Differential gene expression during floral transition in pineapple.
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ABSTRACT: Pineapple (Ananas comosus var. comosus) and ornamental bromeliads are commercially induced to flower by treatment with ethylene or its analogs. The apex is transformed from a vegetative to a floral meristem and shows morphological changes in 8 to 10 days, with flowers developing 8 to 10 weeks later. During eight sampling stages ranging from 6 hours to 8 days after treatment, 7,961 genes were found to exhibit differential expression (DEG) after the application of ethylene. In the first 3 days after treatment, there was little change in ethylene synthesis or in the early stages of the ethylene response. Subsequently, three ethylene response transcription factors (ERTF) were up-regulated and the potential gene targets were predicted to be the positive flowering regulator CONSTANS (CO), a WUSCHEL gene, two APETALA1/FRUTFULL (AP1/FUL) genes, an epidermal patterning gene and a jasmonic acid synthesis gene. We confirm that pineapple has lost the flowering repressor FLOWERING LOCUS C. At the initial stages, the SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) was not significantly involved in this transition. Another WUSCHEL gene and a PHD homeobox transcription factor, though not apparent direct targets of ERTF, were up-regulated within a day of treatment, their predicted targets being the up-regulated CO, auxin response factors, SQUAMOSA, and histone H3 genes with suppression of abscisic acid response genes. The FLOWERING LOCUS T (FT), TERMINAL FLOWER (TFL), AGAMOUS-like APETELAR (AP2) and SEPETALA (SEP) increased rapidly within 2 to 3 days after ethylene treatment. Two FT genes were up-regulated in the apex and not the leaf bases after treatment, suggesting that transport did not occur. These results indicated that the ethylene response in pineapple and possibly most bromeliads acts directly to promote the vegetative to flower transition via APETALA1/FRUITFULL (AP1/FUL) and its interaction with SPL, FT, TFL, SEP and AP2.
Project description:Pineapple (Ananas comosus var. comosus) and ornamental bromeliads are commercially induced to flower by treatment with ethylene or its analogs. The apex is transformed from a vegetative to a floral meristem and shows morphological changes in 8 to 10 days, with flowers developing 8 to 10 weeks later. During eight sampling stages ranging from 6 hours to 8 days after treatment, 7,961 genes were found to exhibit differential expression (DEG) after the application of ethylene. In the first 3 days after treatment, there was little change in ethylene synthesis or in the early stages of the ethylene response. Subsequently, three ethylene response transcription factors (ERTF) were up-regulated and the potential gene targets were predicted to be the positive flowering regulator CONSTANS (CO), a WUSCHEL gene, two APETALA1/FRUTFULL (AP1/FUL) genes, an epidermal patterning gene and a jasmonic acid synthesis gene. We confirm that pineapple has lost the flowering repressor FLOWERING LOCUS C. At the initial stages, the SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) was not significantly involved in this transition. Another WUSCHEL gene and a PHD homeobox transcription factor, though not apparent direct targets of ERTF, were up-regulated within a day of treatment, their predicted targets being the up-regulated CO, auxin response factors, SQUAMOSA, and histone H3 genes with suppression of abscisic acid response genes. The FLOWERING LOCUS T (FT), TERMINAL FLOWER (TFL), AGAMOUS-like APETELAR (AP2) and SEPETALA (SEP) increased rapidly within 2 to 3 days after ethylene treatment. Two FT genes were up-regulated in the apex and not the leaf bases after treatment, suggesting that transport did not occur. These results indicated that the ethylene response in pineapple and possibly most bromeliads acts directly to promote the vegetative to flower transition via APETALA1/FRUITFULL (AP1/FUL) and its interaction with SPL, FT, TFL, SEP and AP2.
Project description:Floral transition and flower development are regulated by numerous environmental and endogenous signals, which are integrated at a relatively small number of floral integrators, such as FLOWERING LOCUS T (FT) and SUPPRESSOR OF CONSTANS OVEREXPRESSION 1 (SOC1). Of the environmental factors, photoperiod is regarded the most important one in promoting floral transition in Arabidopsis thaliana and most labstrains will flower earlier under long day (LD) conditions than under short day (SD) conditions. Arabidopsis is therefore considered a facultative LD plant. To monitor gene expression changes during floral transition and early flower development plants were grown under SD (9 hr light, 15 hr dark) for 30 days. Plants were then shifted to LD (16 hr light, 8 hr dark) conditions to induce flowering. RNA was isolated from micro-dissected apical tissue harvested 0, 3, 5, and 7 days after the shift to LD and double-stranded cDNA was synthesized. Biotinylated cRNA probes were prepared and hybridized to the Affymetrix ATH1 array in duplicate (biological replicates). To study floral transition, we not only analyzed response of wildtype Landsberg erecta (Ler) plants, but also the effect of mutants in the flowering time genes CONSTANS (CO; co-2) and FT (ft-2). Early flower development was analyzed by comparing Col-0 wildtype plants with the meristem identity mutant lfy-12 (Col-0).
