Effect of Heat Stress on Assimilate Metabolism in Tomato Flower Buds

Assimilate import by flower buds in two cultivars of tomato(Lycopersicon esculentum Mill.) was inhibited by heat stress.With increasing temperature, levels of sucrose in the sourceorgan increased while levels of starch decreased. The transportof radioactive carbon was correlated with the starch contentof the flower buds. In Saladette, a heat-tolerant cultivar,conversion of the imported carbon to starch occurred to a greaterextent than in Roma VF, a heat-sensitive cultivar. Uptake ofsucrose from agar medium by detached flower buds was negativelycorrelated with their internal ratio of sucrose to hexoses.Glucose uptake from agar medium by detached flower buds decreasedwith increasing temperatures. Sucrose hydrolysis was negativelyaffected by high temperatures, and this was more pronouncedin the heat-sensitive than in the heat-tolerant cultivar. Theeffect of heat stress on assimilate translocation from the leavesto the sink organ is discussed. Lycopersicon esculentum Mill., starch, sucrose, heat stress     

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     

Effect of Heat Stress on Assimilate Partitioning in Tomato

Differences in assimilate partitioning in response to heat stresswere observed between the heat-sensitive tomato cultivar, RomaVF and the heat-tolerant cultivar, Saladette. Transport of carbonto the trusses and apex was inhibited in both cultivars, particularlyin Roma VF. Basipetal transport to the roots was inhibited inRoma VF only. Assimilate partitioning within the young trusswas also affected by heat stress in both cultivars, but wasmore pronounced in Roma VF: at higher temperatures more ¹⁴Cwas found in the peduncle and in the older flower buds thanin the younger flower buds. The youngest flower buds were moresensitive than other parts of the truss to heat stress. Theuptake of [¹⁴C]sucrose by detached flower buds from agar mediumwas lower at higher temperatures. Alteration of assimilate partitioningwas also observed following the application of GA₃+Kinetin tothe first truss. The possible relationship between flower-setin tomato and carbohydrate stress is discussed. Lyeopersicon esculentum Mill., carbon translocation, starch     

Effects of Leaf Age and Position on the Shoot Axis on Potassium Absorption by Tomato Leaf Slices

The effect of leaf age on K (⁸⁶Rb) influx into tomato (Lycopersiconesculentum Mill.) leaf lamina slices was determined for leaves5, 9 and 13 counting acropetally. Potassium influx rates expressedon a leaf fresh weight basis declined rapidly during leaf elongationat external KCI concentrations between 0.5 and 20.0 mM. In fullyexpanded leaves, K influx rates declined more slowly with age.The onset of senescence in mature leaves did not result in alarge loss in K uptake capability. Leaf position on the shootaxis and the stage of whole plant development had little influenceon K influx into leaf cells. It is suggested that the rapiddecrease in K influx in growing leaves is related to a dilutionin the concentration of K transporter sites resulting from anincrease in cell volume and weight. Lycopersicon esculentum Mill, tomato, free space, potassium, influx rate, ion uptake, leaf slices, leaf age leaf ontogeny     

Evidence for the involvement of sucrose phosphate synthase in the pathway of sugar accumulation in sucrose-accumulating tomato fruits

To better understand the mechanism of sugar unloading and sugar concentration in hexose- and sucrose-accumulating tomato fruits (Lycopersicon chmielewskii and L. esculentum, respectively) and to determine the causes of the late accumulation of sucrose present in sucrose-accumulating tomato fruits, the assimilation of [³H](fructosyl)-sucrose was studied. Key enzymes involved in carbohydrate metabolism were also assayed. The results demonstrated that the low level of sucrose present in young fruits accumulates directly without undergoing hydrolysis, suggesting a symplastic pathway for sucrose unloading. By contrast, the large quantity of the sucrose present in ripe sucrose-accumulating fruits originates from hydrolysis and resynthesis, suggesting an apoplastic pathway for sucrose unloading. The increase in sucrose level observed in sucrose-accumulating fruits is associated with a gradual decline in invertase activity and an increase in sucrose phosphate synthase activity. This latter enzyme seems to play a key biochemical role in the accumulation of sucrose and the establishment of a high sugar content in tomato fruits.     

