Fig 1.
Flower morphology and developmental stages of Conandron ramondioides, Hemiboea bicornuta, and Lysionotus pauciflorus.
(A) Three developmental stages (early, middle, and late) of C. ramondioides—Early Stage: petal length < sepal length; Middle Stage: petal length > sepal length; Late Stage: flower at anthesis. (B) Floral diagram: Five sepals (sep, black); five petals (from p1 to p5, blue); five stamens (st, yellow); gynoecium (gyn, red). (C) Three developmental stages of H. bicornuta—Early Stage: petal length < sepal length; Middle Stage: petal length > sepal size; Late Stage: flower at anthesis. (D) Floral diagram: sepals (sep, black); dorsal petals (dp, blue); lateral petals (lp, yellow); ventral petal (vp, orange); staminodes (sta, blue); ventral stamens (vst, orange); gynoecium (gyn, light blue). (E) Three development stages of L. pauciflorus—Early Stage: petal length < sepal length; Middle Stage: petal length > sepal size; Late Stage: flower at anthesis. (F) Floral diagram: sepals (sep, black); dorsal petals (dp, blue); lateral petals (lp, yellow); ventral petal (vp, orange); staminodes (sta, blue); ventral stamens (vst, orange); gynoecium (gyn, light blue).
Fig 2.
Branch models used in PAML analyses.
(A) Schematic representation of the tree used in PAML. Arrowheads indicate the duplication event. Post duplication denotes all branches arising after the duplication event. Before divergence denotes the first branches following immediately the duplication event and post divergence denotes all subsequent branches. (B) One-ratio model, assumes equal ω across the tree. (C) Two-ratio model, assumes that ω changes pre- and post- duplication of GCYC1C and GCYC1D. Light blue line denotes same ω on branches post-duplication. (D) Three-ratio model, assumes ω varies before divergence (light blue lines), post divergence (dark blue lines) and outgroups (black lines). (E) Four-ratio model, assumes ω varies among branches before divergence (light blue lines), branches in GCYC1C clade (dark blue lines), branches in GCYC1D clade (dark blue triangle) and outgoups (black line).
Fig 3.
Genealogy and duplication events of GCYC in Gesneriaceae.
Phylogram derived from a Maximum Likelihood (ML) analysis showing average branch lengths. Numbers along branches are BI, ML approximate likelihood value (aLRT), and ML bootstrap values. (--) indicates branch support value lower than 0.7 either in either BI, aLRT or ML bootstrap. GCYC copies of species used for expression studies are colored: Conandron ramondioides (light green); Hemiboea bicornuta (orange); Lysionotus pauciflorus (light blue). Dup1 to Dup5 indicate inferred duplication events.
Fig 4.
Expression pattern of GCYC in floral parts of Conandron ramondioides, Hemiboea bicornuta, and Lysionotus pauciflorus.
(A) Conandron ramondioides. (B) Hemiboea bicornuta. (C) Lysionotus pauciflorus. E: Early Stage flower bud, M: Middle Stage flower bud, L: flower bud at anthesis; n denotes no-template control; dp: dorsal petals, lp: lateral petals vp: ventral petal; p1 to p5: pooled C. ramondioides petals; st: pooled stamens (C. ramondioides); vst: ventral stamens, sta: staminodes (H. bicornuta, L. pauciflorus); gyn: gynoecium; sep: pooled sepals (C. ramondioides). A very faint band appears sometimes in no-template controls in C. ramondioides and represent primer dimer artefacts (see S5 Fig).
Fig 5.
GCYC1 phylogenetic tree with summarized GCYC1C and GCYC1D expression pattern in tribe Trichosporeae species.
BI cladogram based on GCYC1 sequences of species in tribe Trichosporeae. Numbers above branches and below branches are approximate likelihood and posterior probability values obtaining from ML and BI analyses respectively. GCYC1 expression patterns are shown in color by organ: D: dorsal petal, L: lateral petal, V: ventral petal; red squares represent presence and white squares absence of gene expression in the respective flower part; square with ‘?’ denote unknown expression. Expression pattern were compiled from this work and previous studies ([27, 28, 31]). GCYC genes isolated from actinomorphy species in boxes. Light green font: Conandron ramondioides, dark green font: Oreocharis dinghushanensis, light blue font: Lysionotus pauciflorus, Orange font: Hemiboea bicornuta, Brown font: Primulina heterotricha, Black font: Oreocharis leiophylla.
Table 1.
Parameter estimates under branch and branch-site models of GCYC1C and GCYC1D in tribe Trichosporeae.
Fig 6.
Reconstructed ECE-CYC2 evolution emphasizing shifts of GCYC expression pattern after gene duplication in correlation to floral symmetry transitions among selected Asiatic Trichosporeae species.