bremer. 2000) there is a grade with Gentianales as sister to the rest followed by Solanales.. Gentianales comprise five families and the five representatives show totally resolved and well supported interrelationships. However.. Boraginaceae. 2001. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 Fig. 2001a) there is still no jackknife support for the interrelationships of these taxa. In the consensus tree from the 3-genes analyses of rbcL/atpB/18S rDNA (Soltis et al.. In investigations of rbcL and atpB data (Savolainen et al. and further Boraginaceae and Vahliaceae. without any jackknife support for these interrelationships. Lamiales. and unfortunately the same holds also for our combined analysis. Cornales. (A) Outgroups. 2000. and Solanales. the taxon sampling is small and our different 3-markers analyses yield partly different results compared to that of the combined analysis. and Lamiales. Genera in bold have a new family placement and families in bold a new position compared to APG (1998)..

’s (2000a) analysis all five families are included but they do not constitute a clade. 2. (2000) or Albach et al. In Savolainen et al. (2000). The close relationship between Solanaceae and Convolvulaceae has been long known and here receives 100% jackknife support. 2001a. Sphenoclea of Sphenocleaceae and Kaliphora of the former Kaliphoraceae were not included by Olmstead et al. A study with more taxa but only two genes did not resolve the interrelationships (Backlund et al. Soltis et al.. 2000). In the earlier analyses of four families Montiniaceae were sister to Hydroleaceae (Albach et al. In our tree. (continued ) these two together as sister to the pair of Gelsemiaceae and Loganiaceae. (2001a. and Loganiaceae.. Soltis et al. We here include representatives of all five Solanales families and for the first time show that they are supported as a monophyletic group (90%). 2000). with Gelsemiaceae at the base followed by Apocynaceae. Gentianaceae. Disregarding Sphenoclea and Kaliphora that were absent from these earlier analyses this relationship is congruent with our results but here we also show that Hydroleaceae are closer to Sphenocleaceae. Bremer et al.b). 2000. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 283 Fig. the two families together form the sister to the strongly supported family . Olmstead et al.B. while in the non-coding analysis Loganiaceae have shifted placed with Apocynaceae compared to the result of the coding analysis). part of which is only weakly supported... Interrelationships among these four families are different in our 3-marker analyses (in the coding analysis there is a grade. In other analyses only four of the families have been included.

2001. 1999). Joly et al. Plocospermataceae are sister group to the rest of the Lamiales followed by Oleaceae as sister to the rest (Olmstead et al. 2001). 1999).. then Tetrachondraceae as sister to the rest (Oxelman et al. with 95% support.. Oxelman et al. Scrophulariaceae are recircumscribed to include Myoporum and Buddleja and other genera of Myoporaceae and Buddlejaceae (Olmstead et al. 2. The position of the Montiniaceae in Solanales has been disputed and ontogenetic and anatomical data point more to an affinity to Escalloniaceae..) but only with low support. 2000) alone and by Albach et al. Within the order the basal branches are strongly supported as in previously published results.. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 Fig. 2000. so also in our study. Olmstead et al. Olmstead et al.. Grevea. respectively (cf. 2001a. The Lamiales currently comprises 23 families (APGII. in prep. (2000). 1999). 2001. Plantaginaceae include also Globularia and Antirrhinum.. 2001... The latter position of Gesneriaceae is also supported by ndhF data (Olmstead et al.’s (2001a. 2000).284 B. and Kaliphora (following APGII. in prep. Most molecular analyses have identified Lamiales as a large clade of asterid families (Albach et al. formerly of Globulariaceae and Scrophulariaceae.. (2000).b) 4-genes analysis but not so by the 3-genes analysis of Soltis et al. Soltis et al. while in the coding analysis Selago and Scrophularia are sister taxa. Bremer et al. 1999). (continued ) Montiniaceae including all the three genera Montinia. Oxelman et al. There is strong support for the monophyly of Plantaginaceae and Scrophulariaceae in new circumscriptions. 2000. 2001. We have not been able to trace the reason for this incongruency but a close relationship between .. with 88% support.. according to Decraene et al. and Selago are different in our 3-markers analyses (in the non-coding analysis Buddleja is sister taxon to Selago. Interrelationships among the three genera Buddleja.). Oxelman et al. and subsequently Gesneriaceae as sister to the rest of the order (here including also Peltanthera and Sanango. Scrophularia..

. Recent analyses indicate that the single genus Hydrostachys is nested in Hydrangeaceae (Soltis et al. (2000) Marcgraviaceae Myrsinaceae (Pellicieraceae—included in Tetrameristaceae) Pentaphylacaceae—transferred from the list of families of uncertain position Polemoniaceae Primulaceae Roridulaceae Sapotaceae Sarraceniaceae Sladeniaceae—transferred from the list of families of uncertain position Styracaceae—expanded to include Halesiaceae Symplocaceae Ternstroemiaceae Tetrameristaceae—expanded to include Pellicieraceae Theaceae Theophrastaceae LAMIIDS Boraginaceae (Plocospermataceae—transferred to Lamiales) Icacinaceae—transferred from the campanulids Oncothecaceae—transferred from Garryales Vahliaceae Garryales (Aucubaceae—included in Garryaceae) Eucommiaceae Garryaceae—expanded to include Aucubaceae (Oncothecaceae—transferred to the lamiids without order) Gentianales Apocynaceae Gelsemiaceae Gentianaceae Loganiaceae Rubiaceae Lamiales Acanthaceae (Avicenniaceae—included in Acanthaceae) 285 Bignoniaceae (Buddlejaceae—included in Scrophulariaceae) Byblidaceae Calceolariaceae—re-established from Scrophulariaceae by Olmstead et al. (2000a) but not included in this study (Cyclocheilaceae—included in Orobanchaceae) Gesneriaceae Lamiaceae Lentibulariaceae (Myoporaceae—included in Scrophulariaceae) Martyniaceae—re-established from synonymy of Pedaliaceae Oleaceae Orobanchaceae—expanded to include Cyclocheilaceae Paulowniaceae Pedaliaceae Phrymaceae Plantaginaceae Plocospermataceae—transferred from the lamiids without order Schlegeliaceae Scrophulariaceae—expanded to include Buddlejaceae and Myoporaceae Stilbaceae Tetrachondraceae Verbenaceae Solanales Convolvulaceae Hydroleaceae Montiniaceae—expanded to include Kaliphoraceae from the list of families of uncertain position Solanaceae Sphenocleaceae CAMPANULIDS (Adoxaceae—transferred to Dipsacales) Bruniaceae (Carlemanniaceae—transferred to Lamiales) Columelliaceae Eremosynaceae Escalloniaceae (Icacinaceae—transferred to the lamiids without order) Paracryphiaceae—transferred from the list of families of uncertain position Polyosmaceae Sphenostemonaceae—not included in this study Tribelaceae Apiales Apiaceae Araliaceae Aralidiaceae Griseliniaceae Melanophyllaceae Pennantiaceae—circumscribed by K arehed (2001) but not included in this study Pittosporaceae Torricelliaceae Aquifoliales Aquifoliaceae Cardiopteridaceae—transferred from the list of families with uncertain position Helwingiaceae Phyllonomaceae Stemonuraceae—described by K arehed (2001) but not included in this study .B. 2000) Loasaceae Ericales Actinidiaceae Balsaminaceae Clethraceae Cyrillaceae Diapensiaceae Ebenaceae—expanded to include Lissocarpaceae from the list of families of uncertain position. Bremer et al. Ericaceae Fouquieriaceae (Halesiaceae—included in Styracaceae) Lecythidaceae Maesaceae—described by Anderberg et al. (2001) but not included in this study Carlemanniaceae—transferred to Lamiales by Savolainen et al. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 Table 2 Classification of asterids following APG (1998) with commented changes ASTERIDS Cornales Cornaceae Grubbiaceae Hydrangeaceae Hydrostachyaceae—not included in this study.

