Firmness is an indicator of fruit freshness and a main component of tomato (Solanum lycopersicum) fruit texture. In this work, the genetic variability in fruit firmness and stiffness was analyzed in pre- and postharvest periods and underlying anatomical and biochemical traits were identified. Three tomato contrasted parental lines and six derived quantitative trait loci (QTL)-NILs harboring texture QTL on chromosome 4 (QTL4) and 9 (QTL9) were analyzed; the seasonal variability was assessed on two distant trusses. Firmness and stiffness were measured by compression and puncture tests at harvest and after 7-day storage at 20 A degrees C. QTL4 poorly influenced the textural variables, on the contrary to QTL9 which increased firmness measured by puncture test and had similar effects in the two genetic backgrounds. According to this test, firmness increased along the season, but ranks among genotypes and QTL effects were hardly affected. Only some of the QTL effects were still significant after storage and firmness losses were not predicted by firmness at harvest. Fruit firmness and stiffness measured by puncture tests correlated with both morphological (locular number, R = -0.89), histological (cell size, R < -0.80) and biochemical (dry matter (R > 0.82) and soluble sugar content (R < -0.74)) fruit traits. In contrast, compression test values hardly correlated with any of the measured traits. This work provided an original comprehensive approach to analyse fleshy fruit firmness and paves the way for a future predictive model.