Petaloid Stamens Research Articles

Selection of Stable Reference Genes for QRT-PCR in Tree Peony 'Doulv' and Functional Analysis of PsCUC3.

(1) Background: Tree peonies display extensive cultivar diversity due to widespread hybridization, resulting in a complex genetic architecture. This complexity complicates the selection of universal reference genes across different cultivars for qRT-PCR analyses. Paeonia suffruticosa 'Doulv', notable for its unique green blooms in China, exhibits chlorosis post-flowering and features petaloid stamens and pistils. (2) Methods: Based on published literature and RNA-seq data from 'Doulv', nine candidate reference genes-ACT (Actin), TUB (β-Tubulin), UBC (Ubiquitin Conjugating Enzyme), UBQ (Ubiquitin), UPL (Ubiquitin Protein Ligase), PP2A (Protein Phosphatase 2A), PP2C (Protein Phosphatase 2C), MBF1A (Multiprotein Bridging Factor 1A), and GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase)-were selected. Their expression stability was assessed across various tissues and developmental stages of 'Doulv' flowers using qRT-PCR, with evaluations conducted via GeNorm_v3.5, NormFinder_v20, and BestKeeper_v1.0. Gene cloning and expression analyses of PsCUC3, including its subcellular localization, were performed. (3) Results: GAPDH and ACT were identified as the most stable reference genes in petaloid stamens across various developmental stages of 'Doulv', whereas UBC and MBF1A were optimal across different tissues. Notably, specific conserved amino acids in PsCUC3 from 'Doulv' diverged from those in NAM/CUC3 proteins of other species, impacting its protein structure. PsCUC3 expression analysis revealed no correlation with chlorophyll content in petaloid stamens but an association with petaloid organ development. Furthermore, PsCUC3 was predominantly localized in the nucleus. (4) Conclusions: This study comprehensively evaluated suitable reference genes using GeNorm_v3.5, NormFinder_v20, and BestKeeper_v1.0 software, establishing a robust qRT-PCR detection system for 'Doulv' peony. These results provide a solid experimental foundation for further research on 'Doulv' peony. Building on this experimental foundation, the functional analysis of the PsCUC3 gene was conducted. The findings suggest a potential association between the PsCUC3 gene and floral morphology alterations in 'Doulv', identifying PsCUC3 as crucial for understanding the molecular mechanisms influencing floral structure in tree peonies.

The transcription factor RhMYB17 regulates the homeotic transformation of floral organs in rose (Rosa hybrida) under cold stress.

Low temperatures affect flower development in rose (Rosa hybrida), increasing petaloid stamen number and reducing normal stamen number. We identified the low-temperature-responsive R2R3-MYB transcription factor RhMYB17, which is homologous to Arabidopsis MYB17 by similarity of protein sequences. RhMYB17 was up-regulated at low temperatures, and RhMYB17 transcripts accumulated in floral buds. Transient silencing of RhMYB17 by virus-induced gene silencing decreased petaloid stamen number and increased normal stamen number. According to the ABCDE model of floral organ identity, class A genes APETALA 1 (AP1) and AP2 contribute to sepal and petal formation. Transcription factor binding analysis identified RhMYB17 binding sites in the promoters of rose APETALA 2 (RhAP2) and APETALA 2-LIKE (RhAP2L). Yeast one-hybrid assays, dual-luciferase reporter assays, and electrophoretic mobility shift assays confirmed that RhMYB17 directly binds to the promoters of RhAP2 and RhAP2L, thereby activating their expression. RNA sequencing further demonstrated that RhMYB17 plays a pivotal role in regulating the expression of class A genes, and indirectly influences the expression of the class C gene. This study reveals a novel mechanism for the homeotic transformation of floral organs in response to low temperatures.

Genome-Wide Identified MADS-Box Genes in Prunus campanulata 'Plena' and Theirs Roles in Double-Flower Development.