Project description:MicroProteins are short, single domain proteins that act by sequestering larger, multidomain proteins into non-functional complexes. MicroProteins have been identified in plants and animals, where they are mostly involved in the regulation of developmental processes. Here we show that two Arabidopsis thaliana microProteins, miP1a and miP1b, physically interact with CONSTANS (CO) a potent regulator of flowering time. The miP1a/b-type microProteins evolved in dicotyledonous plants and have an additional carboxy-terminal PF(V/L)FL motif. This motif enables miP1a/b microProteins to interact with TOPLESS/TOPLESS-RELATED (TPL/TPR) proteins. Interaction of CO with miP1a/b/TPL causes late flowering due to a failure in the induction of FLOWERING LOCUS T (FT) expression under inductive long day conditions. Both miP1a and miP1b are expressed in vascular tissue, where CO and FT are active. Genetically, miP1a/b act upstream of CO thus our findings unravel a novel layer of flowering time regulation via microProtein-inhibition. RNA-Seq transcriptome analysis of four biological samples were analysed with two technical replicates. Columbia wildtype plants Col-0, constans mutant plants co-SAIL, and two transgenic lines overexpressing a microProtein (miP1a and miP1b) were sequenced.
Project description:Wild type and mutanat Arabiposis plants grown in short days (9L:15D) for 30 days at 21°C, then shifted to long days (16L:8D). Genotypes: Columbia wild type (Col-0) Landsberg erecta (Ler) leafy-12 (lfy-12, in Col-0) constans-2 (co-2, in Ler) flowering locus T-2 (ft-2, in Ler) Time points: 0, 3, 5, and 7 days after shift to long days Keywords = flowering Keywords: time-course
Project description:Wild type and mutanat Arabiposis plants grown in short days (9L:15D) for 30 days at 21°C, then shifted to long days (16L:8D).,Genotypes:,Columbia wild type (Col-0),Landsberg erecta (Ler),leafy-12 (lfy-12, in Col-0),constans-2 (co-2, in Ler),flowering locus T-2 (ft-2, in Ler),Time points:,0, 3, 5, and 7 days after shift to long days
Project description:The circadian clock represents a critical regulatory network, which allows plants to anticipate environmental changes as inputs and promote plant survival by regulating various physiological outputs. Here, we examine the function of the clock-regulated transcription factor, CYCLING DOF FACTOR 6 (CDF6), during cold stress in Arabidopsis thaliana. We found that the clock gates CDF6 transcript accumulation in the vasculature during cold stress. CDF6 mis-expression results in an altered flowering phenotype during both ambient and cold stress. A genome-wide transcriptome analysis links CDF6 to genes associated with flowering and seed germination during cold and ambient temperatures, respectively. Analysis of key floral regulators indicates that CDF6 alters flowering during cold stress by repressing photoperiodic flowering components, FLOWERING LOCUS T (FT), CONSTANS (CO), and BROTHER OF FT (BFT). Gene ontology enrichment further suggests that CDF6 regulates circadian and developmental associated genes. These results provide insight into how the clock-controlled CDF6 modulates plant development during moderate cold stress.
Project description:MicroProteins are short, single domain proteins that act by sequestering larger, multidomain proteins into non-functional complexes. MicroProteins have been identified in plants and animals, where they are mostly involved in the regulation of developmental processes. Here we show that two Arabidopsis thaliana microProteins, miP1a and miP1b, physically interact with CONSTANS (CO) a potent regulator of flowering time. The miP1a/b-type microProteins evolved in dicotyledonous plants and have an additional carboxy-terminal PF(V/L)FL motif. This motif enables miP1a/b microProteins to interact with TOPLESS/TOPLESS-RELATED (TPL/TPR) proteins. Interaction of CO with miP1a/b/TPL causes late flowering due to a failure in the induction of FLOWERING LOCUS T (FT) expression under inductive long day conditions. Both miP1a and miP1b are expressed in vascular tissue, where CO and FT are active. Genetically, miP1a/b act upstream of CO thus our findings unravel a novel layer of flowering time regulation via microProtein-inhibition.
