Flower Organ Development Research Articles

Down-regulation of AUXIN RESPONSE FACTORS 6 and 8 by microRNA 167 leads to floral development defects and female sterility in tomato.

Auxin regulates the expression of diverse genes that affect plant growth and development. This regulation requires AUXIN RESPONSE FACTORS (ARFs) that bind to the promoter regions of these genes. ARF6 and ARF8 in Arabidopsis thaliana are required to promote inflorescence stem elongation and late stages of petal, stamen, and gynoecium development. All seed plants studied thus far have ARF6 and ARF8 orthologues as well as the microRNA miR167, which targets ARF6 and ARF8. Whether these genes have broadly conserved roles in flower development is not known. To address this question, the effects of down-regulation of ARF6 and ARF8 were investigated through transgenic expression of Arabidopsis MIR167a in tomato, which diverged from Arabidopsis before the radiation of dicotyledonous plants approximately 90-112 million years ago. The transgenic tomato plants overexpressing MIR167a exhibited reductions in leaf size and internode length as well as shortened petals, stamens, and styles. More significantly, the transgenic plants were female-sterile as a result of failure of wild-type pollen to germinate on the stigma surface and/or to grow through the style. RNA-Seq analysis identified many genes with significantly altered expression patterns, including those encoding products with functions in 'transcription regulation', 'cell wall' and 'lipid metabolism' categories. Putative orthologues of a subset of these genes were also differentially expressed in Arabidopsis arf6 arf8 mutant flowers. These results thus suggest that ARF6 and ARF8 have conserved roles in controlling growth and development of vegetative and flower organs in dicots.

Overexpression of a novel chrysanthemum SUPERMAN-like gene in tobacco affects lateral bud outgrowth and flower organ development

Previous studies have shown that the SUP genes play important roles in flower development and plant growth and morphogenesis. In this study, we isolated and characterized a SUPERMAN-like gene DgSZFP from chrysanthemum. DgSZFP contains one conserved Cys2/His2-type zinc finger motifs in the N-terminal region and an EAR-box in C-terminus. Its expression was significantly higher in nodes, flower buds, disc stamens, and petals than in the other tissues. Overexpression of DgSZFP in tobacco resulted in enhanced branching, reduced plant height, increased the width of petal tubes, produced the staminoid petals and petaloid stamens in flowers, and enhanced the seed weight and size. In addition, DgSZFP-overexpression tobacco plants accumulated high concentrations of cytokinin and chlorophyll. These results suggest that DgSZFP may be the candidate gene for regulating branching and floral organ development in chrysanthemum.

Identification of floral genes for sex determination in Calamus palustris Griff. by using suppression subtractive hybridization.

Calamus palustris Griff. is an economically important dioecious rattan species in Southeast Asia. However, dioecy and onset of flowering at 3-4 years old render uncertainties in desired female:male seedling ratios to establish a productive seed orchard for this rattan species. We constructed a subtractive library for male floral tissue to understand the genetic mechanism for gender determination in C. palustris. The subtractive library produced 1536 clones with 1419 clones of high quality. Reverse Northern screening showed 313 clones with differential expression, and sequence analyses clustered them into 205 unigenes, including 32 contigs and 173 singletons. The subtractive library was further validated with reverse transcription-quantitative polymerase chain reaction analysis. Homology identification classified the unigenes into 12 putative functional proteins with 83% unigenes showing significant match to proteins in databases. Functional annotations of these unigenes revealed genes involved in male flower development, including MADS-box genes, pollen-related genes, phytohormones for flower development, and male flower organ development. Our results showed that the male floral genes may play a vital role in sex determination in C. palustris. The identified genes can be exploited to understand the molecular basis of sex determination in C. palustris.

