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Gene expression analysis on Hydrangea macrophylla sepal pigmentation due to aluminum accumulation

ABSTRACT: Most blue color in flowers is due to anthocyanin, and considerable proportion of blue coloration can be attributed to metal-complexed anthocyanins. Recently, we reported vacuolar localized iron-transporter in blue petal cells of Tulipa gesneriana. However the mechanism of another metal ion transporters and subsequent flower color development has yet to be fully explored. In Hydrangea macrophylla, Al3+ is involved in blue coloration and the anthocyanin is formed Al3+-complex in vacuoles. To identify the molecular mechanism of blue coloration in hydrangea flowers, we tried to isolate the related genes transporting metal ion into vacuoles. From the sepal cDNA library we read the sequences of ca. 12000 genes, then a microarray analysis was carried out. From the sequences information, we chose several genes that might localize vacuolar membrane and transport Al3+. By using Al3+-sensitive yeast strain, we could identify the gene transporting Al3+ into vacuole. From the functional similarity and predicted localization, we could also identify the gene transporting Al3+ into cytosol. We will report the Al3+ mobilization from out of cell into vacuole in the sepal of Hydrangea macrophylla.

ORGANISM(S): Hydrangea macrophylla

PROVIDER: GSE26665 | GEO | 2012/12/20

SECONDARY ACCESSION(S): PRJNA136409

REPOSITORIES: GEO

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Gene expression analysis on Hydrangea macrophylla sepal pigmentation due to aluminum accumulation

Project description:Most blue color in flowers is due to anthocyanin, and considerable proportion of blue coloration can be attributed to metal-complexed anthocyanins. Recently, we reported vacuolar localized iron-transporter in blue petal cells of Tulipa gesneriana. However the mechanism of another metal ion transporters and subsequent flower color development has yet to be fully explored. In Hydrangea macrophylla, Al3+ is involved in blue coloration and the anthocyanin is formed Al3+-complex in vacuoles. To identify the molecular mechanism of blue coloration in hydrangea flowers, we tried to isolate the related genes transporting metal ion into vacuoles. From the sepal cDNA library we read the sequences of ca. 12000 genes, then a microarray analysis was carried out. From the sequences information, we chose several genes that might localize vacuolar membrane and transport Al3+. By using Al3+-sensitive yeast strain, we could identify the gene transporting Al3+ into vacuole. From the functional similarity and predicted localization, we could also identify the gene transporting Al3+ into cytosol. We will report the Al3+ mobilization from out of cell into vacuole in the sepal of Hydrangea macrophylla. Three sepal pigmentation stages were chosen: S1=no pigmentation-sepal closed; S2=started pigmentation-sepal opening; and S3=pigmentation complete-sepal opened. One pooled sepal sample per stage was prepared and gene expression pattern was analyzed by custom-designed Hydrangea oligo DNA microarray (CombiMatrix 12K). The genes that were expressed >2-fold or <0.5-fold in S3 compared with S1 and S2 were selected as potential players involved in sepal pigmentation due to aluminum accumulation.

2012-12-20 | E-GEOD-26665 | biostudies-arrayexpress

Identification of plant vacuolar transporters mediating phosphate storage

Project description:Plant vacuoles serve as the primary intracellular compartments for inorganic phosphate (Pi) storage. Passage of Pi across vacuolar membranes plays a critical role in buffering the cytoplasmic Pi level against fluctuations of external Pi and metabolic activities. Here we demonstrate that the SPX-MFS proteins, designated as Phosphate Transporter 5 family (PHT5), also named Vacuolar Phosphate Transporter (VPT), function as vacuolar Pi transporters. Based on 31P-magnetic resonance spectroscopy analysis, Arabidopsis pht5;1 loss-of-function mutants accumulate less Pi and exhibit a lower vacuolar-to-cytoplasmic Pi ratio than controls. Conversely, overexpression of PHT5 leads to massive Pi sequestration into vacuoles and altered regulation of Pi starvation-responsive genes. Furthermore, we show that heterologous expression of OsSPX-MFS1, the rice PHT5 homolog, mediates Pi influx to yeast vacuoles. Our findings uncover a group of Pi transporters in vacuolar membranes that regulate the cytoplasmic Pi homeostasis required for the fitness of plant growth. 10-day seedlings grown on Pi-sufficient medium or followed by 1-day or 3-day Pi starvation, shoot RNA and root RNA were extracted separately