The MADS-box gene family plays key roles in flower induction, floral initiation, and floral morphogenesis in flowering plants. To understand their functions in the double-flower formation of Prunus campanulata 'Plena' (hereafter referred to as PCP), which is an excellent flowering cherry cultivar, we performed genome-wide identification of the MADS-box gene family. In this study, 71 MADS-box genes were identified and grouped into the Mα, Mβ, Mγ and MIKC subfamilies according to their structures and phylogenetic relationships. All 71 MADS-box genes were located on eight chromosomes of PCP. Analysis of the cis-acting elements in the promoter region of MADS-box genes indicated that they were associated mainly with auxin, abscisic acid, gibberellin, MeJA (methyl jasmonate), and salicylic acid responsiveness, which may be involved in floral development and differentiation. By observing the floral organ phenotype, we found that the double-flower phenotype of PCP originated from petaloid stamens. The analysis of MIKC-type MADS-box genes in PCP vegetative and floral organs by qRT-PCR revealed six upregulated genes involved in petal development and three downregulated genes participating in stamen identity. Comparative analysis of petaloid stamens and normal stamens also indicated that the expression level of the AG gene (PcMADS40) was significantly reduced. Thus, we speculated that these upregulated and downregulated genes, especially PcMADS40, may lead to petaloid stamen formation and thus double flowers. This study lays a theoretical foundation for MADS-box gene identification and classification and studying the molecular mechanism underlying double flowers in other ornamental plants.

Transcriptomic and proteomic analyses reveal changes in the metabolic pathways of Paeonia lactiflora petaloid stamens

Paeonia lactiflora or herbaceous peony, a famous traditional Chinese flower, is popular around the world for its cut flowers. Petaloid stamens are a common form of double flowers, but their underlying molecular mechanisms during development are still unclear. In this study, using the petaloid stamens of P. lactiflora ‘Lian Tai’ as research material, transcriptomic and proteomic analyses of flower buds at three developmental stages (stamen primordium, stamen primordium partly petaloid, and stamen primordium completely petaloid) were performed. A total of 81,971 unigenes were identified in the transcriptome, 8,288 proteins were identified in the proteome, and 8,152 genes and proteins were jointly identified in the transcriptome and proteome. GO annotation and KEGG enrichment analysis of differentially expressed proteins indicated that the metabolic pathways related to polysaccharides, carbohydrates, starch, sucrose, fructose, mannose, and photosynthesis were significantly enriched in petaloid stamens. After removing duplicates, 85 differentially expressed genes/proteins were identified in these metabolic pathways. These significantly enriched metabolic pathways and differentially expressed genes/proteins may play an important role in physiological and developmental processes associated with P. lactiflora petaloid stamens. This study provides additional transcriptomic and proteomic reference points for further exploring the molecular mechanisms during the development of P. lactiflora petaloid stamens.

Morphogenesis of Stamens and Petaloid Stamens in Lilium hybrid ‘Red Twin’ under Different Temperatures and the Expression Characteristics of Two AGAMOUS-like Genes Linked to These Processes

The double-flowered lily ‘Red Twin’, in which stamens are transformed into petaloid organs, sometimes exhibits normal stamens owing to an unknown mechanism and thus greatly affecting its commercial quality. In this study, the morphogenesis of stamens and petaloid stamens in this cultivar grown under different temperature treatments were investigated. Two AGAMOUS-like genes were isolated and their expression levels were analyzed. The results showed that relatively high temperatures induced the morphogenesis of stamens, while relatively low temperatures promoted petaloidy in ‘Red Twin’. The stage with 1–6 mm flower buds was identified as the critical development period for stamen morphogenesis; furthermore, keeping the flower buds under relatively low temperatures in the 1–3 or 3–6 mm stages would be sufficient for the formation of petaloid stamens to a high degree. In addition, LrtAG1 and LrtAG2 showed the highest expression level in whorls 3 and 4 of 3–6 mm flower buds, respectively. LrtAG1 showed a higher reduction ratio than LrtAG2 when the flower buds were transferred to low temperature, implying that the decreased LrtAG1 expression level is associated with low temperature and might be correlated with the petaloidy of the stamens.

