Early Flower Development Research Articles

STUDIES ON NURSERY MANAGEMENT IN PAPAYA (CARICA PAPAYA L.) VAR. CO2

Considering the need for optimizing the number of seedling per bag, an attempt was made with different organic potting mixtures and number of seedlings per polythene bag on growth and development of papaya var. CO2 in the nursery and their effect on first flowering height and first bearing height in the main field. The study revealed the superiority of single seedling grown with potting mixture treated with vermicompost by registering higher seedling height (28.50 cm), seedling diameter (0.639 cm), petiole length (6.410 cm), petiole diameter (0.186 cm), number of leaves (11.65), short dry weight (1.254 g), root length (29.715 cm) root dry weight (0.392 g) and root shoot ratio (0.340). In main field condition, three seedlings per polybag grown with potting mixture and vermicompost was found to have early flowering development (86.69 days) and first bearing height (96.695 cm) with minimum plant height (90.930 cm).

GREENHOUSE SAFFRON CULTURE – TEMPERATURE EFFECTS ON FLOWER EMERGENCE AND VEGETATIVE GROWTH OF THE PLANTS

The physiology of flower initiation and early flower development of the saffron plant (Crocus sativus L.) is well characterised. However, the optimal conditions for anthesis have not been ascertained. In this work we determined the influence of temperature after flower initiation on flowering and saffron spice production, as well as the influence of the environmental conditions in the greenhouse on plant growth. To this aim, saffron corms which had been stored at 25°C during 105 days, were sprouted at different temperature regimes in growth chambers in the light, with an 16/8 h photoperiod and a photon fluence rate of 20-40 μmol m -2 s -1 . One month after the completion of flowering the plants were transplanted to the open field, and their CO 2 assimilation rates were compared to that of plants coming from corms planted directly in the open. In plants grown at a constant temperature, this parameter, in the range 13-20°C, had no significant effect on the number of sprouts nor that of flowers formed per corm. Flowering occurred earlier at 17°C than at higher and lower temperatures. The flowers formed at 20°C had a wilt aspect. The size of the styles was directly related to temperature and therefore was highest at 20°C. in these plants, however, the styles were difficult to separate from the flower. Growing the plants at a 20/10°C term period hastened flowering and resulted in the formation of normal flowers, but also reduced style size. Growth in the chambers resulted in a 90% reduction of the maximum photosynthetic CO 2 fixation as compared with plants grown in the open. This loss was recovered in part (about 50%) several weeks after transferring the plants to the open field.

A defensin from tomato with dual function in defense and development

Defensins are antimicrobial peptides that are part of the innate immune system, contributing to the first line of defense against invading pathogens. Defensins and defensin-like peptides are functionally diverse, disrupting microbial membranes and acting as ligands for cellular recognition and signaling. Here we show that the tomato defensin DEF2 is expressed during early flower development. Defensin mRNA abundance, peptide expression and processing are differentially regulated in developing flowers. Antisense suppression or constitutive overexpression of DEF2 reduces pollen viability and seed production. Furthermore, overexpression of DEF2 pleiotropically alters the growth of various organs and enhances foliar resistance to the fungal pathogen Botrytis cinerea. Partially purified extracts from leaves of a DEF2-overexpressing line inhibited tip growth of B. cinerea. Besides providing insights into regulation of defensin expression, these data demonstrate that plant defensins, like their animal counterparts, can assume multiple functions related to defense and development.

Influence of clavata3-2 mutation on early flower development in Arabidopsis thaliana: quantitative analysis of changing geometry.

