Floral Determinacy Research Articles

The SEPALLATA-like CaSEP5 gene regulates flower sepal, pedicel, and fruit development in pepper (Capsicum annuum L.)

Flowers are reproductive plant organs that determine final fruit or seed yield. SEPALLATA-like MADS-box genes participate in floral determination and/or flower organ specification. Nevertheless, their molecular mechanisms in bell pepper have not yet been elucidated. In this study, we identified CaSEP5, which encodes a duplicated SEPALLATA-like MADS-box protein grouped in the FBP9/23-subclade. CaSEP5 genes were detected in all whorls of floral organs, and were relatively highly expressed in the sepals and petals. Subcellular localization revealed that CaSEP5 was in the nucleus. This finding aligns with transcription factor characteristics. The physical interactions of CaSEP5 with B-, C-, and other CaSEPs indicated that CaSEP5 act as glue to form protein complexes to regulate flower development by altering the hormones level and distribution. CaSEP5-silenced plants had elongated, leaf-like sepals, rigid, straight pedicels, reduced fruit size, altered expression patterns of other floral homeotic genes, and varied hormone levels. We proposed that CaSEP5 plays an important role in pepper flower and fruit development by means of a complex hormone and genetic network.

Heat stress impairs floral meristem termination and fruit development by affecting the BR-SlCRCa cascade in tomato

Floral meristem termination is a key step leading to carpel initiation and fruit development. The frequent occurrence of heat stress due to global warming often disrupts floral determinacy, resulting in defective fruit formation. However, the detailed mechanism behind this phenomenon is largely unknown. Here, we identify CRABS CLAW a (SlCRCa) as a key regulator of floral meristem termination in tomato. SlCRCa functions as an indispensable floral meristem terminator by suppressing SlWUS activity through the TOMATO AGAMOUS 1 (TAG1)–KNUCKLES (SlKNU)–INHIBITOR OF MERISTEM ACTIVITY (SlIMA) network. A direct binding assay revealed that SlCRCa specifically binds to the promoter and second intron of WUSCHEL (SlWUS). We also demonstrate that SlCRCa expression depends on brassinosteroid homeostasis in the floral meristem, which is repressed by heat stress via the circadian factor EARLY FLOWERING 3 (SlELF3). These results provide new insights into floral meristem termination and the heat stress response in flowers and fruits of tomato and suggest that SlCRCa provides a platform for multiple protein interactions that may epigenetically abrogate stem cell activity at the transition from floral meristem to carpel initiation.

Variation in floral form of CRISPR knock-outs of the poplar homologs of LEAFY and AGAMOUS after FT heat-induced early flowering.

Plant migration and gene flow from genetically modified or exotic trees to nearby lands or by crossing with wild relatives is a major public and regulatory concern. Many genetic strategies exist to mitigate potential gene flow; however, the long delay in onset of flowering is a severe constraint to research progress. We used heat-induced FT overexpression to speed assessment of the expected floral phenotypes after CRISPR knockout of poplar homologs of the key floral genes, LEAFY and AGAMOUS. We selected events with previously characterized CRISPR-Cas9 induced biallelic changes then re-transformed them with the Arabidopsis thaliana FLOWERING LOCUS T (AtFT) gene under control of either a strong constitutive promoter or a heat-inducible promoter. We successfully obtained flowering in both a male and female clones of poplar, observing a wide range of inflorescence and floral forms among flowers, ramets, and insertion events. Overall, flowers obtained from the selected LFY and AG targeted events were consistent with what would be predicted for loss-of-function of these genes. LFY-targeted events showed small catkins with leaf-like organs, AG-targeted events had nested floral organs consistent with reduction in floral determinacy and absence of well-formed carpels or anthers. These findings demonstrate the great developmental plasticity of Populus flowers during genetically accelerated flowering, which may be of horticultural value. They also provide an informative early view of floral phenotypes and apparent sterility from knockouts of both these gene targets.

A model worker: Multifaceted modulation of AUXIN RESPONSE FACTOR3 orchestrates plant reproductive phases.