Soltis et al. 2000). One clade with medium support (69%) includes Eremosynaceae. 2001. Eremosynaceae.. contains the three well defined and strongly supported (100%) orders Apiales.. 1983). Cyclocheilon. Sphenostemon is not included in our analyses and Quintinia remained in an unresolved position in Gustafsson et al. 1998) is nested within Orobanchaceae and we consequently here include it in that family. Scrophulariaceae.. as well as a number of families without order (APG. and Orobanchaceae.. Martyniaceae. Paracryphia appears as sister to Sphenostemon in Savolainen et al. Bremer et al. 2001). pers. Olmstead et al.. Albach et al. Recently.. Asterales. the support values from our combined analysis (Fig. Byblidaceae. Campanulids in this study and in most other molecular studies have been demonstrated to have a basal split between Aquifoliales and the rest of the campanulids (K arehed. The relationship between the four families of Aquifoliales is fully resolved and strongly supported. Soltis et al. 1B) are not high enough to establish interrelationships among. Polyosmaceae.g.. McDade. Stilbaceae.. 2000). The former was in APG (1998) listed as a family Paracryphiaceae with uncertain position in the system. 2000a. 1992. 2000. From a morphological point of view it seems more plausible that Phyllonomaceae and Helwingiaceae are sister taxa. Earlier studies including some of these taxa have also failed to give any clear indication of where they belong within the campanulids (Savolainen et al. 2000b. Paulowniaceae. The three ndhF sequences we have used for these taxa were from GenBank (Olmstead et al. and Phyllonomaceae (APG. 2001a. 2000). Olmstead et al. A new and strongly supported sister group relationship (99%) is that between Paracryphia and Quintinia. and Dipsacales. 1997) no molecular analyses have shown these taxa to be sister groups (e. 2000). 1998) but now shown to belong in Aquifoliales. Polyosmaceae. The relationships among these families and the three orders are in most parts still unclear. Bremer et al. Savolainen et al. 2000. Soltis et al. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 Asterales Alseuosmiaceae Argophyllaceae Asteraceae Calyceraceae Campanulaceae (Carpodetaceae—included in Rousseaceae) Donatiaceae Goodeniaceae Menyanthaceae Pentaphragmataceae Phellinaceae Rousseaceae—expanded to include Carpodetaceae Stylidiaceae Dipsacales Adoxaceae—transferred from the campanulids without order Caprifoliaceae Diervillaceae Dipsacaceae Linnaeaceae Morinaceae Valerianaceae Selago and Scrophularia is supported in a more detailed study of Selago and close relatives (Kornhall et al. Earlier investigations have indicated the same supported relationship between four of these families (Olmstead et al. Bruniaceae. Lamiaceae and Verbenaceae are here sister taxa with medium support (90%) in agreement with pre-molecular systematics (Cronquist. Dahlgren. 2000). 1998). a family formerly of uncertain position (APG. Here for the first time a totally resolved and well supported phylogeny for these seven families is shown. Here we also show that among the remaining families there is one supported group of families comprising Phrymaceae.. Acanthaceae. The major clade of the campanulids. for example. Escalloniaceae.). Wagstaff and Olmstead. Columelliaceae...286 Table 2 (continued) B. The mangrove genus Avicennia (Avicenniaceae) is sister to Acanthus of the Acanthaceae and with a more extended sampling of Acanthaceae it turns out that Avicennia is nested inside the Acanthaceae such that Avicenniaceae should be reduced to synonymy (B. Helwingiaceae. Escalloniaceae. formerly in a separate family Cyclocheilaceae (APG. This last relationship is different from what has been found by a few other studies in which Aquifoliaceae and Helwingiaceae are sister taxa (Olmstead et al. 1998) but also some former Icacinaceae genera (Soltis et al. the sister group to Aquifoliales. Lentibulariaceae. and L. The Apiales are basally split in two branches. they share the presence of epiphyllous inflorescences. namely. In the Apiales.. and Tribelaceae. K arehed (2001) has proposed that many former Icacinaceae belong in Cardiopteridaceae. 1981. Oxelman et al. In our data. and Tribelaceae. comm.. five of the six genes support a close relationship between Phyllonoma and Helwingia and only ndhF data indicate Ilex as sister to Helwingia. Cardiopteridaceae are sister to the rest with Aquifoliaceae as sister to Phyllonomaceae and Helwingiaceae together. Most other relationships between the families are unclear. 2000. one contain Aralidiaceae as sister to Melanophyllaceae and . However. we have investigated taxa representing all seven families of the APG (1998) system. 1999. The latter is a genus of Escalloniaceae.. unpublished data. 2000). (1996). Soltis et al. 2000) with strong support. after the move of several taxa from Verbenaceae to Lamiaceae (Cantino. This example of incongruency may represent a case of mix-up of sequences or misidentification and has to be investigated. Olmstead et al. it has been shown that Aquifoliales contain not only Aquifoliaceae..’s (2000a) rbcL analysis. and R. Bignoniaceae. Both clades receive 100% support.. and Lamiaceae + Verbenaceae. Orobanchaceae. Pedaliaceae..

Goodeniaceae. Remaining problems not addressed in this study are the circumscriptions and delimitations of Apiaceae. Dipsacales are expanded relative to the APG (1998) classification by inclusion of Adoxaceae (Bremer et al. the number of nodes with medium to high support ð P 67%Þ is almost the same in the non-coding analysis and in the coding analysis. however only with low support (61%). 2000a) there is bootstrap support for a sister group relationship between Calyceraceae and Goodeniaceae. the non-coding analysis is even . however. as the total jackknife support in the tree divided by the number of aligned nucleotides. rbcL. All nodes except one are strongly supported (100%). The fraction of parsimony-informative characters to aligned characters (nc/na in Table 1) is somewhat higher for the coding matrix (51. An earlier recognised (Cosner et al.99).. 91 nodes with P 67% and 64 nodes with P 95% jackknife support. 1999). 2000). 4. and Phellinaceae (K arehed et al. including 10 orders and >100 families. 2000) or rbcL and ndhF data combined (K arehed et al.. Calyceraceae. Therefore strongly supported interrelationships among Linnaea- ceae. Michaels et al. Our study has shown that at least for the asterids.67. Backlund and Pyck (1998) suggest that Morinaceae are sister to Dipsacaceae and Valerianaceae. 1993) and here strongly supported group contains Asteraceae.. e. but even the higher number is very small compared to what you may obtain in an analysis of 132 taxa. 78 and 79. Gustafsson et al. There is strong support for Menyanthaceae as sister to the other three families. 61 versus 41. 1996. and Goodeniaceae have been uncertain in earlier analyses. All earlier analyses of asterids. and Pittosporaceae (e. 2001).g. respectively) and this relationship is corroborated by our results (88%). Even if available. On the other hand. ndhF.. Araliaceae. Combining all data in the combined analysis yielded. Calyceraceae. even more well supported nodes. 1997). respectively).1.g. Viburnum representing the latter family is here with 100% support placed as sister group of the Dipsacales as circumscribed by APG (1998). 2001). The number of strongly supported nodes ð P 95%Þ is somewhat higher for the coding analysis than for non-coding analysis.... Linnaeaceae and Morinaceae are sister groups with 64% support only. and the same relationship holds for the 3-genes analysis of rbcL/atpB/18S rDNA (Soltis et al.. respectively (Table 1). atpB. Our results have Alseuosmiaceae and Phellinaceae as sister groups with medium support (87%) but in K arehed et al. Another supported (94%) group of families comprises Argophyllaceae. The number of equally parsimonious trees is considerably higher in the non-coding analysis than in the coding analysis (7452 versus 24). considerably lower (TJ/na ¼ 0. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 287 Torricelliaceae and the other branch contain Griseliniaceae as sister to the rest followed by Araliaceae as sister to Pittosporaceae and Apiaceae. the noncoding markers are almost as good as the coding markers.. The relationships among Asteraceae.4%) and the sum of all jackknife support values (TJ in Table 1) is also somewhat higher for the coding results (8550) than for the non-coding results (8009). the high support they refer to comes from a still unpublished analysis. the non-coding analysis actually scored higher than the coding analysis (TJ/na ¼ 1. including large samples of taxa. Carpodetus and Roussea are strongly supported sister taxa and classified together as Rousseaceae (Lundberg.B. respectively. Supported resolution is the goal of phylogenetic reconstruction and at least in our study the non-coding data thus proved more useful than the coding data when considered in relation to the number of aligned positions. Bremer et al. 1996. i. Plunkett and Lowry. probably due to a preconceived assumption that only coding genes are informative for studies above family level...4%) than for the non-coding matrix (47. however.’s analysis based on rbcL and ndhF data Argophyllaceae and Phellinaceae are sister groups also with medium support (78%). With ndhF data (Olmstead et al. If the strength of the results is measured in relation to the amount of input data. and the strict consensus tree was not very much collapsed. 1999). when the total jackknife support is compared to the number of aligned characters (TJ/na in Table 1). The total support in relation to the number of aligned characters was. 2001). With somewhat different sampling. 1997). Furthermore.. non-coding DNA has not been used. 1994.91 and 1. The interrelationships among these three families remain somewhat unclear. In e.. Comparison of coding and non-coding sequences Comparison between the three different analyses shows that even at this higher taxonomic level the phylogenetic utility of the non-coding markers is fully comparable to that of the coding genes. Asteraceae and Calyceraceae are sister groups (98% and 99%. All families are included in our analysis and the interrelationships are completely resolved and in agreement with the first comprehensive rbcL analysis of the order (Backlund and Bremer. However. and Menyanthaceae.. Morinaceae. They are sister to the rest of the order. have been based on coding DNA. Soltis and Soltis (1998) the taxonomic level of utility for introns and spacers is given as population to family level with a note that these markers may work in some groups within orders. Savolainen et al.e.g. Asteraceae and Calyceraceae may appear as sister groups with rbcL data alone (Gustafsson and Bremer. as expected. We have investigated 14 species representing all families of the Asterales included in the APG (1998) system. and Dipsacaceae + Valerianaceae remain to be demonstrated. and 18S rDNA. Alseuosmiaceae. With rbcL data alone (Gustafsson et al.

