Dataset Information

Arabidopsis seeds translation regulation

ABSTRACT: This study explores translational regulation during germination by comparing two Arabidopsis thaliana accessions with contrasting dormancy phenotypes: Columbia (Col), which germinates readily, and Cape Verde Islands (Cvi), which exhibits deep dormancy. Using sucrose gradient centrifugation, we isolated monosomal and polysomal fractions from freshly harvested (FH), after-ripened (AR), and imbibed (IM) seeds. In this study, we have compared the mRNAs and associated proteins stored in monosomes or polysomes at the stages of seed maturity, post-ripening and early germination using two Arabidopsis genotypes differing in dormancy level

INSTRUMENT(S):

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Seed

SUBMITTER: Václav Pustka

LAB HEAD: Zbynek Zdrahal

PROVIDER: PXD060515 | Pride | 2025-12-15

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	5118_TIMS1_RSLC-104m_DIA_01P_1_RB7_1_9792.d.zip	Other
	5118_TIMS1_RSLC-104m_DIA_02M_1_RB1_1_9827.d.zip	Other
	5118_TIMS1_RSLC-104m_DIA_04M_1_RB2_1_9883.d.zip	Other
	5118_TIMS1_RSLC-104m_DIA_06P_1_RC1_1_9835.d.zip	Other
	5118_TIMS1_RSLC-104m_DIA_07P_1_RC2_1_9800.d.zip	Other

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Publications

Translational and epitranscriptomic regulation of seed germination in Arabidopsis thaliana genotypes with contrasting dormancy phenotypes.

Balarynová J J Klčová B B Čegan R R Raabe K K Krejčí P P Bednář P P Potěšil D D Pustka V V Tarkowská D D Turečková V V Zdráhal Z Z Honys D D Smýkal P P

Plant molecular biology 20251204 6

Seed germination represents the crucial transition from dormancy to active growth, orchestrated by environmental signals and endogenous regulatory networks. This study investigates translational regulation during germination in two Arabidopsis thaliana accessions with contrasting dormancy phenotypes: Columbia (Col-0), which germinates readily, and Cape Verde Islands (Cvi-0), which exhibits deep dormancy. Hormonal profiling revealed dynamic changes in abscisic acid and gibberellins, central regul ...[more]

PMID: 41342975

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Project description:Seed dormancy is the inability for seeds to germinate even under favorable conditions. In the Arabidopsis Landsberg erecta (Ler) ecotype, 2 weeks of dry storage, called after-ripening, is sufficient to relieve seed dormancy. Such seed is referred to as after-ripened (AR) and has a high rate of germination when imbibed. While widespread transcriptome changes have been previously observed with seed dormancy loss, this experiment was designed to characterize transcriptional changes associated with the increased seed dormancy and dormancy loss of the gibberellin (GA) hormone-insensitive sleepy1-2 (sly1-2) mutant. The SLY1 gene encodes the F-box subunit of an SCF E3 ubiquitin ligase needed for GA-triggered proteolysis of DELLA repressors of seed germination. In the sly1-2 mutant, GA-directed DELLA proteolysis cannot occur leading to DELLA protein accumulation and increased dormancy. sly1-2 mutant seeds are fully dormant at 2 weeks of dry storage (0% germination), but germinate well with very long after-ripening (51% germination after 19 months). sly1-2 seed germination can also be rescued by overexpression of the GA receptor, GA-INSENSITIVE DWARF1b (GID1b-OE), which resulted in 74% germination at 2 weeks of dry storage. In this experiment, we compared seeds of wild-type Ler at 2 weeks of dry storage (non-dormant), dormant sly1-2 (2 weeks of dry storage; sly1-2(D)), long after-ripened sly1-2 (non-dormant, 19 months of dry storage; sly1-2(AR)), and sly1-2 GID1b-OE (non-dormant, 2 weeks of dry storage). Samples were collected at two imbibition timepoints: 1) a 0h timepoint after 4 days at 4°C, and 2) a 12h timepoint after 4 days at 4°C followed by 12 hours in the light at 22°C. These timepoints were selected to capture the transcriptomes at an early and late time in Phase II of imbibition. Using this experimental design we were able to determine transcriptome differences associated with seed dormancy in the sly1-2 mutation (Ler wt vs sly1-2(D)), and changes associated with sly1-2 dormancy loss through dry after-ripening (sly1-2(AR) vs sly1-2(D)) or through GID1b-overexpression (sly1-2 GID1b-OE vs sly1-2(D)). Seeds for Ler wt, sly1-2(D), and sly1-2 GID1b-OE were grown alongside each other under the same conditions and after-ripened for 2 weeks. Seeds from sly1-2(AR) were grown under the same conditions in advance of the other lines to allow for the long after-ripening requirement. RNA was extracted using a phenol-chloroform-based extraction from three biological replicates per treatment.

Project description:Plant embryos can survive years in a desiccated, quiescent state within seeds. In many species, seeds are dormant and unable to germinate at maturity. They acquire the capacity to germinate through a period of dry storage called after-ripening (AR), a biological process that occurs at 5-15% moisture when most metabolic processes cease. Because stored transcripts will be among the first proteins translated upon water uptake, they likely impact germination potential. Transcriptome changes associated with the increased seed dormancy of the GA-insensitive sly1-2 mutant, and with dormancy loss through sly1-2 after-ripening or constitutive overexpression of the GA receptor (GID1b) were characterized in dry seeds. This experiment used the same seed batches as a previous experiment (E-MTAB-4782) to characterize transcriptional changes associated with the increased seed dormancy and dormancy loss in imbibing seeds. The SLY1 gene encodes the F-box subunit of an SCF E3 ubiquitin ligase needed for GA-triggered proteolysis of DELLA repressors of seed germination. In the sly1-2 mutant, GA-directed DELLA proteolysis cannot occur leading to DELLA protein accumulation and increased dormancy. sly1-2 mutant seeds are fully dormant at 2 weeks of dry storage (0% germination), but germinate well with very long after-ripening (51% germination after 19 months). sly1-2 seed germination can also be rescued by overexpression of the GA receptor, GA-INSENSITIVE DWARF1b (GID1b-OE), which resulted in 74% germination at 2 weeks of dry storage. In this experiment, we sampled dry seeds of wild-type Ler at 2 weeks of dry storage (non-dormant), dormant sly1-2 (2 weeks of dry storage; sly1-2(D)), long after-ripened sly1-2 (non-dormant, 19 months of dry storage; sly1-2(AR)), and sly1-2 GID1b-OE (non-dormant, 2 weeks of dry storage). This experimental design allowed comparison between these transcriptomes in dry seeds to determine if dry seed stored mRNA differences contribute to the dormancy phenotypes observed once seeds are imbibed. Seeds for Ler wt, sly1-2(D), and sly1-2 GID1b-OE were grown alongside each other under the same conditions and after-ripened for 2 weeks. Seeds from sly1-2(AR) were grown under the same conditions in advance of the other lines to allow for the long after-ripening requirement. RNA was extracted using a phenol-chloroform-based extraction from three biological replicates per treatment.