Updated publication reference for PubMed record(s): 33643342. Mitochondria play important roles in the plant stress responses and the detoxification of the reactive oxygen species generated in the electron-transport chain. Expression of genes encoding stress-related proteins such as the mitochondrial small heat shock proteins (M-sHSP) is upregulated in response to different abiotic stresses. In Arabidopsis thaliana, three M-sHSPs paralogous genes were identified, although their function under physiological conditions remains elusive. Here, we analyzed the phenotype, proteomic and metabolic profiles of the loss-of-function mutants of M-sHSPs (single, double and triple mutants) during normal plant growth. The triple mutant showed the most prominent altered phenotype at vegetative and reproductive stages without any externally applied stress. They displayed chlorotic leaves, growth arrest and low seed production. Concomitantly, they exhibited increased levels of sugars, free amino acids such as proline, citric and ascorbic acid, among other metabolites. Single and double mutants displayed intermediate phenotype suggesting a redundant function of these proteins. All single, double and triple mutants showed alteration of proteins involved in photosynthesis, mitochondrial metabolism and antioxidant defense compared to the wild-type plants. Overall, depletion of M-sHSPs causes severe impact in fundamental metabolic processes, localized in different cell compartments, leading to alterations in the correct plant growth and development.