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Signalling events triggered in response to gibberellins during potato tuber sprouting

ABSTRACT: The aim of this study is to analize the genes that are up or down-regulated in response to sprouting promoting effectors such as GAs and light, in order to elucidate the signalling cascades triggered during this process. Half sections of certified pathogen tested Spunta tubers were put into sterile MS-agar (0.7%) containing flasks at 16„aC and subjected to different treatments: Dark controls: Tuber sections were incubated for 10 or 35 days in complete darkness. Light control: Tuber sections were incubated for 10 days under continuous light conditions. Treatments performed in complete darkness during 35 days: a) GA treatment: 5 uM gibberellic acid (C19H22O6) = GA3 (Sigma) b) Sucrose treatment: 1% sucrose c) GA & sucrose treatment: 5 uM GA3 (Sigma) + 1% sucrose Treatments performed in light (10 days)d) GA treatment : 5 uM GA3 (Sigma). Three independent biological replicates of each condition (3 controls and 4 treatments) wereperformed. At the moment of harvest (10 or 35 days) sprout length was measured and photographs of the different samples were taken. Sprouts were excised and frozen in liquid Nitrogen. Total RNA was obtained using the Trizol reagent (Invitrogen) according to the manufacturer¡¦s instructions. RNAs were quantified by OD: 260/280 and checked in agarose gels. Keywords: Loop design

ORGANISM(S): Solanum tuberosum

PROVIDER: GSE10492 | GEO | 2008/02/13

SECONDARY ACCESSION(S): PRJNA107947

REPOSITORIES: GEO

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Project description:Cold storage (CS) is widely used to extend fruit postharvest. In peach, chilling injuries may cause intense juice loss leading to a dry âwoollyâ texture of the fruit flesh. The disturbance, named woolliness, is associated to abnormal pectin metabolism and results in anatomical and physiological alterations. Application of gibberellic acid (GA) at the initial stages of pit hardening has been shown to impair woolliness incidence, however the mechanisms controlling the response remain unknown. We have employed genome wide transcription analyses to investigate the effects of GA application and CS of peaches. Approximately half (48.26%, 13846) of the investigated genes exhibited significant differential expression in response to the treatments. Gene ontology classes associated to cellular and developmental processes were overrepresented among the differentially regulated genes, whereas sequences classified in cell death and immune response categories were underrepresented. Gene set enrichment analyses demonstrated a predominant role of CS in repressing the transcription of genes associated to cell wall metabolism. In contrast, genes involved in hormone metabolism and signaling exhibited a more complex transcriptional response to the factors, indicating an extensive network of crosstalk between GA and low temperatures. Time course transcriptional profiling analyses also confirmed the involvement of cell wall metabolism genes in woolliness onset in peach. Overall, our results provide further insights on the mechanisms controlling the complex phenotypes associated to postharvest textural changes in peach. Four samples (CONT, CONTcs, GA3, GA3cs), each with three biological replicates (R1, R2 and R3), were analyzed. Control samples (CONT and CONTcs) consist of peach mesocarp not treated with GA3 at pit hardening, and either assayed at harvest (CONT) or after 15 days of cold storage (CONTcs). GA3 samples (GA3 and GA3cs) consist of peach mesocarp treated with GA3 at pit hardening, and either assayed at harvest (GA3) or after 15 days of cold storage (GA3cs).

Project description:The proper transition between the skotomorphogenic and photomorphogenic developmental programs is key for seedling survival and it is therefore tightly regulated. Besides light, which is the major cue that triggers this transition, several hormone pathways participate in this control, mainly antagonizing the light effect. Two of these hormones are gibberellins (GA) and brassinosteroids (BR). Both hormones promote the skotomorphogenesis and prevent photomorphogenesis, as evidenced by the partially de-etiolated phenotype caused by GA or BR deficiency, or by a blockage in the respective signaling pathways. We have previously shown that BR mediate GA activity in the control of this transition. For instance, treatment of dark-grown, GA-deficient seedlings with BRs was able to partially restore morphological phenotypes such as hypocotyl growth, and importantly to restore completely the molecular phenotype of CAB2 and RbcS gene expression. On the contrary, GA-treatment did not alter the same phenotypes in the BR deficient det2 mutant. Thus, the aim of this study was to investigate to what extent BRs mediate GA activity in dark-grown seedlings, and for that purpose we have the examined global changes in gene expression caused by treatment with GA and BRs in BR- and GA-deficient seedlings, respectively. We performed two sets of experiments, each one including three different samples. The first experiment included dark-grown, WT Col-0 mock-treated seedlings, or seedlings treated with either the GA biosynthesis inhibitor paclobutrazol (PAC) or with PAC+epibrassinolide (EBR). In this experiment, PAC samples were labelled with Cy3 and compared to WT and PAC+EBR samples, which were labelled with Cy5. Two biological repeast were performed. The second experiment included dark-grown, WT Col-0 seedlings and the det2-1 mutant, which was either mock-treated or treated with GA3. Two biological replicates were performed, in the first one the WT and det2-1+GA3 were labelled with Cy3 and the det2-1 sample with Cy5. The reverse labelling was used in the second replicate.

Project description:Background: Grape is highly sensitive to gibberellin (GA), which is crucial during seed and berry development (SBD) either by itself or by interacting with other hormones, such as auxin, Abscisic acid (ABA), and Cytokinin (CK). However, no systematic analysis of GA metabolic and signal transduction (MST) pathway has been undertaken in grapevine. Results: In this study, total endogenous GA3 content significantly decreased during SBD, and a total of 48 known genes in GA metabolic (GAM; 31) and signal transduction (ST; 17) pathways were identified in this process. In the GAM pathway, out of 31 genes, VvGA20ox1-1, VvGA3ox4-1, and VvGA2ox1-1 may be the major factors interacting at the green-berry stage (GBS) accompanied with higher accumulation rate. GA biosynthesis was greater than GA inactivation at GBS, confirming the importance of seeds in GA synthesis. The visible correlation between endogenous GA3 content and gene expression profiles suggested that the transcriptional regulation of GA biosynthesis pathway genes was a key mechanism of GA accumulation at the stone-hardening stage (SHS). Interestingly, we observed a negative feedback regulation between VvGA3oxs-VvGAI1-4, VvGA2oxs-VvGAI1-4, and VvGID1B-VvGAI1-4 in maintaining the balance of GA3 content in berries. Moreover, 11 miRNAs may be involved in the modulation of GA MST pathway by mediating their target genes, such as VvGA3ox, VvGID1B, and VvGAMYB. Many genes in auxin, ABA, and CK MST pathways were further identified and found to have a special pattern in the berry, and the crosstalk between GA and these hormones may modulate the complex process during SBD through the interaction gene network of the multihormone pathway. Lastly, based on the expression characterization of multihormone MST pathway genes, a proposed model of the GA-mediated multihormone regulatory network during SBD was proposed. Conclusions: Our results provided novel insights into GA-mediated regulatory networks during SBD in grape. The complexity of GA-mediated multihormone ST in SBD was also elucidated, thereby providing valuable information for future functional characterizations of specific genes in grape.

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Signalling events triggered in response to gibberellins during potato tuber sprouting

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