Person:

Kramer, Elena

Shifts in flower symmetry have occurred frequently during the diversification of angiosperms, and it is thought that such shifts play important roles in plant–pollinator interactions. In the model developmental system Antirrhinum majus (snapdragon), the closely related genes CYCLOIDEA (CYC) and DICHOTOMA (DICH) are needed for the development of zygomorphic flowers and the determination of adaxial (dorsal) identity of floral organs, including adaxial stamen abortion and asymmetry of adaxial petals. However, it is not known whether these genes played a role in the divergence of species differing in flower morphology and pollination mode. We compared A. majus with a close relative, Mohavea confertiflora (desert ghost flower), which differs from Antirrhinum in corolla (petal) symmetry and pollination mode. In addition, Mohavea has undergone a homeotic-like transformation in stamen number relative to Antirrhinum, aborting the lateral and adaxial stamens during flower development. Here we show that the patterns of expression of CYC and DICH orthologs have shifted in concert with changes in floral morphology. Specifically, lateral stamen abortion in Mohavea is correlated with an expansion of CYC and DICH expression, and internal symmetry of Mohavea adaxial petals is correlated with a reduction in DICH expression during petal differentiation. We propose that changes in the pattern of CYC and DICH expression have contributed to the derived flower morphology of Mohavea and may reflect adaptations to a pollination strategy resulting from a mimetic relationship, linking the genetic basis for morphological evolution to the ecological context in which the morphology arose.

FLOWERING LOCUS C (FLC) is a major regulator of flowering responses to seasonal environmental factors. Here, we document that FLC also regulates another major life-history transition-seed germination, and that natural variation at the FLC locus and in FLC expression is associated with natural variation in temperature-dependent germination. FLC-mediated germination acts through additional genes in the flowering pathway (FT, SOC1, and AP1) before involving the abscisic acid catabolic pathway (via CYP707A2) and gibberellins biosynthetic pathway (via GA20ox1) in seeds. Also, FLC regulation of germination is largely maternally controlled, with FLC peaking and FT, SOC1, and AP1 levels declining at late stages of seed maturation. High FLC expression during seed maturation is associated with altered expression of hormonal genes (CYP707A2 and GA20ox1) in germinating seeds, indicating that gene expression before the physiological independence of seeds can influence gene expression well after any physical connection between maternal plants and seeds exists. The major role of FLC in temperature-dependent germination documented here reveals a much broader adaptive significance of natural variation in FLC. Therefore, pleiotropy between these major life stages likely influences patterns of natural selection on this important gene, making FLC a promising case for examining how pleiotropy influences adaptive evolution.

Aquilegia is an emerging model organism that is phylogenetically intermediate between the core eudicot and monocot models, Arabidopsis and Oryza. In this study, we have used a comparative genomics approach to identify 45 Aquilegia microRNAs that comprise 20 separate plant microRNA families. We have predicted 84 targets of these newly identified Aquilegia microRNAs including transcription factors and loci involved in metabolism, stress responses, transport, and auxin signaling. microRNA families from 16 plant species and the newly identified microRNAs from Aquilegia were analyzed in a phylogenetic context revealing 40 distantly conserved microRNA families. In addition to these highly conserved plant microRNA families, several families with disjointed phylogenetic distribution were identified. This study provides a phylogenetically important dataset for plant microRNA evolution studies. The current study is the first to identify miRNAs in a lower eudicot in which comprehensive genomic resources are becoming available.

Background: The evolution of type II MADS box genes has been extensively studied in angiosperms. One of the best-understood subfamilies is that of the Arabidopsis gene APETALA3 (AP3). Previous work has demonstrated that the ancestral paleoAP3 lineage was duplicated at some point within the basal eudicots to give rise to the paralogous TM6 and euAP3 lineages. This event was followed in euAP3 orthologs by the replacement of the C-terminal paleoAP3 motif with the derived euAP3 motif. It has been suggested that the new motif was created by an eight-nucleotide insertion that produced a translational frameshift. Results: The addition of 25 eudicot AP3 homologs to the existing dataset has allowed us to clarify the process by which the euAP3 motif evolved. Phylogenetic analysis indicates that the euAP3/TM6 duplication maps very close to the base of the core eudicots, associated with the families Trochodendraceae and Buxaceae. We demonstrate that although the transformation of paleoAP3 into euAP3 was due to a frameshift mutation, this was the result of a single nucleotide deletion. The use of ancestral character state reconstructions has allowed us to demonstrate that the frameshift was accompanied by few other nucleotide changes. We further confirm that the sequence is evolving as coding region. Conclusion: This study demonstrates that the simplest of genetic changes can result in the remodeling of protein sequence to produce a kind of molecular 'hopeful monster.' Moreover, such a novel protein motif can become conserved almost immediately on the basis of what appears to be a rapidly generated new function. Given that the existing data on the function of such C-terminal motifs are somewhat disparate and contradictory, we have sought to synthesize previous findings within the context of the current analysis and thereby highlight specific hypotheses that require further investigation before the significance of the euAP3 frameshift event can be fully understood.