Genomic basis of the giga-chromosomes and giga-genome of tree peony Paeonia ostii

Tree peony (Paeonia ostii) is an economically important ornamental plant native to China. It is also notable for its seed oil, which is abundant in unsaturated fatty acids such as α-linolenic acid (ALA). Here, we report chromosome-level genome assembly (12.28 Gb) of P. ostii. In contrast to monocots with giant genomes, tree peony does not appear to have undergone lineage-specific whole-genome duplication. Instead, explosive LTR expansion in the intergenic regions within a short period (~ two million years) may have contributed to the formation of its giga-genome. In addition, expansion of five types of histone encoding genes may have helped maintain the giga-chromosomes. Further, we conduct genome-wide association studies (GWAS) on 448 accessions and show expansion and high expression of several genes in the key nodes of fatty acid biosynthetic pathway, including SAD, FAD2 and FAD3, may function in high level of ALAs synthesis in tree peony seeds. Moreover, by comparing with cultivated tree peony (P. suffruticosa), we show that ectopic expression of class A gene AP1 and reduced expression of class C gene AG may contribute to the formation of petaloid stamens. Genomic resources reported in this study will be valuable for studying chromosome/genome evolution and tree peony breeding.

Candidate genes screening based on phenotypic observation and transcriptome analysis for double flower of Prunus mume

BackgroundPrunus mume is an early spring flower of Rosaceae, which owns high application value in gardens. Being an excellent ornamental trait, the double flower trait has always been one of the important breeding goals of plant breeders. However, the key regulatory genes of double flower traits of P. mume are still unclear at present.ResultsThe floral organs’ morphological differences of 20 single and 20 double flower cultivars of P. mume were compared firstly. And it was found that double flower trait of P. mume were often accompanied by petaloid stamen, multiple carpels and an increase in the total number of floral organs. Then, transcriptome sequencing of two representative cultivars P. mume ‘Danban Lve’ and P. mume ‘Xiao Lve’ were conducted at 3 Stage of flower bud development with distinct morphological differentiation. 3256 differentially expression genes (DEGs) were detected, and 20 candidate genes for double flower trait of P. mume were screened out including hub genes PmAP1–1 and PmAG-2 based on DEGs function analysis and WGCNA analysis. And it was found that epigenetic and hormone related genes may also play an important role in the process of double flower.ConclusionsThis study suggested that the double flower trait of P.mume is more like accumulation origin based on morphological observation. 20 genes and co-expression network related to the formation of double flower P. mume were preliminarily screened through transcriptomics analysis. The results provided a reference for further understanding of the molecular mechanism of double flower trait in P. mume.

Disruption of transcription factor RhMYB123 causes the transformation of stamen to malformed petal in rose (Rosa hybrida)

We find that the R2R3 MYB transcription factor RhMYB123 has a novel function to regulate stamen-petal organ specification in rose. Rose is one of the ornamental plants with economic importance worldwide. Malformed flower seriously affects the ornamental value and fertility of rose. However, the regulatory mechanism is largely unknown. In this work, we identified a R2R3 MYB transcription factor RhMYB123 from rose, the expression of which significantly decreased from flower differentiation stage to floral organ development stage. Phylogenetic analysis indicated that it belongs to the same subgroup as MYB123 of Arabidopsis and located in nucleus. In addition, RhMYB123 was confirmed to have transcriptional activation function by dual luciferase assay. Silencing RhMYB123 using Virus-Induced Gene Silencing (VIGS) in rose could increase the number of malformed petaloid stamen. Furthermore, we identified 549 differential expressed genes (DEGs) in TRV-RhMYB123 lines compared to TRV controls by RNA-seq of floral buds (flower differentiation stage). Among of those genes, expression of 3 MADS box family genes related to floral organ development reduced in TRV-RhMYB123 lines, including AGAMOUS (RhAG), AGAMOUS LIKE 15 (RhAGL15), and SHATTERPROOF 1 (RhSHP1). Given, previous studies have shown that auxin plays a crucial role in floral meristem initiation and stamen-petal organ specification. We also found 6 DEGs were involved in auxin signal transduction, of which five were reduced expression in TRV-RhMYB123. Taken together, our findings suggested that RhMYB123 may govern the development of malformed petaloid stamen by regulating the expressions of some MADS box family members and auxin signaling pathway elements.