Enzymic Components of Sucrose Accumulation in the Wild Tomato Species Lycopersicon peruvianum

Sugar and soluble solids content and invertase (EC 3.2.1.26), sucrose synthase (EC 2.4.1.13), and sucrose phosphate synthase (EC 2.4.1.14) enzyme activities were measured throughout fruit development in tomato (Lycopersicon esculentum Mill.) and the green fruited species Lycopersicon peruvianum. Fruit of L. peruvianum accumulated predominantly sucrose, in contrast with hexose accumulation, which is characteristic of L. esculentum. The percentage of soluble solids in ripe L. peruvianum fruit was more than twice that present in L. esculentum and attributed primarily to the high level of sucrose accumulated in L. peruvianum. Low levels of invertase and sucrose synthase activity were associated with the period of significant sucrose accumulation and storage in L. peruvianum. Increased sucrose phosphate synthase activity was observed during the latter stages of fruit development in sucrose-accumulating fruit but was not coincident with maximum rates of sucrose accumulation.     

Control of Flower Number in the First Inflorescence of Tomato (Lycopersicon esculentum Mill.): The Role of Gibberellins

The number of flowers in the first inflorescence of tomato plantswas increased by low temperatures and reduced by the applicationof GA³. The effect of GA³, was greater in a low temperatureregime (12 °C minimum) than at normal temperatures (16 °Cminimum). Increases in flower number could be produced by theremoval of young developing leaves but the treatment was nolonger effective if plants wen grown at low temperatures orwere treated with GA³. Young developing leaves were shown to be sources of diffusiblegibberellin-like substances. Leaves from plants grown in a normaltemperature regime yielded greater amounts of gibberellin-likesubstances than leaves from plants grown in the low temperatureregime. It is suggested that high levels of endogenous gibberellinsact to reduce the number of flowers formed in the first inflorescence,and that leaf removal and low temperatures influence flowernumbers by lowering levels of diffusible gibberellins in theplants. Lycopersicon esculentum, tomato, flower number, gibberellins, temperature     

Effect of Leaf Age on Potassium Efflux and Net Flux in Tomato Leaf Slices

The effects on leaf age on K (⁸⁶Rb) efflux, influx and net fluxinto lamina slices from leaf 7 on a tomato plant (Lycopersiconesculentum Mill.) were determined. The ontogenetic trend inK efflux was dependent on the external K concentration. At externalKCI concentrations between 0.5 and 10.0 mM, K efflux rates increasedduring leaf elongation. Only a small increase in efflux occurredin mature leaves with increasing age. It is suggested that thetonoplast retains its structural integrity through the initialstages of leaf senescence. In fully expanded leaves, a zeronet K flux (a balance between influx and efflux) was achievedat external KCI concentrations between 1.0 and 3.5 mM. The Kcontent of lamina slices from leaves 5 and 13 remained constantwhen bathed in a solution containing 2 to 3 mM K. It is suggestedthat the decline in K concentration in mature tomato leaf tissueis due to a decline in leaf free space K concentrations below1 to 3 mM which would result in a net efflux out of leaf cells. Lycopersicon esculentum Mill., tor ato, free space, ion fluxes, leaf age, leaf ontogeny, potassium     

Ornithine decarboxylase and arginine decarboxylase activities in meristematic tissues of tomato and potato plants

Ornithine decarboxylase and arginine decarboxylase activities were measured in roots and buds of tomato (Lycopersicon esculentum Mill. cv. Pearson ms-35) and potato (Solanum tuberosum cv. Desire) plants. In both tomato and potato, the activity of ornithine decarboxylase was the highest at the root tip, decreasing proximally. The same was true for potato buds. In vegetative buds of tomato, the highest activity was found in the youngest leaves. The older the leaf, the lower was orithine decarboxylase activity. Arginine decarboxylase, on the other hand, did not display a similar gradient. These findings are in accordance with the suggestion that in tomato and potato elevated ornithine decarboxylase activity is associated with intense mitotic activity.     