The key phytohormone auxin is involved in practically every aspect of plant growth and development. Auxin regulates these processes by controlling gene expression through functionally distinct AUXIN RESPONSE FACTORs (ARFs). As a noncanonical ARF, ARF3/ETTIN (ETT) mediates auxin responses to orchestrate multiple developmental processes during the reproductive phase. The arf3 mutation has pleiotropic effects on reproductive development, causing abnormalities in meristem homeostasis, floral determinacy, phyllotaxy, floral organ patterning, gynoecium morphogenesis, ovule development, and self-incompatibility. The importance of ARF3 is also reflected in its precise regulation at the transcriptional, posttranscriptional, translational, and epigenetic levels. Recent studies have shown that ARF3 controls dynamic shoot apical meristem (SAM) maintenance in a non-cell autonomous manner. Here, we summarize the hierarchical regulatory mechanisms by which ARF3 is regulated and the diverse roles of ARF3 regulating developmental processes during the reproductive phase.

Effects of Water-deficit Stress and Gibberellic Acid on Floral Gene Expression and Floral Determinacy in ‘Washington’ Navel Orange

Effects of water-deficit stress and foliar-applied gibberellic acid (GA3) on ‘Washington’ navel orange (Citrus sinensis) floral gene expression and inflorescence number were quantified. Trees subjected to 8 weeks of water-deficit stress [average stem water potential (SWP) −2.86 MPa] followed by 3 weeks of re-irrigation (SWP recovered to > −1.00 MPa) produced more inflorescences in week 11 than trees well-irrigated (SWP > −1.00 MPa) for the full 11 weeks (P < 0.001). After 8 weeks of water-deficit stress, bud expression of flowering locus t (FT), suppressor of overexpression of constans1 (SOC1), leafy (LFY), apetala1 (AP1), apetala2 (AP2), sepallata1 (SEP1), pistillata (PI), and agamous (AG) increased during the re-irrigation period (weeks 9 and 10), but only AP1, AP2, SEP1, PI, and AG expression increased to levels significantly greater than that of well-irrigated trees. Foliar-applied GA3 (50 mg·L−1) in weeks 2 through 8 of the water-deficit stress treatment did not reduce bud FT, SOC1, or LFY expression, but prevented the upregulation AP1, AP2, SEP1, PI, and AG expression that occurred during re-irrigation in water-deficit stressed trees not treated with GA3. Applications of GA3 to water-deficit stressed trees reduced inflorescence number 95% compared with stressed trees without GA3. Thus, GA3 inhibited citrus (Citrus sp.) floral development in response to water-deficit stress through downregulating AP1 and AP2 expression, which likely led to the failed activation of the downstream floral organ identity genes. The results reported herein suggest that bud determinacy and subsequent floral development in response to water-deficit stress in ‘Washington’ navel orange are controlled by AP1 and AP2 transcript levels, which regulate downstream floral organ identity gene activity and the effect of GA3 on citrus flower formation. The water-deficit stress floral-induction pathway provides an alternative to low-temperature induction that increases the potential for successful flowering in citrus trees grown in areas experiencing warmer, drier winters due to global climate change.

Tomato CRABS CLAW paralogues interact with chromatin remodelling factors to mediate carpel development and floral determinacy.

Summary CRABS CLAW (CRC) orthologues play a crucial role in floral meristem (FM) determinacy and gynoecium formation across angiosperms, the key developmental processes for ensuring successful plant reproduction and crop production. However, the mechanisms behind CRC mediated FM termination are far from fully understood.Here, we addressed the functional characterization of tomato (Solanum lycopersicum) paralogous CRC genes. Using mapping‐by‐sequencing, RNA interference and CRISPR/Cas9 techniques, expression analyses, protein–protein interaction assays and Arabidopsis complementation experiments, we examined their potential roles in FM determinacy and carpel formation.We revealed that the incomplete penetrance and variable expressivity of the indeterminate carpel‐inside‐carpel phenotype observed in fruit iterative growth (fig) mutant plants are due to the lack of function of the S. lycopersicum CRC homologue SlCRCa. Furthermore, a detailed functional analysis of tomato CRC paralogues, SlCRCa and SlCRCb, allowed us to propose that they operate as positive regulators of FM determinacy by acting in a compensatory and partially redundant manner to safeguard the proper formation of flowers and fruits.Our results uncover for the first time the physical interaction of putative CRC orthologues with members of the chromatin remodelling complex that epigenetically represses WUSCHEL expression through histone deacetylation to ensure the proper termination of floral stem cell activity.