QnAs with Detlef Weigel

In his scientific career, National Academy of Sciences member Detlef Weigel has gone from studying how fruit flies and flowers develop to investigating how plants adapt to different environments and the genomic variation that allows them to do so. Director of the Department of Molecular Biology at the Max Planck Institute for Developmental Biology, Weigel has also made crucial contributions to studies of the evolution of the plant immune system and the role of microRNAs in plant development. Weigel’s use of genomics technologies to study the model plant Arabidopsis thaliana has led to a nuanced understanding of the variation in plant genomes, and could help prevent diseases and increase yield in crop plants. Detlef Weigel. Image courtesy of Anne Faden, MPI for Developmental Biology. > PNAS:What led you to study so many different subjects during your career? > Weigel:When I discover a new problem that I haven’t thought of before, I often end up pursuing it, especially when few others are already working on it. When I initially began studying Arabidopsis the idea was to look at the development of flower organs, but instead I became fascinated with the step before that, the actual initiation of the flower. After I set up my …

Phylogenetic Analysis of Brassica rapa MATH-Domain Proteins

The MATH (meprin and TRAF-C homology) domain is a fold of seven anti-parallel β-helices involved in protein-protein interaction. Here, we report the identification and characterization of 90 MATH-domain proteins from the Brassica rapa genome. By sequence analysis together with MATH-domain proteins from other species, the B. rapa MATH-domain proteins can be grouped into 6 classes. Class-I protein has one or several MATH domains without any other recognizable domain; Class-II protein contains a MATH domain together with a conserved BTB (Broad Complex, Tramtrack, and Bric-a-Brac ) domain; Class-III protein belongs to the MATH/Filament domain family; Class-IV protein contains a MATH domain frequently combined with some other domains; Class-V protein has a relative long sequence but contains only one MATH domain; Class-VI protein is characterized by the presence of Peptidase and UBQ (Ubiquitinylation) domains together with one MATH domain. As part of our study regarding seed development of B. rapa, six genes are screened by SSH (Suppression Subtractive Hybridization) and their expression levels are analyzed in combination with seed developmental stages, and expression patterns suggested that Bra001786, Bra03578 and Bra036572 may be seed development specific genes, while Bra001787, Bra020541 and Bra040904 may be involved in seed and flower organ development. This study provides the first characterization of the MATH domain proteins in B. rapa

Phylogenetic Analysis of Brassica rapa MATH-Domain Proteins

The MATH (meprin and TRAF-C homology) domain is a fold of seven anti-parallel β-helices involved in protein-protein interaction. Here, we report the identification and characterization of 90 MATH-domain proteins from the Brassica rapa genome. By sequence analysis together with MATH-domain proteins from other species, the B. rapa MATH-domain proteins can be grouped into 6 classes. Class-I protein has one or several MATH domains without any other recognizable domain; Class-II protein contains a MATH domain together with a conserved BTB (Broad Complex, Tramtrack, and Bric-a-Brac ) domain; Class-III protein belongs to the MATH/Filament domain family; Class-IV protein contains a MATH domain frequently combined with some other domains; Class-V protein has a relative long sequence but contains only one MATH domain; Class-VI protein is characterized by the presence of Peptidase and UBQ (Ubiquitinylation) domains together with one MATH domain. As part of our study regarding seed development of B. rapa, six genes are screened by SSH (Suppression Subtractive Hybridization) and their expression levels are analyzed in combination with seed developmental stages, and expression patterns suggested that Bra001786, Bra03578 and Bra036572 may be seed development specific genes, while Bra001787, Bra020541 and Bra040904 may be involved in seed and flower organ development. This study provides the first characterization of the MATH domain proteins in B. rapa.

