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Next-Generation Sequencing Facilitates Quantitative Analysis of Wild Type and abi5 Transcriptomes

ABSTRACT: It has been known that ABA INSENSITIVE 5 (ABI5) plays a vital role in regulating seed germination. In the present experiment, we showed that 3-amino-1,2,4-triazole (3-AT) inhibits seed germination of the loss-of-function mutant abi5-1, but promotes seed germination of the ABI5-overexpression transgenic line, and that ABI5 affects reactive oxygen species (ROS) homeostasis. RNA-sequencing analysis indicates that the transcription of the genes involved in ROS metabolic process or genes responsive to ROS stress is impaired in abi5-1 seeds. Together, all the findings suggest that ABI5 regulates seed germination at least partly by affecting ROS homeostasis.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE90004 | GEO | 2017/07/11

SECONDARY ACCESSION(S): PRJNA354330

REPOSITORIES: GEO

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NUP1 modulates proteasome based degradation of ABI5 in Nuclear Pore Complex region to regulate Arabidopsis seed germination

Project description:NUCLEOPORIN 1 (NUP1), a member of the Nuclear Pore Complex (NPC), is located on the inner side of the nuclear membrane. It is highly expressed in seeds; however, its role in seeds including germination has not been explored yet. Here, we identified an ABA hypersensitive phenotype of nup1 during germination. ABA treatment drastically changes the expression pattern of thousands of genes in nup1, including the major transcription factors (TFs) involved in germination, ABI3, ABI4, and ABI5. Double mutant analysis of NUP1 and these ABA-related genes showed that mutations in ABI5 can rescue the phenotype of nup1, suggesting that NUP1 acts upstream of ABI5 to regulate seed germination. ABI5, a negative regulator of germination was previously reported to be predominantly expressed in nucleus. Our subnuclear localization study indicates ABI5 is mostly localized in nucleoplasm. It is abundant in dry seeds and rapidly degraded during germination, but the details of its degradation site is unclear. Here, we found that NUP1 is physically associated with ABI5 and the 19S regulatory particle (RP) of the 26S proteasome near NPC. Mutation in NUP1 delayed the ABI5 degradation through its post-translation retention in nucleolus after abiotic stress treatment. Taken together, we propose, NUP1 anchors the proteasome to NPC and modulates Arabidopsis seed germination through proteasome-mediated degradation of ABI5 in the vicinity of NPC in nucleoplasm.

2023-08-14 | GSE189513 | GEO

The COP9 Signalosome regulates seed germination by facilitating protein degradation of RGL2 and ABI5

Project description:The control of seed germination and seed dormancy are critical for the successful propagation of plant species, and are important agricultural traits. Seed germination is tightly controlled by the balance of gibberellin (GA) and abscisic acid (ABA), and is influenced by environmental factors. The COP9 Signalosome (CSN) is a conserved multi-subunit protein complex that is best known as a regulator of the Cullin-RING family of ubiquitin E3 ligases (CRLs). Multiple viable mutants of the CSN showed poor germination, except for csn5b-1. Detailed analyses showed that csn1-10 has a stronger seed dormancy, while csn5a-1 mutants exhibit retarded seed germination in addition to hyperdormancy. Both csn5a-1 and csn1-10 plants show defects in the timely removal of the germination inhibitors: RGL2, a repressor of GA signaling, and ABI5, an effector of ABA responses. We provide genetic evidence to demonstrate that the germination phenotype of csn1-10 is caused by over-accumulation of RGL2, a substrate of the SCF (CRL1) ubiquitin E3 ligase, while the csn5a-1 phenotype is caused by over-accumulation of RGL2 as well as ABI5. The genetic data are consistent with the hypothesis that CSN5A regulates ABI5 by a mechanism that may not involve CSN1. Transcriptome analyses suggest that CSN1 has a more prominent role than CSN5A during seed maturation, but CSN5A plays a more important role than CSN1 during seed germination, further supporting the functional distinction of these two CSN genes. Our study delineates the molecular targets of the CSN complex in seed germination, and reveals that CSN5 has additional functions in regulating ABI5, thus the ABA signaling pathway.

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2013-10-30 | E-GEOD-51830 | biostudies-arrayexpress

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Project description:Abscisic acid (ABA) plays a key role in plant development and responses to abiotic stress. A wide number of regulatory mechanisms on ABA perception and signaling are known, including transcriptional regulation and post-translational regulations; however, less to know about the post-transcriptional regulations. In this work, we have found that SKIP, a splicing factor in spliceosome, positively regulates ABA signal transduction. Mutation of SKIP, skip-1, confers ABA insensitive phenotype in seed germination, root growth, and ABA-responsive gene expression. Transformation of SKIP genomic DNA to skip-1 is able to recover its defects. SKIP binds to the pre-mRNA of genes in ABA signaling, such as PYL8, to regulate their splicing. The alternative splicing of PYL7, PYL8, ABF2, and ABI5 in ABA pathway is disrupted by skip-1, reducing the mRNA levels of them. The abnormal splicing of PP2Cs activates their transcription by skip-1, suggesting there is a feedback regulation for PP2Cs caused by skip-1. The down-regulation of PYL ABA receptors, ABF2 and ABI5 ABA response transcription factors, and the up-regulation of PP2Cs through alternative splicing reduce the plant responses to ABA in skip-1. SKIP is required for ABA-mediated genome-wide alternative splicing. ABA treatment enhances the novel splicing events in WT genome-widely. The novel alternative splicing events are increased by skip-1 in the absence and present of ABA. Our results first reveal a principle on how a splicing factor affects the ABA signaling.

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Project description:A transcriptome analysis of developing abi5 and wild type (R108) seeds from Medicago truncatula was performed to decipher the role of ABI54 in the regulation of late seed maturation and seed longevity.

2016-09-24 | GSE87313 | GEO

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2013-08-01 | E-GEOD-44165 | biostudies-arrayexpress

affy_sunflower_2010_13-ROS and hormones in sunflower seed dormancy.

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2019-12-31 | GSE25451 | GEO

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