Transcriptome profiling during double-flower development provides insight into stamen petaloid in cultivated <i>Lilium</i>

Asiatic hybrid lilies (<italic>Lilium</italic> spp.), as the biggest cultivar groups of the lily variety are an ornamental plant with elegant floral patterns and bright colors. We discovered a double-flower Asiatic hybrid cultivar ‘Annemarie’s Dream’ whose stamens convert into petaloid stamens in varying degrees. Double-flower is a significant ornamental trait of the flower organ. However, the molecular mechanism of stamen petaloid formation has not been widely studied in Asiatic hybrid lily. Therefore, we used RNA-seq to contrast transcriptomes of stamen, petaloid stamen and inner tepal at two developmental stages. In total, 190,488 unigenes were obtained and 37,549 differentially expressed genes (DEGs) were identified. We focused on DEGs involved in phytohormone signaling and transcription factor regulation, especially the MADS-box genes (A-class gene<italic> LiAP1</italic>; B-class gene <italic>LiPI</italic>; C-class gene <italic>LiAG</italic>; E-class gene <italic>LiAGL6</italic>, <italic>LiSEP3</italic>). Furthermore, we performed weight gene co-expression network analysis (WGCNA) and identified two co-expressed MADS-box homeotic genes (<italic>LiAG</italic>, CL14315.Contig2_All; <italic>LiAP1</italic>, CL10060.Contig3_All) as hubs. We also found that CL3014.Contig2_All (<italic>PYL</italic>) and CL5627.Contig1_All (<italic>GID2</italic>) as phytohormone-related genes may participate in the regulation of the stamen petaloid during double-flower development. In summary, our findings provide an insight into the molecular regulatory network underlying stamen petaloid and thereby offering a theoretical basis for double-flower breeding in <italic>Lilium</italic>.

Expression level of B- and C-class MADS-box genes is associated with the petaloidy of stamens in cultivated amaryllis (Hippeastrum hybridum)

ABSTRACT One of the most important attributes of Hippeastrum hybridum (amaryllis) flower is its double-flower trait, which is partly derived from the petaloidy of its stamens. Three types of floral organs in whorl 3 of double flowers from H. hybridum cv. Jewel, namely normal stamens, partly petaloid stamens, and completely petaloid stamens, were sampled for RNA-seq analysis to identify candidate genes related to the petaloidy of stamens. 12,584 differentially expressed genes between normal stamens and petaloid stamens were obtained, in which several MADS-box genes and a series of genes related to morphogenesis, pigment, and cell wall development were identified. Quantitative real-time PCR analysis of nine MADS-box genes clustered to the ABCE-class was performed in three cultivars ‘Jewel’, ‘Merry Christmas’, and ‘Red Nymph’. Consistent with the transcriptome data, higher expression of the B-class genes HhAP3.2 and HhPI1, but lower expression of the C-class gene HhAG1 was observed in petaloid stamens than those in normal stamens, and this trend was enhanced with an increase in the degree of petaloidy. These results suggest that the expression level of B- and C-class MADS-box genes is associated with the petaloidy of stamens in H. hybridum, which provide candidate genes for double-flower breeding in H. hybridum.

Expression and Functional Analyses of Nymphaea caerulea MADS-Box Genes Contribute to Clarify the Complex Flower Patterning of Water Lilies.