The plant-endogenous Fe(II)-chelator nicotianamine restricts the ferrochelatase activity of tomato chloroplasts

Stripped chloroplasts were prepared from young leaves of a tomatowild type (Lycopersicon esculentum Mill.) and its mutant chloronerva.Several morphological and biochemical abnormalities of thismutant are caused by the total lack of the plant-endogenousFe²⁺ chelator nicotianamine (NA). The ferrochelatase activitywas estimated by determination of ⁵⁹Fe incorporated into haem.A mercaptoethanol concentration of 250 mM was necessary to maintainfull enzyme activity. The reducing agent supported the reducedstate of the active site of the enzyme more than that of theiron as revealed by use of ferrous and ferric ionproviding compoundsas substrates. Chloroplasts of both genotypes exhibited a similar enzyme activity.NA inhibited this activity by nearly 100% depending on the concentrationapplied. On the basis of the formation constant of the Fe(ll)–NAcomplex and the concentrations of iron and NA in the enzymeassay as well as in the tomato shoot apex region it is proposedthat ferrochelatase acts in vivo with an iron level at the attomolarrange which is provided by NA. Key words: Ferrochelatase activity, ferrous ion concentration, nicotianamine, tomato chloroplasts, substrate limitation     

The Relationship between Sucrose Supply, Sucrose-cleaving Enzymes and Flower Abortion in Pepper

Abortion of pepper flowers depends on the light intensity perceivedby the plant and on the amounts of sucrose taken up by the flower(Aloni B, Karni L, Zaidman Z, Schaffer AA. 1996.Annals of Botany78: 163–168). We hypothesize that changes in the activityof sucrose-cleaving enzymes within the flower ovary might beresponsible for the changes in flower abortion under differentlight conditions. In the present study we report that the activityof sucrose synthase, but not of cytosolic acid invertase, increasesin flowers of pepper plants which were exposed, for 2 d, toincreasing photosynthetically active radiation (PAR) in therange of 85–400 µmol m^-2s^-1at midday. Sucrose synthaseactivity increased in parallel with the increasing concentrationsof starch in the flower ovary. Feeding flower explants, preparedfrom 3-d-predarkened plants, with 100 mM sucrose for 24 h, causeda 23% increase in reducing sugars and a 2.5-fold increase instarch concentration, compared with explants fed with buffer.Likewise, feeding explants of pepper flowers with sucrose, glucose,fructose and also mannitol increased the sucrose synthase activityin the ovaries. Concomitantly, sucrose, glucose and fructose,but not mannitol, reduced the abortion of flower explants. Itis suggested that sucrose entry into the flower increases theflower sink activity by inhibiting abscission and inducing metabolicchanges, thus enhancing flower set. Pepper; Capsicum annuum L.; abscission; light; pepper flowers; sucrose; glucose; fructose; starch; acid invertase; sucrose synthase     

Antisense acid invertase (TIV1) gene alters soluble sugar composition and size in transgenic tomato fruit.

Invertase (beta-fructosidase, EC 3.2.1.26) hydrolyzes sucrose to hexose sugars and thus plays a fundamental role in the energy requirements for plant growth and maintenance. Transgenic plants with altered extracellular acid invertase have highly disturbed growth habits. We investigated the role of intracellular soluble acid invertase in plant and fruit development. Transgenic tomato (Lycopersicon esculentum Mill.) plants expressing a constitutive antisense invertase transgene grew identically to wild-type plants. Several lines of transgenic fruit expressing a constitutive antisense invertase gene had increased sucrose and decreased hexose sugar concentrations. Each transgenic line with fruit that had increased sucrose concentrations also had greatly reduced levels of acid invertase in ripe fruit. Sucrose-accumulating fruit were approximately 30% smaller than control fruit, and this differential growth correlated with high rates of sugar accumulation during the last stage of development. These data suggest that soluble acid invertase controls sugar composition in tomato fruit and that this change in composition contributes to alterations in fruit size. In addition, sucrose-accumulating fruit have elevated rates of ethylene evolution relative to control fruit, perhaps as a result of the smaller fruit size of the sucrose-accumulating transgenic lines.     

Sucrose Phosphate Synthase, Sucrose Synthase, and Invertase Activities in Developing Fruit of Lycopersicon esculentum Mill. and the Sucrose Accumulating Lycopersicon hirsutum Humb. and Bonpl

