Characterization and Experimental Host Range of a Brazilian Tomato Isolate of Tomato severe rugose virus

We report the complete molecular characterization of the DNA‐A and DNA‐B of a Brazilian tomato isolate of Tomato severe rugose virus (ToSRV) and the experimental host range of the virus determined using whitefly transmission tests. Genome analysis showed that ToSRV has a close evolutionary relationship with Tomato rugose mosaic virus. Of 33 plants species inoculated with viruliferous Bemisia tabaci biotype B, 13 species were susceptible to ToSRV, nine asymptomatically. Therefore, ToSRV disease management strategy should include the control of infected weeds close to tomato fields.     

Regulation and manipulation of ABA biosynthesis in roots

Overexpression of 9-cis-epoxycarotenoid dioxygenase (NCED) is known to cause abscisic acid (ABA) accumulation in leaves, seeds and whole plants. Here we investigated the manipulation of ABA biosynthesis in roots. Roots from whole tomato plants that constitutively overexpress LeNCED1 had a higher ABA content than wild-type (WT) roots. This could be explained by enhanced in situ ABA biosynthesis, rather than import of ABA from the shoot, because root cultures also had higher ABA content, and because tetracycline (Tc)-induced LeNCED1 expression caused ABA accumulation in isolated tobacco roots. However, the Tc-induced expression led to greater accumulation of ABA in leaves than in roots. This demonstrates for the first time that NCED is rate-limiting in root tissues, but suggests that other steps were also restrictive to pathway flux, more so in roots than in leaves. Dehydration and NCED overexpression acted synergistically in enhancing ABA accumulation in tomato root cultures. One explanation is that xanthophyll synthesis was increased during root dehydration, and, in support of this, dehydration treatments increased beta-carotene hydroxylase mRNA levels. Whole plants overexpressing LeNCED1 exhibited greatly reduced stomatal conductance and grafting experiments from this study demonstrated that this was predominantly due to increased ABA biosynthesis in leaves rather than in roots. Genetic manipulation of both xanthophyll supply and epoxycarotenoid cleavage may be needed to enhance root ABA biosynthesis sufficiently to signal stomatal closure in the shoot.     

Spider mites (Tetranychus urticae) perform poorly on and disperse from plants exposed to methyl jasmonate

Jasmonates are plant hormones involved in wound and defense responses against herbivorous arthropods. Methyl jasmonate (MeJA) is used experimentally to induce defense responses in plants. In experiments outlined here we utilized a novel preference assay with unwounded plants that allowed us to study the impact of a MeJA spray on subsequent Tetranychus urticae Koch (Acari: Tetranychidae) proliferation and preference. Spraying plants with 100 μm MeJA 1 day before infestation caused mites to disperse within 2 days from treated impatiens [Impatiens wallerana Hook f., ‘Super Elfin Pink’ (Balsaminaceae)], pansy [Viola × wittrockiana Gams, ‘Imperial Beaconsfield’ (Violaceae)], and tomato [Solanum lycopersicum L., ‘Big Boy’ (Solanaceae)] plants. In addition, MeJA application reduced mite proliferation rate on impatiens and pansy by 60% (measured 22–34 days after infestation). Proteinase inhibitor (PI) assays suggested that MeJA‐induced PIs alone were not responsible for the observed results in pansy and impatiens but may have been a factor in tomato. Implications of these results in the context of MeJA‐induced resistance responses and possible directions for future research and application are discussed.     

Two Erysiphe species associated with recent outbreak of soybean powdery mildew: results of molecular phylogenetic analysis based on nuclear rDNA sequences

 Serious outbreaks of powdery mildew by a fungus belonging to the mitosporic genus Oidium subgenus Pseudoidium have been reported on soybean (Glycine max) in a wide area of eastern Asia since 1998. The taxonomic and phylogenetic placement of the causal fungus has not yet been determined because of lack of the perfect stage. We found ascomata having mycelioid appendages on a single leaf of soybean infested by powdery mildew. Molecular phylogenetic analysis was conducted based on a total of 14 sequences of the rDNA internal transcribed spacer (ITS) region from 13 soybean and wild soybean (Glycine soja) materials collected in Japan, Korea, Vietnam, and the United States, combined with 47 sequence data obtained from the DNA databases. It was revealed that two Erysiphe species were associated with the outbreak of soybean powdery mildew. There was 16% difference between the two species in genetic divergence of the ITS sequence. One species with perfect stage has an ITS sequence identical to that of Erysiphe glycines on Amphicarpaea and is identified as Erysiphe glycines based on the ITS sequence and morphology of ascomata. The second species, without the perfect stage, is likely to be Erysiphe diffusa (= Microsphaera diffusa), known as the fungus causing soybean powdery mildew in the United States, because the ITS sequences are identical to those from materials collected in the United States. However, we need materials having ascomata of E. diffusa to confirm the species name. Received: March 15, 2002 / Accepted: May 22, 2002     

Virus‐induced gene silencing reveals control of reactive oxygen species accumulation and salt tolerance in tomato by γ‐aminobutyric acid metabolic pathway

