Nitrogen fixation in tomato (Lycopersicon esculentum Mill ‘Pusa Ruby’)

Nitrogen fixation (acetylene reduction) was found in intact tomato (Lycopersicon esculentum Mill ‘Pusa Ruby’) plants in the field, in pots and also in aseptic cultures. The unsterilized as well as sterilized rhizoplane and phylloplane of the plant when assayed separately also responded to the test. From root bits of tomato sterilized upto 20 minutes with 0.1% mercuric chloride, growth of the bacterium from the interior of the root into the medium was observed thereby indicating their presence within the endorhizosphere. Phase contrast and electron microscopic studies of the root system of tomato revealed the presence of bacterial colonies in the epidermis, cortex and vascular bundles. Bacterial numbers in the endosphere, of root and leaf were 30×10⁴ and 12×10⁴, respectively, per gram fresh weight of tissue. The bacteria were predominantly rod-shaped 1.4–4.8×0.9–1.95 μm in 24-h-old cultures, pleomorphic, polar or bipolary flagellated having β-hydroxy butyrate granules. The bacterium has been identified as a new species of Azospirillum.     

Tomato (Lycopersicon esculentum Mill., cv. 'Better Bush') Plant Response to Root Restriction

Observations presented here describe changes in the growth ofLycopersicon esculentum Mill., cv. ‘Better Bush’,at the onset of root-restriction stress. Plants were grown ineither 1500 cm³ (control) or 25 cm³ (treated) containers ina flow-through hydroponic culture system (FTS). Seeds were imbibed,germinated, held for 13 d in sterile germination bags, thenplaced into the FTS and designated day zero plants. Plants grownin the FTS were harvested, every 7 d, from day 11 to day 46.Plant height, root and shoot weights, leaf areas, leaf lengths,branching and flowering were measured to illustrate morphologicalchanges that occurred in response to root-restriction stress.Plants restricted to a 25 cm³ root volume experienced a markedstress after 18 d in the FTS. A reduction in leaf elongationwas evident and preceded the time (prior to day 25) when maximumdry weights were attained by restricted root systems. Controlplants had greater leaf areas and plant dry weights than thoseof root-restricted plants after day 18. Treated plants showedno significant increase in dry weights from day 32 to the endof the experiment. Between day 18 and 25 the root: shoot ratiofor the treated plants dropped below that of the controls. Adventitiousrooting was evident in treated plants by day 25. Roots of thetreated plants showed an apparent waterlogging indicated bybrowning of roots by day 32. Root turnover followed as the primaryroot system was replaced by adventitious roots. The number ofbranches or flowers did not differ between control and treatedplants. The timing of the events described here can be usedto further the study of specific physiological responses oftomato plants to root-restriction stress.     

Chromoplast-Targeted Proteins in Tomato (Lycopersicon esculentum Mill.) Fruit

The chloroplast to chromoplast transition during tomato (Lycopersicon esculentum Mill.) fruit ripening is characterized by a dramatic change in plastid structure and function. We have asked whether this process is mediated by an increase in the steady-state level of RNA for plastid targeted proteins. Assays for import of radiolabeled translation products into isolated pea (Pisum sativum L.) chloroplasts were used to monitor levels of chromoplast-targeted proteins at four stages of tomato fruit development. We have found striking increases during development in levels of translatable RNA for two such proteins. Additionally, the import of in vitro translation products was examined for seven individual cDNA clones known to encode RNA that increase during fruit ripening. Three of these clones produced in vitro translation products that were imported into pea chloroplasts. This implies that there is synthesis and import of new proteins during the transition from chloroplast to chromoplast and that the plastid conversion is an active developmental program rather than a simple decline in synthesis of the photosynthetic apparatus. Furthermore, our results demonstrate the utility of this method for identification of structural genes involved in plastid morphogenesis.     

