Michaels, H. J., Scott, K. M., Olmstead, R. G., Szaro, T. M
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Interfamilial Relationships of the Asteraceae: Insights from rbcL Sequence Variation Author(s): Helen J. Michaels, Kathy M. Scott, Richard G. Olmstead, Tim Szaro, Robert K. Jansen, Jeffrey D. Palmer Reviewed work(s): Source: Annals of the Missouri Botanical Garden, Vol. 80, No. 3 (1993), pp. 742-751 Published by: Missouri Botanical Garden Press Stable URL: http://www.jstor.org/stable/2399858 . Accessed: 30/11/2011 19:22 Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at . http://www.jstor.org/page/info/about/policies/terms.jsp JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms of scholarship. For more information about JSTOR, please contact [email protected]. Missouri Botanical Garden Press is collaborating with JSTOR to digitize, preserve and extend access to Annals of the Missouri Botanical Garden. http://www.jstor.org INTERFAMILIAL HelenJ. Michaels,2 THE KathyM. Scott,3 RELATIONSHIPS OF Richard G. Olmstead,4 ASTERACEAE:INSIGHTS Tim Szaro, Robert K. Jansen,6 FROM rbcL SEQUENCE and JeffreyD. Palmer3 VARIATION' ABSTRACT Nucleotidesequences of the chloroplastgene rbcL were analyzed to examine relationshipsamong the large, distinctivefamily Asteraceae and eight putativelyclosely related families.Phylogenetic analysis of a total of 24 sequences of rbcL identifieda lineage consistingof two families,the Goodeniaceae and Calyceraceae, as the sister group to the Asteraceae. In addition,a stronglysupported major monophyleticclade consistingof Asteraceae, Goodeniaceae,Calyceraceae, Corokia (Cornaceae sensuCronquist), Menyanthaceae, Lobeliaceae, and Campanulaceae was found.These resultsclearly distance from the Asteraceaecertain groups previously considered closely related; moreover,the results support alternative hypotheses of affinity that were based uponfloral and inflorescencemorphology, biogeography,pollen morphology, chemistry, and pollen-presentationmechanisms. The angiospermfamily Asteraceae has longbeen the Asteraceae,he suggestedconvergence as the recognizedas one of the large, "natural" families basisfor this and identifiedthe Campanulales (Cam- withwell-established limits defined by several spe- panulaceae and Lobeliaceae) as theclosest relative. cialized floralcharacteristics and distinctivesec- Aspects of the distinctivechemistry of the Aster- ondarychemistry. Many recentstudies have illu- aceae (e.g., alkaloids,polyacetylenes, terpenes, in- minatednumerous phylogenetic controversies within ulin forcarbohydrate storage) have been notedin the family(Bremer, 1987; Bremeret al., 1992; several membersof the Campanulales(Campanu- Jansenet al., 1990, 1991a, b; Jansen& Palmer, laceae, Lobeliaceae, Goodeniaceae, Stylidiaceae), 1987a, b, 1988; Karis et al., 1992; Keeley & whileother chemical evidence has pointedto the Jansen,1991; Kim et al., 1992; Watson et al., Apiaceae and Araliaceae(Hegnauer, 1964, 1977). 1991). However, relationshipsamong the Aster- Cronquist(1955) advocatedthe Rubialesas closest aceae and otherfamilies have remainedobscure, relativesof the Asteraceae(grouped with the Gen- due to considerableparallel and convergentevo- tianales in the systemof Takhtajan, 1980), but lutionof conventionalcharacters used to inferaf- also acknowledgedstrong similarities in floraland finities,lack of recent studiesemploying modern inflorescencemorphology between the Asteraceae methodsof phylogeneticanalysis, and substantial and the Calyceraceae (as did Takhtajan, 1980). confusionas to relationshipsamong the various An associationwith the Calyceraceae is also sup- familieswithin the subclassAsteridae itself (but see ported by biogeographyand capitular structure Olmsteadet al., 1992). (Turner,1977) and pollenmorphology (Skvarla et At least 12 familieshave been proposedas clos- al., 1977). Others(reviewed in Skvarlaet al., 1977) est relativesof the Asteraceae based on a variety have noted a palynologicalresemblance of Vale- of traditionaltaxonomic characters. Although rianaceae, Goodeniaceaeand Brunoniaceaeto As- Hutchinson(1969) notedthe superficialsimilarity teraceae. In a morphologicalcladistic study of tribal of the inflorescenceof the Dipsacales (Caprifolia- relationshipswithin the Asteraceae, stylar mor- ceae, Valerianaceae, and Dipsacaceae) to that of phology,chemical characters,and pollen-presen- 1 We thankD. Morganand D. Soltisfor providing an unpublishedrbcL sequence of Corokia. We are gratefulto M. Chase, D. Soltis,and T. Lammersfor comments on the manuscript.This researchhas been supportedby Ohio Board of RegentsResearch Challenge and NSF grantsBSR-8700195 to H. Michaelsand BSR-8717600 to J. Palmer. 