The green-fruited Lycopersicon hirsutum Humb. and Bonpl. accumulated sucrose to concentrations of about 118 micromoles per gram fresh weight during the final stages of development. In comparison, Lycopersicon esculentum Mill. cultivars contained less than 15 micromoles per gram fresh weight of sucrose at the ripe stage. Glucose and fructose levels remained relatively constant throughout development in L. hirsutum at 22 to 50 micromoles per gram fresh weight each. Starch content was low even at early stages of development, and declined further with development. Soluble acid invertase (EC 3.2. 1.26) activity declined concomitant with the rise in sucrose content. Acid invertase activity, which was solubilized in 1 molar NaCl (presumably cell-wall bound), remained constant throughout development (about 3 micromoles of reducing sugars (per gram fresh weight) per hour. Sucrose phosphate synthase (EC 2.4.1.14) activity was present at about 5 micromoles of sucrose (per gram fresh weight) per hour even at early stages of development, and increased sharply to about 40 micromoles of sucrose (per gram fresh weight) per hour at the final stages of development studied, parallel to the rise in sucrose content. In comparison, sucrose phosphate synthase activity in L. esculentum remained low throughout development. The possible roles of the sucrose metabolizing enzymes in determining sucrose accumulation are discussed.     

Effects of Heat Stress on Carbon Transport from Tomato Leaves

Export of radioactive carbon from two cultivars of tomato, (Lycopersiconesculentum Mill.) leaves was inhibited in response to heat stress.Increasing temperatures resulted in a marked decrease in leafstarch levels. The depletion of starch concentration in theleaves was primarily due to hydrolysis and an inhibition ofstarch formation. At high temperatures, starch hydrolysis wasinhibited in Roma VF, a heat sensitive cultivar, while Saladette,a heat tolerant cultivar was not similarly affected. Calloseformation was found on phloem sieve tube plates of leaf petiolesexposed to 72 h of high temperatures. More sieve tube plateswere covered with a thicker callose layer in Roma VF than inSaladette. Lycopersicon esculentum (Mill.), tomato, carbon translocation, starch hydrolysis, callose, heat stress     

Partitioning of [14C]sucrose and Acid Invertase Activity in Reproductive Organs of Pepper Plants in Relation to Their Abscission Under Heat Stress

The effect of heat stress on processes related to carbohydratepartitioning was investigated in young bell pepper (Capsicumannum L. cv. Maor) plants in relation to abscission of theirreproductive organs at different stages of development. None of the reproductive organs abscised after 5 d in a normalday/night temperature regime (25/18°C). With a temperatureregime of 35°C day, 25°C night, abscission occurredin only a small portion of the flower buds and none of the flowersand fruitlets. However, when temperatures in the day and nightwere reversed (25/35°C, day/night) all the buds and someof the flowers abscised during that time period. The young fruitat the first node did not abscise under any temperature regime.The abscission rate of the flower buds was reduced under heatstress if the developing fruit at the first node had been removed. High temperature during either the light or dark periods reducedthe export of [¹⁴C]sucrose from the source leaf (fed for 48h with [¹⁴C]sucrose). Both heat stress and fruit presence reduced the relative amountof [¹⁴C]sucrose which was exported to the flower buds, flowersand roots. Likewise, these treatments reduced the concentrationof reducing sugars in the reproductive organs. Concomitantly,the heat stress and fruit presence on the first node reducedthe activity of soluble acid invertase in the flower buds andthe roots, but not in young leaves. Overall, the results show that heat stress causes alternationin sucrose distribution in the plant, but may also have specificeffects on metabolic activities related to sucrose import andutilization in flower buds and flowers which in turn may enhancetheir abscission. Bell pepper, (Capsicum annuum L. cv. Maor), abscission, acidinvertase, heat stress, reproductive organs, sink leaves. Bell pepper, (Capsicum annuum L. cv. Maor), abscission, acid invertase, heat stress, reproductive organs, sink leaves.     

Partitioning of [14C]sucrose and Acid Invertase Activity in Reproductive Organs of Pepper Plants in Relation to Their Abscission Under Heat Stress

The effect of heat stress on processes related to carbohydratepartitioning was investigated in young bell pepper (Capsicumannum L. cv. Maor) plants in relation to abscission of theirreproductive organs at different stages of development. None of the reproductive organs abscised after 5 d in a normalday/night temperature regime (25/18 °C). With a temperatureregime of 35 °C day, 25 °C night, abscission occurredin only a small portion of the flower buds and none of the flowersand fruitlets. However, when temperatures in the day and nightwere reversed (25/35 °C, day/night) all the buds and someof the flowers abscised during that time period. The young fruitat the first node did not abscise under any temperature regime.The abscission rate of the flower buds was reduced under heatstress if the developing fruit at the first node had been removed. High temperature during either the light or dark periods reducedthe export of [¹⁴C]sucrose from the source leaf (fed for 48h with [¹⁴C]sucrose). Both heat stress and fruit presence reduced the relative amountof [¹⁴C]sucrose which was exported to the flower buds, flowersand roots. Likewise, these treatments reduced the concentrationof reducing sugars in the reproductive organs. Concomitantly,the heat stress and fruit presence on the first node reducedthe activity of soluble acid invertase in the flower buds andthe roots, but not in young leaves. Overall, the results show that heat stress causes alternationin sucrose distribution in the plant, but may also have specificeffects on metabolic activities related to sucrose import andutilization in flower buds and flowers which in turn may enhancetheir abscission. Bell pepper, (Capsicum annuum L. cv. Maor), abscission, acid invertase, heat stress, reproductive organs, sink leaves     

