Floral organ identity: 20 years of ABCs

One of the early successes of the application of molecular genetics to study plant development was the discovery of a series of genes that act together, in an apparently simple combinatorial model, to specify the identity of the different organs of a flower. Widely known as the ABC model, this framework for understanding has been investigated and modified over the course of the last two decades. The cast list of genes has been defined and, as other chapters in this volume will show, great progress has been made in understanding how they are regulated, how they act together, what they do and how they have contributed to the evolution of the flower in its varied forms. In this introductory review to the volume we will review the derivation and elaboration of the most current version of the ABC model, highlighting the modifications that have been necessary to ensure it fits the available experimental data. We will highlight the remaining difficulties in fitting the current model to the experimental data and propose a further modification to enable it to regain its applicability.     

SUPERWOMAN1 and DROOPING LEAF genes control floral organ identity in rice

We analyzed recessive mutants of two homeotic genes in rice, SUPERWOMAN1 (SPW1) and DROOPING LEAF (DL). The homeotic mutation spw1 transforms stamens and lodicules into carpels and palea-like organs, respectively. Two spw1 alleles, spw1-1 and spw1-2, show the same floral phenotype and did not affect vegetative development. We show that SPW1 is a rice APETALA3 homolog, OsMADS16. In contrast, two strong alleles of the dl locus, drooping leaf-superman1 (dl-sup1) and drooping leaf-superman2 (dl-sup2), cause the complete transformation of the gynoecium into stamens. In these strong mutants, many ectopic stamens are formed in the region where the gynoecium is produced in the wild-type flower and they are arranged in a non-whorled, alternate pattern. The intermediate allele dl-1 (T65), results in an increase in the number of stamens and stigmas, and carpels occasionally show staminoid characteristics. In the weakest mutant, dl-2, most of the flowers are normal. All four dl alleles cause midrib-less drooping leaves. The flower of the double mutant, spw1 dl-sup, produces incompletely differentiated organs indefinitely after palea-like organs are produced in the position where lodicules are formed in the wild-type flower. These incompletely differentiated organs are neither stamens nor carpels, but have partial floral identity. Based on genetic and molecular results, we postulate a model of stamen and carpel specification in rice, with DL as a novel gene controlling carpel identity and acting mutually and antagonistically to the class B gene, SPW1.     

Genetic basis for innovations in floral organ identity

Of the many innovations associated with the radiation of the angiosperms, the evolution of a petal identity program is among the best understood from a genetic standpoint. Although the existing data do indicate that similar genetic mechanisms control petal development across diverse taxa, there is also considerable evidence for variability in petal identity programs, likely due to a number of factors. These points are illustrated through a review of our current knowledge on the subject, integrating phylogenetic, morphological, and genetic studies. Comparative studies of petal identity highlight the complex nature of homology in plants and stand as a cautionary tale for the interpretation of gene expression data.     

Development of floral organ identity: stories from the MADS house

Recent studies on AGAMOUS-LIKE2-, DEFICIENS- and GLOBOSA-like MADS-box genes in diverse seed plant species have provided novel insights into the mechanisms by which the identity of the different floral organs is specified during flower development. These advances in understanding may lead to major refinements in the classical ABC model of floral organ identity.     

New members of the floral organ identity AGAMOUS pathway

The Arabidopsis floral organ identity gene AGAMOUS (AG) specifies stamen and carpel development as well as floral determinacy. Recent reports suggest that the HUA1, HUA2, HEN1 and HEN2 genes function redundantly as components of the AG pathway. The HUA1, HUA2, HEN1 and HEN2 genes encode nuclear proteins that perhaps play a role in RNA metabolism. The HUA and HEN genes function not only on the AG pathway, but also in vegetative development.     