Project description:Phalaenopsis amabilis, one of the most important flowers in the current international flower market, is a plant that undergoes vernalization and requires low temperature treatment for flowering. There have been few reports on the proteomic analysis of the development of flower buds. In this study, by using isobaric tags for relative and absolute quantification (iTRAQ), 4096 differentially expressed proteins were identified in P. amabilis under low temperature treatment, of which 42 were associated with early floral induction, and 18 were verified by mass spectrometry multi-reaction monitoring (MRM). Among the proteins associated with the vernalization pathway, PEQU_11434 (glycine-rich RNA-binding protein GRP1A-like) and PEQU_11045 (UDP-N-acetylglucosamine diphosphorylase) were upregulated compared to their expression in control flower buds. It was therefore inferred that O-GlcNAc glycosylation was involved in the posttranscriptional modification of VRN1 (API homolog) and that the GRP2 protein (glycine-rich RNA-binding protein) was glycosylated to relieve binding to the VRN1 mRNA precursor to promote the expression of VRN1, which initiates floral development. Furthermore, phytochromes A (PEQU_13449, PEQU_35378), B (PEQU_09249) and C (PEQU_41401) were downregulated under low temperature treatment compared to their expression in control flower buds, suggesting that they could repress the expression of the VRN2 gene and thus release its repression of VRN3 to enable the high-level expression of FTPEQU_19304 (FT, VRN3 homolog), which promotes VRN1 expression and then stimulates flowering.
Project description:In plants, endogenous and environmental signals such as light control the timing of the transition to flowering . Two phytochrome B-interacting transcription factors, VASCULAR PLANT ONE–ZINC FINGER1 (VOZ1) and VOZ2 redundantly promote flowering in Arabidopsis thaliana. In the voz1 voz2 mutant the expression of FLOWERING LOCUS C (FLC) was up-regulated and expression of FLOWERING LOCUS T (FT) was down-regulated, which was proposed to be the cause of late flowering in voz1 voz2. However, the detailed mechanism by which the VOZ genes promote flowering is not well understood. Here, we show that neither the reduced FT-expression nor the late-flowering phenotype of voz1 voz2 is suppressed in the voz1 voz2 flc triple mutant . Genetic interaction experiments between voz1 voz2 and constans-2 (co-2) mutants reveal that the VOZs and CO work in the same genetic pathway. Using in vitro pull-down, electrophoretic mobility shift assays and bimolecular fluorescence complementation assays, we show that VOZ1 and VOZ2 interact with CO. The voz1 voz2 35S::CO:YFP plants show suppression of the early-flowering phenotype induced by CO-overexpression, showing that CO requires VOZ for induction of flowering. Determination of the VOZ consensus binding site followed by genome-wide sequence analysis failed to identify any VOZ-binding sites near known flowering-time genes. Together, these results indicate that the VOZ genes regulate flowering primarily through the photoperiod pathway, independent of FLC, and suggest that VOZs modulate CO function to promote flowering.
Project description:Senescence represents the last stage of flower development. Phosphorylation is one of the key post-translational modifications that regulate protein functions in diverse biological pathways and contexts. Generally, kinases may be more required than phosphatases during plant growth and development. However, little is known about global phosphorylation change during flower senescence. In this work, we quantitatively investigated the petunia phosphoproteome following ethylene or air treatment. In total, 2170 phosphosites in 1184 protein groups were identified, among which 2059 sites in 1124 proteins were quantified. Treatment with ethylene resulted in 711 down-regulated and only 117 up-regulated phosphosites using a 1.5-fold threshold (P<0.05), showing that ethylene negatively regulates global phosphorylation levels and that phosphorylation of lots of proteins was not necessary during flower senescence. Our results show that protein dephosphorylation may play an important role of in ethylene-induced corolla senescence in petunia and that phosphatases may be more required than kinases during flower senescence. In addition, our results show that ethylene regulates ethylene and ABA signaling transduction pathways via phosphorylation level, and plant mRNA splicing machinery was a major target of ethylene-induced dephosphorylation. Moreover, ethylene treatment increases the number of alternative splicing of precursor RNAs in petunia corollas.