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.     

Cellular Growth in Roots of a Gibberellin-Deficient Mutant of Tomato (Lycopersicon esculentum Mill.) and its Wild-type

The role of gibberellins in regulating the growth of tomatoroots was investigated by comparing various cellular parametersin cultured roots of the gibberellin-deficient mutant gib-l/gib-lwith those in roots of the near-isogenic wild-type. In addition,wild-type roots treated with 0?1 µM 2S,3S paclobutrazol,an inhibitor of gibberellin biosynthesis, and mutant roots treatedwith 0?1 µM GA₃ were also compared: the former roots constitutea phenocopy of the mutant, whereas the latter roots appear tobe ‘normalized’ and similar to wild-type. The elongationof mutant and phenocopied roots were similar, their maximumelongation rates being about half or two-thirds that of wild-typeor GA₃-treated mutant roots, respectively. These rates wereinterpreted in terms of the numbers and lengths of cells withinthe meristematic and non-meristematic portions of the elongationzone. Mean meristem length tended to be shorter in both themutant and the 2S,3S paclobutrazol-treated wild-type roots thanin the other two types of root. A major difference between thetwo pairs of mutant and normal roots was their mean final celllengths: mean lengths of cortical cells of the mutant and 2S,3Spaclobutrazol-treated roots were, respectively, 39% and 25%shorter than the mean length of wild-type roots. Final celllength in the GA³-treated mutant roots were similar to wild-type.By contrast, the diameters of mature cortical cells of the mutantand phenocopy were about 20% greater than the diameters of equivalentwild-type or ‘normalized’ mutant cells. The meanvolumes of cortical cells in all four types of roots showedno significant differences. Knowledge of the distribution ofcortical cell lengths, widths and volumes along the root axis,together with information about the rate of root elongation,permitted comparisons of the relative elemental growth ratesof each of these three cellular parameters. The available evidence suggests that the level of endogenousgibberellins in mutant roots is lower than in wild-type roots.The present results, therefore, suggest that endogenous gibberellinsare necessary for normal growth of cultured tomato roots andthat they regulate the relative amounts of growth at the longitudinaland transverse walls of the cells which, in turn, affects theshape of the elongating cells. Key words: Cell growth, cultured roots, gibberellin, gib-l mutant, Lycopersicon esculentum, 2S,3S paclobutrazol, relative elemental growth rate, root meristem     

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.     

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.     

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.     

Invertase Activity, Soluble Carbohydrates and Inflorescence Development in the Tomato (Lycopersicon esculentum Mill.)

The concentration of reducing sugars in the developing firstinflorescence of the tomato (Lycopersicon esculentum Mill.)increased steadily between the macroscopic appearance of theflower buds and the initial stages of fruit expansion. Overthis period sucrose concentrations remained relatively constant.The rise in reducing sugar concentration was accompanied byan increase in the activity of an acid invertase. In individualflower buds invertase activity rose to a maximum shortly beforeanthesis and declined sharply as the anthers dehisced. Increased planting densities and removal of source leaves reducedthe rate of dry matter accumulation by the first inflorescenceand increased the incidence of flower bud abortion. These changeswere correlated with reductions in reducing sugar concentrations,in reducing sugar/sucrose ratios and in acid invertase levels.Removal of young leaves at the shoot apex significantly increasedthe relative growth rate of the inflorescence and led to a substantialincrease in its invertase content. These treatments had relativelylittle effect on sucrose concentration in the inflorescence. The data are consistent with the operation of an invertase-mediatedunloading mechanism for transported sucrose at sinks in theflower buds. It is suggested that the retarded development ofthe first inflorescence and the high incidence of flower budabortion observed under conditions of reduced photoassimilateavailability are causally related to the decline in invertaseproduction in the flower buds. Possible mechanisms for the regulationof invertase synthesis in the flowers are discussed. Lycopersicon esculentum Mill, tomato, inflorescence development, invertase, sink activity     