Early development of the flower primordium has been studied in Arabidopsis thaliana clavata3-2 (clv3-2) plants with the aid of sequential in vivo replicas and longitudinal microtome sections. Sequential replicas show that, although there is no regular phyllotaxis in the clv3-2 inflorescence shoot apex, the sites of new primordium formation are, to a large extent, predictable. The primordium always appears in a wedge-like region of the meristem periphery flanked by two older primordia. In general, stages of primordium development in clv3-2 are similar to the wild type, but quantitative geometry analysis shows that the clv3-2 primordium shape is affected even before the CLAVATA/WUSCHEL regulatory network would start to operate in the wild-type primordium. The shape of the youngest primordium in the mutant is more variable than in the wild type. In particular, the shape of the adaxial primordium boundary varies and seems to be related to the shape of the space available for the given primordium formation, suggesting that physical constraints play a significant role in primordium shape determination. The role of physical constraints is also manifested in that the shape of the primordium in the later stages, as well as the number and position of sepals, are adjusted to the available space. Longitudinal sections of clv3-2 apices show that the shape of surface cells of the meristem and young primordium is different from the wild type. Moreover, there is only one tunica layer in both the meristem and in the primordium until it becomes a bulge that is distinctly separated from the meristem. Starting from this stage, the anticlinal divisions predominate in subprotodermal cells, suggesting that the distribution of periclinal and anticlinal cell divisions in the early development of the flower primordium is not directly affected by the clv3-2 mutation.

RETARDED PALEA1 Controls Palea Development and Floral Zygomorphy in Rice

Poaceae, one of the largest flowering plant families in angiosperms, evolved distinct inflorescence and flower morphology diverging from eudicots and other monocots. However, the mechanism underlying the specification of flower morphology in grasses remains unclear. Here we show that floral zygomorphy along the lemma-palea axis in rice (Oryza sativa) is partially or indirectly determined by the CYCLOIDEA (CYC)-like homolog RETARDED PALEA1 (REP1), which regulates palea identity and development. The REP1 gene is only expressed in palea primordium during early flower development, but during later floral stages is radially dispersed in stamens and the vascular bundles of the lemma and palea. The development of palea is significantly retarded in the rep1 mutant and its palea has five vascular bundles, which is similar to the vascular pattern of the wild-type lemma. Furthermore, ectopic expression of REP1 caused the asymmetrical overdifferentiation of the palea cells, altering their floral asymmetry. This work therefore extends the function of the TCP gene family members in defining the diversification of floral morphology in grasses and suggests that a common conserved mechanism controlling floral zygomorphy by CYC-like genes exists in both eudicots and the grasses.

Analyses of the floral organ morphogenesis and the differentially expressed genes of an apetalous flower mutant in Brassica napus

The floral organ morphogenesis of the apetalous flower mutant Apet33-10 in Brassica napus was investigated and the result showed that all the floral organ morphogenesis was normal except that petal primordium was not observed during flower development. Eighteen genes were found to be down regulated in early floral buds (less than 200 mum in length) of Apet33-10 at the stage of floral organ initiation by means of suppressive subtraction hybridization (SSH) and RT-PCR. These genes were involved in petal identity, calcium iron signal transduction, mRNA processing, protein synthesis and degradation, construction of cytoskeleton, hydrogen transportation, nucleic acid binding, alkaloid biosynthesis and unknown function. Three overall coding region cDNAs of APETALA3 (AP3) gene, BnAP3-2, BnAP3-3 and BnAP3-4 were obtained by RT-PCR, respectively. Real-time quantitative PCR analysis showed that the expression ratio among BnAP3-2, BnAP3-3 and BnAP3-4 was 3.67:3.68:1 in early floral buds of wild type Pet33-10. The expression level of BnAP3-2, BnAP3-3 and BnAP3-4 in early floral buds of Apet33-10 was down-regulated to 36.6, 28.3 and 66.8% with the comparison of that of wild type, respectively, and the overall expression level of AP3 genes in apetalous mutant amounted to 45.0% of that in wild type. The difference in the expression level of each AP3 gene in stamen between apetalous and wild type lines was not significant. It is suggested that lower abundant expression of AP3 genes during the early flower development might be enough for stamen primordium initiation, but not enough for petal primordium initiation in the apetalous line Apet33-10.