1998) unplaced families. Ericales. Resolution among orders within lamiids and campanulids. implying that it should be useless at higher taxonomic levels. supporting the same groups. Lissocarpaceae. Montiniaceae. Sennblad taxonomic levels (e.g. For any analysis. 2000). The three noncoding markers proved almost equally useful as the three coding genes in phylogenetic reconstruction at the high level of orders and families in asterids. Curtis and Clegg. 5. Plocospermataceae. however. and different functions. Acknowledgments We thank Jan-Eric Mattsson. one would expect saturation of multiple hits and concealment of multiple hit indels in any non-coding region. and Per Kornhall and Bengt Oxelman for the use of some unpublished primers. Our results indicate that there are no major differences in the utility of non-coding and coding sequences (given that alignment is possible). and require further investigation. . Within the three large orders Ericales. Instead. Ebenaceae.. Kaliphoraceae. Vincent Savolainen. which have been shown to be informative at higher € et al. respectively. as measured by the consistency and retention indices. respectively). Orobanchaceae. respectively). and Paracryphiaceae among campanulids. Gordon McPherson. Kelchner. and Douglas Soltis for providing plant material or DNA. Constrained DNA markers could be conserved enough to be informative at higher level. Lamiales. and Dipsacales. Ericales. Apiales. demonstrated the utility of non-coding DNA also at higher levels. there is no reason to suspect these to be more problematic for non-coding DNA than for coding regions. probably results of slipped-strand mispairing). Anderberg and in part by NSF grants (DEB-9509804 and DEB-9727025) to Richard G. 1987). Several families of euasterids.. we excluded all parts where we felt uncertain about the alignment (poly-N-sequences. However. Tetrameristaceae. Kelchner’s (2000) recommendation that non-coding data should or must be ‘‘corrected’’ by consideration of evolutionary mechanisms in order to be useful in phylogenetic analyses is an interesting approach. we conclude that non-coding DNA are just as useful without a priori corrections. decreasing its utility as a phylogenetic tool. and Adoxaceae (in Ericales. and to the placement of hitherto (APG. Our data also show that the level of homoplasy is even lower in the non-coding data. 1998. remains partly unclear. 1994. Suhua Shi. Cynthia Morton. and campanulids. Bengt Oxelman for comments on the manuscript. Bo 1984. Hence it is reasonable to assume that non-coding DNA consists of both independently and randomly evolving parts as well as more constrained parts. and contributed to ordinal classification of several families of asterids. but we do know that there are secondary structures. unclassified to order. independently of taxonomic level. e. e. Solanales.. regulating sequences. Our analysis has contributed also to reclassification of several families.g.’’ However. and Myoporaceae + Buddlejaceae. randomly and inde€hle et al. K€ are Bremer and Arne A. remain.. Halesiaceae. Cyclocheilaceae. Cardiopteridaceae. if there are structural/functional constraints one could just as well argue for the opposite. The latter may well be much more conserved and useful also for high taxonomic levels. Lamiales. Aquifoliales. one must have sufficient numbers of variable and informative characters.. Earlier it was assumed that non-coding DNA is more or less free from constraints and rapidly evolving. and Asterales. David Boufford.288 B. lamiids. that all cause constraints on the DNA (e. Garryales. Since the results from non-coding DNA are almost fully congruent with those from coding DNA. pendently (e. and Dipsacales.g. with very large data set as ours (of more than five hundred thousand bases in the noncoding matrix) this is not possible to do manually. Joel Jeremie. family interrelationships remain partly unclear. and Scrophulariaceae (by inclusion of Pellicieraceae. K€ allersjo and Bremer. We know very little about non-coding DNA evolution. James Solomon. Olmstead. We submit that there is no logical ground for a preconceived assumption that non-coding DNA is less informative at higher taxonomic level. Johannes Lundberg and Staffan Lid en for technical assistance. Gentianales.g. The allegedly randomly evolving non-coding DNA is comparable to third position data in coding DNA. Sladeniaceae. Sebsebe Demissew.. In conclusion This study has provided increased support for resolution within the asterids. Styracaceae. Peter Linder. Palmer. For possible mutational ‘‘hot spots’’ and the problem of multiple hits leading to homoplasy. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 better. and in relation to the number of aligned positions the non-coding markers were even more effective. The study was supported by Swedish Research Council grants to Birgitta Bremer. Family interrelationships have been fully or almost fully resolved with medium to strong support in Cornales. Bremer et al. HaiNing Qin. 2000). Aquifoliales. We have been able to resolve with strong support the basal interrelationships among Cornales.. Pentaphylacaceae.g. at least not for our rather high taxonomic level. Being free from constraints it was assumed that non-coding DNA comparatively rapidly attained saturation of mutations. For non-coding DNA Kelchner (2000) argued that there are structural constraints and mechanisms that will make these data less useful and he concluded ‘‘if taxonomic level is too high. especially within the campanulids.

(1992) L01885 AJ429124 AJ429370 AJ429321 AJ011982 X91760 AF130206 X98730 AJ429128 L11166 AJ429125 AJ429126 AF299087 X87379 AJ238335 AJ429379 AJ429380 AF097517 L39383 AF130210 AJ429280 Z80197 AJ429328 AF102647 AF102631 AJ429308 L36393 L11680 AJ429664 AJ431019 AJ430896 AJ429681 AJ431039 AJ430914 AJ429641 AJ430993 AJ430870 AJ429726 AJ429727 AJ431092 AJ431093 AJ430966 AJ430967 AJ429371 U58627 AJ429717 AJ429718 AJ431082 AJ431083 AJ430957 AJ430083 AJ429376 AJ429722 AJ431088 AJ430962 AJ429716 AJ429674 AJ431081 AJ431032 AJ430956 AJ430907 AF060157 X87377 AJ429378 AJ429725 AJ431091 AJ430965 L12592 AJ429327 AJ429116 U28868 AJ236238 L01882 AJ429279 AJ429391 X87398 AF027273 AJ429640 AJ429736 AJ430992 AJ431103 AJ430869 AJ430979 AJ429680 AJ431038 AJ430913 rbcL ndhF matK AJ429326 AJ429115 trnV AJ429679 rps16 AH431037 trnL AJ430912 . Citation/voucher Erixon and Bremer 44 (UPS) Harris 1931 (K) Hedr en et al. Aralidium pinnatifidum Miq.H. with sequence accession numbers and references. Viburnum rhytidophyllum Graebn. Viburnum rhytidophyllum Graebn. (2001a. Anderson Avicennia marina Vierh. Alstonia scholaris (L. Impatiens biflora Walt. Borago officinalis L.) R. Helianthus annuus L. Alstonia scholaris (L. Helianthus annuus L. Gentry Borago officinalis L. (1997) Saleh s. Alseuosmia macrophylla A. (1996) Oxelman et al. (1992) Erixon and Bremer 26 (UPS) Backlund 271 (UPS) Gustafsson et al. (1999) Sennblad and Bremer (1996) Olmstead et al.n.) Radlk. (1999) Kim and Jansen (1995) Olmstead et al.b) Albert et al. Actinidia arguta Miq. unpublished Gustafsson et al. Apium graveolens L. (2000) Manen et al. (1999) Gustafsson et al. Aralidium pinnatifidum Miq. Don Jacaranda sparrei A. Br. Anderson Acanthus montanus T.) R. Br. (2000) Erixon and Bremer 12 (UPS) Morton et al. Cunn. (1995) Bremer 3061 (UPS) Bruhl 1283 (NE) Wagstaff and Olmstead (1997) Albach et al. (1997) Erixon and Bremer 28 (UPS) Spangler and Olmstead (1999) Erixon and Bremer 11 (UPS) Olmstead and Reeves (1995) Olmstead et al. Cunn. Alseuosmia macrophylla A. Aralia spinosa L. (UPS) Roels. for new sequences voucher information is given Family Acanthaceae Acanthaceae Acanthaceae Acanthaceae Acanthaceae Acanthaceae Actinidiaceae Actinidiaceae Actinidiaceae Adoxaceae Adoxaceae Adoxaceae Alseuosmiaceae Alseuosmiaceae Alseuosmiaceae Apiaceae Apiaceae Apocynaceae Apocynaceae Apocynaceae Aquifoliaceae Aquifoliaceae Aquifoliaceae Araliaceae Araliaceae Aralidiaceae Aralidiaceae Aralidiaceae Argophyllaceae Argophyllaceae Argophyllaceae Asteraceae Asteraceae Asteraceae Balsaminaceae Balsaminaceae Balsaminaceae Bignoniaceae Bignoniaceae Boraginaceae Boraginaceae Boraginaceae Species name with author Acanthus longifolius Host Acanthus montanus T. (1999) Telford 5462 (CBG) Bergqvist 56 (S) Kanevski et al.B. Impatiens repens Moon Jacaranda mimosaefolia D. Bremer et al. Ilex sp. Cunn. (1993) Erixon and Bremer 6 (UPS) Plunkett et al. (1992) Erixon and Bremer 45 (UPS) Fanning 212 (FTG) Oxelman et al.) R. Avicennia nitida Jacq. Impatiens capensis Meerb. (KEP) Xiang and Soltis. Alseuosmia macrophylla A. (1996) Mackinder s. Viburnum rhytidophyllum Graebn. Apium graveolens L. Aralidium pinnatifidum Miq. Alstonia scholaris (L. Borago officinalis L. Ilex mitis (L. (1996) K arehed et al. Actinidia kolomikta Maxim. Argophyllum sp. Argophyllum sp.n. (1998) Erixon and Bremer 52 (UPS) Chase et al. Aralia spinosa L. Avicennia marina Vierh. Ilex crena ta Thunb. unpublished Albert et al. Br. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 289 Appendix A List of investigated taxa. Helianthus annuus L. Actinidia chinensis Planch. Argophyllum sp.