2 Departmentof BiologicalSciences, BowlingGreen State University,Bowling Green, Ohio 43403, U.S.A. 3Department of Biology,Indiana University,Bloomington, Indiana 47405, U.S.A. 4Department of E.P.O. Biology,University of Colorado,Boulder, Colorado 80309, U.S.A. 5 Departmentof Plant Pathology,University of California,Berkeley, California 94720, U.S.A. 6 Departmentof Botany,University of Texas at Austin,Austin, Texas 78713, U.S.A. ANN. MISSOURI BOT. GARD. 80: 742-751. 1993. Volume 80, Number3 Michaels et al. 743 1993 InterfamilialRelationships of Asteraceae tationmechanism were identified by Bremer (1987) ofputatively closely related families in the subclass as potentialsynapomorphies linking the Lobeli- Asteridae.For thesesequences, fresh leaf material aceae and Asteraceae. Finally,strong similarities obtainedeither from the fieldor seedlingswas used to the highlyspecialized secondary pollen-presen- to isolateDNA as purifiedchloroplast DNA by the tation mechanismof the Asteraceae have been sucrosegradient method (Palmer, 1986) or as total documentedin Goodeniaceae,Brunoniaceae, Cam- cellularDNA by a modifiedCTAB procedure(Doyle panulaceae,and Lobeliaceae(Leins & Erbar,1990). & Doyle, 1987) followedby CsCl gradientpurifi- As notedin manyof the above attemptsto resolve cation. The rbcL gene was isolatedfor cloning by this controversy,most of the morphologicalor one of two methods.(1) For mosttaxa, fragments chemicalcharacters that supporta particularhy- containingthe entirerbcL gene were gel-isolated pothesisof ancestryare oftenalso foundin several fromeither Sac I or Sac I/BamHI digests and related groups,and some must certainlybe the ligatedinto the plasmidvector Bluescript (Strata- productof parallelevolution. gene,Inc., LaJolla,California). Recombinant, rbcL- Several of the recent molecular advances in containingcolonies were confirmedby Southern elucidatingphylogenetic relationships within the hybridizationto cloned rbcL fragmentsfrom peas. familyhave employedrestriction site analysis of The coding regionwas sequenced froma single- chloroplastDNA. However,this approach is gen- strandedtemplate by the dideoxy chain termination erallyunsuitable at the interfamiliallevel because method(Sanger et al., 1977) using a series of thehomology of site changes becomes doubtful due primersbased on rbcL sequences frommaize and to increased levels of both nucleotidesequence spinach (obtainedfrom G. Zurawski,DNAX). (2) divergence(causing multiple "hits" withinrestric- Sequences fromPentas and Boopis wereobtained tion sites) and lengthvariation causing problems followingamplification and cloning of a double- in alignmentof sites (Downie & Palmer, 1992b; strandedfragment using the polymerasechain re- Palmer et al., 1988). At highertaxonomic levels, action followingthe methodsof Olmstead et al. restrictionsite analysis of onlythe moreconserved (1992). invertedrepeat region of chloroplastDNA may be Preliminaryanalyses included 14 newsequences used to circumventthese problems(e.g., Downie (see Table 1 and Appendixto thisissue) and one & Palmer, 1992a), but fewercharacters are gen- previouslypublished sequence from Flaveria in the erated than in whole genome surveys. DNA se- Asteraceae(Hudson et al., 1990) and, to serve as quence analysisof the slowlyevolving chloroplast outgroups,sequences fromSpinacia in the Car- gene rbcL and nuclear rRNA genes has proven yophyllidae(Zurawski et al., 1981) and Nicotiana highlyeffective in resolvinghigher-level relation- in the Asteridae(Shinozaki et al., 1988). These ships in plants (Chase et al., 1993; Hamby & analyses (Michaels & Palmer, 1990) on only a Zimmer, 1992; Palmer et al., 1988; Ritland & subsetof the data reportedhere identifieda closest Clegg, 1987; Zurawski& Clegg, 1987). In par- sistergroup identical to thepresent expanded anal- ticular,recent studies by a numberof researchers ysis. The results of concurrentstudies of rbcL employingcomparative sequencing of the chloro- sequencesin theSaxifragaceae (Soltis et al., 1990), plast gene encodingthe large subunitof the pho- Asteridae(Olmstead et al., 1992), and 499 angio- tosyntheticenzyme ribulose-1,5-bisphosphate car- sperms(see Chase et al., 1993) have motivated boxylase (rbcL) indicate an appropriatesize and the inclusionof data fromother outgroupsand rate of evolutionfor providing a sufficientnumber fromtaxa notpreviously suspected to be associated ofcharacters for phylogenetic studies at thefamilial withthe Asteraceae. Data forHeuchera (Soltiset and ordinallevels (Donoghueet al., 1992; Kim et al., 1990), Magnolia (Golenberget al., 1990), al., 1992; Olmstead et al., 1992; Soltis et al., Villarsia, Menyanthes,Hedera, and Coriandrum 1990). In this paper we analyze nucleotidese- (Olmsteadet