Polycomb proteins control floral determinacy by H3K27me3-mediated repression of pluripotency genes in Arabidopsis thaliana.

Polycomb group (PcG) protein-mediated histone methylation (H3K27me3) controls the correct spatiotemporal expression of numerous developmental regulators in Arabidopsis. Epigenetic silencing of the stem cell factor gene WUSCHEL (WUS) in floral meristems (FMs) depends on H3K27me3 deposition by PcG proteins. However, the role of H3K27me3 in silencing of other meristematic regulator and pluripotency genes during FM determinacy has not yet been studied. To this end, we report the genome-wide dynamics of H3K27me3 levels during FM arrest and the consequences of strongly depleted PcG activity on early flower morphogenesis including enlarged and indeterminate FMs. Strong depletion of H3K27me3 levels results in misexpression of the FM identity gene AGL24, which partially causes floral reversion leading to ap1-like flowers and indeterminate FMs ectopically expressing WUS and SHOOT MERISTEMLESS (STM). Loss of STM can rescue supernumerary floral organs and FM indeterminacy in H3K27me3-deficient flowers, indicating that the hyperactivity of the FMs is at least partially a result of ectopic STM expression. Nonetheless, WUS remained essential for the FM activity. Our results demonstrate that PcG proteins promote FM determinacy at multiple levels of the floral gene regulatory network, silencing initially floral regulators such as AGL24 that promotes FM indeterminacy and, subsequently, meristematic pluripotency genes such as WUS and STM during FM arrest.

A Novel Embryo Phenotype Associated With Interspecific Hybrid Weakness in Rice Is Controlled by the MADS-Domain Transcription Factor OsMADS8.

A novel hybrid weakness gene, DTE9, associated with a dark tip embryo (DTE) trait, was observed in CR6078, an introgression line derived from a cross between the Oryza sativa spp. japonica “Hwayeong” (HY) and the wild relative Oryza rufipogon. CR6078 seeds exhibit protruding embryos and flowers have altered inner floral organs. DTE9 was also associated with several hybrid weakness symptoms including decreased grain weight. Map-based cloning and transgenic approaches revealed that DTE9 is an allele of OsMADS8, a MADS-domain transcription factor. Genetic analysis indicated that two recessive complementary genes were responsible for the expression of the DTE trait. No sequence differences were observed between the two parental lines in the OsMADS8 coding region; however, numerous single nucleotide polymorphisms were detected in the promoter and intronic regions. We generated overexpression (OX) and RNA interference (RNAi) transgenic lines of OsMADS8 in HY and CR6078, respectively. The OsMADS8-OX lines showed the dark tip embryo phenotype, whereas OsMADS8-RNAi recovered the normal embryo phenotype. Changes in gene expression, including of ABCDE floral homeotic genes, were observed in the OsMADS8-OX and OsMADS8-RNAi lines. Overexpression of OsMADS8 led to decreased expression of OsEMF2b and ABA signaling-related genes including OsVP1/ABI3. HY seeds showed higher ABA content than CR6078 seeds, consistent with OsMADS8/DTE9 regulating the expression of genes related ABA catabolism in CR6078. Our results suggest that OsMADS8 is critical for floral organ determination and seed germination and that these effects are the result of regulation of the expression of OsEMF2b and its role in ABA signaling and catabolism.

Cloning, Expression, and Tobacco Overexpression Analyses of a PISTILLATA/GLOBOSA-like (OfGLO1) Gene from Osmanthus fragrans.