NADPH Oxidase Genes from Tomato (Lycopersicon esculentum) and Curly-Leaf Pondweed (Potamogeton crispus)

Abstract: The oxidative burst is an integral component of plant resistance to pathogens. There is accumulating evidence that the oxidative burst is catalyzed by an enzyme with similarities to the phagocyte NADPH oxidase. We have cloned a full length homolog of the gp91 ( phox ) subunit of the plasma membrane NADPH oxidase complex from tomato named LeRBOM. The predicted protein contains 989 amino acids. The large N-terminal domain contains two EF hand calcium binding motifs and one conserved glycosylation site. Six putative membrane spans are present in the C-terminal half of the predicted protein. Extensive homology with the human gp91 ( phox ) subunit was found including conservation of amino acid residues important for heme coordination and substrate binding. We have also isolated partial genomic clones from tomato and from the aquatic plant Potamogeton crispus. These species serve as models for studies of signal transduction leading to NADPH oxidase activation. In tomato, LeRBOH1 expression was too low to be detected on Northern blots. RT-PCR indicated that LeRBOH1 was expressed in all tissues tested.     

Effects of Low O(2) Root Stress on Ethylene Biosynthesis in Tomato Plants (Lycopersicon esculentum Mill cv Heinz 1350)

Low O₂ conditions were obtained by flowing N₂ through the solution in which the tomato plants (Lycopersicon esculentum Mill cv Heinz 1350) were growing. Time course experiments revealed that low O₂ treatments stimulated 1-aminocyclopropane-1-carboxylate (ACC) synthase production in the roots and leaves. After the initiation of low O₂ conditions, ACC synthase activity and ACC content in the roots increased and reached a peak after 12 and 20 hours, respectively. The conversion of ACC to ethylene in the roots was inhibited by low levels of O₂, and ACC was apparently transported to the leaves where it was converted to ethylene. ACC synthase activity in the leaves was also stimulated by low O₂ treatment to the roots, reaching a peak after 24 hours. ACC synthase levels were enhanced by cobalt chloride and aminooxyacetic acid (AOA), although they inhibited ethylene production. Cobalt chloride enhanced ACC synthase only in combination with low O₂ conditions in the roots. Under aeration, AOA stimulated ACC synthase activity in both the roots and leaves. However, in combination with low O₂ conditions, AOA caused a stimulation in ACC synthase activity in the leaves and no effect in the roots.     

Tissue Culture Studies of Tomato (Lycopersicon esculentum)

Tomato is a major vegetable crop that has achieved tremendous popularity over the last century. It is grown in almost every country of the world. Development of protocols for in vitro selection can provide new advances for the production of stress tolerant cultivars. Techniques have been optimised for the production of haploids and somatic hybrids. Attempts have also been made to transfer the higher regenerative ability of wild varieties to cultivated tomatoes. Although, some information is available on the morphogenesis of tomato, the techniques have not been developed to a level at which they can be utilised in large-scale multiplication of commercially important cultivars. The morphogenesis response seems to be highly dependent PGRs used in the media, which is again cultivar and genotypic specific. Somatic embryogenesis in tomato is still at its infancy, and efficient procedures for large-scale production via somatic embryogenesis are yet to be developed. Genetic stability of the tissue culture raised tomato plants also needs to be addressed. The use of a combination of molecular and conventional breeding techniques could be the option for the development of cultivars resistant to biotic and abiotic stresses. This paper reviews the advances made in various aspects of tissue culture in tomato. It also discusses the issues that still need to be addressed to utilise the full potential of plant tissue culture techniques in genetic improvement and mass propagation of tomato.     

Mannosyl- and Xylosyl-Containing Glycans Promote Tomato (Lycopersicon esculentum Mill.) Fruit Ripening

The oligosaccharide glycans mannosylα1-6(mannosylα1-3)mannosylα1-6(mannosylα1-3) mannosylβ1-4-N-acetylglucosamine and mannosylα1-6(mannosylα1-3)(xylosylβ1-2) mannosylβ1-4-N-acetylglucosaminyl(fucosylα1-3) N-acetylglucosamine were infiltrated into mature green tomato fruit (Lycopersicon esculentum Mill., cv Rutgers). Coinfiltration of 1 nanogram per gram fresh weight of the glycans with 40 micrograms per gram fresh weight galactose, a level of galactose insufficient to promote ripening, stimulated ripening as measured by red coloration and ethylene production.     