Nymphaeaceae are early diverging angiosperms with large flowers characterized by showy petals and stamens not clearly whorled but presenting a gradual morphological transition from the outer elements to the inner stamens. Such flower structure makes these plant species relevant for studying flower evolution. MADS-domain transcription factors are crucial components of the molecular network that controls flower development. We therefore isolated and characterized MADS-box genes from the water lily Nymphaea caerulea. RNA-seq experiments on floral buds have been performed to obtain the transcript sequences of floral organ identity MADS-box genes. Maximum Likelihood phylogenetic analyses confirmed their belonging to specific MADS-box gene subfamilies. Their expression was quantified by RT-qPCR in all floral organs at two stages of development. Protein interactions among these transcription factors were investigated by yeast-two-hybrid assays. We found especially interesting the involvement of two different AGAMOUS-like genes (NycAG1 and NycAG2) in the water lily floral components. They were therefore functionally characterized by complementing Arabidopsis ag and shp1 shp2 mutants. The expression analysis of MADS-box genes across flower development in N. caerulea described a complex scenario made of numerous genes in numerous floral components. Their expression profiles in some cases were in line with what was expected from the ABC model of flower development and its extensions, while in other cases presented new and interesting gene expression patterns, as for instance the involvement of NycAGL6 and NycFL. Although sharing a high level of sequence similarity, the two AGAMOUS-like genes NycAG1 and NycAG2 could have undergone subfunctionalization or neofunctionalization, as only one of them could partially restore the euAG function in Arabidopsis ag-3 mutants. The hereby illustrated N. caerulea MADS-box gene expression pattern might mirror the morphological transition from the outer to the inner floral organs, and the presence of transition organs such as the petaloid stamens. This study is intended to broaden knowledge on the role and evolution of floral organ identity genes and the genetic mechanisms causing biodiversity in angiosperm flowers.

Elucidating Mitochondrial DNA Markers of Ogura-Based CMS Lines in Indian Cauliflowers (Brassica oleracea var. botrytis L.) and Their Floral Abnormalities Due to Diversity in Cytonuclear Interactions.

Mitochondrial markers can be used to differentiate diverse mitotypes as well as cytoplasms in angiosperms. In cauliflower, cultivation of hybrids is pivotal in remunerative agriculture and cytoplasmic male sterile lines constitute an important component of the hybrid breeding. In diversifying the source of male sterility, it is essential to appropriately differentiate among the available male sterile cytoplasms in cauliflower. PCR polymorphism at the key mitochondrial genes associated with male sterility will be instrumental in analyzing, molecular characterization, and development of mitotype-specific markers for differentiation of different cytoplasmic sources. Presence of auto- and alloplasmic cytonuclear combinations result in complex floral abnormalities. In this context, the present investigation highlighted the utility of organelle genome-based markers in distinguishing cytoplasm types in Indian cauliflowers and unveils the epistatic effects of the cytonuclear interactions influencing floral phenotypes. In PCR-based analysis using a set of primers targeted to orf-138, 76 Indian cauliflower lines depicted the presence of Ogura cytoplasm albeit the amplicons generated exhibited polymorphism within the ofr-138 sequence. The polymorphic fragments were found to be spanning over 200–280 bp and 410–470 bp genomic regions of BnTR4 and orf125, respectively. Sequence analysis revealed that such cytoplasmic genetic variations could be attributed to single nucleotide polymorphisms and insertion or deletions of 31/51 nucleotides. The cytoplasmic effects on varying nuclear-genetic backgrounds rendered an array of floral abnormalities like reduction in flower size, fused flowers, splitted style with the exposed ovule, absence of nonfunctional stamens, and petaloid stamens. These floral malformations caused dysplasia of flower structure affecting female fertility with inefficient nectar production. The finding provides an important reference to ameliorate understanding of mechanism of cytonuclear interactions in floral organ development in Brassicas. The study paves the way for unraveling developmental biology of CMS phenotypes in eukaryotic organisms and intergenomic conflict in plant speciation.