GLOBOSA (GLO), a B-class MADS-box gene, is involved in floral organ determination but has rarely been studied in Osmanthus fragrans, which is a very popular ornamental tree species in China. Here, the full-length cDNA of a homologous GLO1 gene (named OfGLO1) was cloned from a flower bud of O. fragrans using the RACE technique. The OfGLO1 has a 645 bp open reading frame, encoding 214 amino acids. Similar to other PI/GLO proteins, OfGLO1 has two conserved domains, MADS MEF2-like and K-box, and a 16-amino-acid PI motif in the C terminal region. Our phylogeny analysis classified OfGLO1 as a PI-type member of the B-class MADS-box gene family. The qRT-PCR assay showed that the expression of OfGLO1 in O. fragrans was continuously upregulated from the tight bud stage to the full flowering stage but barely expressed in the pistils, sepals, and non-floral organs, such as root, leaf, and stem. The genetic effect of OfGLO1 was assayed by ectopic expression in tobacco plants. Compared with the wild-type, OfGLO1 transformants showed reduced plant size, earlier flowering, shorter stamens, and lower seed setting rates. Furthermore, some stamens were changed into petal-like structures. These findings indicate that OfGLO1 plays an important role in the regulation of flower development. This study improved our understanding of class B gene function in woody plants.

Promoting virus-induced gene silencing of pepper genes by a heterologous viral silencing suppressor.

Pepper is an important vegetable cultivated worldwide. Pepper fruits accumulate unique metabolites, capsanthin and capsaicin, which are important raw materials for natural pigment and medicine. Pepper plants are rich in genetic diversity and are attractive subjects for fruit developmental studies. The release of pepper reference genomes provided key genetic information for dissecting function of pepper genes underlying various interesting phenotypes (Kim et al., 2014; Qin et al., 2014). However, due to the difficulty in transformation, dissecting genetic mechanisms in pepper remained technically challenging.

SlGT11 controls floral organ patterning and floral determinacy in tomato

BackgroundFlower development directly affects fruit production in tomato. Despite the framework mediated by ABC genes have been established in Arabidopsis, the spatiotemporal precision of floral development in tomato has not been well examined.ResultsHere, we analyzed a novel tomato stamenless like flower (slf) mutant in which the development of stamens and carpels is disturbed, with carpelloid structure formed in the third whorl and ectopic formation of floral and shoot apical meristem in the fourth whorl. Using bulked segregant analysis (BSA), we assigned the causal mutation to the gene Solanum lycopersicum GT11 (SlGT11) that encodes a transcription factor belonging to Trihelix gene family. SlGT11 is expressed in the early stages of the flower and the expression becomes more specific to the primordium position corresponding to stamens and carpels in later stages of the floral development. Further RNAi silencing of SlGT11 verifies the defective phenotypes of the slf mutant. The carpelloid stamen in slf mutant indicates that SlGT11 is required for B-function activity in the third whorl. The failed termination of floral meristem and the occurrence of floral reversion in slf indicate that part of the C-function requires SlGT11 activity in the fourth whorl. Furthermore, we find that at higher temperature, the defects of slf mutant are substantially enhanced, with petals transformed into sepals, all stamens disappeared, and the frequency of ectopic shoot/floral meristem in fourth whorl increased, indicating that SlGT11 functions in the development of the three inner floral whorls. Consistent with the observed phenotypes, it was found that B, C and an E-type MADS-box genes were in part down regulated in slf mutants.ConclusionsTogether with the spatiotemporal expression pattern, we suggest that SlGT11 functions in floral organ patterning and maintenance of floral determinacy in tomato.