The Quiescent Centre and Rates of Mitosis in the Root Meristem of Allium sativum

The root apices of Allium sativum have been examined by continuous-and pulse-labelling with tritiated thymidine and by colchicinetreatment to measure the time parameters of the mitotic cyclein various parts of the meristem. There is a quiescent centre of 30–50 cells whose averagerate of mitosis is low because the G₁ period is extended toabout 140 h compared with about 4 h in the othe regions of themeristem. The stele just above the quiescent centre and at 200microns above it and the cap initials just below the quiescentcentre are very similar in their mitotic cycles, the total lengthsof which are about 30 h of which nearly half is taken up byDNA synthesis. Allium thus differs from Zea in having root capinitials whose mitotic cycle is not telescoped by the eliminationof the G₁ phase. These facts are discussed in relation to theradiosensitivity of the meristem.     

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.     

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.     

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.     

Characterization of the Stimulation of Ethylene Production by Galactose in Tomato (Lycopersicon esculentum Mill.) Fruit

We have characterized the stimulation of ethylene production by galactose in tomatoes (Lycopersicon esculentum Mill.). The effect of concentration was studied by infiltrating 0, 4, 40, 100, 200, 400, or 800 micrograms galactose for each gram of fresh fruit weight into mature green `Rutgers' fruit. Both 400 and 800 micrograms per gram fresh weight consistently stimulated a transient increase in ethylene approximately 25 hours after infiltration; the lower concentrations did not. Carbon dioxide evolution of fruit infiltrated with 400 to 800 micrograms per gram fresh weight was greater than that of lower concentrations. The ripening mutants, rin and nor, also showed the transient increase in ethylene and elevated CO₂ evolution by 400 micrograms per gram fresh weight galactose. 1-Aminocyclopropane-1-carboxylic acid (ACC) content and ACC-synthase activity increased concurrently with ethylene production. However, galactose did not stimulate ACC-synthase activity in vitro. The infiltrated galactose in pericarp tissue was rapidly metabolized, decreasing to endogenous levels within 50 hours. Infiltrated galacturonic acid, dulcitol, and mannose stimulated transient increases in ethylene production similar to that of galactose. The following sugars produced no response: sucrose, fructose, glucose, rhamnose, arabinose, xylose, raffinose, lactose, and sorbitol.     

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.     

番茄"白化"突变体果实的几个生理生化特性检测

在番茄“白化”突变体果实发育过程中,总叶绿素、叶绿素a、叶绿素b的含量均小于正常植株的果实;果肉细胞内含有白色体;果实在贮藏期间的硬度较大。而果实品质,如糖酸比、维生素C和可溶性固形物的含量与正常果实差异不大。     

Patterns of Assimilate Distribution and Source--sink Relationships in the Young Reproductive Tomato Plant (Lycopersicon esculentum Mill.)

Patterns of distribution of ¹⁴C were determined in 47-day-oldtomato plants (Lycopersicon esculentum Mill.) 24 h after theapplication of [¹⁴C]sucrose to individual source leaves fromleaves 1–10 (leaf 1 being the first leaf produced abovethe cotyledons). The first inflorescence of these plants wasbetween the ‘buds visible’ and the ‘firstanthesis’ stages of development. The predominant sink organs in these plants were the root system,the stem, the developing first inflorescence and the shoot ‘apex’(all tissues above node 10). The contribution made by individualsource leaves to the assimilate reaching these organs dependedupon the vertical position of the leaf on the main-stem axisand upon its position with respect to the phyllotactic arrangementof the leaves about this axis. The root system received assimilateprincipally from leaf 5 and higher leaves, and the stem apexfrom the four lowest leaves. The developing first inflorescencereceived assimilates mainly from leaves in the two orthostichiesadjacent to the radial position of the inflorescence on thevertical axis of the plant; these included leaves which weremajor contributors of ¹⁴C to the root system (leaves 6 and 8)and to the shoot apex (leaves 1 and 3). This pattern of distributionof assimilate may explain why root-restriction treatments andremoval of young leaves at the shoot apex can reduce the extentof flower bud abortion in the first inflorescence under conditionsof reduced photoassimilate availability. Lycopersicon esculentum Mill, tomato, assimilate distribution, source-sink relationships     

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.