Brunia albiflora Phillips Brunia albiflora Phillips Brunia albiflora Phillips Byblis liniflora Salisb. Boopis anthemoides Juss. (1993) DeVore 1442 (OS) Erixon and Bremer 49 (UPS) Kim and Jansen (1995) Michaels et al. Acicarpha tribuloides Juss. Cornus mas L. (2002) Anderberg 7389 (S) Beier 122 (UPS) Anderberg et al.290 B. (2002) Kron and Chase (1983) Erixon and Bremer 39 (UPS) Bremer et al. Campanula ramosissima Sibth. Columellia oblonga Ruiz and Pav. Cornus mas L. (1994) rbcL AJ428894 Y10674 ndhF Missing matK AJ429309 trnV Missing rps16 AJ431020 trnL AJ430897 AJ429361 AF060159 L01891 Missing AJ429129 X87376 AJ429354 AJ429381 AJ429707 AJ431072 AJ430948 AJ429533 AJ429728 AJ431070 AJ431094 AJ430941 AJ430968 AJ430969 L39384 L13860 AJ429382 AJ430387 L39387 L13861 AJ430196 X87389 AF027274 AJ312963 AJ402936 Y08461 AJ429383 AJ238336 AJ236242 AJ429281 L12609 Y10675 AF060160 Z29670 AJ011988 AJ429362 AJ429363 AJ429355 L14400 U08918 AJ429275 AF060161 L11216 L01900 AF421051 AJ429282 AJ429283 AF421052 L12612 AJ429392 Z29672 AJ429738 AJ431105 AJ430981 AJ429527 Missing AJ430995 Missing AJ430872 AJ430873 AJ429636 AJ430988 AJ430866 AJ429708 AJ429709 AJ429702 AJ431073 AJ431074 AJ431071 AJ430949 AJ430950 AJ430942 AJ429526 AJ430994 AJ430871 AJ429535 Missing AJ430971 AJ429310 AJ429665 AJ431021 AJ430898 AJ429737 AJ431104 AJ430980 AJ429729 AJ430391 AJ431095 AJ430970 . unpublished Xiang et al. Desfontainia spinosa Ruiz and Pav. Boopis anthemoides Juss. Acicarpha tribuloides Juss. (1993) Backlund 267 (UPS) Gustafsson et al. Citation/voucher Colwell CAP76P-2 (MO) Backlund and Bremer (1997) Gustafsson 239 (UPS) Roels. Boopis graminea Phil. (1992) Qiu 95128 (IND) Gustafsson 207 (UPS) Gustafsson et al. (UPS) K arehed et al. (1993) Albert et al. (2001a. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 Appendix A (continued ) Family Boraginaceae Bruniaceae Bruniaceae Bruniaceae Byblidaceae Byblidaceae Calyceraceae Calyceraceae Calyceraceae Calyceraceae Calyceraceae Calyceraceae Campanulaceae Campanulaceae Campanulaceae Caprifoliaceae Caprifoliaceae Caprifoliaceae Cardiopteridaceae Cardiopteridaceae Carpodetaceae Carpodetaceae Carpodetaceae Clethraceae Clethraceae Clethraceae Columelliaceae Columelliaceae Columelliaceae Columelliaceae Columelliaceae Columelliaceae Convolvulaceae Convolvulaceae Convolvulaceae Cornaceae Cornaceae Cornaceae Cyrillaceae Cyrillaceae Cyrillaceae Diapensiaceae Diapensiaceae Diapensiaceae Diervillaceae Diervillaceae Species name with author Pholisma arenarium Nutt. (1993) Olmstead et al. Cardiopteris quinqueloba Hassk. (1996) Oxelman et al. Carpodetus serratus Forst. Ipomoea coccinea L. Ipomoea coccinea L. Columellia oblonga Ruiz and Pav. (1993) Bremer 3318 (UPS) Roels. (1999) Meebold 16830 (S) Savolainen et al. (1999) Albach et al. (2000a) Gustafsson and Bremer (1997) Cameron s. (1999) Bremer et al. Bremer et al. Lonicera orientalis Lam. Campanula elatines L. unpublished Albert et al. unpublished Bremer et al. Lonicera orientalis Lam. and Sm. (1992) Anderberg et al. Carpodetus serratus Forst. Boopis anthemoides Juss. Cardiopteris quinqueloba Hassk. Diervilla rivularis Gatt. Byblis liniflora Salisb.b) Erixon and Bremer 5 (UPS) Kron and Chase (1983) Backlund and Bremer (1997) Roels. Clethra alnifolia L. Desfontainia spinosa Ruiz and Pav. Desfontainia spinosa Ruiz and Pav. Cyrilla racemiflora L. Diapensia lapponica L. Cyrilla racemiflora L. Clethra alnifolia L. Diapensia lapponica L.n. Campanula ramosissima Sibth. Ipomoea batatas (L. and Sm. Carpodetus serratus Forst. Diapensia lapponica L. (1996) Hunziker 25258 (CORD) Kim and Jansen (1995) Michaels et al. Columellia oblonga Ruiz and Pav. Cyrilla racemiflora L. Lonicera orientalis Lam. Cornus mas L. Clethra alnifolia L. Diervilla sessilifolia Buckl. 3374 (UPS) Bremer 2739 (UPS) Erixon and Bremer 18 (UPS) Olmstead et al. (1994) Oxelman et al.) Lam.

Garrya elliptica Dougl. (1993) Struwe 1009 (UPS) Oxelman et al. Sanago sp. Garrya elliptica Dougl. Perrier Androya decaryi H. Aucuba japonica Thunb. Gelsemium sempervirens Ait. Fouquieria splendens Engelm. unpublished Xiang et al. Erica carnea L. Fouquieria splendens Engelm. Johnst. ex Lindl. Escallonia x langleyensis Vilm. (2000) Annels and Hearn 4795 (UWA) Hibsch-Jetter et al.b) Olmstead et al. 3352 (UPS) Oxelman et al. Backlund. (1993) Olmstead et al.. Sanago sp. (1993) Albert et al. Bremer et al. Muell Quintinia verdonii F. Peltanthera floribunda Benth. (2000) Gustafsson et al. (1999) Philipson et al. (1992) Olmstead 97-11 (WTU) Erixon and Bremer 25 (UPS) Albach et al. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 291 Appendix A (continued ) Family Diervillaceae Dipentodontaceae Dipsacaceae Dipsacaceae Dipsacaceae Donatiaceae Donatiaceae Donatiaceae Ebenaceae Ebenaceae Ebenaceae Eremosynaceae Eremosynaceae Ericaceae Ericaceae Escalloniaceae Escalloniaceae Escalloniaceae Escalloniaceae Escalloniaceae Escalloniaceae Eucommiaceae Eucommiaceae Eucommiaceae Fouquieriaceae Fouquieriaceae Fouquieriaceae Garryaceae Garryaceae Garryaceae Garryaceae Garryaceae Garryaceae Gelsemiaceae Gelsemiaceae Gelsemiaceae Gentianaceae Gentianaceae Gentianaceae Gesneriaceae Gesneriaceae Gesneriaceae Gesneriaceae Gesneriaceae Gesneriaceae Gesneriaceae Gesneriaceae Species name with author Diervilla sessilifolia Buckl. Donatia fascicularis Forst. Eremosyne pectinata Endl. Diospyros kaki Thunb.. (2000) Rancho Santa Ana Bot. Donatia fascicularis Forst. ex Lindl. (1998) Swenson 301 (UPS) Erixon and Bremer 19 (UPS) Morton et al. Sanago sp. Gelsemium sempervirens Ait. (1999) Bremer et al. (1997) Olmstead et al.M. Streptocarpus caulescens Vatke Citation/voucher Roels. Diospyros kaki Thunb. Diospyros texana Scheele Eremosyne pectinata Endl. (2001a.B. Erica australis L. Muell Eucommia ulmoides Oliver Eucommia ulmoides Oliver Eucommia ulmoides Oliver Fouquieria digueti I. Muell Quintinia verdonii F. 11829. unpublished Kron and Chase (1983) Erixon and Bremer 32 (UPS) Morgan and Soltis (1993) A. no voucher Bremer 3026 (UPS) Oxelman et al. Perrier Peltanthera floribunda Benth. (1993) Olmstead and Reeves (1995) Olmstead et al. ex Lindl. (1992) Olmstead et al. Garrya elliptica Dougl. 3747 (MO) Hammel 19855 (MO) Oxelman et al. (1996) Olmstead 97-141 (WTU) Albert et al. (1999) Erixon and Bremer 35 (UPS) rbcL ndhF AF060164 AJ429102 matK trnV rps16 trnL AJ428890 AJ429273 AJ429393 AJ429634 AJ429739 AJ430986 AJ431106 AJ430865 AJ430982 L13864 AF130190 X87385 AJ225074 AJ429384 AJ430197 Z80185 AF130213 AJ429120 L47969 L12617 AJ429105 L11183 AJ429365 AF060165 AJ429366 AJ238344 X87394 AJ429113 L01917 AJ429317 AJ429285 AJ236249 L11675 AJ429318 AF060158 L11210 L01919 AF147714 AJ429319 AJ429322 AJ011984 L14397 L36400 L14398 AJ429323 AJ001756 AF027276 AJ429329 AJ429330 AJ001762 AF027281 Missing AJ001763 AF027283 AJ429331 AJ429684 AJ431043 AJ430918 AJ429683 AJ431042 AJ430917 AJ429530 AJ429682 AJ431040 AJ431041 AJ430915 AJ430916 AJ429676 AJ431034 AJ430909 AJ429529 AJ429675 AJ431030 AJ431033 Missing AJ430908 AJ429672 AJ431029 AJ430906 Missing AJ429643 AJ431028 AJ430998 AJ430905 AJ430876 AJ429712 AJ431077 AJ430953 AJ429711 AJ431076 AJ430952 AJ429284 Missing AJ430997 AJ430875 AJ429364 AJ429710 AJ431075 AJ430951 Missing AJ429642 AJ431096 AJ430996 AJ430972 AJ430874 . and Bois Quintinia verdonii F. Gard. Escallonia coquimbensis Remy in Gay Escallonia rubra Pers. Gelsemium sempervirens Ait. no voucher Roels. (1996) Laurent et al. (1999) Oxelman et al. Androya decaryi H. (1999) Gustafsson et al. Aucuba japonica Thunb. (1999) Olmstead et al. Dipentodon sinicus Dunn Dipsacus sativus Garsault Dipsacus sativus Garsault Dipsacus sativus Garsault Donatia fascicularis Forst. Aucuba japonica Thunb. unpublished Tsai 58398 (A) Erixon and Bremer 46 (UPS) Michaels et al. unpublished Telford 3244 (CBG) K arehed et al. Gentiana procera Holm Gentiana procera Holm Gentiana purpurea L. (1992) Erixon and Bremer 30 (UPS) Roels.