Comparative transcriptomics and WGCNA reveal candidate genes involved in petaloid stamens in Paeonia lactiflora

ABSTRACT Petaloid stamens in Paeonia lactiflora constitute an important developmental phenomenon that explain double flower formation. Although research on stamen development in model plants has progressed considerably, the underlying molecular mechanism in P. lactiflora petaloid stamens is still unclear. In this study, acomparative transcriptomic analysis was performed on two cultivars of P. lactiflora (‘Fen Yu Nu’ and ‘Lian Tai’) with different stamen developmental patterns. Following transcriptome sequencing, 89,393 unigenes were identified in P. lactiflora. Trend analysis and weighted gene co-expression network analysis (WGCNA) indicated that 18 candidate genes were likely involved in the formation and development of petaloid stamens, including seven MADS-box genes, PlAP3, PlDEFA, PlPI2, PlAG-1, PlSEP3, PlSEP1-1, and PlSEP1-2, and 11 other transcription factors (TFs), PlTCP2, PlTCP4, PlTCP9, PlbHLH36, PlICE1, PlLBD38, PlNAC083, PlBLH11, PlPDF2, PlGBF1, and PlIIIA. Using these 18 selected candidate genes, we propose ahypothetical model of gene expression network that regulates the development of petaloid stamens. Our results provide acollection of candidate genes that are useful for the analysis of P. lactiflora petaloid stamens, allowing for in-depth studies of the development pattern of P. lactiflora flower organs, and providing atheoretical basis for related research on petaloid stamens of flowers in other herbaceous plants.

Homeotic transformation from stamen to petal in Eriobotrya japonica is associated with hormone signal transduction and reduction of the transcriptional activity of EjAG.

Double-flower loquat (Eriobotrya japonica) is a new germplasm with homeotic transformation of stamen into petal in whorl 3. However, little information is available on the molecular mechanism of this transformation. Herein, we analyzed the transcriptome, candidate genes and endogenous hormones to investigate the mechanisms underlying this homeotic transformation. Some transcription factors, such as MADS-box, TCP and MYB, were significantly differentially expressed. Importantly, we confirmed that one of these (DN39625_c0_g1), which encoded a C-class floral homeotic protein referred to as AGAMOUS ortholog (EjAG), was significantly downregulated. Subcellular localization of EjAG was found to be in the nucleus. Ectopic expression of EjAG rescued the development of stamens and carpels from the double-flower phenotype in an Arabidopsis ag mutant, suggesting that EjAG expression is associated with double-flower formation. Meanwhile, enrichment analyses showed that the differentially expressed genes (DEGs) were mainly involved in the metabolic pathways of hormone signal transduction. The DEGs of auxin, gibberellin A (GA) and cytokinin signaling pathways were mainly upregulated. However, the DEGs of abscisic acid (ABA) and the ethylene signaling pathway were mainly downregulated. Accordingly, the concentrations of indoleacetic acid, kinetin and GA3 were high at the petaloid stamen stage, but the ABA concentration remained low. The identified genes and pathways provide abundant sequence resources for studying the mechanisms underlying the homeotic transformation in loquat and other Rosaceae species.

Transcriptome analysis during floral organ development provides insights into stamen petaloidy in Lagerstroemia speciosa

As one of the most popular woody species that blooms in summer, Lagerstroemia speciosa has been used abundantly in urban landscape for its excellent floral beauty. For the first time, we discovered a double-flower variant with all petaloid stamens. To understand the molecular basis of this variation, we contrasted the transcriptomes of single- and double-flower buds at three stamen development stages. In total, 73,536 unigenes were mapped and 30,714 differently expressed genes (DEGs) were identified in the tissues. We focused on the DEGs expressing in both phenotypes and investigated the association of their expression profiles with their functions in transcription pathways. Furthermore, we performed WGCNA and identified co-expressed genes with four floral homeotic genes as hubs (MADS16, Unigene0026169; AP2, Unigene0042732; SOC1, Unigene0046314; AG, Unigene0056437). The expression of these hub genes has been conserved across the three developmental stages but significantly different between the two floral phenotypes. As a result, the robust transcriptional regulation of stamen petaloidy in double flowers was deduced. These findings will help to unravel the regulatory mechanisms of several specific genes, thereby providing a basis to study double-flower molecular breeding in L. speciosa.

