Indirect ABA-dependent regulation of seed storage protein genes by FUSCA3 transcription factor in Arabidopsis.
ABSTRACT The key transcription factors that control seed maturation, ABSCISIC ACID INSENSITIVE3 (ABI3) and FUSCA3 (FUS3), share homologous DNA-binding domains. Regulation of seed storage protein genes At2S3 and CRC by ABI3 and FUS3 was investigated using transgenic plants in which ABI3 and FUS3 could be ectopically induced by steroid hormones. Like ABI3, the presence of FUS3 led to expression of At2S3 and CRC in vegetative tissues. FUS3-mediated induction of CRC was completely dependent on exogenous abscisic acid (ABA), while At2S3 was weakly induced without ABA but strongly enhanced with ABA. This ABA dependency of FUS3-induced CRC and At2S3 expression was similar to that observed for ABI3. However, kinetic analysis revealed distinctions between the mechanisms of ABA-dependent CRC regulation by FUS3 or ABI3, and between target genes. While At2S3 activation by FUS3 was rapid, CRC induction by FUS3 in the presence of ABA, and by ABA followed by the presence of FUS3, took a significantly longer time (24-36 h). This suggested the involvement of an indirect mechanism requiring the ABA- and FUS3-dependent synthesis of intermediate regulatory factor(s). A chimeric protein composed of the FUS3 B3 domain, and a heterologous activation domain and nuclear localization signal exhibited a tight coupling with ABA regulation as observed for wild-type FUS3. Simultaneous induction of FUS3 and ABI3 did not result in the synergistic activation of CRC and At2S3. Based on these results, similarities and differences in the mechanisms of seed storage protein gene regulation by FUS3 and ABI3 are discussed.
Article: The role of the Arabidopsis FUSCA3 transcription factor during inhibition of seed germination at high temperature.[show abstract] [hide abstract]
ABSTRACT: Imbibed seeds integrate environmental and endogenous signals to break dormancy and initiate growth under optimal conditions. Seed maturation plays an important role in determining the survival of germinating seeds, for example one of the roles of dormancy is to stagger germination to prevent mass growth under suboptimal conditions. The B3-domain transcription factor FUSCA3 (FUS3) is a master regulator of seed development and an important node in hormonal interaction networks in Arabidopsis thaliana. Its function has been mainly characterized during embryonic development, where FUS3 is highly expressed to promote seed maturation and dormancy by regulating ABA/GA levels. In this study, we present evidence for a role of FUS3 in delaying seed germination at supraoptimal temperatures that would be lethal for the developing seedlings. During seed imbibition at supraoptimal temperature, the FUS3 promoter is reactivated and induces de novo synthesis of FUS3 mRNA, followed by FUS3 protein accumulation. Genetic analysis shows that FUS3 contributes to the delay of seed germination at high temperature. Unlike WT, seeds overexpressing FUS3 (ML1:FUS3-GFP) during imbibition are hypersensitive to high temperature and do not germinate, however, they can fully germinate after recovery at control temperature reaching 90% seedling survival. ML1:FUS3-GFP hypersensitivity to high temperature can be partly recovered in the presence of fluridone, an inhibitor of ABA biosynthesis, suggesting this hypersensitivity is due in part to higher ABA level in this mutant. Transcriptomic analysis shows that WT seeds imbibed at supraoptimal temperature activate seed-specific genes and ABA biosynthetic and signaling genes, while inhibiting genes that promote germination and growth, such as GA biosynthetic and signaling genes. In this study, we have uncovered a novel function for the master regulator of seed maturation, FUS3, in delaying germination at supraoptimal temperature. Physiologically, this is important since delaying germination has a protective role at high temperature. Transcriptomic analysis of seeds imbibed at supraoptimal temperature reveal that a complex program is in place, which involves not only the regulation of heat and dehydration response genes to adjust cellular functions, but also the activation of seed-specific programs and the inhibition of germination-promoting programs to delay germination.BMC Plant Biology 01/2012; 12:15. · 3.45 Impact Factor
Article: Gene coexpression clusters and putative regulatory elements underlying seed storage reserve accumulation in Arabidopsis.[show abstract] [hide abstract]
ABSTRACT: In Arabidopsis, a large number of genes involved in the accumulation of seed storage reserves during seed development have been characterized, but the relationship of gene expression and regulation underlying this physiological process remains poorly understood. A more holistic view of this molecular interplay will help in the further study of the regulatory mechanisms controlling seed storage compound accumulation. We identified gene coexpression networks in the transcriptome of developing Arabidopsis (Arabidopsis thaliana) seeds from the globular to mature embryo stages by analyzing publicly accessible microarray datasets. Genes encoding the known enzymes in the fatty acid biosynthesis pathway were found in one coexpression subnetwork (or cluster), while genes encoding oleosins and seed storage proteins were identified in another subnetwork with a distinct expression profile. In the triacylglycerol assembly pathway, only the genes encoding diacylglycerol acyltransferase 1 (DGAT1) and a putative cytosolic "type 3" DGAT exhibited a similar expression pattern with genes encoding oleosins. We also detected a large number of putative cis-acting regulatory elements in the promoter regions of these genes, and promoter motifs for LEC1 (LEAFY COTYLEDON 1), DOF (DNA-binding-with-One-Finger), GATA, and MYB transcription factors (TF), as well as SORLIP5 (Sequences Over-Represented in Light-Induced Promoters 5), are overrepresented in the promoter regions of fatty acid biosynthetic genes. The conserved CCAAT motifs for B3-domain TFs and binding sites for bZIP (basic-leucine zipper) TFs are enriched in the promoters of genes encoding oleosins and seed storage proteins. Genes involved in the accumulation of seed storage reserves are expressed in distinct patterns and regulated by different TFs. The gene coexpression clusters and putative regulatory elements presented here provide a useful resource for further experimental characterization of protein interactions and regulatory networks in this process.BMC Genomics 06/2011; 12:286. · 4.07 Impact Factor