Subcellular Proteomics in Conifers: Purification of Nuclei and Chloroplast Proteomes

Lamelas, Laura; García, Lara; Cañal, María Jesús; Meijón, Mónica

doi:10.1007/978-1-0716-0528-8_5

Cited by 5 publications

(4 citation statements)

References 11 publications

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“…Nuclear and chloroplast fractions for proteomics analyses were purified as described in (Lamelas, García, et al, 2020). In brief, samples were homogenised, incubated in their respective cellular lysis Weckwerth, 2014).…”

Section: Proteomics Analysismentioning

confidence: 99%

Nucleus and chloroplast: A necessary understanding to overcome heat stress in Pinus radiata

Lamelas

Valledor

López-Hidalgo

et al. 2021

Plant Cell & Environment

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The recovery and maintenance of plant homeostasis under stressful environments are complex processes involving organelle crosstalk for a coordinated cellular response. Here, we revealed through nuclear and chloroplast subcellular proteomics, biochemical cell profiles and targeted transcriptomics how chloroplasts and nuclei developed their responses under increased temperatures in a long-lived species (Pinus radiata). Parallel to photosynthetic impairment and reactive oxygen species production in the chloroplast, a DNA damage response was triggered in the nucleus followed by an altered chromatin conformation. In addition, in the nuclei, we found several proteins, such as HEMERA or WHIRLY, which change their locations from the chloroplasts to the nuclei carrying the stress message. Additionally, our data showed a deep rearrangement of RNA metabolism in both organelles, revealing microRNAs and AGO1 as potential regulators of the acclimation mechanisms.Altogether, our study highlights the synchronisation among the different stages required for thermotolerance acquisition in P. radiata, pointing out the role of chromatin conformation and posttranscriptional gene regulation in overcoming heat stress and assuring plant survival for the following years.

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Section: Proteomics Analysismentioning

confidence: 99%

Nucleus and chloroplast: A necessary understanding to overcome heat stress in Pinus radiata

Lamelas

Valledor

López-Hidalgo

et al. 2021

Plant Cell & Environment

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“…Chloroplast enriched fraction for proteomics analyses was purified as previously described (Lamelas, García, et al, 2020). In brief, samples were homogenized, incubated in cellular lysis buffer, and then filtrated.…”

Section: Chloroplast Isolation and Protein Extractionmentioning

confidence: 99%

Like mother like son: Transgenerational memory and cross‐tolerance from drought to heat stress are identified in chloroplast proteome and seed provisioning in Pinus radiata

Lamelas,

López‐Hidalgo,

Valledor

et al. 2024

Plant Cell & Environment

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How different stressors impact plant health and memory when they are imposed in different generations in wild ecosystems is still scarce. Here, we address how different environments shape heritable memory for the next generation in seeds and seedlings of Pinus radiata, a long‐lived species with economic interest. The performance of the seedlings belonging to two wild clonal subpopulations (optimal fertirrigation vs. slightly stressful conditions) was tested under heat stress through physiological profiling and comparative time‐series chloroplast proteomics. In addition, we explored the seeds conducting a physiological characterization and targeted transcriptomic profiling in both subpopulations. Our results showed differential responses between them, evidencing a cross‐stress transgenerational memory. Seedlings belonging to the stressed subpopulation retained key proteins related to Photosystem II, chloroplast‐to‐nucleus signalling and osmoprotection which helped to overcome the applied heat stress. The seeds also showed a differential gene expression profile for targeted genes and microRNAs, as well as an increased content of starch and secondary metabolites, molecules which showed potential interest as biomarkers for early selection of primed plants. Thus, these finds not only delve into transgenerational cross‐stress memory in trees, but also provide a new biotechnological tool for forest design.

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“…The last decade has witnessed a huge advance in pine proteomics research that has continued the transition from gel-based to gel-free systems based on mass spectrometry thanks to important methodological developments, such as those of databases ( Nystedt et al., 2013 ; Zimin et al., 2014 ; Stevens et al., 2016 ; Zimin et al., 2017 ) and their use ( Romero-Rodríguez et al., 2014 ). In addition, the development of new protein isolation protocols ( Valledor et al., 2014 ; Colina et al., 2020 ) and fractionation methods has allowed studying subcellular proteomes ( Alegre et al., 2016 ; Lamelas et al., 2020a ). Noteworthy, proteomic analysis has been very commonly performed in combination with other omics, like transcriptomics or metabolomics, which has needed new computational strategies and algorithms to allow the integration of different omic layers effectively, and comprehensively ( Escandón et al., 2020 ; Sundararaman et al., 2020 ).…”

Section: Proteomics Survey (2012-2022): Where Are We Now?mentioning

confidence: 99%

Proteomics research in forest trees: A 2012-2022 update

et al. 2023

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This review is a compilation of proteomic studies on forest tree species published in the last decade (2012-2022), mostly focused on the most investigated species, including Eucalyptus, Pinus, and Quercus. Improvements in equipment, platforms, and methods in addition to the increasing availability of genomic data have favored the biological knowledge of these species at the molecular, organismal, and community levels. Integration of proteomics with physiological, biochemical and other large-scale omics in the direction of the Systems Biology, will provide a comprehensive understanding of different biological processes, from growth and development to responses to biotic and abiotic stresses. As main issue we envisage that proteomics in long-living plants will thrive light on the plant responses and resilience to global climate change, contributing to climate mitigation strategies and molecular breeding programs. Proteomics not only will provide a molecular knowledge of the mechanisms of resilience to either biotic or abiotic stresses, but also will allow the identification on key gene products and its interaction. Proteomics research has also a translational character being applied to the characterization of the variability and biodiversity, as well as to wood and non-wood derived products, traceability, allergen and bioactive peptides identification, among others. Even thought, the full potential of proteomics is far from being fully exploited in forest tree research, with PTMs and interactomics being reserved to plant model systems. The most outstanding achievements in forest tree proteomics in the last decade as well as prospects are discussed.

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Subcellular Proteomics in Conifers: Purification of Nuclei and Chloroplast Proteomes

Cited by 5 publications

References 11 publications

Nucleus and chloroplast: A necessary understanding to overcome heat stress in Pinus radiata

Nucleus and chloroplast: A necessary understanding to overcome heat stress in Pinus radiata

Like mother like son: Transgenerational memory and cross‐tolerance from drought to heat stress are identified in chloroplast proteome and seed provisioning in Pinus radiata

Proteomics research in forest trees: A 2012-2022 update

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