Cassinopsis ilicifolia (Hochst. Pinguicula sp. (1993) Erixon and Bremer 4 (UPS) Morgan and Soltis (1993) Olmstead et al.-Ham. Dietrich Helwingia japonica (Thunb. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 Appendix A (continued ) Family Gesneriaceae Gesneriaceae Goodeniaceae Goodeniaceae Goodeniaceae Griseliniaceae Griseliniaceae Griseliniaceae Griseliniaceae Grubbiaceae Grubbiaceae Helwingiaceae Helwingiaceae Helwingiaceae Hydrangeaceae Hydrangeaceae Hydrangeaceae Hydroleaceae Hydroleaceae Hydroleaceae Icacinaceae Icacinaceae Icacinaceae Icacinaceae Icacinaceae Icacinaceae Icacinaceae Lamiaceae Lamiaceae Lamiaceae Lecythidaceae Lecythidaceae Lecythidaceae Lentibulariaceae Lentibulariaceae Linnaeaceae Linnaeaceae Linnaeaceae Species name with author Streptocarpus holstii Engl. Lamium purpureum L. Dietrich Helwingia japonica (Thunb. (2000a. and G.) Kurz Barringtonia asiatica (L.) Kurz Pinguicula caerulea Walt. (1992) Erixon and Bremer 54 (UPS) A.) Ser. Bremer et al. Forst Griselinia lucida (J.b) Erixon and Bremer 15 (UPS) Kaufmann and Wink (1996) Wagstaff and Olmstead (1997) Anderberg et al. (1993) Lundberg 55 (UPS) Tibell NZ3 (UPS) Olmstead et al. ex Murray) F. ex Bernh. ex D. Citation/voucher Olmstead and Reeves (1995) Olmstead et al. (1997) Z80174 Albert et al. Forst) G. Krause Scaevola sp. Krause Scaevola frutescens K. unpublished L01942 Missing AJ429333 AJ429394 AJ429686 AJ429740 AJ431045 AJ431107 AJ430920 AJ430983 AJ428899 AF060166 . Lamium album L. Forst) G. ex Murray) F. Don Hydrangea macrophylla (Thunb.R. ex Murray) F. (UPS) Bremer 3770 (UPS) Bremer 3769 (UPS) Jongkind 2012 (UPS) Pettersson 3026 (UPS) Bremer 3795 (UPS) K arehed (2001) Savolainen et al. Backlund.n. (1996) Olmstead et al. Scaevola frutescens K.R. (2000) Peng et al. Forst. Grubbia tomentosa (Thunb. Pyrenacantha malvaefolia Engl. Forst. Streptocarpus holstii Engl. Hydrolea ovata Nutt. Barringtionia asiatica (L. Forst) G. (1993) Chase 5704 (K) Morton et al. Hydrolea ovata Nutt. Dietrich Hydrangea aspera Buch. Lamium purpureum L. Linnaea borealis L. Forst Grubbia rosmarinifolia Berg.) Harms Helwingia japonica (Thunb. (1993) Kim and Jansen (1995) Michaels et al. (WTU) Roels. Pyrenacantha grandifolia Engl. Icacina senegalensis Juss.) Ser. Forst Griselinia lucida (J.) Kurz Barringtonia asiatica (L. (2002) AJ402995 AJ429332 Z37403 U78694 AF421044 AJ429286 AJ429644 AJ430999 AJ430877 AJ429685 AJ431044 AJ430919 L11187 AF130218 AF013999 L14293 AJ428895 AJ428896 AJ429109 AJ429110 AJ429111 AJ428897 AJ429313 AJ429314 AJ312952 AJ429668 AJ429669 AJ431024 AJ431025 AJ430901 AJ430902 AJ429356 AJ429311 AJ429312 AJ429703 AJ429666 AJ429667 Missing AJ431022 AJ431023 AJ430943 AJ430899 AJ430900 L11226 AJ429277 AJ429638 AJ430990 Missing L11225 AJ429104 Z83141 AF130207 AJ430195 AJ429723 AJ431089 AJ430963 AJ429276 AJ429637 AJ430989 AJ430867 AF130205 U58628 rbcL ndhF L36415 L14409 L39385 L13932 AJ429385 AJ429372 AJ429730 AJ429719 AJ431097 AJ431084 AJ430973 AJ430958 matK trnV rps16 trnL Chung and Anderberg 1417 (HAST) Morton et al. Linnaea borealis L. and G.) Sleumer Icacina senegalensis Juss. (2000) Ferguson (1998) Olmstead et al. (1997) Xiang et al.R. Apodytes dimidiata E. 17408 (S) Xiang et al. no voucher Olmstead s. Hydrangea macrophylla (Thunb. Linnaea borealis L. Hydrolea ovata Nutt. Mey. (1993) Torrey s. Griselinia littoralis (Raoul) Raoul Griselinia lucida (J. Pyrenacantha malvaefolia Engl. Forst.n. (2000) Plunkett et al. and G.292 B.

(2001a. Menyanthes trifoliata L. Montinia caryophyllacea Thunb. Loasa vulcanica Andr e Logania sp. Maesa myrsinoides Leveille Maesa tene ra Mez Maesa tene ra Mez Marcgravia rectiflora Triana and Planch. f Kaliphora madagascariensis Hook. (1992) Albach et al. Olea europaea L. Bonpl. Muell.b) Morton et al.b) Albert et al.b) Plunkett et al. (S) Erixon and Bremer 41 (UPS) Albach et al. Citation/voucher Anderberg et al. 10282 (UPS) Kim and Jansen (1995) No voucher Olmstead et al.) Thell. (2000) Backlund and Bremer (1997) Erixon and Bremer 38 (UPS) Anderberg et al.) Decne. (1998) € et al. s. Myrsine africana L. Bremer et al. (2000a) Schatz et al. Melanophylla alnifolia Baker Melanophylla alnifolia Baker Melanophylla sp. Proboscidea louisianica (Mill.) Thell. 8364 (UPS) Wagstaff and Olmstead (1997) Thulin 8079 (UPS) Y10706 AJ429130 U96652 AF213751 AJ429290 AJ429335 AJ001766 AF027288 AJ429114 AJ429647 AJ429687 AJ431002 AJ431047 AJ430880 AJ430922 AJ429395 AJ429741 AJ431108 AJ430984 L11194 AF130178 L01946 AJ236244 U50254 AJ429373 L39388 AJ429386 L14006 AJ428898 AJ402963 AJ430426 AJ429357 AJ429731 AJ429704 AJ431098 AJ431066 AJ430974 AJ430944 AJ430390 AJ431085 AJ430959 U17876 AJ429278 AF060167 AJ235837 Z68826 AJ429324 Z80203 AJ429288 AF213750 AJ236263 Z83148 AJ429289 AJ429334 AJ236267 AJ429646 AJ430388 AJ431001 AJ431046 AJ430879 AJ430921 Missing AJ431000 AJ430878 AJ429677 AJ431035 AJ430910 AJ429639 AJ430991 AJ430868 rbcL AF421094 ndhF AF421062 AJ429287 AJ429645 Missing Missing matK trnV rps16 trnL AJ431206 AJ429358 AJ429359 AJ429705 AJ429706 AJ431067 AJ431068 AJ430945 AJ430946 AJ429320 AJ429673 AJ431031 AJ430529. Montinia caryophyllacea Thunb. Morina coulteriana Royle Morina longifolia Wall. Logania vaginalis F. (2001a. Myrsine africana L. Cyclocheilon somaliense Oliver Cyclocheilon somaliense Oliver Lindenbergia sp. (1996) Anderberg s. Olea europaea L. Myrsine africana L. unpublished Backlund et al. (S) € et al. (2000a. (1995) Erixon and Bremer 42 (UPS) Roels. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 293 Appendix A (continued ) Family Lissocarpaceae Lissocarpaceae Loasaceae Loasaceae Loasaceae Loganiaceae Loganiaceae Loganiaceae Maesaceae Maesaceae Maesaceae Marcgraviaceae Marcgraviaceae Marcgraviaceae Martyniaceae Martyniaceae Martyniaceae Melanophyllaceae Melanophyllaceae Melanophyllaceae Menyanthaceae Menyanthaceae Menyanthaceae Montiniaceae Montiniaceae Montiniaceae Montiniaceae Montiniaceae Montiniaceae Montiniaceae Morinaceae Morinaceae Myrsinaceae Myrsinaceae Myrsinaceae Oleaceae Oleaceae Oncothecaceae Oncothecaceae Orobanchaceae Orobanchaceae Orobanchaceae Species name with author Lissocarpa guianensis Gleason Lissocarpa guianensis Gleason Loasa loxensis Humb. Menyanthes trifoliata L. (1997) St ahl s. AJ430530 AJ430923 AJ131950 AJ429117 U28871 AJ429337 AJ429689 AJ431049 AJ430924 AJ429336 AJ429688 AJ431048 . Marcgravia rectiflora Triana and Planch. Oncotheca balansae Baill.B. (UPS) Savolainen et al. f Montinia caryophyllacea Thunb. Grevea sp. (1999) Jaffre 3238 (NOU) Savolainen et al.n. f Kaliphora madagascariensis Hook. (2000) Bremer 1996 Bremer 3013 (UPS) Morton et al. (2001a. (2000) K€ allersjo Erixon and Bremer 17 (UPS) Erixon and Bremer 34 (UPS) Oxelman et al. Proboscidea louisianica (Mill. Muell. and Kunth Loasa triphylla Juss.b) Thulin et al. Marcgravia sp. (2002) Maas and Westra 3999 (S) Hempel et al.n. Kaliphora madagascariensis Hook. 4081-B81 (UPS) Bremer 3521 (UPS) Morgan and Soltis (1993) Olmstead et al.n. 3568 (MO) Bremer et al. (1996) Thulin et al. Menyanthes trifoliata L. Logania vaginalis F. Oncotheca balansae Baill. (2000) K€ allersjo Albach et al. (1993) Thulin et al. Proboscidea fragrans (Lindl.