γ‐Aminobutyric acid (GABA) accumulates in many plant species in response to environmental stress. However, the physiological function of GABA or its metabolic pathway (GABA shunt) in plants remains largely unclear. Here, the genes, including glutamate decarboxylases (SlGADs), GABA transaminases (SlGABA‐Ts) and succinic semialdehyde dehydrogenase (SlSSADH), controlling three steps of the metabolic pathway of GABA, were studied through virus‐induced gene silencing approach in tomato. Silencing of SlGADs (GABA biosynthetic genes) and SlGABA‐Ts (GABA catabolic genes) led to increased accumulation of reactive oxygen species (ROS) as well as salt sensitivity under 200 mm NaCl treatment. Targeted quantitative analysis of metabolites revealed that GABA decreased and increased in the SlGADs‐ and SlGABA‐Ts‐silenced plants, respectively, whereas succinate (the final product of GABA metabolism) decreased in both silenced plants. Contrarily, SlSSADH‐silenced plants, also defective in GABA degradation process, showed dwarf phenotype, curled leaves and enhanced accumulation of ROS in normal conditions, suggesting the involvement of a bypath for succinic semialdehyde catabolism to γ‐hydroxybutyrate as reported previously in Arabidopsis, were less sensitive to salt stress. These results suggest that GABA shunt is involved in salt tolerance of tomato, probably by affecting the homeostasis of metabolites such as succinate and γ‐hydroxybutyrate and subsequent ROS accumulation under salt stress.     

Elevated carbon dioxide plus chronic warming causes dramatic increases in leaf angle in tomato,which correlates with reduced plant growth

Limited evidence indicates that moderate leaf hyponasty can be induced by high temperatures or unnaturally high CO₂. Here, we report that the combination of warming plus elevated CO₂ (eCO₂) induces severe leaf hyponasty in tomato (Solanum lycopersicum L.). To characterize this phenomenon, tomato plants were grown at two levels of CO₂ (400 vs. 700 ppm) and two temperature regimes (30 vs. 37°C) for 16–18 days. Leaf hyponasty increased dramatically with warming plus eCO₂ but increased only slightly with either factor alone and was slowly reversible upon transfer to control treatments. Increases in leaf angle were not correlated with leaf temperature, leaf water stress, or heat‐related damage to photosynthesis. However, steeper leaf angles were correlated with decreases in leaf area and biomass, which could be explained by decreased light interception and thus in situ photosynthesis, as leaves became more vertical. Petiole hyponasty and leaf‐blade cupping were also observed with warming + eCO₂ in marigold and soybean, respectively, which are compound‐leaved species like tomato, but no such hyponasty was observed in sunflower and okra, which have simple leaves. If severe leaf hyponasty is common under eCO₂ and warming, then this may have serious consequences for food production in the future.     

The cell cycle-associated protein kinase WEE1 regulates cell size in relation to endoreduplication in developing tomato fruit

Tomato fruit size results from the combination of cell number and cell size which are respectively determined by cell division and cell expansion processes. As fruit growth is mainly sustained by cell expansion, the development of pericarp and locular tissues is characterized by the concomitant arrest of mitotic activity, inhibition of cyclin-dependent kinase (CDK) activity, and numerous rounds of endoreduplication inducing a spectacular increase in DNA ploidy and mean cell size. To decipher the molecular basis of the endoreduplication-associated cell growth in fruit, we investigated the putative involvement of the WEE1 kinase (Solly;WEE1). We here report a functional analysis of Solly;WEE1 in tomato. Impairing the expression of Solly;WEE1 in transgenic tomato plants resulted in a reduction of plant size and fruit size. In the most altered phenotypes, fruits displayed a reduced number of seeds without embryo development. The reduction of plant-, fruit- and seed size originated from a reduction in cell size which could be correlated with a decrease of the DNA ploidy levels. At the molecular level downregulating Solly;WEE1 in planta resulted in the increase of CDKA activity levels originating from a decrease of the amount of Y15-phosphorylated CDKA, thus indicating a release of the negative regulation on CDK activity exerted by WEE1. Our data indicated that Solly;WEE1 participates in the control of cell size and/or the onset of the endoreduplication process putatively driving cell expansion.     

Metabolite annotations based on the integration of mass spectral information

A large number of metabolites are found in each plant, most of which have not yet been identified. Development of a methodology is required to deal systematically with unknown metabolites, and to elucidate their biological roles in an integrated 'omics' framework. Here we report the development of a 'metabolite annotation' procedure. The metabolite annotation is a process by which structures and functions are inferred for metabolites. Tomato ( Solanum lycopersicum cv. Micro-Tom) was used as a model for this study using LC-FTICR-MS. Collected mass spectral features, together with predicted molecular formulae and putative structures, were provided as metabolite annotations for 869 metabolites. Comparison with public databases suggests that 494 metabolites are novel. A grading system was introduced to describe the evidence supporting the annotations. Based on the comprehensive characterization of tomato fruit metabolites, we identified chemical building blocks that are frequently found in tomato fruit tissues, and predicted novel metabolic pathways for flavonoids and glycoalkaloids. These results demonstrate that metabolite annotation facilitates the systematic analysis of unknown metabolites and biological interpretation of their relationships, which provide a basis for integrating metabolite information into the system-level study of plant biology.