2016-03-28 | E-GEOD-74856 | biostudies-arrayexpress

Identification of plant vacuolar transporters mediating phosphate storage

2016-03-28 | GSE74856 | GEO

TRIM21 innate immune detection of GBP1-positive Toxoplasma gondii is essential for host survival

Project description:Interferon gamma (IFN) is the major pro-inflammatory cytokine conferring resistance to the intracellular vacuolar pathogen Toxoplasma gondii. Autophagy along with immunity-related GTPases (IRGs), guanylate binding proteins (GBPs) and the E3 ubiquitin ligase TNF receptor-associated factor 6 (TRAF6) mediate clearance of Toxoplasma in IFN-stimulated cells. With the exception of inflammasomes, no host innate immune mediators impacting host survival have been identified at disrupted parasitophorous vacuoles. We show that IFN drives recruitment of the E3 ubiquitin ligase TRIM21 to GBP1-positive avirulent Toxoplasma vacuoles. This led to Lys63-linked ubiquitination of the vacuole and secretion of pro-inflammatory cytokines. GBPs were ubiquitinated during infection and TRIM21 controlled their expression levels and the efficient recruitment of GBP1 to the vacuole. TRIM21 deficiency led to an enhanced early replication of Toxoplasma without interfering with vacuolar disruption. TRIM21-/- mice were highly susceptible to Toxoplasma infection, exhibiting decreased levels of pro-inflammatory cytokines and higher parasite burden in the brain. This study identifies TRIM21 as a previously unknown modulator of Toxoplasma gondii resistance thereby extending host innate immune recognition of eukaryotic pathogens to include E3 ubiquitin ligases.

2018-03-12 | PXD003217 | Pride

Transcriptome analysis of apple peel samples at different time points in the process of changing from green to red under light conditions after unbagging

Project description:Purpose:The red coloration of apple (Malus × domestica Borkh.) is due to the accumulation of anthocyanins in the fruit peel. Light is essential for anthocyanin biosynthesis in apple.Apple peel can quickly turn red under light conditions after unbagging. Therefore, the implementation of transcriptome sequencing to find genes that promote anthocyanin accumulation in response to light signals is necessary to clarify the mechanism of light-induced anthocyanin accumulation in apple peel.

2021-10-02 | GSE185089 | GEO

Comparison of expression profiles in response to blue, UV-A, and UV-B in the swollen hypocotyls of turnip

Project description:UV-A mediated regulation of anthocyanin biosynthesis was investigated in swollen hypocotyls of the red turnip ‘Tsuda’. The shaded swollen hypocotyls which contained negligible anthocyanin were exposed to artificial light sources including low fluence UV-B, UV-A, blue, red, far-red, red plus UV-A, far-red plus UV-A, and blue plus red. Among these lights, only UV-A induced anthocyanin biosynthesis and co-irradiation of red or far-red with UV-A did not affect the extent of UV-A induced anthocyanin accumulation. The expression of phenylalanine ammonia lyase (PAL; EC 4.3.1.5), chalcone synthase (CHS; EC 2.3.1.74), flavanon 3-hydrocylase (F3H; EC 1.14.11.9), dihydroflavonol 4-reductase (DFR; EC 1.1.1.219) and anthocyanidin synthase (ANS; EC 1.14.11.-) genes were increased with time during a 24 hour exposure of UV-A. In contrast, irradiation of red, blue, UV-B, and a combination of blue with red failed to induce CHS expression. Microarray analysis showed that only a few genes, including chalcone synthase and flavanon 3-hydroxylase were induced significantly by UV-A, while a separate set of many genes was induced by low fluence UV-B. The UV-A specific induction of anthocyanin biosynthesis and the unique gene expression profile upon UV-A irradiation as compared with blue and UV-B demonstrated that the observed induction of anthocyanin biosynthesis in red turnips was mediated by a distinct UV-A specific photoreceptor, but not by phytochromes, UV-A/blue photoreceptors, or UV-B photoreceptors. Keywords: light response

2007-02-22 | GSE6876 | GEO

Transcriptome analysis reveals candidate genes related to anthocyanin biosynthesis in different carrot genotypes and tissues

Project description:Taproots of different carrot genotypes were used to identify the candidate genes related to anthocyanin synthesis with particular focus on R2R3MYB, bHLH transcription factors, and glutathione S-transferase gene (GST). The RNA-sequencing analysis (RNA-Seq) showed that DcMYB6 and DcMYB7 had genotypic dependent expressed and they are likely involved in the regulation of anthocyanin biosynthesis. They were specifically upregulated in solid black taproots, including both black phloem and xylem. DcbHLH3 (LOC108204485) was upregulated in all black samples compared with the orange ones. We also found that GST1 (LOC108205254) might be an important anthocyanin transporter, and its upregulated expression resulted in the increasing of vacuolar anthocyanin accumulation in black samples.