A gain-of-function mutation in IAA8 alters Arabidopsis floral organ development by change of jasmonic acid level

Auxin regulates a variety of physiological processes via its downstream factors included Aux/IAAs. In this study, one of these Aux/IAAs, IAA8 is shown to play its role in Arabidopsis development with transgenic plants expressing GFP-mIAA8 under the control of IAA8 promoter, in which IAA8 protein was mutated by changing Pro170 to Leu170 in its conserved domain II. These transgenic dwarfed plants had more lateral branches, short primary inflorescence stems, decreased shoot apical dominance, curled leaves and abnormal flower organs (short petal and stamen, and bent stigmas). Further experiments revealed that IAA8::GFP-mIAA8 plants functioned as gain-of-function mutation to increase GFP-mIAA8 amount probably by stabilizing IAA8 protein against proteasome-mediated protein degradation with IAA8::GFP-IAA8 plants as control. The searching for its downstream factors indicated its interaction with both ARF6 and ARF8, suggesting that IAA8 may involve in flower organ development. This was further evidenced by analyzing the expression of jasmonic acid (JA) biosynthetic genes and JA levels because ARF6 and ARF8 are required for normal JA production. These results indicated that in IAA8::GFP-mIAA8 plants, JA biosynthetic genes including DAD1 (AT2G44810), AOS (AT5G42650) and ORP3 (AT2G06050) were dramatically down-regulated and JA level in the flowers was reduced to 70% of that in wild-type. Furthermore, exogenous JA application can partially rescue short petal and stamen observed IAA8::GFP-mIAA8 plants. Thus, IAA8 plays its role in floral organ development by changes in JA levels probably via its interaction with ARF6/8 proteins.

Characterization of the procera Tomato Mutant Shows Novel Functions of the SlDELLA Protein in the Control of Flower Morphology, Cell Division and Expansion, and the Auxin-Signaling Pathway during Fruit-Set and Development

procera (pro) is a tall tomato (Solanum lycopersicum) mutant carrying a point mutation in the GRAS region of the gene encoding SlDELLA, a repressor in the gibberellin (GA) signaling pathway. Consistent with the SlDELLA loss of function, pro plants display a GA-constitutive response phenotype, mimicking wild-type plants treated with GA₃. The ovaries from both nonemasculated and emasculated pro flowers had very strong parthenocarpic capacity, associated with enhanced growth of preanthesis ovaries due to more and larger cells. pro parthenocarpy is facultative because seeded fruits were obtained by manual pollination. Most pro pistils had exserted stigmas, thus preventing self-pollination, similar to wild-type pistils treated with GA₃ or auxins. However, Style2.1, a gene responsible for long styles in noncultivated tomato, may not control the enhanced style elongation of pro pistils, because its expression was not higher in pro styles and did not increase upon GA₃ application. Interestingly, a high percentage of pro flowers had meristic alterations, with one additional petal, sepal, stamen, and carpel at each of the four whorls, respectively, thus unveiling a role of SlDELLA in flower organ development. Microarray analysis showed significant changes in the transcriptome of preanthesis pro ovaries compared with the wild type, indicating that the molecular mechanism underlying the parthenocarpic capacity of pro is complex and that it is mainly associated with changes in the expression of genes involved in GA and auxin pathways. Interestingly, it was found that GA activity modulates the expression of cell division and expansion genes and an auxin signaling gene (tomato AUXIN RESPONSE FACTOR7) during fruit-set.

The effect of sucrose on the expression of the VvTFL1 and VFL genes during flower development in the “Xiangfei” grapevine

VFL and VvTFL1 genes expression patterns and the effects of sucrose on the expression of VFL and VvTFL1 genes in different organs of the “Xiangfei” grapevine ( Vitis vinifera L.) were investigated. VvTFL1 gene expression was detected in the meristem of the apical bud and lateral bud, but was not detected during inflorescence differentiation and flower organ development. After sucrose treatment, VvTFL1 gene expression increased in the apical bud, but decreased in the lateral bud. These results suggested that the VvTFL1 gene might be mainly involved in the apical growth process of shoots, and exogenous sucrose had an effect on the VvTFL1 gene by increasing shoot apical meristem initiation of apical buds. The VFL gene was expressed primarily during inflorescence differentiation and early flower organ development, but it gradually reduced in later flower development. After sucrose treatment, VFL gene expression increased in the inflorescence and small or middle flower, but a little change was seen in the large flower. These results suggested that the VFL gene plays important roles in the initiation of inflorescence meristems and the morphological formation of flower organs. Exogenous sucrose had an effect on VFL gene expression at the early stage of flower development.

