Dataset Information

Proteomic Insight into Early Establishment of Seed Dormancy in Xanthium Strumarium

ABSTRACT: Along with the inherent complexities of seed and dormancy, the use of different genotypes or mutants to study the molecular mechanisms underlying seed dormancy leads to potential biases and data misinterpretation. To provide a comprehensive insight into the actual protein activities involved in seed dormancy establishment, a SWATH-MS proteomics was performed on dormant and non-dormant developing seeds of Xanthium strumarium at five consecutive time intervals including three, 10, 20, and 30 days after burr emergence and full maturation. The amount of approximately 3.5% differentially abundant proteins (DAPs) with a ~94% stage-specificity supports considerable proteome overlap in the two seed types. More than 38% of all differentially abundant proteins were observed at the first stage, supporting the importance of this stage of seed development for seed fate determination. Rapid overrepresentation of proteins responsible for cell wall biosynthesis, cytokinesis, and seed development were detected for non-dormant seed at the first stage, while dormancy-associated proteins showed less abundance. In the middle of seed development, we identified DAPs involved in seed maturation and ABA signaling. Interestingly, higher abundant proteins in the mature non-dormant seed were mainly involved in the facilitation of seed germination. Taken together, the temporal pattern of the accumulated proteins demonstrated a delay in the initiation of active cell division, enriched response to ABA, and defect in the seed maturation in developing dormant seeds. Moreover, stored proteins in the mature dormant seed are responsible for delaying germination but not dormancy induction. Finally, assume that dormancy may be established at a stage of seed development earlier than previously thought.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Xanthium Strumarium

TISSUE(S): Seed

SUBMITTER: Sara Hamzelou

LAB HEAD: Paul A. Haynes

PROVIDER: PXD052504 | Pride | 2024-08-22

REPOSITORIES: Pride

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			Action	DRS
	181123SeedSWATH1.wiff	Wiff
	181123SeedSWATH1.wiff.scan	Wiff
	181123SeedSWATH10.wiff	Wiff
	181123SeedSWATH10.wiff.scan	Wiff
	181123SeedSWATH11.wiff	Wiff

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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.

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.

Dataset Information

Proteomic Insight into Early Establishment of Seed Dormancy in Xanthium Strumarium

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