Inflorescence and Early Flower Development in Loteae (Leguminosae) in a Phylogenetic and Taxonomic Context

Molecular phylogeny shows that the temperate legume tribe Loteae is close to the mostly tropical Robinieae and monogeneric Sesbanieae, but comparative morphological studies of these groups are limited. Unusual patterns of inflorescence symmetry and calyx development have been described in some Loteae, but taxon sampling was low. We studied these features with scanning electron microscopy in 25 species of Loteae plus in three Robinia species. Phylogenetic trees of Loteae based on nuclear ribosomal ITS sequences and 77 morphological characters are constructed. Our data show that whorled flower arrangement is a synapomorphy of Loteae; joint initiation of the two adaxial sepals is a synapomorphy of a clade containing Hippocrepis, Scorpiurus, and Coronilla; floral buds bent backward early in development are a synapomorphy of Coronilla; bilateral umbel symmetry and the presence of a single whorl of flowers are probably primitive within Loteae. Inflorescences of Robinia show no special similarities with those of Loteae. Developmental data support homologies between sterile bracts in all Loteae. Even if the sterile bract is situated at the top of the peduncle, it is morphologically the first leaf on the peduncle. Monosymmetric umbels of Loteae (including the model legume Lotus japonicus) could be useful for investigating genetic control of symmetry in structures of hierarchic levels higher than flowers.

Response of Chickpea Yield to High Temperature Stress during Reproductive Development

Minimizing the exposure of an annual crop to abiotic stresses may increase seed yield. A study was conducted to determine the effect of high temperature stress during reproductive development on pod fertility, seed set, and seed yield of chickpea (Cicer arietinum L). ‘Myles’ desi and ‘Xena’ kabuli chickpea were grown in a controlled environment under 20/16°C day/night air temperatures (control). High (35/16°C) and moderate (28/16°C) temperature stresses were imposed for 10 d during early flowering and pod development. Compared to the control, the early flower high temperature stress decreased (P < 0.01) pod production by 34% for Myles and 22% for Xena, whereas high temperature stress during pod development decreased (P < 0.05) seeds per plant by 33% for Myles and 39% for Xena. Consequently, the high temperature stress during pod development decreased (P < 0.01) seed yield by 59% for Myles and 53% for Xena. Yield reduction was greater due to the stress during pod development compared to the stress during early flowering. Plants recovered to a greater degree from the early flower stress compared to the pod development stress. The Myles desi produced 40 seeds per plant and the Xena kabuli produced 15 seeds per plant, whereas the Myles had smaller individual seed size than the Xena. Consequently, the Myles desi produced 26% greater seed yield than the Xena kabuli under the same conditions. Minimizing the exposure of chickpea to high temperature stress during pod development will increase pod fertility, seed set, and seed yield of the crop.

Genome-Wide Analysis of Gene Expression during Early Arabidopsis Flower Development

Detailed information about stage-specific changes in gene expression is crucial for the understanding of the gene regulatory networks underlying development. Here, we describe the global gene expression dynamics during early flower development, a key process in the life cycle of a plant, during which floral patterning and the specification of floral organs is established. We used a novel floral induction system in Arabidopsis, which allows the isolation of a large number of synchronized floral buds, in conjunction with whole-genome microarray analysis to identify genes with differential expression at distinct stages of flower development. We found that the onset of flower formation is characterized by a massive downregulation of genes in incipient floral primordia, which is followed by a predominance of gene activation during the differentiation of floral organs. Among the genes we identified as differentially expressed in the experiment, we detected a significant enrichment of closely related members of gene families. The expression profiles of these related genes were often highly correlated, indicating similar temporal expression patterns. Moreover, we found that the majority of these genes is specifically up-regulated during certain developmental stages. Because co-expressed members of gene families in Arabidopsis frequently act in a redundant manner, these results suggest a high degree of functional redundancy during early flower development, but also that its extent may vary in a stage-specific manner.

Cloning and characterization of a novel chalcone synthase gene from Phalaenopsis hybrida orchid flowers

Chalcone synthase (CHS, EC 2.3.1.74) is the key enzyme involved in flavonoid and anthocyanin biosynthesis. A complete DNA sequence of chalcone synthase gene designated Pchs1 was isolated by means of usual and then inverse polymerase chain reactions from genomic DNA of an orchid, Phalaenopsis hybrida, cv. Formosa rose. Nucleotide sequence analysis based on alignment with published Phalaenopsis chs cDNA revealed that Pchs1 contained an intact open reading frame of 1173-bp with one 109-bp intron at the conserved site. The deduced polypeptide (PCHS1) from Pchs1 comprised 390 amino acids with a predicted mol wt of 42.5 kD. PCHS1 showed 61–65% identities with CHS from other plants and retained most of the conserved residues. Some putative cis-regulatory elements were present at the 5′ and 3′ flanking regions of Pchs1. Southern blot analysis predicted at least four chs-like genes, thus indicating the presence of a small multigene chs family in P. hybrida. Relative quantitative RT-PCR showed that Pchs1 is expressed in petals at early flower development as well as in lip tissue when the flower has just opened.