Molecular Cloning and Characterization of a Novel Gossypium barbadense L. RAD-Like Gene

Cotton fiber development is controlled by MYB family transcription factors. Here, we identify a SANT/MYB transcription factor from Gossypium barbadens, designated as \GbRL1. The full-length cDNA of GbRL1 contains an open-reading frame of 243 bp, encoding a protein of 81 amino acids with a calculated molecular mass of 9.32 kDa and an isoelectric point of 6.72. Sequence alignment shows that GbRL1 has high homology with other single SANT/MYB domain-containing genes, especially the RADIALIS (RAD) genes in Antirrhinum majus and Bournea leiophylla. Expression analysis reveals that at the seedling stage, GbRL1 is strongly expressed in roots but weakly in leaves and stamens. In 0-day post-anthesis (DPA) flowers, the expression of GbRL1 is strong in the petals and ovules, moderate in stamens. In ovules, during early fiber development, GbRL1 has high expression level at fiber initiation stage (−3 and 0 DPA) but low level at fiber elongation stage (3, 5, and 8 DPA). In addition, we also detect the expression of GbRL1 in elongating fibers (8 DPA). Promoter analysis indicates that the expression of GbRL1 may be controlled by genes involved in flower organ development and flowering time control, such as AGAMOUS-like genes and FLC gene. The overexpression of GbRL1 in Arabidopsis causes similar developmental alterations as overexpression of RAD. Together, our results suggest that GbRL1 may not only control flower development as RAD, but also plays an important role in early ovule development and/or fiber initiation in cotton.

Conserved Functions of Arabidopsis and Rice CC-Type Glutaredoxins in Flower Development and Pathogen Response

Glutaredoxins (GRXs) are ubiquitous oxidoreductases that play a crucial role in response to oxidative stress by reducing disulfides in various organisms. In planta, three different GRX classes have been identified according to their active site motifs. CPYC and CGFS classes are found in all organisms, whereas the CC-type class is specific for higher land plants. Recently, two Arabidopsis CC-type GRXs, ROXY1 and ROXY2, were shown to exert crucial functions in petal and anther initiation and differentiation. To analyze the function of CC-type GRXs in the distantly related monocots, we isolated and characterized OsROXY1 and OsROXY2-two rice homologs of ROXY1. Both genes are expressed in vegetative and reproductive stages. Although rice flower morphology is distinct from eudicots, OsROXY1/2 floral expression patterns are similar to their Arabidopsis counterparts ROXY1/2. Complementation experiments demonstrate that OsROXY1 and OsROXY2 can fully rescue the roxy1 floral mutant phenotype. Overexpression of OsROXY1, OsROXY2, and ROXY1 in Arabidopsis causes similar vegetative and reproductive plant developmental defects. ROXY1 and its rice homologs thus exert a conserved function during eudicot and monocot flower development. Strikingly, overexpression of these CC-type GRXs also leads to an increased accumulation of hydrogen peroxide levels and hyper-susceptibility to infection from the necrotrophic pathogen Botrytis cinerea, revealing the importance of balanced redox processes in flower organ development and pathogen defence.