Descriptive Physiology and Biochemistry of the Abnormally Ripening Tomato Fruit (Lycopersicon esculentum) cv. Snowball

Aspergillus tamarii Kita grew and sporulated best at 30°C. The best pH for growth and sporulation were 5.5 and 6.5, respectively. Among the carbon sources employed, glucose supported the highest growth and sporulation. Best growth was obtained with sodium nitrate as nitrogen source and best sporulation with ammonium tar-tarate. When glucose was used as carbon source, the carbohydrates found in the mycelium included myoinositol. dulcitol, fructose, arabinose and ribose.     

Metabolism of Indole-3-Acetic Acid by Pericarp Discs from Immature and Mature Tomato (Lycopersicon esculentum Mill)

[1′-¹⁴C, ¹³C₆]Indole-3-acetic acid was infiltrated into immature pericarp discs from fruits of tomato (Lycopersicon esculentum Mill., cv Moneymaker). After a 24-h incubation period the discs were extracted with methanol and the partially purified extract was analyzed by reversed-phase high-performance liquid chromatography-radiocounting. Five metabolite peaks (1-5) were detected and subsequently analyzed by combined high-performance liquid chromatography-frit-fast atom bombardment-mass spectrometry. The metabolite 4 fraction was found to contain [¹³C₆]-indole-3-acetylaspartic acid, and analysis of metabolite 5 identified [¹³C₆]indole-3-acetyl-β-d-glucose. The other metabolites could not be identified, but alkaline hydrolysis studies and gel permeation chromatography indicated that metabolites 1 and 3 were both amide conjugates with a molecular weight of approximately 600. Studies with radiolabeled indole-3-acetic acid, indole-3-acetylaspartic acid, and indole-3-acetyl-β-d-glucose demonstrated that in immature pericarp indole-3-acetic acid is deactivated primarily via metabolism to indole-3-acetylaspartic acid, which is further converted to metabolites 1, 2, and 3. In mature, pink pericarp discs, indole-3-acetic acid is converted more extensively to its glucosyl conjugate. Conjugation of indole-3-acetic acid to indole-3-acetylaspartic acid appears to be dependent upon protein synthesis because it is inhibited by cycloheximide. In contrast, cycloheximide has little effect on the further conversion of indole-3-acetylaspartic acid to metabolites 1, 2, and 3.     

Growth and Water Relations of Wilty Mutants of Tomato (Lycopersicon esculentum Mill.)

In contrast to some previous reports on the growth of the ABA-deficientwilty mutants of tomato, growth was at least as rapid in themutants as in the wild type, as long as an adequate plant waterstatus was maintained by growing the plants under mist. Moreover,shoot extension was greater and the rate of leaf productionmore rapid in the mutants. Stomatal changes in response to environmentand to time in the light-dark cycle were generally similar inboth wilty mutants and the wild type, though the wild-type weregenerally more closed. Grafting experiments confirmed that thegenotype of the shoot was dominant in determining stomatal aperture,though wild-type rootstocks could cause a slight reduction inthe stomatal conductance of mutant leaves. The effect on plantwater relations of draughting only part of the root system wasinvestigated in a ‘split-root’ experiment. Withholdingwater from only part of the root system was found to lower significantlythe mean leaf water potential, even though the potential evaporationrate was kept very small. Key words: Abscisic acid, stomata, tomato     

Compartments and Permeability for Potassium in Developing Fruits of Tomato (Lycopersicon esculentum Mill.)

Permeabilities of plasmalemma and tonoplast, and the distributionof potassium between free space, cytoplasm, and vacuole, wereestimated from the kinetics of the efflux of potassium fromslices of tomato pericarp. The permeabilities of plasmalemmaand tonoplast did not change during development, but the proportionof potassium in the cytoplasm increased during ripening. Itis suggested that the effects of ethylene, and the changes takingplace during ripening are not caused by changing permeabilitiesof membranes. Increasing activities of ions in the cytoplasm,rather than increasing membrane permeabilities, may explainprevious observations that the efflux of solutes from fruittissues increases during ripening.     

Studies on two Gibberellin-like Substances in Young Shoots of Tomato (Lycopersicon esculentum Mill.)

Two gibberellin-like substances were found in the acidic fractionof shoot extracts of the tomato (Lycopersicon esculentum Mill.,cultivar Potentate). These were resolved by paper chromotographywith iso-propanol/ammonia/water (10:1:1) as the developing solventbut not with n-butanol/1.5 N ammonia (3:1). Both substanceswere active in the dwarf maize bioassay on mutants d-1, d-2,d-3, and d-5, and appeared to be more active on d-5 than d-1.Neither was active in the Meteor Pea assay. Neutral and basicfractions were inactive. The relative amounts of these two substances varied accordingto the age of the tissues from which they were extracted andthis feature is discussed in relation to future studies on thephysiology of gibberellin-like substances in vivo.     