Olmstead et al. Bremer 3865 (UPS) Plantaginaceae Globularia cordifolia L. (2000) ex Ait. (1999) Phellinaceae Phelline lucida Vieill. Phellinaceae Phelline comosa Labill. (1996) ex Ait. Olmstead et al. (2000a. ceae Poulsen (UPS) Pentaphylacaceae Pentaphylax euryoides Tang 20001210 (UPS) Gardn. Steenis Paracryphiaceae Paracryphia alticola (Schltr. Wolfe and dePamphilis (1997) Paeoniaceae Paeonia anomala L. Polemoniaceae Polemonium caeruleum L. Wagstaff and Olmstead (1997) Phyllonomaceae Phyllonoma laticuspis Engl. Plocospermataceae Plocosperma buxifolium Oxelman et al.b) Paeoniaceae Paeonia veitchii Lynch Bremer and Bremer 4095 (UPS) Paracryphiaceae Paracryphia alticola (Schltr. Oxelman et al. Erixon and Bremer 43 (UPS) PentaphragmataPentaphragma ellipticum Cosner et al. (1996) Benth. Anderberg et al. (1999) Phellinaceae Phelline lucida Vieill. Pittosporaceae Pittosporum undulatum Vent. (1999) Erixon and Bremer 47 (UPS) Orobanchaceae Orobanche ramosa L. Pittosporaceae Pittosporum tobira Dryand. Plocospermataceae Plocosperma buxifolium Salinas 8050 (MEXU) Benth. Olmstead et al. (2000a. Plunkett et al. (1995) Pedaliaceae Sesamum indicum L.) McPherson 1999 (MO) v. (1999) Benth.) Steud. Olmstead and Reeves (1995) Plantaginaceae Antirrhinum majus L. K arehed et al. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 Appendix A (continued ) Family Orobanchaceae Orobanchaceae Species name with author Lindenbergia sp. (1994) ceae Poulsen PentaphragmataPentaphragma ellipticum Ex Singapore Bot. Pittosporaceae Pittosporum tobira Dryand. and Champ. Asplund 10683 (UPS) ex Schult. Olmstead et al. (1999) Plantaginaceae Plantago argentea Chaix Erixon and Bremer 14 (UPS) Plantaginaceae Plantago lanceolata L. Savolainen et al.) Steud. Erixon and Bremer 10 (UPS) Plantaginaceae Antirrhinum majus L. (1992) Plantaginaceae Globularia cordifolia L. Olmstead and Reeves (1995) Pedaliaceae Sesamum indicum L.294 B. Cantino 1376 (BHO) Phrymaceae Phryma leptostachya L. (2000) Paeoniaceae Paeonia suffruticosa Andr. Erixon and Bremer 36 (UPS) Plantaginaceae Antirrhinum majus L. Morgan and Soltis (1993) Phyllonomaceae Phyllonoma laticuspis Engl. (2000) Phyllonomaceae Phyllonoma ruscifolia Willd. (UPS) Paulowniaceae Paulownia tomentosa Olmstead and Reeves (Thunb. Steenis Paulowniaceae Paulownia tomentosa Erixon and Bremer 22 (Thunb. van Balgooy 7034 (L) Phrymaceae Phryma leptostachya L. Olmstead et al. ex Baill.b) v. (1993) Pedaliaceae Sesamum orientale L. Orobanche hederae Duby Citation/voucher rbcL AJ001768 Missing U73971pseudogene AF130223 AJ402982 AJ430198 AJ429121 AJ402983 AJ429339 L36447 L36406 L36413 L14408 AJ429340 L18794 AJ429387 AJ428891 AJ429106 AJ238342 AJ238347 AJ429118 U28881 L11201 AF130208 AJ429377 AF130201 U50261 AJ429374 AJ429342 L36392 L11688 AJ429343 AJ001764 AF027282 AJ429344 L36454 Z68829 AJ011985 AJ429315 AF421070 AJ429670 AJ431026 AJ430903 L36408 AJ429695 AJ431056 AJ430931 AJ429694 AJ431055 AJ430930 AJ429720 AJ429693 AJ431086 AJ431054 AJ430960 AJ430929 AJ429724 AJ431090 AJ430964 AJ429388 AJ429341 AJ429733 AJ429692 AJ431100 AJ431053 AJ430976 AJ430928 AJ429291 AJ429732 AJ429648 AJ431099 AJ431003 AJ430975 AJ430881 AJ429691 AJ431052 AJ430927 AJ429690 AJ431051 AJ430926 AJ429367 AJ430199 AJ429713 AJ430201 AJ431078 AJ430200 AJ430392 ndhF AF027286 Missing matK trnV rps16 trnL Oxelman et al. Gard. (2002) AJ429338 Missing AJ431050 AJ430925 . K arehed et al. ex Baill. Bremer et al. Olmstead and Reeves (1995) Plocospermataceae Plocosperma buxifolium Endress et al.) Savolainen et al.

