SUMMARYSuccessful plant survival depends upon the proper integration of information from the environment with endogenous cues to regulate growth and development. We have investigated the interplay between ambient temperature and hormone action during the regulation of hypocotyl elongation, and we have found that gibberellins (GAs) and auxin are quickly and independently recruited by temperature to modulate growth rate, whereas activity of brassinosteroids (BRs) seems to be required later on. Impairment of GA biosynthesis blocked the increased elongation caused at higher temperatures, but hypocotyls of pentuple DELLA knockout mutants still reduced their response to higher temperatures when BR synthesis or auxin polar transport were blocked. The expression of several key genes involved in the biosynthesis of GAs and auxin was regulated by temperature, which indirectly resulted in coherent variations in the levels of accumulation of nuclear GFP-RGA (repressor of GA1) and in the activity of the DR5 reporter. DNA microarray and genetic analyses allowed the identification of the transcription factor PIF4 (phytochrome-interacting factor 4) as a major target in the promotion of growth at higher temperature. These results suggest that temperature regulates hypocotyl growth by individually impinging on several elements of a pre-existing network of signaling pathways involving auxin, BRs, GAs, and PIF4.

SummaryTomato (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.

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 5 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.

Plants undergo two different developmental programs depending on whether they are growing in darkness (skotomorphogenesis) or in the presence of light (photomorphogenesis). It has been proposed that the latter is the default pathway followed by many plants after germination and before the seedling emerges from soil. The transition between the two pathways is tightly regulated. The conserved COP1-based complex is central in the light-dependent repression of photomorphogenesis in darkness. Besides this control, hormones such as brassinosteroids (BRs), cytokinins, auxins, or ethylene also have been shown to regulate, to different extents, this developmental switch. In the present work, we show that the hormone gibberellin (GA) widely participates in this regulation. Studies from Arabidopsis show that both chemical and genetic reductions of endogenous GA levels partially derepress photomorphogenesis in darkness. This is based both on morphological phenotypes, such as hypocotyl elongation and hook and cotyledon opening, and on molecular phenotypes, such as misregulation of the light-controlled genes CAB2 and RbcS. Genetic studies indicate that the GA signaling elements GAI and RGA participate in these responses. Our results also suggest that GA regulation of this response partially depends on BRs. This regulation seems to be conserved across species because lowering endogenous GA levels in pea (Pisum sativum) induces full de-etiolation in darkness, which is not reverted by BR application. Our results, therefore, attribute an important role for GAs in the establishment of etiolated growth and in repression of photomorphogenesis.One of the most dramatic changes in plant growth and development occurs during the transition from life in the dark just after germination, to life in a light environment when the seedling emerges from soil. Development in darkness is referred to as skotomorphogenesis, whereas development in the light is referred to as photomorphogenesis. Skotomorphogenesis is characterized by an etiolated appearance of seedlings with a fast-growing hypocotyl or epicotyl, presence of an apical hook, and small and closed cotyledons or primary leaves. Moreover, these seedlings present etioplasts instead of chloroplasts, and the expression of genes that are normally lightregulated is repressed or kept at low, basal levels. When light triggers the photomorphogenic development, growth of hypocotyl or epicotyl is slowed down, cotyledons or primary leaves open and expand, etioplasts develop into chloroplasts, and the expression of light-controlled genes is up-regulated (Neff et al., 2000).After germination, it is extremely important for plants to be able to maintain the skotomorphogenic development before reaching the light to preserve and protect both the shoot apical meristem and cotyledons or primary leaves. In many plants, photomorphogenesis is the default developmental pathway after germination (Wei et al., 1994). This is based on the fact that many lower plants lack an etiolated growth phase, and the new program...