An APETALA2 Homolog, RcAP2, Regulates the Number of Rose Petals Derived From Stamens and Response to Temperature Fluctuations.

Rosa chinensis, which is a famous traditional flower in China, is a major ornamental plant worldwide. Long-term cultivation and breeding have resulted in considerable changes in the number of rose petals, while most wild Rosaceae plants have only one whorl consisting of five petals. The petals of double flowers reportedly originate from stamens, but the underlying molecular mechanism has not been fully characterized. In this study, we observed that the number of petals of R. chinensis ‘Old Blush’ flowers increased and decreased in response to low- and high-temperature treatments, respectively, similar to previous reports. We characterized these variations in further detail and found that the number of stamens exhibited the opposite trend. We cloned an APETALA2 homolog, RcAP2. A detailed analysis of gene structure and promoter cis-acting elements as well as RcAP2 temporospatial expression patterns and responses to temperature changes suggested that RcAP2 expression may be related to the number of petals from stamen origin. The overexpression of RcAP2 in Arabidopsis thaliana transgenic plants may induce the transformation of stamens to petals, thereby increasing the number of petals. Moreover, silencing RcAP2 in ‘Old Blush’ plants decreased the number of petals. Our results may be useful for clarifying the temperature-responsive mechanism involved in petaloid stamen production, which may be relevant for the breeding of new rose varieties with enhanced flower traits.

Functional Conservation of an AGAMOUS Orthologous Gene Controlling Reproductive Organ Development in the Gymnosperm Species Taxus chinensis var. mairei

Arabidopsis AGAMOUS (AG) has roles in specifying reproductive organ (stamens and carpels) identity, floral meristem determinacy, and repression of A-function. To investigate possible roles of AG orthologous genes in gymnosperm species and evolution of C function, we isolated and identified AG orthologous gene TcAG from Taxus chinensis var. mairei (family Taxaceae, order Coniferales), a member of the last divergant lineage from higher Conifer that sisters to Gnetales. Sequence alignment and phylogenetic analysis grouped TcAG into the gymnosperm AG lineage. TcAG was expressed in both developing male and female cones, but there was no expression in juvenile leaves. Ectopic expression of TcAG in an Arabidopsis ag mutant produced flowers with the third whorl petaloid stamen and fourth whorl normal carpel, but failed to convert first whorl sepals into carpeloid organs and second whorl petals into stamenoid organs. A 35S::TcAG transgenic Arabidopsis ag mutant had very early flowering, and produced a misshapen inflorescence with a shortened floral axis. Our results suggest that establishment of the complete C-function occurred gradually during AG lineage evolution even in gymnosperms.

RNA-Seq analysis reveals the distinctive adaxial-abaxial polarity in the asymmetric one-theca stamen of Canna indica.