Cloning and expression analysis of an E-class MADS-box gene from Populus deltoides

An E-class MADS-box gene, PdMADS2, was isolated from Populus deltoides Bartr. ex Marsh male floral buds by RT-PCR. As shown by relative–quantitative real-time polymerase chain reaction analysis, the expression of PdMADS2 was high in apical and floral buds, intermediate in immature xylem and roots and low in mature leaves of adult male P. deltoides. In developing male floral buds, PdMADS2 expression was high and remained constant from July to September, increased significantly on February 6 and quickly fell to a very low level with the maturation of the flowers. In male inflorescences, PdMADS2 expression was abundant in the perianth cups and inflorescence peduncles; it was not detected in mature pollen. In female inflorescences, PdMADS2 was highly expressed in both ovaries and inflorescence peduncles. Our results suggest that PdMADS2 plays an important role in the development of inflorescence meristems and flower organs in poplar.

A MADS-box gene of Populus deltoides expressed during flower development and in vegetative organs

A MADS-box gene (PdPI) was isolated from a cDNA library constructed from male flower buds of Populus deltoides Bartr. ex Marsh. According to an analysis of genomic DNA structure and putative protein structure, and a phylogenetic study, PdPI is an ortholog of the Arabidopsis PI gene. Relative-quantitative real-time polymerase chain reaction analysis showed that PdPI has a broader expression pattern than PI in Arabidopsis. PdPI was strongly expressed in floral buds and roots and weakly expressed in immature xylem, leaves and apical buds of the male P. deltoides tree. In male inflorescences, PdPI expression was abundant in the perianth and anther, and weak in the peduncle and mature pollen. The large differences in PdPI expression at various phases of male floral bud development were closely related to the development of flower organs (perianth and stamen) and pollen. PdPI was also expressed in female inflorescences. Our results suggest that PdPI has multiple functions in the development of P. deltoides.

High temperatures applied at fall forcing disturb ovule development in Syringa vulgaris L. “Mme Florent Stepman”

Under natural conditions, Syringa vulgaris blooms in May but it may be forced to produce blooming panicles in autumn, winter and early spring. To break winter dormancy, plants require high temperatures, the range of which depends on the forcing date. Forcing in November requires exposure to temperatures of ∼37°C. Such an exposure may affect the development of both male and female flower organs. In this work we have followed the anatomical changes in the ovule development in buds collected from plants forced in autumn, and compared them to those in plants blooming under natural conditions in May. From the earliest identifiable stages, in flowers forced in November the ovule development was delayed relative to the normal development pattern. Ultimately, most of the ovules (83%) degenerated before the differentiation of the embryo sacs was completed. We believe that it is the high temperatures required during autumn forcing that affect the development of female gametophytes, leading to the formation of defective structures which often degenerate. This in turn may reduce the esthetic appeal of the inflorescences and reduce their commercial value.

イネの花器官形成における二つのクラスG MADS-box 遺伝子の機能

イネの花器官形成における二つのクラスG MADS-box 遺伝子の機能

The effect of the cytoplasms of Brassica napus and B. juncea on some characteristics of B. carinata, including flower morphology

This study was conducted to assess the cytoplasm effects of Brassica napus and B. juncea on the some characteristics of B. carinata, as well as the phylogenetic distances separating the three species. Alloplasmic lines of B. carinata were developed from B. napus × B. carinata and B. juncea × B. carinata hybrids by recurrent backcrossing to the BC7 generation. Sixteen populations from three generations were compared for a number of characteristics. Plants with the cytoplasm of B. napus flowered later, had shorter filaments and longer pistils, lower pollen amount, lower seed set, lower petal length and width and different petal color; plants with the cytoplasm of B. juncea had shorter pistils and filaments, and lower petal length and width than their corresponding euplasmic sibs, respectively. The results suggest that the cytoplasm is involved in the development of flower organs. The natural species, B. carinata showed a balance between the nucleus and cytoplasm. The cytoplasm from B. napus showed a stronger disturbing effect than that of B. juncea, suggesting that B. carinata might be genetically closer to B. juncea than to B. napus. The significant difference in the alloplasmic effect of the cytoplasms of B. napus and B. juncea also suggests that in B. carinata the B genome may play a greater role than the C genome.