Meristem Genes in the Highly Reduced Endoparasitic Pilostyles boyacensis (Apodanthaceae)

The family Apodanthaceae comprises two genera (Apodanthes and Pilostyles) and 14 endoparasitic species, all of them lacking root and shoot apical meristems, stems and leaves. Their vegetative phase is reduced to a mycelium-like endophyte formed by strands of parenchyma cells that are in close contact to the host vasculature. These plants become apparent only when their tiny gregarious flowers emerge breaking through the host cortex. The lack of vegetative meristems in these plants sharply contrasts to the typical formation of floral meristems. Our target species, Pilostyles boyacensis, provides a suitable system to investigate the evolution of meristem –related genes in an endoparasitic flowering plant without a typical vegetative shoot apical meristem. We have generated transcriptomes from two different developmental stages of the parasite (emerged floral buds and fruits), as well as a mixed sample that comprises the endophyte and growing floral buds of the parasite and its host, Dalea cuatrecasasii (Fabaceae). We specifically assessed copy number and domain conservation for the WUSCHEL HOMEOBOX, LRR-RLK-IX-a, and CLE families related to the maintenance of the shoot apical meristem (SAM) as well as the ARF and LFY families responsible for floral meristem identity. Five out of the 11 canonical gene families responsible for SAM maintenance and floral fate determinacy targeted in this study were found in Pilostyles. P. boyacensis shows at least one transcript with all functional domains conserved for BAM1, WUS, WOX9, ARF7 and LFY. Other genes implicated in the canonical regulatory network could not be found, including ARF5, ARF19, BAM2, BAM3, CLV1, and CLV3. In conclusion, the endoparasitic lifestyle of Apodanthaceae appears to correlate with a substantial reduction in the transcriptomic machinery linked to SAMs, whereas the genes involved in flower fate have remained intact.

PWR/HDA9/ABI4 Complex Epigenetically Regulates ABA Dependent Drought Stress Tolerance in Arabidopsis.

Drought stress adversely affects plant growth and development and significantly reduces crop productivity and yields. The phytohormone abscisic acid (ABA) rapidly accumulates in response to drought stress and mediates the expression of stress-responsive genes that help the plant to survive dehydration. The protein Powerdress (PWR), which interacts with Histone Deacetylase 9 (HDA9), has been identified as a critical component regulating plant growth and development, flowering time, floral determinacy, and leaf senescence. However, the role and function of PWR and HDA9 in abiotic stress response had remained elusive. Here we report that a complex of PWR and HDA9 interacts with ABI4 and epigenetically regulates drought signaling in plants. T-DNA insertion mutants of PWR and HDA9 are insensitive to ABA and hypersensitive to dehydration. Furthermore, the expression of ABA-responsive genes (RD29A, RD29B, and COR15A) is also downregulated in pwr and hda9 mutants. Yeast two-hybrid assays showed that PWR and HDA9 interact with ABI4. Transcript levels of genes that are normally repressed by ABI4, such as CYP707A1, AOX1a and ACS4, are increased in pwr. More importantly, during dehydration stress, PWR and HDA9 regulate the acetylation status of the CYP707A1, which encodes a major enzyme of ABA catabolism. Taken together, our results indicate that PWR, in association with HDA9 and ABI4, regulates the chromatin modification of genes responsible for regulation of both the ABA-signaling and ABA-catabolism pathways in response to ABA and drought stress.

Tomato locule number and fruit size controlled by natural alleles of lc and fas.

Improving yield by increasing the size of produce is an important selection criterion during the domestication of fruit and vegetable crops. Genes controlling meristem organization and organ formation work in concert to regulate the size of reproductive organs. In tomato, lc and fas control locule number, which often leads to enlarged fruits compared to the wild progenitors. LC is encoded by the tomato ortholog of WUSCHEL (WUS), whereas FAS is encoded by the tomato ortholog of CLAVATA3 (CLV3). The critical role of the WUS‐CLV3 feedback loop in meristem organization has been demonstrated in several plant species. We show that mutant alleles for both loci in tomato led to an expansion of the SlWUS expression domain in young floral buds 2–3 days after initiation. Single and double mutant alleles of lc and fas maintain higher SlWUS expression during the development of the carpel primordia in the floral bud. This augmentation and altered spatial expression of SlWUS provided a mechanistic basis for the formation of multilocular and large fruits. Our results indicated that lc and fas are gain‐of‐function and partially loss‐of‐function alleles, respectively, while both mutations positively affect the size of tomato floral meristems. In addition, expression profiling showed that lc and fas affected the expression of several genes in biological processes including those involved in meristem/flower development, patterning, microtubule binding activity, and sterol biosynthesis. Several differentially expressed genes co‐expressed with SlWUS have been identified, and they are enriched for functions in meristem regulation. Our results provide new insights into the transcriptional regulation of genes that modulate meristem maintenance and floral organ determinacy in tomato.