(1999) Myoporum mauritianum A. Roridula gorgonias Planch.b) Albert et al. (1992) Anderberg et al.b) Sladenia celastrifolia Kurz Anderberg et al. Bremer (1996) L36403 AJ429347 AJ429348 AJ429698 AJ429699 AJ431059 AJ431060 AJ430934 AJ430935 L36450 L36412 AJ429349 AJ429531 AJ431061 AJ430936 L36449 L36411 AJ429297 AJ429654 AJ431009 AJ430081 AJ403004 AF421081 L14953 Z00044 L18798 AJ429119 Z00044 Z00044 Z00044 Z00044 AJ429360 AJ429534 AJ431069 AJ430947 AF027287 AJ429350 Z68827 AJ429532 AJ431062 AJ430937 . Roridula gorgonias Planch. Bremer 3927 (UPS) Selago thomsoni Rolfe Bremer 3095 (UPS) ex Oliver Selago thunbergii Choisy Olmstead and Reeves (1995) Scrophularia arguta Sol. Sphenoclea zeylanica Madsen 5986 (S) Gaertn. Olmstead and Reeves (1995) Sladenia celastrifolia Kurz Bartholomen et al. Morr Primula veris L. 1636 (A) Sladenia celastrifolia Kurz Savolainen et al. (1994) Gaertn. Shinozaki et al. (1995) Myoporum montanum R.n. (2000a. Buddleja davidii Franch. Luculia grandifolia Ghose Luculia gratissima Sweet Luculia gratissima Sweet Manilkara zapota (L. Roussea simplex Sm. (1998) Erixon and Bremer 33 (UPS) Albach et al. Mey.) van Royen Sarracenia flava L.) Monach. Oxelman et al. (1999) Stilbe ericoides L.) van Royen Manilkara zapota (L. Schlegelia parviflora (Oerst. Thulin and Gifri 8633 ex Ait.B. Bremer 3731 (UPS) Stilbe vestita Berg. Sphenoclea zeylanica Thulin et al. (2001a. Polyosma cunninghamii Benn. Bremer et al. no voucher From Soltis (MAU) Koontz and Soltis (1999) Lundberg (2001) Bremer et al. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 295 Appendix A (continued ) Family Polemoniaceae Polemoniaceae Polyosmaceae Polyosmaceae Polyosmaceae Primulaceae Primulaceae Primulaceae Roridulaceae Roridulaceae Roridulaceae Rousseaceae Rousseaceae Rousseaceae Rubiaceae Rubiaceae Rubiaceae Sapotaceae Sapotaceae Sapotaceae Sarraceniaceae Sarraceniaceae Schlegeliaceae Schlegeliaceae Scrophulariaceae Scrophulariaceae Scrophulariaceae Scrophulariaceae Scrophulariaceae Scrophulariaceae Scrophulariaceae Scrophulariaceae Scrophulariaceae Sladeniaceae Sladeniaceae Sladeniaceae Solanaceae Solanaceae Sphenocleaceae Sphenocleaceae Sphenocleaceae Stilbaceae Stilbaceae Stilbaceae Species name with author Polemonium pulcherrimum Hook. Oxelman et al. (2000a.b) € et al. Br. (2000) K€ allersjo Anderberg et al. Olmstead and Reeves DC. (UPS) Gaertn. no voucher Oxelman et al. Schlegelia parviflora (Oerst. (1992) Linder. (1992) Erixon and Bremer 21 (UPS) Gentry 14221 (MO) AJ402992 AF213757 U96657 AJ429293 AJ236270 L01950 AJ429294 AJ429389 AF084477 AJ277384 X83648 AJ429325 AJ011987 L01932 AF213732 AJ429295 L01952 Missing AJ429296 AJ429345 L36448 L36410 AJ429346 L36394 AJ001757 L36445 AJ429697 AJ431058 AJ430933 AJ429653 AJ429696 AJ431008 AJ431057 AJ430886 AJ430932 AJ429652 AJ431007 AJ430885 AJ429678 AJ431036 AJ430911 AJ429651 AJ429734 AJ431006 AJ431101 AJ430884 AJ430977 AJ429650 AJ431005 AJ430883 L11687 AJ429122 AJ429368 AJ429714 AJ431079 AJ430954 rbcL ndhF matK AJ429292 trnV AJ429649 rps16 AJ431004 trnL AJ430882 Olmstead and Reeves (1995) Bremer 3500 (UPS) Olmstead and Reeves (1995) Buddleja davidii Franch. Morr Primula sieboldi E. (1992) Bremer 3941 (UPS) Cejie 22179 (UNSW) Savolainen et al.) Monach.) van Royen Manilkara zapota (L. Polemonium reptans L. s. Citation/voucher Erixon and Bremer 8 (UPS) Olmstead et al. Olmstead et al. (UPS) Scrophularia sp. Roussea simplex Sm. cult. Polyosma cunninghamii Benn. (1995) CONN 870064. Primula sieboldi E.. (2002) Erixon and Bremer 23 (UPS) Albert et al. (1986) Sphenoclea zeylanica Cosner et al. (2002) Nicotiana tabacum L. (1999) Albert et al. Sarracenia purpurea L. Roridula gorgonias Planch. (1993) Nicotiana tabacum L. Buddleja asiatica Lour. Roussea simplex Sm. Polyosma cunninghamii Benn. Stilbe albiflora E.

and Champ. (2002) Harling et al. (1999) Anderberg et al. Stylidium calcaratum R. (1999) Struwe 1000 (UPS) Wagstaff and Olmstead (1997) Martinsson and Swenson 314 (UPS) Oxelman et al. Tribeles australis Phil.-L. 586 (K) Coode 7925 (K) Morton et al. Schima superba Gardn. (2000) K€ allersjo Xiang and Soltis. and Champ. Styrax officinale L. Pelliciera rhizophoreae Planch. Schima superba Gardn. Halesia carolina L. Torricellia tiliaefolia DC. (1997) Oxelman et al. and Champ. Ternstroemia gymnanthera Sprague Ternstroemia gymnanthera Sprague Ternstroemia stahlii Krug and Urb. Styrax americanum Lam. Theophrasta americana L.n. (1998) Erixon and Bremer 24 (UPS) € et al. (1997) Erixon and Bremer 20 (UPS) Anderberg et al. Vahlia capensis Thunb. Valeriana fauriei Briquet Valeriana hirtella Kunth Citation/voucher Bremer and Gustafsson 44 (UPS) Laurent et al. (2000) Lundqvist 15364 (S) Anderberg et al.b) Bremer 3785 (UPS) Morgan and Soltis (1993) Bremer 3316 (UPS) Olmstead et al. Halesia carolina L. Symplocos bogotensis Brand Symplocos bogotensis Brand Symplocos costata Choisy ex Zoll. Tetrachondra patagonica Skottsb. (2000) Kron and Chase (1983) Erixon and Bremer 40 (UPS) Anderberg et al. (2002) Morton et al. Tetramerista sp. (2000) Erixon and Bremer 37 (UPS) Kron and Chase (1983) Olmstead et al. Theophrasta americana L. Tetrachondra hamiltonii Petrie Tetrachondra patagonica Skottsb. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 Appendix A (continued ) Family Stylidiaceae Stylidiaceae Styracaceae Styracaceae Styracaceae Styracaceae Styracaceae Styracaceae Styracaceae Symplocaceae Symplocaceae Symplocaceae Ternstroemiaceae Ternstroemiaceae Ternstroemiaceae Tetrachondraceae Tetrachondraceae Tetrachondraceae Tetrachondraceae Tetrachondraceae Tetrameristaceae Tetrameristaceae Tetrameristaceae Tetrameristaceae Theaceae Theaceae Theaceae Theaceae Theaceae Theaceae Theophrastaceae Theophrastaceae Theophrastaceae Torricelliaceae Torricelliaceae Tribelaceae Tribelaceae Vahliaceae Vahliaceae Valerianaceae Valerianaceae Valerianaceae Species name with author Stylidium bulbiferum Benth.296 B. Torricellia tiliaefolia DC. (2000a. (2000) Backlund and Bremer (1997) U96649 AJ429307 AF213762 AF299089 AJ429127 AJ429123 AJ403010 AJ429112 L11208 AJ429316 AJ429375 AJ429369 AJ429663 AJ431018 AJ430895 AJ429721 AJ429715 AJ431087 AJ431080 AJ430961 AJ430955 AJ429671 AJ431027 AJ430904 AJ431680 AF130192 Y10699 . 26476 (S) Morton et al. Camellia japonica L. (1997) Z80208 Anderberg et al. Halesia tetraptera L. Tribeles australis Phil. unpublished Y. Valeriana officinalis L. Tu 1991 s. Pterostyrax hispidum Sieber and Zucc. Bremer et al. Camellia japonica L. Br. and Triana Tetramerista sp. Styrax americanum Lam. (1997) Olmstead et al. Vahlia capensis Thunb. no herbarium listed Gardner and Knees 3879 (K) Savolainen et al. (2002) Pennington et al. Theophrasta americana L. Polypremum procumbens L. (1997) Olmstead et al. Camellia sinensis Kuntze Schima superba Gardn. and Triana Pelliciera rhizophoreae Planch. Polypremum procumbens L. (2002) AJ428893 AJ429108 Z80199 AF130216 L12602 AJ429305 AF421073 AJ429306 AJ429662 AJ431017 AJ430894 AJ429661 AJ431016 AJ430893 Z80211 AJ011989 U28885 AJ429352 AF027272 AF421069 AJ429303 AJ429304 AJ429660 AJ429528 AJ431014 AJ431015 AJ430891 AJ430892 AJ430389 AJ431064 AJ430939 AJ011986 AJ429351 AJ429700 AJ431063 AJ430938 rbcL ndhF matK AJ429390 AJ225053 AJ225079 AJ429298 Z80190 AJ428892 L12623 AF130215 AJ429300 AF421074 AJ429301 Z80192 AJ429302 AF421076 AJ429659 AJ431013 AJ430890 AJ429658 AJ431012 AJ430889 AJ429657 AJ431011 AJ430888 AF130214 AJ429107 AJ429299 AJ429656 Missing AJ430887 AJ429655 AJ431010 AJ430082 trnV AJ429735 rps16 AJ431102 trnL AJ430978 Chung and Anderberg 1410 (HAST) Morton et al. (1998) Mullens and Rodgers 64036 (S) Morton et al.

