1. ITS 797 bp long, 161 parsimony informative characters Branch and bound search using parsimony criterion found 15 trees of length 419 and consistency index of 0.8258. Bootstrap Analysis Monophyly of sections Auricula (98%) and Aleuritia (100%) is strongly supported. The monophyly of section Crystallophlomis (55%) is less well supported. Monophyly of subgenera Auriculastratum (98%) and Aleuritia (100%) is well supported. Monophyly of clades based upon chromosome number 2n=22 (100%), 2n=18 (100%) and 2n=66 (98%) is strongly supported. Intrasectional relationships were not well resolved. matK 872 bp long, 79 parsimony informative characters Branch and bound search using parsimony criterion found 3 trees of length 147 and consistancy index of 0.939. Boostrap Analysis Monophyly of section Aleuritia (100%), subgenera Aleuritia (100%) and the clades based on chromosome number 2n=18 (100%) and 2n=22 (100%) are well supported. Other clades suggested by the ITS topology are similar but are not well supported. Evaluate congruence of chloroplast and nuclear phylogenies Test molecular support of traditional sectional classifications Test Kelsos hybridization hypotheses Explore distribution of breeding systems and chromosome number Preliminary phylogenetic analyses of ITS and matK sequences support: Monophyly of sections Aleuritia and Auricula Base chromosome number as a useful taxonomic character Preliminary support of Kelsos hypotheses homostylous species grouped with proposed heterostylous progenitors P.eximia with P. tschuktchorum P. stricta with P. farinosa P. incana and P. laurentiana with P. mistassinica P. borealis with P. modesta need increased matK and ITS sampling to further resolve the relationships equal sample sizes would allow statistical tests of congruence Traditional Classification Over 400 species worldwide in 37 sections (Richards 1993) Based on 15 taxonomic characters, the most informative include: Chromosome number Presence of multicellular hairs Pollen type Breeding system Molecular techinques can provide many more informative characters that can improve resolution of relationships. Breeding Systems Three breeding systems, according to Richards (1993) Primary homostyly Self-fertile populations with a single morphology and mating type Heterostyly Populations a mix of two flower morphs Morphology and a sporophytic self incompatibility system prevent intramorph fertilization Secondary Homostyly Self-fertile with a single floral morphology Develop when hybridization, and most often polyploidy occurs and recombination disables the heterostyly supergene Hybridization Hypothesis An association between secondary homostyly, polyploidy, and extreme arctic or alpine conditions has been reported in Primula. Kelso (1991, 1987) proposed an explanation based on the secondary contact hypothesis (Stebbins 1985). Evidence of hybridization may be evaluated by statistical tests of congruence between nuclear and chloroplast trees P. nutans (2n) P. mistassinica (2n) P. anvilensis (2n) P. farinosa (2n) P. modesta (2n) P. tschuktchorum (2n) P. egaliksensis (4n) P. incana (6n) P. laurentia (8n) P. scotia (6n) P. scandinavica (8n) P. borealis (4n) Heterostyle P. eximia (2n) P. stricta ?Heterostyles Secondary Homostyles Implications for sectional circumscription, relationships and evolution of breeding systems Suring, Erik,* 1 Elena Conti 1, Sylvia Kelso 2 1 University of Alaska Fairbanks 2 Colorado College Reciprocal herkogamy found in heterostylic primulas. Compatible pollinations are indicated by the arrows. Polymorphisms associated with distyly include anther and stigma length, pollen size, and stigmatic papillae size (from Barret and Cruzan, 1994). ITS Maximum Parsimony Tree. The numbers on top of the branches are bootstrap values and the numbers below are distance values. Sectional, and subgeneric groupings are shown. Chromosomal and breeding system distributions are shown. Highlighted taxa are secondary homostyles. P.angustifolia P.glutinosa P.minima P.spectabilis P.clusiana P.palinuri P.nutans P.secundiflora P.tschuktschorum P.eximia.1 P.eximia.2 P.borealis P.modesta P.laurentiana P.mistassinica P.incana P.farinosa P.stricta P.frondosa P.scandinavica 98 100 72 100 88 55 58 100 89 82 72 36 21 14 22 8 10 9 17 15 45 43 18 13 22 5 4 1 0 0 46 10 5 9 0 15 2 4 7 10 6 6 4 Section Crystallophlomis Section Auricula Section Aleuritia Armerina Sikkimensis Subgenus Aleuritia Subgenus Auriculastrum 2n=66 2n=22 2n=18 2n=72 2n=126 2n=54 2n=72 2n=36 2n=198 ITS A diploid, heterostylic taxon of Primula occupied a contiguous area Glacial retreat allowed separated and partially differentiated diploid taxa to come into contact and hybridize, giving rise to polyploid taxa Recombination in the heterostyly supergene gave rise to secondary homostylous taxa Diploid, heterostylous populations of Primula became isolated because of habitat fragmentation caused by climate changes, for example Quaternary glacial peaks Secondary homostylous taxa successfully colonized the new habitats opened by glacial retreat. 100 99 85 100 85 54 50 7 20 44 4 0 5 3 16 0 21 10 0 0 11 2 1 1 2 0 Section Crystallophlomis Section Auricula Section Aleuritia Armerina Subgenus Aleuritia Subgenus Auriculastrum 2n=66 2n=22 2n=18 Sikkimensis P.angustifolia P.borealis.1 P.borealis.2 P.stricta P.laurentiana P.nutans P.eximia P.tschuktschorum P.secundiflora P.glutinosa P.palinuri P.minima 2n=36 2n=126 2n=72 matK Maximum Parsimony Tree. The numbers on top of the branches are bootstrap values and the numbers below are distance values. Sectional, and subgeneric groupings are shown. Chromosomal and breeding system distributions are shown. Highlighted taxa are secondary homostyles. matK Regions Sequenced Internal Transcribed Spacer (ITS) Nuclear sequence Non-coding region maturase K gene (matK) Chloroplast gene Maternally inherited Both have been used in other intrageneric plant studies From Johnson & Soltis 1994Boxed areas represent coding regions Leu F C26A R ITS 1F ITS 2R ITS 4R ITS 3F P. veris P. farinosa Funding was provided by the UAF Office of Arctic Research Undergraduate Research Opportunities Program and The Research Foundation.