RNA interference suppression of AGAMOUS and SEEDSTICK alters floral organ identity and impairs floral organ determinacy, ovule differentiation, and seed-hair development in Populus.

Summary The role of the floral homeotic gene AGAMOUS (AG) and its close homologues in development of anemophilous, unisexual catkins has not previously been studied.We transformed two RNA interference (RNAi) constructs, PTG and its matrix‐attachment‐region flanked version MPG, into the early‐flowering female poplar clone 6K10 (Populus alba) to suppress the expression of its two duplicate AG orthologues.By early 2018, six out of 22 flowering PTG events and 11 out of 12 flowering MPG events showed modified floral phenotypes in a field trial in Oregon, USA. Flowers in catkins from modified events had ‘carpel‐inside‐carpel’ phenotypes. Complete disruption of seed production was observed in seven events, and sterile anther‐like organs in 10 events. Events with strong co‐suppression of both the two AG and two SEEDSTICK (STK) paralogues lacked both seeds and associated seed hairs. Alterations in all of the modified floral phenotypes were stable over 4 yr of study. Trees from floral‐modified events did not differ significantly (P < 0.05) from nonmodified transgenic or nontransgenic controls in biomass growth or leaf morphology. AG and STK genes show strong conservation of gene function during poplar catkin development and are promising targets for genetic containment of exotic or genetically engineered trees.

REBELOTE, a regulator of floral determinacy in Arabidopsis thaliana, interacts with both nucleolar andnucleoplasmic proteins.

The nucleoplasm and nucleolus are the two main territories of the nucleus. While specific functions are associated with each of these territories (such as mRNA synthesis in the nucleoplasm and ribosomal rRNA synthesis in the nucleolus), some proteins are known to be located in both. Here, we investigated the molecular function of REBELOTE (RBL), an Arabidopsis thaliana protein previously characterized as a regulator of floral meristem termination. We show that RBL displays a dual localization, in the nucleolus and nucleoplasm. Moreover, we used direct and global approaches to demonstrate that RBL interacts with nucleic acid‐binding proteins. It binds to the NOC proteins SWA2, AtNOC2 and AtNOC3 in both the nucleolus and nucleoplasm, and also to OBE1 and VFP3/ENAP1. Taking into account the identities of these RBL interactors, we hypothesize that RBL acts both in ribosomal biogenesis and in the regulation of gene expression.

Effects of low temperature and gibberellic acid on floral gene expression and floral determinacy in ‘Washington’ navel orange (Citrus sinensis L. Osbeck)

Abstract Low temperature (LT) and gibberellic acid (GA3) were used, respectively, to promote and inhibit flowering in ‘Washington’ navel orange to identify relationships between floral gene expression and floral intensity. All trees were maintained under warm temperature (WT) (24/19 °C, day/night) from April to September. For trees receiving 11 additional weeks of WT, buds expressed low levels of FLOWERING LOCUS T (FT), SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1), LEAFY (LFY), APETALA1 (AP1) and APETALA2 (AP2), but never expressed SEPALLATA1 (SEP1), PISTILLATA (PI) and AGAMOUS (AG) and did not flower. In contrast, trees transferred to LT (15/10 °C, day/night) for 8 weeks followed by WT through week 11 had significantly greater bud expression of FT and SOC1 in week 2, AP1 and AP2 during weeks 8 through 10, and SEP1, PI and AG after transfer to WT; 84% of buds flowered. Foliar-applied GA3 (50 mg L−1) in weeks 2 through 8 of LT treatment did not affect FT, SOC1 or LFY expression, but significantly reduced transcripts of AP1 by week 8, AP2, SEP1, PI and AG after transfer to WT and inflorescence number to values equal to 11-week WT-treated trees. Delaying GA3 application to weeks 4 through 8 of LT treatment increased AP1, AP2, SEP1, PI and AG expression equal to LT-treated trees and significantly greater than WT-treated trees; inflorescence number was significantly greater than trees in the WT treatment or receiving seven GA3 applications, but less than LT-treated trees. The results provide evidence suggesting that in adult citrus buds determinacy and subsequent floral development are controlled by AP1 and AP2 transcript levels, which regulate downstream floral organ identity gene activity and the effect of GA3 on citrus flower formation.