The basal eudicot Aquilegia (columbine) has an unusual floral structure that includes two morphologically distinct whorls of petaloid organs and a clearly differentiated fifth organ type, the staminodium. In this study, we have sought to determine how Aquilegia homologs of the B class genes APETALA3 (AP3) and PISTILLATA (PI) contribute to these novel forms of organ identity. Detailed expression analyses of the three AP3 paralogs and one PI homolog in wild-type and floral homeotic mutant lines reveal complex patterns that suggest that canonical B class function has been elaborated in Aquilegia. Yeast two-hybrid studies demonstrate that the protein products of Aquilegia's AP3 and PI homologs can form heterodimers, much like what has been observed for their core eudicot homologs. Downregulation of AqvPI using virus-induced gene silencing indicates that in addition to petal and stamen identity, this locus is essential to staminodial identity but may not control the identity of the petaloid sepals. Our findings show that preexisting floral organ identity programs can be partitioned and modified to produce additional organ types. In addition, they indicate that some types of petaloid organs are not entirely dependent on AP3/PI homologs for their identity.

Our recently acquired understanding of the ABC program, which controls floral organ identity in model plant species such as Arabidopsis thaliana and Antirrhinum majus, has provided a new set of characters with which to evaluate floral evolution. What is still lacking, however, is a clear assessment of the actual degree of conservation of this genetic program across the angiosperms. To this end, we have begun to investigate the evolution of members of the B class gene lineages, which are known to control petal and stamen identity in the higher eudicots, and to analyze their expression patterns in selected species from the lower eudicots and basal angiosperms. The B class genes comprise the homologues of the A. thaliana genes APETALA3 (AP3) and PISTILLATA (PI), which are closely related paralogues encoding MADS box-containing DNA-binding proteins. This study has uncovered many examples of gene duplication and divergence in both the AP3 and PI lineages as well as complex and variable patterns of gene expression. These findings indicate that although some aspects of the ABC program are conserved, others display a high degree of plasticity and may not have become fixed until later in angiosperm evolution.

It is increasingly clear that additional "model" systems are needed to elucidate the genetic and developmental basis of organismal diversity. Whereas model system development previously required enormous investment, recent advances including the decreasing cost of DNA sequencing and the power of reverse genetics to study gene function are greatly facilitating the process. In this review, we consider two aspects of the development of new genetic model systems: first, the types of questions being advanced using these new models; and second, the essential characteristics and molecular tools for new models, depending on the research focus. We hope that researchers will be inspired to explore this array of emerging models and even consider developing new molecular tools for their own favorite organism.

Petaloid organs are a major component of the floral diversity observed across nearly all major clades of angiosperms. The variable morphology and development of these organs has led to the hypothesis that they are not homologous but, rather, have evolved multiple times. A particularly notable example of petal diversity, and potential homoplasy, is found within the order Ranunculales, exemplified by families such as Ranunculaceae, Berberidaceae, and Papaveraceae. To investigate the molecular basis of petal identity in Ranunculales, we used a combination of molecular phylogenetics and gene expression analysis to characterize APETALA3 (AP3) and PISTILLATA (PI) homologs from a total of 13 representative genera of the order. One of the most striking results of this study is that expression of orthologs of a single AP3 lineage is consistently petal-specific across both Ranunculaceae and Berberidaceae. We conclude from this finding that these supposedly homoplastic petals in fact share a developmental genetic program that appears to have been present in the common ancestor of the two families. We discuss the implications of this type of molecular data for long-held typological definitions of petals and, more broadly, the evolution of petaloid organs across the angiosperms.

To understand the developmental basis of a novel anatomical feature, we present a comparative developmental study of an ecologically significant novelty in fruit morphology. Most members of the tribe Brassiceae have heteroarthrocarpic fruits, in contrast to the unsegmented fruits of many Brassicaceae. Heteroarthrocarpy is characterized by a joint that bisects fruits into heteromorphic segments and by partial or complete indehiscence. In order to better understand the development of heteroarthrocarpic characteristics and their relationships to typical siliques, we studied carpel and fruit development in two closely related species of the Brassiceae, Erucaria erucarioides and Cakile lanceolata. Our results indicate that proximal segments of heteroarthrocarpic fruits correspond to valves of typical siliques, regardless of whether these segments are dehiscent. Indehiscent distal segments are composed of both stylar and ovary elements, although the ovary wall of this segment does not differentiate into valve tissue. The joint itself comprises the distal extent of the valve margin and an internal proliferation of the mesocarp. Additionally, Cakile fruits form a transverse dehiscence zone through the joint, allowing the segments to separate. Heteroarthrocarpy entails modifications of lignification patterns and alterations of relationships between valve, style, ovary, and mesocarp. Possible genetic mechanisms underlying these modifications are discussed in reference to what is known about silique development in Arabidopsis.

It has been proposed that the diversification of the MADS-box gene family of transcription factors has played a major role in the radiation of land plants. This suggestion is based on the critical roles that these genes play in plant development and the apparent coincidence of key duplication events with major radiations, such as the establishment of the B and C lineages concurrent with the evolution of the seed plants. On a more recent scale, it is also possible that subsequent duplication events have contributed to later morphological diversifications. In order to investigate this possibility, we are studying the evolution of homologs of the petal and stamen identity genes APETALA3 (AP3) and PISTILLATA (PI) in the Ranunculaceae. In this family, the AP3 and PI lineages have undergone many duplication events at every phylogenetic level. Early duplications gave rise to three paralogous AP3 lineages, which are found throughout the family. In contrast, numerous duplications have occurred relatively recently in the PI lineage. We outline a hypothesis that these duplications have played a role in the evolution of the unique types of petaloid organs in the Ranunculaceae and present preliminary expression data supporting such a scenario.