Dataset Information

Targeted Proteomics Allows Quantification of Ethylene Receptors and Reveals SlETR3 Accumulation in Never-Ripe Tomatoes.

ABSTRACT: Ethylene regulates fruit ripening and several plant functions (germination, plant growth, plant-microbe interactions). Protein quantification of ethylene receptors (ETRs) is essential to study their functions, but is impaired by low resolution tools such as antibodies that are mostly nonspecific, or the lack of sensitivity of shotgun proteomic approaches. We developed a targeted proteomic method, to quantify low-abundance proteins such as ETRs, and coupled this to mRNAs analyses, in two tomato lines: Wild Type (WT) and Never-Ripe (NR) which is insensitive to ethylene because of a gain-of-function mutation in ETR3. We obtained mRNA and protein abundance profiles for each ETR over the fruit development period. Despite a limiting number of replicates, we propose Pearson correlations between mRNA and protein profiles as interesting indicators to discriminate the two genotypes: such correlations are mostly positive in the WT and are affected by the NR mutation. The influence of putative post-transcriptional and post-translational changes are discussed. In NR fruits, the observed accumulation of the mutated ETR3 protein between ripening stages (Mature Green and Breaker + 8 days) may be a cause of NR tomatoes to stay orange. The label-free quantitative proteomics analysis of membrane proteins, concomitant to Parallel Reaction Monitoring analysis, may be a resource to study changes over tomato fruit development. These results could lead to studies about ETR subfunctions and interconnections over fruit development. Variations of RNA-protein correlations may open new fields of research in ETR regulation. Finally, similar approaches may be developed to study ETRs in whole plant development and plant-microorganism interactions.

SUBMITTER: Chen Y

PROVIDER: S-EPMC6727826 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Publications

Targeted Proteomics Allows Quantification of Ethylene Receptors and Reveals SlETR3 Accumulation in Never-Ripe Tomatoes.

Chen Yi Y Rofidal Valérie V Hem Sonia S Gil Julie J Nosarzewska Joanna J Berger Nathalie N Demolombe Vincent V Bouzayen Mondher M Azhar Beenish J BJ Shakeel Samina N SN Schaller G Eric GE Binder Brad M BM Santoni Véronique V Chervin Christian C

Frontiers in plant science 20190829

Ethylene regulates fruit ripening and several plant functions (germination, plant growth, plant-microbe interactions). Protein quantification of ethylene receptors (ETRs) is essential to study their functions, but is impaired by low resolution tools such as antibodies that are mostly nonspecific, or the lack of sensitivity of shotgun proteomic approaches. We developed a targeted proteomic method, to quantify low-abundance proteins such as ETRs, and coupled this to mRNAs analyses, in two tomato l ...[more]

PMID: 31555314

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Project description:Consumer complaints about the flavor of fresh tomato fruits (Solanum lycopersicum L.) have increased in the past few decades, and numerous studies have been done on the flavor of tomatoes and how it is influenced. However, it has not yet been taken into account how consumer handling affects the flavor when considering the complete post-harvest chain-from retailer (distributor) to retail to consumer. In this study, the impact of two household storage regimes on the volatile profile and important flavor-related compounds were examined, considering the entire post-harvest handling. New breeding lines (n = 2) and their parental cultivars (n = 3) were evaluated. Fruits were harvested ripe and stored at 12.5°C for 1 day, at 20°C for 2 days, and afterward at either 20 or 7°C for another 4 days. The aroma volatile profile was measured using GC-MS and GC-FID. A trained panel was used to characterize the sensory attributes of the fruits. In both storage regimes, the relative amount of hexanal increased during the storage period in three of the five cultivars/breeding lines while benzaldehyde was the only volatile compound that decreased significantly in four cultivars/breeding lines. The relative concentration of the precursors of lipid-derived volatiles-linoleic (C18:2) and linolenic (C18:3) acid-did not change in both storage regimes. The lycopene and ?-carotene contents increased slightly during storage (20 and 7°C), as the carotenoid-derived volatile 6-methyl-5-hepten-2-one did. The fructose and glucose concentrations did not vary significantly, while the content of total soluble solids increased during both storage regimes. No significant difference could be found between the fruits stored at 20 or 7°C for 4 days after the post-harvest handling in all the parameters analyzed, including the sensory analysis, considering all cultivars/breeding lines. A storage temperature of 7°C is not detrimental for the flavor of ripe fruits under the experimental conditions used. The genetic background of the studied cultivars/breeding lines have a higher impact on the flavor variation than the two common household storage conditions when storing ripe fruits and taking the entire post-harvest handling into account.

Dataset Information

Targeted Proteomics Allows Quantification of Ethylene Receptors and Reveals SlETR3 Accumulation in Never-Ripe Tomatoes.

Publications

Targeted Proteomics Allows Quantification of Ethylene Receptors and Reveals SlETR3 Accumulation in Never-Ripe Tomatoes.

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