2020-03-06 | GSE146419 | GEO

grape berry vacuole proteomic - Quantitative Proteome Analysis Reveals Changes in the Protein Landscape During Grape Berry Development With a Focus on Vacuolar Transport Proteins

Project description:The vacuole occupies a large portion of plant cell volume, it is especially true to fruit tissues. Berry flesh cell vacuole serves as storage organelle for water, sugars, acids, secondary metabolites and others, which largely determining berry quality (Fontes et al., 2011a, b; Shiratake and Martinoia, 2007, Conde et al., 2006). However, the molecular basis of these compartmentation processes is still poorly understood. As in many species, the major bottle neck to study these aspects in grapevine is to obtain highly purified vacuoles with a good yield (Fontes et al., 2010). Up to date, several vacuole or tonoplast proteome researches were applied on a few plants mainly on Arabidopsis thaliana, vacuoles or tonoplast were derived from mesophyll cells (Carter et al., 2004, Endler et al., 2006, Schulze, et al., 2012) or cell culture (Jaquinod et al 2006, Shimaoka et al 2004), cauliflower buds (Schmidt et al., 2007) and sugar beet taproots (Jung et al., 2015). Though the grape berry protoplasts and intact vacuoles were successfully isolated from Cabernet Sauvignon berry suspension-cultured cells (Fontes et al., 2010), the vacuoles isolated from grape berry or different development and ripening stages of grape berry mesocarp tissues were not achieved.

2019-11-12 | PXD010556 | Pride

Yeast TLDc domain-containing proteins control assembly and subcellular localization of the V-ATPase

Project description:Yeast vacuoles, equivalent to lysosomes in other eukaryotes, are important acidic degradative organelles as well as storage compartments and signaling hubs. To perform these functions, they rely on important protein complexes, including the V-ATPase, responsible for organelle acidification. In this study, we used cross-linking mass spectrometry to characterize the protein complexes of isolated vacuoles. We were able to detect many known protein-protein interactions, including known protein complexes, as well as undescribed ones. Among these, we identified the uncharacterized TLDc domain-containing protein Rtc5 as a novel interactor of the V-ATPase. We show that Rtc5 localizes to the vacuole membrane depending on N-myristoylation and on its interactions with the V-ATPase. We further analyzed the influence of this protein, and the second yeast TLDc domain-containing protein, Oxr1, on V-ATPase function. We find that both Rtc5 and Oxr1 promote the disassembly of the vacuolar V-ATPase in vivo, counteracting the role of the assembly chaperone, the RAVE complex. Finally, Oxr1 is necessary for the retention in the late Golgi complex of an organelle-specific subunit of the V-ATPase. Collectively, our results shed light on the in vivo roles of yeast TLDc domain-containing proteins in relation to the V-ATPase, highlighting the multifaceted regulation of this crucial protein complex.

2024-03-11 | PXD046792 | Pride

ESCRTIII-associated protein AtALIX mediates high affinity phosphate transporter trafficking to maintain phosphate homeostasis in Arabidopsis

Project description:Prior to the release of their cargoes into the vacuolar lumen, sorting endosomes mature into multivesicular bodies (MVB) through the action of ENDOSOMAL COMPLEX REQUIRED FOR TRANSPORT (ESCRT) protein complexes. MVB-mediated sorting of high affinity phosphate transporters (PHT1) to the vacuole limits their plasma membrane levels under phosphate sufficient conditions, a process that allows plants to maintain phosphate homeostasis. We describe here AtALIX, a cytosolic protein that associates to MVB by interacting with ESCRT-III subunit SNF7 and enables PHT1;1 trafficking to the vacuole. Thus, we show that the partial loss of function mutant Atalix-1 displays reduced vacuolar degradation of PHT1;1. AtALIX versions containing the Atalix-1 mutation showed reduced interaction with SNF7, providing a simple molecular explanation for impaired cargo trafficking in Atalix-1 mutants. Indeed, Atalix-1 mutation also hampered vacuolar sorting of brassinosteroid receptor BRI1. In line with a presumed broad target spectrum, Atalix-1 mutation is pleiotropic, provoking reduced plant growth, late flowering and altered vacuole morphogenesis, whereas null mutants are lethal, indicating that AtALIX controls diverse processes in plants being essential for their life.

2015-09-09 | PXD002075 | Pride

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