Proper establishment of adaxial-abaxial polarity is essential for the development of lateral organs, while former researches were mostly focused on the polarity regulation in leaves, and little is known in stamens, especially in the asymmetric ones. Canna indica (Zingiberales: Cannaceae) is a widely cultivated ornamental plant and the representative species to study the evolutionary development of Zingiberales. The androecium of Canna indica comprises 3-4 petaloid staminodes and a fertile stamen (FS), which consists of a one-theca anther and a petaloid appendage. The partially petaloid stamen is considered as an intermediate state organ from a two-thecae stamen to a completely petaloid staminode. Using RNA-Seq, we quantified the expressions of the transcripts in anther and petaloid appendage, and detected 64,430 and 57,041 unigenes in these two organs, respectively. 4574 unigenes were down-regulated, and 3525 were up-regulated in petaloid appendage compared with those in anther. GO enrichment analysis indicated that the function of cytokinin is more related to cell differentiation in anther, while auxin is more to cell division in petaloid appendage. B- and C-class floral homeotic genes were expressed in these two androecium parts. Most of the class III HD-ZIP family members, which specify adaxial identity, were expressed lower in petaloid appendage than in anther; while KANADIs and YABBYs, which promote abaxial identity, exhibited opposite expression patterns. In situ hybridization showed that the adaxial marker gene was mainly expressed in the region between the two protrusions of the anther, while the abaxial marker was mainly expressed in petaloid appendage. We hypothesize that the adaxial-abaxial polarity participates in the distinctive anther-petaloid appendage patterning within the asymmetric FS of Canna indica.

Low temperature-induced DNA hypermethylation attenuates expression of RhAG, an AGAMOUS homolog, and increases petal number in rose (Rosa hybrida).

BackgroundFlower development is central to angiosperm reproduction and is regulated by a broad range of endogenous and exogenous stimuli. It has been well documented that ambient temperature plays a key role in controlling flowering time; however, the mechanisms by which temperature regulates floral organ differentiation remain largely unknown.ResultsIn this study, we show that low temperature treatment significantly increases petal number in rose (Rosa hybrida) through the promotion of stamen petaloidy. Quantitative RT-PCR analysis revealed that the expression pattern of RhAG, a rose homolog of the Arabidopsis thaliana AGAMOUS C-function gene, is associated with low temperature regulated flower development. Silencing of RhAG mimicked the impact of low temperature treatments on petal development by significantly increasing petal number through an increased production of petaloid stamens. In situ hybridization studies further revealed that low temperature restricts its spatial expression area. Analysis of DNA methylation level showed that low temperature treatment enhances the methylation level of the RhAG promoter, and a specific promoter region that was hypermethylated at CHH loci under low temperature conditions, was identified by bisulfite sequencing. This suggests that epigenetic DNA methylation contributes to the ambient temperature modulation of RhAG expression.DiscussionOur results provide highlights in the role of RhAG gene in petal number determination and add a new layer of complexity in the regulation of floral organ development.ConclusionsWe propose that RhAG plays an essential role in rose flower patterning by regulating petal development, and that low temperatures increase petal number, at least in part, by suppressing RhAG expression via enhancing DNA CHH hypermethylation of the RhAG promoter.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0623-1) contains supplementary material, which is available to authorized users.

Nectarios septales, superficies nectaríferas, coléteres, y aspectos relacionados con la polinización en "Canna lineata" Ciciarelli ("Cannaceae"-Zingiberales)

This manuscript encompasses part of a study carried out on the structure and anatomy of flowers in C. lineata . Glandular nectariferous epidermal surfaces, nectar guides of androecium pieces, septal nectaries and colleters of the ovary have been described for the first time. Rows of capitate papillae form nectar guides of labellum whereas convexed and flattened papillae constitute staminodes and petaloid stamen. Prenectar has been tested in papillae of all androecium pieces where nectar secretion starts in phloem of amphicribal vascular bundles and associate parenchyma to be secreted through cuticular pores of epidermis. In septal nectaries, 1-3 layers of parenchyma and numerous cylindrical tricomes narrowed in the apex secrete nectar towards the channels. The fluid is stored in the nectar chamber of the floral tube upon the ovary. Analysis of the chemical composition of nectar allowed us to confirm ornitophyly in C. indica and entomophily in C. lineata . Pollination traits are discussed and it is proposed that other species of Canna with nectariferous flowers of wide and flattened labellum which attract both insects and birds, would conform a mixed floral sindrome. External wall of the ovary is composed by many colleters of the claviform type, it is proposed that they would function as protective structures of ovules and subsequently of the developing seeds. Studies have been carried out with fresh flowers under light microscope and SEM.