Analysis of Organ-specific, Expressed Genes in Oncidium Orchid by Subtractive Expressed Sequence Tags Library

The pseudobulb of Oncidium orchid plays a key role in water, carbohydrate, and other nutrition support during floral development, yet a large scale of gene expression analysis involved in the metabolisms have not been evaluated. By subtracting RsaI-digested cDNAs of leaf from those of psuedobulb, an efficient subtractive cDNA library was developed. In total, 1080 subtractive expressed sequence tags (ESTs) were obtained. Analysis revealed approximately 636 unique gene parts, 120 clusters and 516 singles. Of these sequences, 74.8% were annotated on the database of NCBI GenBank. Peroxidase, sodium/dicarboxylate cotransporter, and mannose-binding lectin were highly expressed. Some gene profiles were identified as related to carbohydrate metabolism involved in mannan, pectin, starch and sucrose biosynthesis. A large fraction of the ESTs (35%) were classified into transportation, stress-related, cell cycle, or regulatory functions. Most genes that were differentially expressed are important in early flowering development, carbohydrate metabolism and stress-response physiology. This efficient organ-specific EST library represented an explicit transcriptome profile of Oncidium pseudobulb.

Proanthocyanidin Synthesis and Expression of Genes Encoding Leucoanthocyanidin Reductase and Anthocyanidin Reductase in Developing Grape Berries and Grapevine Leaves

Proanthocyanidins (PAs), also called condensed tannins, can protect plants against herbivores and are important quality components of many fruits. Two enzymes, leucoanthocyanidin reductase (LAR) and anthocyanidin reductase (ANR), can produce the flavan-3-ol monomers required for formation of PA polymers. We isolated and functionally characterized genes encoding both enzymes from grapevine (Vitis vinifera L. cv Shiraz). ANR was encoded by a single gene, but we found two highly related genes encoding LAR. We measured PA content and expression of genes encoding ANR, LAR, and leucoanthocyanidin dioxygenase in grape berries during development and in grapevine leaves, which accumulated PA throughout leaf expansion. Grape flowers had high levels of PA, and accumulation continued in skin and seeds from fruit set until the onset of ripening. VvANR was expressed throughout early flower and berry development, with expression increasing after fertilization. It was expressed in berry skin and seeds until the onset of ripening, and in expanding leaves. The genes encoding LAR were expressed in developing fruit, particularly in seeds, but had low expression in leaves. The two LAR genes had different patterns of expression in skin and seeds. During grape ripening, PA levels decreased in both skin and seeds, and expression of genes encoding ANR and LAR were no longer detected. The results indicate that PA accumulation occurs early in grape development and is completed when ripening starts. Both ANR and LAR contribute to PA synthesis in fruit, and the tissue and temporal-specific regulation of the genes encoding ANR and LAR determines PA accumulation and composition during grape berry development.

The Effects of Conditioning Strawberry Plug Plants under Altered Red/Far-red Light Environments

The effect of altered red/far-red light environment on subsequent field performance of strawberry plug plants was tested. Two wavelength-selective plastic films were compared to neutral shade and full-sun control for conditioning `Chandler' strawberry plug plants before transplanting to a winter production system. The following year, plug plants of `Chandler', `Sweet Charlie', and `Allstar' were conditioned under the same treatments, with the addition of a continuous incandescent light and a short-day photoperiod, and plant performance was followed in the winter production system in Florida, a cold-climate annual hill system in Maryland, and in a low-input greenhouse production system. During the first year, the red light-filtering film slightly advanced fruiting in Florida. However, during the second year, the effect of the red light-filtering film was not significant, and a short-day treatment resulted in a greater reduction in runnering and increased early crown and flower development. For June-bearing strawberry plants maintained above 20 °C, altering the red/far-red environment did not consistently advance flowering.

