Arbuscular mycorrhizal symbiosis is a beneficial association between plant roots and fungi that occurs in approximately 80 % of terrestrial plants and which confers different benefits including mineral nutrient acquisition and enhanced defense capacity. Although mycorrhizal colonization takes place in roots, the symbiosis establishment has systemic effects in other parts of the plant, in processes such as nutrient translocation and systemic resistance. In order to understand the transcriptional changes that occur in leaves of mycorrhizal plants, we used RNA-seq technology to obtain the transcriptomes of leaves from mycorrhizal and nonmycorrhizal tomato plants (Solanum lycopersicum). Four weeks after inoculation with the fungus Rhizophagus irregularis, leaves from mycorrhizal and non-mycorrhizal tomato plants were used for transcriptome sequencing. Of the 21,113 genes expressed in tomato leaves, 742 genes displayed differential expression between the mycorrhizal and nonmycorrhizal conditions. Most of the transcriptional changes occurred in the Bprotein,^BRNA,^Bsignaling,^Btransport,^B biotic and abiotic stresses,^and Bhormone metabolism^categories. Some transcriptional changes also occurred in P, N, and sugar transporters, as would be expected for mycorrhizal colonization. Finally, several differentially expressed genes may be related to systemic defense priming, in agreement with our demonstration that symbiotic plants exhibited mycorrhizainduced resistance against the foliar pathogen Xanthomonas campestris pv. vesicatoria. This is the first study to take on a genome-wide analysis aimed at understanding the expression changes in leaves of mycorrhiza-colonized plants. The results will therefore be valuable to future analyses focused on specific genes, as well as detailed studies of the expression profiles of certain gene families.
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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