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The heat-stable proteome contains a tightly associated LEA protein network during seed germination in Arabidopsis

ABSTRACT: Seed germination involves transition from desiccation tolerance to sensitivity during early stages of seedling establishment accompanied by comprehensive changes of metabolome and proteome. Germination of Arabidopsis seeds was investigated over 72 h with special focus on the heat-stable proteome including late embryogenesis abundant (LEA) proteins together with changes of primary metabolites. Six metabolites in dry seeds important for seed longevity decreased during germination and seedling establishment, while all other metabolites increased simultaneously with activation of growth and development. In the heat stable proteome an almost equal fractioning of ordered and intrinsically disordered proteins (IDP) was discovered. Highly disordered proteins were assigned to functional bins development, protein, RNA and stress. Thermal stable proteins were related to a multitude of active biological processes. The majority of LEA proteins decreased during germination and seedling establishment in parallel to reduced desiccation tolerance. Five germination specific dehydrins were identified most likely providing stress tolerance. Network analysis of common proteins and metabolites over all time points revealed a tight network of LEA proteins with five hub LEAs with high connectivity. Connections to proteins such as Rubisco large subunit and seed storage proteins were discovered. Insights into the heat-stable proteome – metabolome network during seed germination are provided.

INSTRUMENT(S): Q Exactive HF

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

TISSUE(S): Seed

SUBMITTER: Michał Górka

LAB HEAD: Ellen Zuther

PROVIDER: PXD027546 | Pride | 2021-07-29

REPOSITORIES: Pride

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Project description:Seed germination is a complex trait determined by the interaction of hormonal, metabolic, genetic, and environmental components. Variability of this trait in crops has a big impact on seedling establishment and yield in the field. Classical studies of this trait in crops have focused mainly on the analyses of one level of regulation in the cascade of events leading to seed germination. We have carried out an integrative and extensive approach to deepen our understanding of seed germination in Brassica napus by generating transcriptomic, metabolic and hormonal data at different stages upon seed imbibition. Deep phenotyping of different seed germination associated traits in six winter-type B. napus accessions has revealed that seed germination kinetics, in particular seed germination speed, are major contributors to the variability of this trait. Metabolic profiling of these accessions has allowed us to describe a common pattern of metabolic change and to identify the levels of malate and aspartate metabolites as putative metabolic markers to estimate germination performance. Additionally, analysis of seed content of different hormones suggests that hormonal balance between ABA, GA and IAA at crucial time points during this process might underlie seed germination differences in these accessions. In this study, we have also defined the major transcriptome changes accompanying the germination process in B. napus. Furthermore, we have observed that earlier activation of key germination regulatory genes seems to generate the differences in germination speed observed between accessions in B. napus. Finally, we have found that protein-protein interactions between some of these key regulators are conserved in B. napus suggesting a shared regulatory network with other plants species. Altogether, our results provide a comprehensive and detailed picture of seed germination dynamics in oilseed rape. This new framework will be extremely valuable, not only to evaluate germination performance of B. napus accessions, but also to identify key targets for crop improvement in this important process.

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The heat-stable proteome contains a tightly associated LEA protein network during seed germination in Arabidopsis

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