Genome-wide expression profiling and identification of gene activities during early flower development in Arabidopsis

We have used oligonucleotide microarrays to detect Arabidopsis gene expression during early flower development. Among the 22,746 genes represented on the Affymetrix ATH1 chip, approximately 14,660 (approximately 64.5%) genes were expressed with signal intensity at or more than 50 in each of the six organs/structures examined, including young inflorescences (floral stages 1-9), stage-12 floral buds, developing siliques, leaves, stems, and roots. 17,583 genes were expressed with an intensity at or above 50 in at least one tissue, including 12,245 genes that were expressed in all the six tissues. Comparison of genes expressed between young inflorescence or stage-12 floral buds with other tissues suggests that relatively large numbers of genes are expressed at similar levels in tissues that are related morphologically and/or developmentally, as supported by a cluster analysis with data from two other studies. Further analysis of the genes preferentially expressed in floral tissues has uncovered new genes potentially involved in Arabidopsis flower development. One hundred and four genes were determined to be preferentially expressed in young inflorescences, including 22 genes encoding putative transcription factors. We also identified 105 genes that were preferentially expressed in three reproductive structures (the young inflorescences, stage-12 floral buds and developing siliques), when compared with the vegetative tissues. RT-PCR results of selected genes are consistent with the results from these microarrays and suggest that the relative signal intensities detected with the Affymetrix microarray are reliable estimates of gene expression.

Floral MADS box genes and homeotic gender dimorphism in Thalictrum dioicum (Ranunculaceae) – a new model for the study of dioecy

In most dioecious angiosperm species, flowers are initially perfect but abort either stamens or carpels during their development, indicating that sex determination occurs after floral organ identity has been established. Dioecious members of the genus Thalictrum (meadow-rue), however, produce flowers that lack aborted organs. Examination of early flower development of T. dioicum confirms that flowers are male or female from inception, raising the possibility that genetic mechanisms working at or above the level of organ identity promote sex determination through a homeotic-like mechanism. In order to investigate this possibility, we identified homologs of the organ identity genes PISTILLATA (PI), APETALA3 (AP3) and AGAMOUS (AG) from T. dioicum and the hermaphroditic species T. thalictroides. A combination of early and late duplication events was uncovered in these gene lineages and expression analyses indicate that these events are generally associated with divergence in gene regulation. In light of these findings, we discuss the potential of T. dioicum as a new model for the study of sex determination in the basal eudicots.

Genome-Wide Analysis of Gene Expression during Early Arabidopsis Flower Development

Detailed information about stage-specific changes in gene expression is crucial for the understanding of the gene regulatory networks underlying development. Here, we describe the global gene expression dynamics during early flower development, a key process in the life cycle of a plant, during which floral patterning and the specification of floral organs is established. We used a novel floral induction system in Arabidopsis, which allows the isolation of a large number of synchronized floral buds, in conjunction with whole-genome microarray analysis to identify genes with differential expression at distinct stages of flower development. We found that the onset of flower formation is characterized by a massive downregulation of genes in incipient floral primordia, which is followed by a predominance of gene activation during the differentiation of floral organs. Among the genes we identified as differentially expressed in the experiment, we detected a significant enrichment of closely related members of gene families. The expression profiles of these related genes were often highly correlated, indicating similar temporal expression patterns. Moreover, we found that the majority of these genes is specifically up-regulated during certain developmental stages. Because co-expressed members of gene families in Arabidopsis frequently act in a redundant manner, these results suggest a high degree of functional redundancy during early flower development, but also that its extent may vary in a stage-specific manner.