Developmental programmes in floral organ formation

In contrast to animals, organogenesis in plants is continuous, allowing development in response to intrinsic and extrinsic signals. Organs arise from primordia formed on the flanks of meristems. The apical meristem produces primordia that acquire leaf identity, while floral meristems form primordia which develop into four organ types: sepals, petals, stamens and carpels. The production of mature organs involves two distinct processes, the initiation of organ primordia and the establishment of meristem, primordia and cell identities. Here we concentrate on floral organogenesis in Arabidopsis and examine the extent to which these processes utilize similar control mechanisms and regulatory molecules.     

The ABC model and the diversification of floral organ identity

Broad studies of the ABC program across angiosperms have found that interactions between gene duplication, biochemical evolution, shifts in gene expression and modification of existing identity programs have been critical to the evolution of floral morphology. Several themes can be recognized in this context. First, the original concept of “A” function applies only very narrowly to Arabidopsis and its close relatives. Second, while many types of petaloid organs are associated with the expression of AP3/PI homologs, there is growing evidence that there are other genetic mechanisms for producing petaloidy, especially in first whorl organs. Third, pre-existing organ identity programs can be modified to yield novel organ types, often in association with gene duplications. Lastly, there are many aspects of ABC gene function outside the major model systems that remain a mystery, perhaps none more so than the C-terminal amino acid motifs that distinguish specific ABC gene lineages.     

水稻花器官特征决定以及数量控制的分子机制

继双子叶模式植物拟南芥之后, 单子叶模式植物水稻的生殖发育研究受到广泛的重视。随着水稻正向和反向遗传学研究的不断深入, 人们发现了一些调控水稻花器官特征以及花器官数量的重要基因, 使得对水稻花器官发育的调控机制有了更多的了解。本文着重概述和讨论水稻花器官特征决定以及花器官数量控制分子机理研究的最新进展。     

The expression of floral organ identity genes in contrasting water lily cultivars

The floral organs of typical eudicots such as Arabidopsis thaliana are arranged in four characteristic whorls, namely the sepal, petal, stamen and carpel, and the “ABC” floral organ identity model has been based on this arrangement. However, the floral organs in most basal angiosperms are spirally arranged with a gradual transition from the inside to outside, and an alternative model referred to as “fading borders” was developed to take account of this. The flower morphology of the water lily was tested against the “fading borders” model by determining the expression profile of the six primary floral organ identity genes AP2, AGL6, AP3, PI, AG and SEP1 in two cultivars showing contrasting floral morphology. In addition, to get accurate floatation of the genes expression level from outer to inner, we divided the floral organs into eight whorls according to morphological features. All these genes were expressed throughout all whorls of the flower, but their expression level changed gradually from the outside of the flower to its inside. This pattern was consistent with the “fading borders” model.     

Benzylaminopurine induces phenocopies of floral meristem and organ identity mutants in wild-typeArabidopsis plants

Arabidopsis thaliana (L.) Heynh. has been used as a model system to investigate the regulatory genes that control and coordinate the determination, differentiation and morphogenesis of the floral meristem and floral organs. We show here that benzylaminopurine (BAP), a cytokinin, influences flower development inArabidopsis and induces partial phenocopies of known floral homeotic mutants. Application of BAP to wild-type inflorescences at three developmental stages results in: (i) increase in floral organ number; (ii) formation of abnormal floral organs and (iii) induction of secondary floral buds in the axils of sepals. These abnormalities resemble the phenotypes of mutants,clv1 (increase in organ number),ap1,ap2,ap3 (abnormal floral organs) andap1 (secondary floral buds in the axils of first-whorl organs). In addition, BAP induces secondary floral buds in the axils of perianth members ofapt2-6, ap3-1 andag mutants, and accentuates the phenotype of theapt2-1 mutant to resemble theapt2-6 mutant. These observations suggest that exogenous BAP suppresses the normal functioning of the genes for floral meristem identity and thereby affects flower development and the later stages of floral organ differentiation.Abbreviations BAP N6-benzylaminopurine - CK cytokinin     