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Based on its compact habit, Micro-Tom, a dwarf cultivar of tomato (Solanum lycopersicum L.), has been proposed as a preferred variety to carry out molecular research in tomato. This cultivar, however, is poorly characterized. It is shown here that Micro-Tom has mutations in the SELF-PRUNING (SP) and DWARF (D) genes. In addition to this, it is also shown that Micro-Tom harbours at least two independently segregating resistance loci to the plant pathogen Cladosporium fulvum. The presence of the self-pruning mutation in Micro-Tom, that generates a determinate phenotype, was confirmed by crossing and sequence analysis. It was also found that Micro-Tom has a mutation in the DWARF gene (d) that leads to mis-splicing and production of at least two shorter mRNAs. The d mutation is predicted to generate truncated DWARF protein. The d sequence defect co-segregates with dark-green and rugose leaves, characteristics of brassinosteroid biosynthesis mutants. Micro-Tom also carries at least another mutation producing internode length reduction that affects plant height but not active gibberellin (GA) levels, which were similar in dwarf and tall Micro-TomxSeverianin segregants. GAs and brassinosteroids act synergistically in Micro-Tom, and the response to GA depends on brassinosteroids because the elongation of internodes was at least six times higher when GA(3) was applied simultaneously with brassinolide. A novel variety, Micro-0 that is fully susceptible to C. fulvum and almost as dwarf as Micro-Tom, has been generated from the cross of Cf0xMicro-Tom. This line represents a valuable resource for future analysis of Cf resistance genes through breeding or transformation.

We investigated the role of gibberellins (GAs) in the effect of pat-2, a recessive mutation that induces facultative parthenocarpic fruit development in tomato (Lycopersicon esculentum Mill.) using near-isogenic lines with two different genetic backgrounds. Unpollinated wild-type Madrigal (MA/wt) and Cuarenteno (CU/wt) ovaries degenerated, but GA 3 application induced parthenocarpic fruit growth. On the contrary, parthenocarpic growth of MA/pat-2 and CU/pat-2 fruits, which occurs in the absence of pollination and hormone application, was not affected by GA 3 . Pollinated MA/wt and parthenocarpic MA/pat-2 ovary development was negated by paclobutrazol, and this inhibitory effect was counteracted by GA 3 . The main GAs of the early-13-hydroxylation pathway (GA 1 , GA 3 , GA 8 , GA 19 , GA 20 , GA 29 , GA 44 , GA 53 , and, tentatively, GA 81 ) and two GAs of the non-13-hydroxylation pathway (GA 9 and GA 34 ) were identified in MA/wt ovaries by gas chromatography-selected ion monitoring. GAs were quantified in unpollinated ovaries at flower bud, pre-anthesis, and anthesis. In unpollinated MA/pat-2 and CU/pat-2 ovaries, the GA 20 content was much higher (up to 160 times higher) and the GA 19 content was lower than in the corresponding non-parthenocarpic ovaries. The application of an inhibitor of 2-oxoglutarate-dependent dioxygenases suggested that GA 20 is not active per se. The pat-2 mutation may increase GA 20-oxidase activity in unpollinated ovaries, leading to a higher synthesis of GA 20 , the precursor of an active GA.

Gene StGA20ox1 encoding potato GA 20-oxidase is expressed to relatively high levels in leaves and regulated by daylength. To investigate whether this gene is involved in photoperiodic regulation of tuber formation, we have obtained transgenic potato plants expressing sense and antisense copies of the StGA20ox1 cDNA. Over-expression of this cDNA resulted in taller plants that required a longer duration of a short day photoperiod (SD) to tuberize. Tubers from these plants had a decreased time of dormancy and developed sprouts with elongated internodes. Plants expressing antisense copies of the StGA20ox1 cDNA had shorter stems, a decreased length of the internodes and tuberized earlier than control plants, showing increased tuber yields. Antisense inhibition of this gene had no visible effect on the time of dormancy of the tubers, although at the end of dormancy these formed sprouts with shortened internodes. Decreased levels of endogenous GA20 and GA1 were detected in the apex and first leaves of the antisense lines. These results demonstrate the involvement of the GA 20-oxidase activity encoded by StGA20ox1 in the control of stem elongation and in tuber induction but not in tuber dormancy, indicating that the latter may be regulated by another member of the gene family.