AVAG2 is a putative D-class gene from an ornamental asparagus

Members of the AGAMOUS (AG) family of MADS-box genes play important roles in regulating the development of reproductive organs in flowering plants. To elucidate the molecular mechanisms of floral development in Asparagus virgatus, we isolated and characterized an Asparagus AG-homologue, AVAG2. AVAG2 contains an open reading frame that encodes a deduced protein with 234 amino acid residues. Phylogenetic analysis indicated that AVAG2 belongs to the D-lineage of the AG gene family. AVAG2 mRNA was detected in the flower, but not in vegetative organs. Moreover, in in situ hybridization experiments, AVAG2 signals were observed in the stamens and carpels during early flower development, and appeared in the ovule only at later developmental stages. This suggests that the AVAG2 gene is involved in ovule formation. Thus, our expression data support the phylogenetic analysis indicating that AVAG2 belongs to the D-class gene family.

Shape Matters: Hofmeister’s Rule, Primordium Shape, and Flower Orientation

Hofmeister's rule is an empirical heuristic derived from the observation that new leaf primordia are formed in the largest space between the existing flanks of the older primordia. These observations have been repeatedly validated in studies of leaf arrangement, but there has been little attempt to extend them to inflorescence and floral organs. This investigation demonstrates the validity of Hofmeister’s observations to cincinnus and early flower development in Phenakospermum guyannense (Strelitziaceae) and Heliconia latispatha (Heliconiaceae) and relates these results to Paul Green’s work on the biophysics of organ formation. The cincinni of Phenakospermum and Heliconia arise in the axils of primary bracts and produce a prophyll, continuation apex, and flower in regular succession. The shapes and orientations of the apical regions of the cincinni are correlated with the placement of these organs, which in turn effect the positions of the sepals and their sequence of formation. The result is two rows of mirror‐image flowers. The mirror‐image symmetry of the flowers is a direct result of Hofmeister’s rule in connection with the shape of the apical region. These two factors create a self‐sustaining developmental system that produces prophylls, continuation apices, flowers, and sepals in regular succession. Although the cincinni and flowers of these species are built on a common plan, slight differences in apical shape and orientation produce differences in mature floral orientation. Understanding these orientations, and identifying the sequence of sepal formation, allows a proper identification of organ homologies. A study of Green’s results and theories shows that Hofmeister’s rule can be considered as an empirical condensation of the biophysical factors that influence organ position. These biophysical factors are widely applicable to organ formation in many species.

Elements regulatingAGAMOUSexpression are conserved betweenArabidopsis thaliana,Brassica napus, andLinum usitatissimum

To investigate the functional conservation of cis regulatory elements controlling AGAMOUS (AG) expression, we placed the promoter region of AG from Arabidopsis thaliana into a close relative, Brassica napus, and a distantly related species, Linum usitatissimum, and analyzed the subsequent expression patterns in each species. Spatially, the expression patterns in all three species were marginally similar, in that expression was confined primarily to the reproductive organs and nectarium. Within organs however, tissue-specific expression patterns were not conserved between species. Unlike Arabidopsis, the transgenic AG cis elements did not express in the ovules of B. napus and L. usitatissimum. Temporally, the pattern of AG cis-element expression in B. napus was identical to that of Arabidopsis during early development; however, the AG cis elements did not express at all during early flower development in L. usitatissimum. These results suggest that although regulatory factors controlling the generalized local expression of AG have been conserved between these species, those controlling temporal and tissue-specific expression have not.Key words: AGAMOUS, cis elements, regulation, Arabidopsis, Brassica napus, Linum usitatissimum.

FLOWER DEVELOPMENT IN CLOUDBERRY (RUBUS CHAMAEMORUS L.)

The cloudberry (Rubus chamaemorus L.) is a herbaceous dioecious octoploid (2n=56) berry plant that grows on peatland in the circum-polar region. Occasionally, hermaphroditic individuals can be found. The breeding of hermaphroditic commercial varieties is dependent on knowledge of the sex determination system. Here we present an investigation on early flower development of male, female and hermaphroditic flowers in cloudberry using scanning electron microscopy and light microscopy. By examination of the flower buds it is possible to know the sex of an individual the summer before flowering. In order to understand the interactions between the floral homeotic genes in this system we also wanted to identify the gene homologue to SUPERMAN which is involved in the maintenance of the stamen/carpel whorl boundary in Arabidopsis.