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  • Author or Editor: José A. López-Valenzuela x
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Tomato (Solanum lycopersicum) fruit is susceptible to chilling injury (CI), a physiological disorder caused by low, non-freezing temperatures that affects fruit postharvest quality. Little is known about the biochemical basis of CI, and the aim of this study was to identify proteins related to this disorder in ‘Imperial’ tomato fruit. CI and protein expression changes were analyzed during fruit ripening (0, 4, 8, and 12 days at 21 °C) after storage under chilling (5 °C) and non-chilling conditions (21 °C) for 5, 15, and 25 days. The main CI symptoms observed were uneven fruit ripening and color development, pitting, and decay. Protein analysis of two-dimensional gels showed that 6% of the detected spots (≈300) changed their expression in response to cold. The identified proteins are involved in carbon metabolism, oxidative stress, photosynthesis, and protein processing and degradation; two were related to cold stress, showing higher accumulation in non-damaged tissue of chilled fruit: thioredoxin peroxidase (TPxI) and glycine-rich RNA-binding protein (GR-RBP). This is the first report suggesting an important role for TPxI and GR-RBP in cold response during tomato fruit ripening, and they may be acting through redox sensing and regulation of gene expression at low temperature. These enzymes and the other chilling-related proteins might be working together to maintain the cellular homeostasis under cold stress conditions.

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A simplified protocol to obtain transgenic tomato plants was established. The effects of culture media composition and Agrobacterium concentration were evaluated. The highest shoot-forming capacity index (5.6) was observed when leaf explants were cultured for 6 weeks with 2 mg·L−1 zeatin, 0.1 mg·L−1 indoleacetic acid, and 300 mg·L−1 timentin. Shoot elongation and root formation were performed in one step on growth regulator-free media. The highest percentage (82%) of fully developed plantlets was obtained when shoots were cultured for 4 weeks with 0.5× Murashige and Skoog (MS) media and 15 g·L−1 sucrose. A 100% of plant survival rate was observed after 4 weeks of being transplanted to ex vitro conditions followed by fruit production (15 fruits/plant) after 2 more weeks. Transient expression of β-glucuronidase was visualized in 100% of the leaf explants infected with Agrobacterium at an OD600 = 0.5 and cocultured for 48 h with 2 mg·L−1 benzylaminopurine, 0.1 mg·L−1 naphthaleneacetic acid, and 100 μM acetosyringone. Stable transformation was confirmed by histochemical glucuronidase assay and polymerase chain reaction (PCR) analysis with a total efficiency of 19.1%. The complete protocol, from shoot induction to fruit production of soil-adapted transgenic plants can be accomplished in only 4 months, and it seems to be very useful for both micropropagation and genetic transformation purposes.

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