Unknown,Transcriptomics,Genomics,Proteomics

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The embryonic leaf identity gene FUSCA3 also regulates vegetative phase transitions by negatively regulating ethylene action in Arabidopsis

ABSTRACT: The embryonic temporal regulator FUSCA3 (FUS3) plays major roles in the establishment of embryonic leaf identity and regulation of developmental timing. Loss-of-function mutations of this B3-domain transcription factor result in replacement of cotyledons with leaves and precocious germination, while constitutive misexpression causes the conversion of leaves into cotyledon-like organs and delays vegetative and reproductive phase transitions. To identify downstream targets of FUS3 involved in vegetative phase transitions, we performed microarray analysis on seedlings that transiently activate FUS3 using the AtML1:FUS3-GR-DEX inducible system. Using this construct, FUS3 is ectopically expressed in the ML1 or epidermal layer and becomes activated upon addition of dexamethasone. We found that activation of FUS3 after germination dampens the expression of genes involved in the biosynthesis and response to the plant hormone ethylene, while a loss-of-function fus3 mutant shows many phenotypes consistent with increased ethylene signaling. This FUS3-dependent regulation of ethylene signaling also impinges on timing functions outside of embryogenesis. Loss of FUS3 function results in accelerated vegetative phase change and this is again partially dependent on functional ethylene action. This alteration in vegetative phase transition is dependent on both embryonic and vegetative FUS3 function, suggesting that this important transcriptional regulator controls both embryonic and vegetative developmental timing. This study indicates that the embryonic regulator FUS3 not only regulates the embryonic-to-vegetative phase transition through hormonal regulation, but also functions postembryonically to modulate vegetative phase transitions by negatively regulating ethylene action.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Shelley Lumba

PROVIDER: E-MEXP-3465 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The FUSCA3 (FUS3) transcription factor is considered to be a master regulator of seed maturation because a wide range of seed maturation events are impaired in its defective mutant. To comprehensively identify genes under the control of FUS3, two types of microarray experiments were performed. First, transgenic plants in which FUS3 expression could be induced by the application of estrogen (ESTR) were used to identify the genes up-regulated in young seedlings of Arabidopsis in response to ectopic expression of FUS3. Second, transcriptomes were compared between fus3 mutant and wild type developing seeds. Combining the results of these experiments identified genes under relatively immediate and robust control of FUS3 during seed development. The analyses expanded the range of types of genes under the control of FUS3. The genes positively controlled by FUS3 are not confined to previously known seed maturation-related genes and include those for production of secondary metabolites such as glucosinolates, phenylpropanoids and falvonoids, and primary metabolism such as photosynthesis and fatty acid biosynthesis. Furthermore, several different patterns were identified in the manner of ectopic activation by FUS3 including the kinetics and abscisic acid (ABA) requirement of downstream genes depending on their natures of developmental regulation, suggesting mechanistic diversity of gene regulation by FUS3. This Series contains two different types of microarray experiments. First, seven-day-old seedlings of the ESTR-inducible FUS3 transgenic plants (ER-FUS3) were cultured for 3, 6, 12, 24, 36 or 48 h in a standard liquid medium or in that containing ESTR, ABA or both. RNA was prepared from each time point sample of each treatment and used to prepare Cy5-labeled cRNA, which was subsequently probed by DNA array with the reference Cy3-labeled cRNA prepared from the time zero untreated seedlings. Two biological replicates per treatment. Second, transcriptomes were compared between developing seeds of the wild-type (Col-0) and the fus3-3 mutant. This experiment contains two seed developmental stages, 8 days after flowering (DAF; bent cotyledon stage) and at 12 DAF (green cotyledon stage). Sample pairs in three biological replicates were analyzed by the two-color method. Dye-swapped hybridizations were performed in every replicate.

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The embryonic leaf identity gene FUSCA3 also regulates vegetative phase transitions by negatively regulating ethylene action in Arabidopsis

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