Characterization of an Ethylene Overproducing Mutant of Tomato (Lycopersicon esculentum Mill. Cultivar VFN8)

Ethylene production rates and tissue ethylene concentrations were determined for the single-gene, Epinastic (Epi) tomato (Lycopersicon esculentum Mill.) mutant, and its parent, cv VFN8. The Epi phenotype was characterized by severe leaf epinasty, thickened stems and petioles, and a compact growth habit. In 4-day-old seedlings, ethylene production was significantly higher in Epi than in VFN8. Ethylene production rates also were higher for excised root, hypocotyl, cotyledon, and shoot tissue of 14-day-old Epi seedlings as compared with VFN8. The greatest difference in the ethylene production rate was observed in excised Epi shoot tissue, which was more than 2.5 times higher than in VFN8. Tissue ethylene concentrations of 19−, 25−, and 31-day-old Epi plants were 8, 172, and 307% higher than for VFN8, corresponding to increasing expression of the Epi phenotypic characteristics with age. The highest ethylene concentrations occurred in the shoot apex of both genotypes. Higher ethylene concentrations in Epi resulted from greater 1-aminocyclopropane-1-carboxylic acid content rather than increased ethylene-forming enzyme activity. The elevated ethylene levels in Epi did not result from increased auxin sensitivity. The sensitivity of root growth to inhibition by ethylene did not differ between VFN8 and Epi. Although elevated levels of ethylene in Epi plants apparently exacerbate its epinastic growth characteristics, other evidence indicates that this may not be the fundamental lesion. This mutant may provide a unique system for investigating the regulation of ethylene biosynthesis and the role of target cell types in plant development.     

Tomato bushy stunt virus (TBSV) infecting Lycopersicon esculentum

Tomato bushy stunt virus (TBSV) was detected in tomato crop (Lycopersicon esculentum) in Egypt with characteristic mosaic leaf deformation, stunting, and bushy growth symptoms. TBSV infection was confirmed serologically by ELISA and calculated incidence was 25.5%. Basic physicochemical properties of a purified TBSV Egh isolate were identical to known properties of tombusviruses of isometric 30-nm diameter particles, 41-kDa coat protein and the genome of approximately 4800 nt. This is the first TBSV isolate reported in Egypt. Cloning and partial sequencing of the isolate showed that it is more closely related to TBSV-P and TBSV-Ch than TBSV-Nf and TBSV-S strains of the virus. However, it is distinct from the above strains and could be a new strain of the virus which further confirms the genetic diversity of tombusviruses.     

Novel Technique for Measuring Tissue Firmness within Tomato (Lycopersicon esculentum Mill.) Fruit

Developmental changes of tomato (Lycopersicon esculentum) fruit tissues during maturation were analyzed by a physically defined method (stress-relaxation analysis). The tip of a conical probe connected to a load sensor was positioned on the cut surface of a sliced tomato fruit, and the decay of the imposed stress was monitored. Stress-relaxation data thus obtained were used for the calculation of three stress-relaxation parameters. Different zones within tomato fruit harvested at six different ripening stages were analyzed. One of the stress-relaxation parameters, minimum stress-relaxation time (T₀), decreased as the fruits matured. The decrease in T₀ was first found in the core of the carpel junction within the endopericarp at the blossom end during the breaker stage. The decrease in T₀ progressed from the blossom end, through the equatorial region and finally throughout the shoulder, as the fruit matured. In mature green fruit, T₀ values within the placenta and the proximal carpel junction were lower than those by other parts of the fruit. For all measurements the maximum stress-relaxation time was not substantially changed during maturation, nor were their changes observed in different regions of the fruit. The observed relaxation rate was therefore correlated with softening. The results indicate that fruit softening may be physically associated with the stress-relaxation parameter, T₀, and the extent of softening is a function of position within the fruit. Decreases in T₀ value appear to be correlated with the reported regional variation in the appearance of polygalacturonase.     