Phenology of flower and fruit development in Uncaria gambir Species

A phenological study with the aim to elucidate flower and fruit development stages in species of Uncaria gambir was conducted in field trial of agricultural faculty of Andalas University-West Sumatra. Observation during flower and fruit organ development was done in eleven inflorescences. Quantitative and descriptive data were collected during one season of flowering time. Phenology of flower and fruit development could be classified in five stages, namely F0 (flower initiation), F1 (small bud scale), F2 (large bud scale), F3 (anthesis, flower opening) and F4 (fruit development). All these events had average completing time in 112 days, and could be detailed as follows: flower initiation stage (F0) took place in 20 days, small scale bud stage (F1) occurred in 27 days, and large scale bud and anthesis stage (F0 and F3) each took place in 5 days, meanwhile fruit development stage (F4=S0) would complete in 53 days. This result should be useful information especially for creating breeding programme in Uncaria gambir species.Key words: Uncaria gambir, phenology, flower development, fruit development.

Patterns of MADS-box gene expression mark flower-type development in Gerbera hybrida(Asteraceae)

BackgroundThe inflorescence of the cut-flower crop Gerbera hybrida (Asteraceae) consists of two principal flower types, ray and disc, which form a tightly packed head, or capitulum. Despite great interest in plant morphological evolution and the tractability of the gerbera system, very little is known regarding genetic mechanisms involved in flower type specification. Here, we provide comparative staging of ray and disc flower development and microarray screening for differentially expressed genes, accomplished via microdissection of hundreds of coordinately developing flower primordia.ResultsUsing a 9K gerbera cDNA microarray we identified a number of genes with putative specificity to individual flower types. Intrestingly, several of these encode homologs of MADS-box transcription factors otherwise known to regulate flower organ development. From these and previously obtained data, we hypothesize the functions and protein-protein interactions of several gerbera MADS-box factors.ConclusionOur RNA expression results suggest that flower-type specific MADS protein complexes may play a central role in differential development of ray and disc flowers across the gerbera capitulum, and that some commonality is shared with known protein functions in floral organ determination. These findings support the intriguing conjecture that the gerbera flowering head is more than a mere floral analog at the level of gene regulation.

Flower-bud failure in olive and the involvement of amoeboid protists

SummaryFlower bud anomalies in Olea europaea L., ‘Santulhana’ and ‘Conserva de Elvas’ begin as very small microscopic lesions associated with amoeboid bionts, which cause extensive areas of destruction and block flower organ development. In this work, we describe the process of injury in olive flower-buds in parallel with biont development in floral tissues and in the culture medium. Flower-buds were processed for light microscopy (LM) and transmission electron microscopy (TEM). Bionts were isolated from flower-buds in culture medium and processed for LM. Amoeboid bionts in several stages of plasmodial formation, grown in culture medium from olive flower-buds, resembled those observed in sectioned flower-bud tissues. The formation of small plasmodia by the fusion of amoeboid cells and the development of small plasmodia into phaneroplasmodia, suggest the presence of an amoeboid protist of the Myxogastria group. On the other hand, sporulation of plasmodia in hay agar culture medium, gave rise to stalked sporangia containing numerous spores. The morphology and dimensions of the sporangia and spores, as well as the characteristics of the amoebae that germinated from the spores, suggest that the protist Myxogastria belongs to the genus Didymium.

The Role of Gene TEPAL-LIKE BRACT (TLB) in the Separation between the Bracts and Perianth in Fagopyrum esculentum Moench.

The morphological and genetic studies of the tlb mutant of common buckwheat Fagopyrum esculentum from the collection of the All-Russia Research Institute of Legumes and Groat Crops have demonstrated that gene TLB controls an important stage of flower development-it determines the lower boundary of simple perianths. The loss of TLB gene function leads to changes in the structure of the bract from scale-like to tepal-like. Gene TLB is assumed to limit, on the basal side, the region of the expression of genes determining the development of flower organs as petals.