Auxin-induced fruit-set in tomato is mediated in part by gibberellins.

Tomato (Solanum lycopersicum L.) fruit-set and growth depend on gibberellins (GAs). Auxins, another kind of hormone, can also induce parthenocarpic fruit growth in tomato, although their possible interaction with GAs is unknown. We showed that fruit development induced by the auxins indole-3-acetic acid and 2,4-dichlorophenoxyacetic acid (2,4-D) were significantly reduced by the simultaneous application of inhibitors of GA biosynthesis, and that this effect was reversed by the application of GA(3). This suggested that the effect of auxin was mediated by GA. Parthenocarpic fruits induced by 2,4-D had higher levels of the active GA(1), its precursors and metabolites, than unpollinated non-treated ovaries, but similar levels as those found in pollinated ovaries. Application experiments of radioactive-labelled GAs to unpollinated ovaries showed than 2,4-D altered GA metabolism (both biosynthesis and catabolism) in vivo. Transcript levels of genes encoding copalyldiphosphate synthase (SlCPS), SlGA20ox1, SlGA20ox2 and SlGA20ox3, and SlGA3ox1 were higher in unpollinated ovaries treated with 2,4-D. In contrast, transcript levels of SlGA2ox2 (out of the five SlGA2ox genes known to encode this kind of GA-inactivating enzyme) were lower in ovaries treated with 2,4-D. Our results support the idea that auxins induce fruit-set and growth in tomato, at least partially, by enhancing GA biosynthesis (GA 20-oxidase, GA 3-oxidase and CPS), and probably by decreasing GA inactivation (GA2ox2) activity, thereby leading to higher levels of GA(1). The expression of diverse Aux/indole-3-acetic acid (IAA) and auxin response factors, which may be involved in this effect of auxin, was also altered in 2,4-D-induced ovaries.

Role of gibberellins in parthenocarpic fruit development induced by the genetic system pat-3/pat-4 in tomato.

The role of gibberellins (GAs) in the induction of parthenocarpic fruit-set and growth by the pat-3/pat-4 genetic system in tomato (Lycopersicon esculentum Mill.) was investigated using wild type (WT; Cuarenteno) and a near-isogenic line derived from the German line RP75/59 (the source of pat-3/pat-4 parthenocarpy). Unpollinated WT ovaries degenerated but GA3 application induced parthenocarpic fruit growth. On the contrary, parthenocarpic growth of pat-3/pat-4 fruits, which occurs in the absence of pollination and hormone treatment, was not affected by applied GA3. Unpollinated pat-3/pat-4 fruit growth was negated by paclobutrazol, an inhibitor of ent-kaurene oxidase, and this inhibitory effect was negated by GA3. The quantification of the main GAs of the early 13-hydroxylation pathway (GA1, GA8, GA19, GA20, GA29 and GA44) in unpollinated ovaries at 3 developmental stages (flower bud, FB; pre-anthesis, PR; and anthesis, AN), by gas chromatography-selected ion monitoring, showed that the concentration of most of them was higher in pat-3/pat-4 than in WT ovaries at PR and AN stages. The concentration of GA1, suggested previously to be the active GA in tomate, was 2-4 times higher. Unpollinated pat-3/pat-4 ovaries at FB, PR and AN stages also contained relatively high amounts (5-12 ng g-1) of GA3, a GA found at less than 0.5 ng g-1 in WT ovaries. It is concluded that the mutations pat-3/pat-4 may induce natural facultative parthenocarpy capacity in tomato by increasing the concentration of GA1 and GA3 in the ovaries before pollination.

Gibberellin regulation of fruit set and growth in tomato.

The role of gibberellins (GAs) in tomato (Solanum lycopersicum) fruit development was investigated. Two different inhibitors of GA biosynthesis (LAB 198999 and paclobutrazol) decreased fruit growth and fruit set, an effect reversed by GA(3) application. LAB 198999 reduced GA(1) and GA(8) content, but increased that of their precursors GA(53), GA(44), GA(19), and GA(20) in pollinated fruits. This supports the hypothesis that GA(1) is the active GA for tomato fruit growth. Unpollinated ovaries developed parthenocarpically in response to GA(3) > GA(1) = GA(4) > GA(20), but not to GA(19), suggesting that GA 20-oxidase activity was limiting in unpollinated ovaries. This was confirmed by analyzing the effect of pollination on transcript levels of SlCPS, SlGA20ox1, -2, and -3, and SlGA3ox1 and -2, encoding enzymes of GA biosynthesis. Pollination increased transcript content of SlGA20ox1, -2, and -3, and SlCPS, but not of SlGA3ox1 and -2. To investigate whether pollination also altered GA inactivation, full-length cDNA clones of genes encoding enzymes catalyzing GA 2-oxidases (SlGA2ox1, -2, -3, -4, and -5) were isolated and characterized. Transcript levels of these genes did not decrease early after pollination (5-d-old fruits), but transcript content reduction of all of them, mainly of SlGA2ox2, was found later (from 10 d after anthesis). We conclude that pollination mediates fruit set by activating GA biosynthesis mainly through up-regulation of GA20ox. Finally, the phylogenetic reconstruction of the GA2ox family clearly showed the existence of three gene subfamilies, and the phylogenetic position of SlGA2ox1, -2, -3, -4, and -5 was established.

Polyamine metabolism is altered in unpollinated parthenocarpic pat-2 tomato ovaries.

Facultative parthenocarpy induced by the recessive mutation pat-2 in tomato (Lycopersicon esculentum Mill.) depends on gibberellins (GAs) and is associated with changes in GA content in unpollinated ovaries. Polyamines (PAs) have also been proposed to play a role in early tomato fruit development. We therefore investigated whether PAs are able to induce parthenocarpy and whether the pat-2 mutation alters the content and metabolism of PAs in unpollinated ovaries. Application of putrescine, spermidine, and spermine to wild-type unpollinated tomato ovaries (cv Madrigal [MA/wt]) induced partial parthenocarpy. Parthenocarpic growth of MA/pat-2 (a parthenocarpic near-isogenic line to MA/wt) ovaries was negated by paclobutrazol (GA biosynthesis inhibitor), and this inhibition was counteracted by spermidine. Application of alpha-difluoromethyl-ornithine (-Orn) and/or alpha-difluoromethyl-arginine (-Arg), irreversible inhibitors of the putrescine biosynthesis enzymes Orn decarboxylase (ODC) and Arg decarboxylase, respectively, prevented growth of unpollinated MA/pat-2 ovaries. Alpha-difluoromethyl-Arg inhibition was counteracted by putrescine and GA(3), whereas that of alpha-difluoromethyl-Orn was counteracted by GA(3) but not by putrescine or spermidine. In unpollinated MA/pat-2 ovaries, the content of free spermine was significantly higher than in MA/wt ovaries. ODC activity was higher in pat-2 ovaries than in MA/wt. Transcript levels of genes encoding ODC and spermidine synthase were also higher in MA/pat-2. All together, these results strongly suggest that the parthenocarpic ability of pat-2 mutants depends on elevated PAs levels in unpollinated mutant ovaries, which correlate with an activation of the ODC pathway, probably as a consequence of elevated GA content in unpollinated pat-2 tomato ovaries.