Effects of Ethylene on Photosynthesis and Partitioning in Tomato, Lycopersicon esculentum Mill

The extended period of ethylene release from ethephon (2-chloroethylphosphonicacid) after application to intact tomato plants has provideda model system in which the effects of ethylene on photosyntheticmetabolism and carbon partitioning has been studied. Ethylenerelease from leaf tissue after ethephon treatment was 10 timesgreater than that from untreated control leaves. The specificactivity of 14C2H4 released from [14C] ethephon remained constantover several days demonstrating that the ethylene was derivedfrom the applied ethephon. The ethephon-treated plants exhibitedextreme epinasty of the leaves and 24 h after application theflower buds in the first visible cluster had abscised, leafexpansion at the apex had ceased and developing adventitiousroots were visible on the lower stem. Rates of steady-state photosynthesis, respiration, photorespirationand transpiration were the same in treated and control leaves24 h after ethephon application. Both treated and control leavespartitioned similar proportions of newly-fixed 14C from 14CO²into neutral (46.4%), acidic (14.0%), basic (5.0%) and insoluble(34.0%) leaf fractions under steady-state conditions. The speedof 11C-assimilate movement in the stems of control plants (3.62±0.42cm min-1 towards the apex and 4.03±0.15 cm min-1 towardsthe roots) was more rapid than in the ethephon-treated plants(2.90±0.31 cm min-1 upwards and 2.59±0.22 cm min-1downwards). Furthermore, in the control plants 20.0±5.4%of the 14C exported to the plant from the source leaf was transportedtowards the developing flower cluster and young leaves. Twenty-fourhours after ethephon application only 6.5 ±1.7% of theexported 14C was translocated towards the shoot. Contrary tosome reports ethylene did not affect steady-state gas exchangeprocesses while carbon partitioning was significantly alteredindicating that ethylene effects on photosynthetic carbon metabolismare indirect and not due to direct effects on photosyntheticprocesses per se. Key words: Ethylene, photosynthesis, partitioning     

Physiological and biochemical responses resulting from nitrite accumulation in tomato (Lycopersicon esculentum Mill. cv. Ibiza F1)

The sensitivity of hydroponically cultivated tomato (Lycopersicon esculentum Mill. cv. Ibiza F1) submitted to nitrite treatments (0.25-10mM KNO(2)) for 7d was studied. Increasing nitrite levels in the culture medium led to several disruptions of tomato plants, reflected by reductions of both dry matter per plant, chlorophyll concentrations and the appearance of chlorosis symptoms at the leaf surface. This behaviour was accompanied by stimulation of nitrite, nitrate and ammonia ion accumulation, mainly in roots and old leaves. Higher proteolytic and gaiacol peroxidase (GPX, EC. 1.11.1.7) activities and malonyldialdehyde content were also noted. Protein content of the different plant organs was decreased by nitrite treatment. These physiological and biochemical parameters were chosen as they are stress indicators. Taken together, our data partly explain the harmful effects of nitrite ions, when excessive in the culture medium.     

串番茄果实货架期间与耐贮性有关的生理特性的变化

串番茄品种随着货架期延长,果实硬度、果肉硬度、原果胶含量逐渐下降;可溶性果胶含量逐渐上升;多聚半乳糖醛酸酶(PG)活性呈峰值变化,变化幅度小于普通番茄.     

Gibberellins and the Early Disease Syndrome of Aspermy Virus in Tomato (Lycopersicon esculentum Mill.)

A quantitative examination of the development of the diseasesyndrome created by as permyvirus in tomato (Lycopersicon esculentumMill., cultivar Potentate) revealed an early reduction in subapicalmitotic activity which led to the development of smaller internodes.Later symptoms were associated with necrosis of young axillarybuds and the failure of a new vegetative apex to develop onceflower initiation had begun in the primary apical bud. Application of gibberellic acid (GA₃) to the growing point tendedto reverse virus-induced stunting by increasing cell expansionbut the response was less than that in healthy plants. Thiswas due to a reduction in the number of mitoses associated withvirus infection. No consistent difference was found betweenthe endogenous gibberellin levels of healthy and infected plants.It is suggested that the most important single factor in earlysymptom development is a virus-induced reduction in cell divisionwhich cannot be overcome by applying GA₃.