1991 Name of DNA marker and primer rbcL gene FO R W A R D rbcL_50 F rbcL_bs427F rbcL_Z674F rbcL_Z895F RE V E R S E rbcL_Z1020R rbcL_Z674R rbcL_Z895R rbcL_Z1204R rbcL_Z1375R rbcL_30 R ndhF gene FO R W A R D ndhF_IF ()47) ndhF_15F (1) ndhF_3F (396) ndhF_4F (590) ndhF_7F (1201) ndhF_7bF ndhF_9F (1427) ndhF_10F (1600) ndhF_1738F (1658) RE V E R S E ndhF_8dR ndhF_8R (1350R) ndhF_925R (953R) ndhF_2040R (1947R) ndhF_215 OR ndhF_2133R (209R) ndhF_2130R matK gene FO R W A R D matK_1Fb matK_5F Primer sequence from the 50 end Primer position in tobacco (GenBank Z00044) 50 ATG TCA CCA CAA ACA GAA ACT AAA GC 50 GCT TAT ATT AAA ACC TTC CAA GGC CCG CC 50 TTT ATA AAT CAC AAG CCG AAA CTG GTG AAA TC 50 GCA GTT ATT GAT AGA CAG AAA AAT CAT GGT 50 ATC ATC GCG CAA TAA ATC AAC AAA ACC TAA AGT 50 GAT TTC GCC TGT TTC GGC TTG TGC TTT ATA AA 50 ACC ATG ATT CTT CTG CCT ATC AAT AAC TGC 50 CCC TAA GGG TGT CCT AAA GTT TCT CCA CC 50 AAT TTG ATC TCC TTC CAT ATT TCG CA 50 CTT TTA GTA AAA GAT TGG GCC GAG (57595-57620) (58021-58049) (58268-58299) A (58492-58521) A (58650-58618) (58299-58268) (58518-58487) (58829-58801) (58994-58969) (59154-59131) A A A A A 50 AGG TAA GAT CCG GTG AAT CGG AAA C 50 ATG GAA CAG ACA TAT CAA TAY GSR TG 50 TAC TTC CAT GTT GGG ATT AGT TAG TAG 50 TTG GAT AAC GGG GAG TTT CGA ATT T 50 AGG TAC ACT TTC TCT TTG CGG TAT TCC 50 AGG TAC ACT TTC TCT TTG YGG TAT TCC 50 TTC TAT TCA ATA TCT CTA TGG GGT 50 ATC CTT ATG AAT CGG ATA ATA CTAT G 50 TTT GTT CGT TGG ATY YWT AGG AAT 50 GTA AAT AGA TCC GAA ACA TAT AAA ATG 50 ATA GAT CCG ACA CAT ATA AAA TGC GGT TC 50 CCT CTC TTA ATG TCT TTT TGA GCA AGA GCT 50 CTA TGT AAG CMC GAT TAT MYG ACC AA 50 CCA ACY CCA TTY GTA ATT CCA TCA AT 50 CAG GAA CAA GAG GGA TCC ACC GAA 50 CTA CTG ATT TGA TAC CCT CTC CTA (114344-114320) (114292-114267) (113897-112871) (113702-113678) (113093-113067) (113093-113067) (112876-112853) (112692-112667) (112634-112611) B. 3396 (UPS) Olmstead and Reeves (1995) Kaufmann and Wink Z37473 (1996) Erixon and Bremer 9 (UPS) Albert et al.. D ¼ Oxelman et al. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 297 Appendix A (continued ) Family Valerianaceae Verbenaceae Verbenaceae Verbenaceae Vitaceae Vitaceae Species name with author Valeriana hirtella Kunth Verbena bracteata Cav. E ¼ Sang et al.. Vitis aestivalis Michx. DNAX Research Institute. A ¼ Zurawski. Positions of primer corresponding to chloroplast DNA of tobacco (Shinozaki et al. C C C C B. B. Verbena rigida Spreng. All primers except those marked with A–F are constructed at the Department of Systematic Botany. 1999.. B B. Verbena officinalis L. Bremer et al.. B ¼ Kim and Jansen pers. B. Uppsala University.. C B (112939-112965) (112943-112970) B. comm. ex Lag. 1986). C ¼ Oxelman et al. 1997. Vitis vinifera L. and Rodr.B. 1997. C (113340-113369) (112306-112331) (112203-112228) (111860-111883) (112162-112185) 50 TAT ATC CAC TTA TNT TTC AGG AGT 50 GGT ACG GAG TCA AAT KCT AGA AAA T (2603-2580) (2618-2594) . F ¼ Taberlet et al. Citation/voucher rbcL ndhF matK AJ429396 L36418 trnV AJ429742 rps16 AJ431109 trnL Bremer et al.. (1992) L01960 Bremer and Bremer 4091 (UPS) AJ429353 AJ429103 AJ429274 AJ429701 AJ429635 AJ431065 AJ430987 AJ430940 AJ430864 Appendix B Primers used for new sequences in this study.

MO Bot. 211. B. 2002. 763–805..... Williams.. 163–212. Phylogenetic analysis of asterids based on sequences of four genes. C.G..A. 531–553. str. Plant Syst. Phylogenetic Anderberg. Carnivorous plantsphylogeny and structural evolution. 2000. a new Anderberg. 88. St ahl.. 677–687.. N.A. 1997. Pyck.298 B. A. / Molecular Phylogenetics and Evolution 24 (2002) 274–301 Appendix B (continued ) Name of DNA marker and primer matK_3F matK_4F matK_2F matK_1F RE V E R S E matK_1R matK_5R matK_3R matK_6R matK_4R Primer sequence from the 50 end Primer position in tobacco (GenBank Z00044) (3141-3122) E (3157-3135) (3495-3476) E (3727-3707) E (1834-1853)E (2325-2350) (2391-2410) E (2597-2618) (3135-3157) 50 AAG ATG CCT CTT CTT TGC AT 50 CTT CGC TAY TGG GTA AAA GAT GC 50 GTT CAC TAA TTG TGA AAC GT 50 ACT GTA TCG CAC TAT GTA TCA 50 GAA CTA GTC GGA TGG AGT AG 50 GCC AAA GTT CTA GCA CAA GAA ACT CG 50 GAT CCG CTG TGA TAA TGA GA 50 TTC TAG MAT TTG ACT CCG TAC C 50 GCA TCT TTT ACC CAR TAG CGA AG trnT-F intron and intergenic spacers FO R W A R D trnT-F_aF 50 CAT TAC AAA TGC GAT GCT CT trnT-F_a1F 50 ACA AAT GCG ATG CTC TAA CC trnT-F_rF 50 GTT ATA ACT AAT GAG ACA TTC C trnT-F_cF 50 CGA AAT CGG TAG ACG CTA CG trnT-F_eF 50 GGT TCA AGT CCC TCT ATC CC trnT-F_jF 50 GTT CTA ACA AAT GGA GTT GG RE V E R S E T-F_bR trnT-F_iR trnT-F_dR trnT-F_fR trnV intron (Complementary strand) FO R W A R D trnV_1bF trnV_1F trnV_3F trnV_5F RE V E R S E trnV_2R trnV_4R trnV_6bR trnV_6R rps16 intron FO R W A R D rps16_F rps16_2F RE V E R S E rps16_2R rps16_R3 50 TCT ACC GAT TTC GCC ATA TC 50 CCA ACT CCA TTT GTT AGA AC 50 GGG GAT AGA GGG ACT TGA AC 50 ATT TGA ACT GGT GAG ACG AG (48546-48565) F (48550-48469) (48953-48974) (49306-49325) F (49862-49881)F (49493-49512) (49318-49299) F (49508-49495) (49882-49863) F (50299-50280) F 50 GAA CCG TAG ACC TTC TCG GTA AAA CAG ATC 50 TAG GGC TAT ACG GAG TCG AAC CG 50 GTG TAA ACG AGT TGC TCT ACC 50 CAT ACG GCG GGA GTC ATT GGT TC 50 GGT AGA GCA ACT CGT TTA CAC 50 GAA CCA ATG ACT CCC GCC GTA TG 50 GAA GAA ATG ACC TTA AAT CTT TGT G 50 GGA GAG CAA TTT GAA GAA ATG (52772-53798) (53755-53777) (54361-54381) (54622-54644) (54381-54361) (54644-54622) (55290-55266) (55282-55302) 50 GTG GTA GAA AGC AAC GTG CGA CTT 50 GAA GGA CAC GAT CCG YTG TGG AT 50 TCG GGA TCG AAC ATC AAT TGC AAC 50 CGA TAG ACG GCT CAT TGG GAT A (6187-6164) D (6162-6140) (5271-5294) D (5299-5320) References Albach. 93–102..S. Phylogenetic placement of the enigmatic angiosperm Hydrostachys. M. A. Phylogeny of the Asteridae s.E. A. 1998..E. Taxon 47. 183–197.. Bremer. 1992. Chase.. P. J. K€ allersjo tionships in the Primulales inferred from cpDNA rbcL sequence data.C. 207... Plant Syst...l.A. M. B. Soltis. An ordinal classification for the families of flowering plants. Taxon 50. M. Soltis. Ann. A. Evol. M. 89.. Olmstead. D. 1998.A. 85. two new families of caprifolioids.. Backlund.: analyses of molecular data from five genes from the plastid and mitochondrial genomes.. 657–661. Albach.. 225–254. Diervillaceae and Linnaeaceae. €llersjo €. 2001a..l. Bremer et al. 2001b. Ka primuloid family in the order Ericales s. APG.C. Backlund. A. 1491–1495. S. 1998. €. D. Am. B. M.S. Albert. Rydin. Phylogenetic relaAnderberg.W. Gard. Gard. Taxon 49. . with particular reference to the Dipsacales. Maesaceae..W. Science 257. € .E. P. D. St ahl. Chase. R. based on rbcL sequences. D. Soltis. Evol.. Soltis. MO Bot. Ann. K€ allersjo relationships in the order Ericales s. V.. Bot.

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