Functional conservation of MADS-box factors controlling floral organ identity in rice and Arabidopsis

Studies on MADS-box genes in Arabidopsis and other higher eudicotyledonous flowering plants have shown that they are key regulators of flower development. Since Arabidopsis and monocotyledonous rice are distantly related plant species it is interesting to investigate whether the floral organ identity factors have been conserved in their functions, and if not, to understand the differences. Arabidopsis and rice are very suitable for these studies since they are both regarded as models for plant functional genomics. Both their genomes are sequenced and tools are available for the analysis of gene function. These developments have accelerated experiments and increased our knowledge on rice gene function. Therefore it is the right moment to perform a comparative analysis on MADS-box factors controlling floral organ identity as reported in this review.     

The ASK1 gene regulates development and interacts with the UFO gene to control floral organ identity in Arabidopsis.

Normal flower development likely requires both specific and general regulators. We have isolated an Arabidopsis mutant ask1-1 (for -Arabidopsis skp1-like1-1), which exhibits defects in both vegetative and reproductive development. In the ask1-1mutant, rosette leaf growth is reduced, resulting in smaller than normal rosette leaves, and internodes in the floral stem are shorter than normal. Examination of cell sizes in these organs indicates that cell expansion is normal in the mutant, but cell number is reduced. In the mutant, the numbers of petals and stamens are reduced, and many flowers have one or more petals with a reduced size. In addition, all mutant flowers have short stamen filaments. Furthermore, petal/stamen chimeric organs are found in many flowers. These results indicate that the ASK1 gene affects the size of vegetative and floral organs. The ask1 floral phenotype resembles somewhat that of the Arabidopsis ufo mutants in that both genes affect whorls 2 and 3. We therefore tested for possible interactions between ASK1 and UFO by analyzing the phenotypes of ufo-2 ask1-1 double mutant plants. In these plants, vegetative development is similar to that of the ask1-1 single mutant, whereas the floral defects are more severe than those in either single mutant. Interior to the first whorl, the double mutant flowers have more sepals or sepal-like organs than are found in ufo-2, and less petals than ask1-1. Our results suggest that ASK1 interacts with UFO to control floral organ identity in whorls 2 and 3. This is very intriguing because ASK1 is very similar in sequence to the yeast SKP1 protein and UFO contains an F-box, a motif known to interact with SKP1 in yeast. Although the precise mechanism of ASK1 and UFO action is unknown, our results support the hypothesis that these two proteins physically interact in vivo.     

NO APICAL MERISTEM (MtNAM) regulates floral organ identity and lateral organ separation in Medicago truncatula

? The CUP-SHAPED COTYLEDON (CUC)/NO APICAL MERISTEM (NAM) family of genes control boundary formation and lateral organ separation, which is critical for proper leaf and flower patterning. However, most downstream targets of CUC/NAM genes remain unclear. ? In a forward screen of the tobacco retrotransposon1 (Tnt1) insertion population in Medicago truncatula, we isolated a weak allele of the no-apical-meristem mutant mtnam-2. Meanwhile, we regenerated a mature plant from the null allele mtnam-1. These materials allowed us to extensively characterize the function of MtNAM and its downstream genes. ? MtNAM is highly expressed in vegetative shoot buds and inflorescence apices, specifically at boundaries between the shoot apical meristem and leaf/flower primordia. Mature plants of the regenerated null allele and the weak allele display remarkable floral phenotypes: floral whorls and organ numbers are reduced and the floral organ identity is compromised. Microarray and quantitative RT-PCR analyses revealed that all classes of floral homeotic genes are down-regulated in mtnam mutants. Mutations in MtNAM also lead to fused cotyledons and leaflets of the compound leaf as well as a defective shoot apical meristem. ? Our results revealed that MtNAM shares the role of CUC/NAM family genes in lateral organ separation and compound leaf development, and is also required for floral organ identity and development.