Functional Specialization of Duplicated AGAMOUS Homologs in Regulating Floral Organ Development of Medicago truncatula

The C function gene AGAMOUS (AG) encodes for a MADS-box transcription factor required for floral organ identity and floral meristem (FM) determinacy in angiosperms. Unlike Arabidopsis, most legume plants possess two AG homologs arose by an ancient genome duplication event. Recently, two euAGAMOUS genes, MtAGa and MtAGb, were characterized and shown to fulfill the C function activity in the model legume Medicago truncatula. Here, we reported the isolation and characterization of a new mtaga allele by screening the Medicago Tnt1 insertion mutant collection. We found that MtAGa was not only required for controlling the stamen and carpel identity but also affected pod and seed development. Genetic analysis indicated that MtAGa and MtAGb redundantly control Medicago floral organ identity, but have minimal distinct functions in regulating stamen and carpel development in a dose-dependent manner. Interestingly, the stamens and carpels are mostly converted to numerous vexillum-like petals in the double mutant of mtaga mtagb, which is distinguished from Arabidopsis ag. Further qRT-PCR analysis in different mtag mutants revealed that MtAGa and MtAGb can repress the expression of putative A and B function genes as well as MtWUS, but promote putative D function genes expression in M. truncatula. In addition, we found that the abnormal dorsal petal phenotype observed in the mtaga mtagb double mutant is associated with the upregulation of CYCLOIDEA (CYC)-like TCP genes. Taken together, our data suggest that the redundant MtAGa and MtAGb genes of M. truncatula employ a conserved mechanism of action similar to Arabidopsis in determining floral organ identity and FM determinacy but may have evolved distinct function in regulating floral symmetry by coordinating with specific floral dorsoventral identity factors.

REBELOTE, a regulator of floral determinacy in Arabidopsis thaliana, interacts with both nucleolar and nucleoplasmic proteins

REBELOTE, a regulator of floral determinacy in Arabidopsis thaliana, interacts with both nucleolar and nucleoplasmic proteins

The sweet cherry (Prunus avium) FLOWERING LOCUS T gene is expressed during floral bud determination and can promote flowering in a winter-annual Arabidopsis accession.

FT gene is expressed in leaves and buds and is involved in floral meristem determination and bud development in sweet cherry. In woody fruit perennial trees, floral determination, dormancy and bloom, depends on perception of different environmental and endogenous cues which converge to a systemic signaling gene known as FLOWERING LOCUS T (FT). In long-day flowering plants, FT is expressed in the leaves on long days. The protein travels through the phloem to the shoot apical meristem, where it induces flower determination. In perennial plants, meristem determination and flowering are separated by a dormancy period. Meristem determination takes place in summer, but flowering occurs only after a dormancy period and cold accumulation during winter. The roles of FT are not completely clear in meristem determination, dormancy release, and flowering in perennial plants. We cloned FT from sweet cherry (Prunus avium) and analyzed its expression pattern in leaves and floral buds during spring and summer. Phylogenetic analysis shows high identity of the FT cloned sequence with orthologous genes from other Rosaceae species. Our results show that FT is expressed in both leaves and floral buds and increases when the daylight reached 12h. The peak in FT expression was coincident with floral meristem identity genes expression and morphological changes typical of floral meristem determination. The Edi-0 Arabidopsis ecotype, which requires vernalization to flower, was transformed with a construct for overexpression of PavFT. These transgenic plants showed an early-flowering phenotype without cold treatment. Our results suggest that FT is involved in floral meristem determination and bud development in sweet cherry. Moreover, we show that FT is expressed in both leaves and floral buds in this species, in contrast to annual plants.