The influence of chilling on photosynthesis and activities of some enzymes of sucrose metabolism in Lycopersicon esculentum Mill

The effects of chilling stress on leaf photosynthesis and sucrose metabolism were investigated in tomato plants (Lycopersicon esculentum Mill. cultivar Marmande). Twenty-one-day-old seedlings were grown in a growth chamber at 25/23 °C (day/night) (control) and at 10/8 °C (day/night) (chilled) for 7 days. The most evident effect of chilling was the marked reduction of plant growth and of CO₂ assimilation as measured after 7 days, the latter being associated with a decrease in stomatal closure and an increase in Ci. The inhibition in photosynthetic rate was also related to an impairment of photochemistry of photosystem II (PSII), as seen from the slight, but significant change in the ratio of F_v/F_m. The capacity of chilled leaves to maintain higher q_P values with respect to the controls suggests that some protection mechanism prevented excess reduction of PSII acceptors. The results of the determination of starch and soluble sugar content could show that chilling impaired sucrose translocation. The activity of leaf invertase increased significantly in chilled plants, while that of other sucrose-metabolizing enzymes was not affected by growing temperature. Furthermore, the increase in invertase (neutral and acid) activity, which is typical of senescent tissue characterized by reduced growth, seems to confirm that tomato is a plant which is not a plant genetically adapted to low temperatures.     

Sink Metabolism in Tomato Fruit : IV. Genetic and Biochemical Analysis of Sucrose Accumulation

Fruit of domesticated tomato (Lycopersicon esculentum) accumulate primarily glucose and fructose, whereas some wild tomato species, including Lycopersicon chmielewskii, accumulate sucrose. Genetic analysis of progeny resulting from a cross between L. chmielewskii and L. esculentum indicated that the sucrose-accumulating trait could be stably transferred and that the trait was controlled by the action of one or two recessive genes. Biochemical analysis of progeny resulting from this cross indicated that the sucrose-accumulating trait was associated with greatly reduced levels of acid invertase, but normal levels of sucrose synthase. Invertase from hexose-accumulating fruit was purified and could be resolved into three isoforms by chromatofocusing, each with isoelectric points between 5.1 and 5.5. The invertase isoforms showed identical polypeptide profiles on sodium dodecyl sulfate polyacrylamide gel electrophoresis, consisting of a primary 52 kilodalton polypeptide and two lower molecular mass polypeptides that appear to be degradation products of the 52 kilodalton polypeptide. The three invertase isoforms were indistinguishable based on pH, temperature, and substrate concentration dependence. Immunological detection of invertase indicated that the low level of invertase in sucrose-accumulating fruit was due to low levels of invertase protein rather than the presence of an invertase inhibitor. Based on comparison of genetic and biochemical data we speculate that a gene either encoding tomato fruit acid invertase or one required for its expression, plays an important role in determining sucrose accumulation.     

Suppression of Acid Invertase Activity by Antisense RNA Modifies the Sugar Composition of Tomato Fruit

cDNA for an acid invertase (EC 3.2.1.26 [EC] ) of tomato (Lycopersiconesculentum Mill.) fruit was introduced into tomato plants underthe control of the cauliflower mosaic virus 35S promoter inthe antisense orientation. The antisense gene effectively suppressedthe invertase activity in soluble and cell wall fractions fromripening fruits. The sucrose content of fruits of the transformantswas markedly increased, while the hexose content was reduced.These results indicate that acid invertase is one of main determinantsof the sugar composition of tomato fruit. The invertase activityin the cell wall fraction of the leaf tissues of the transformantswas not suppressed to the same extent as that in the solublefraction. Wounding of the control leaf tissues induced invertaseactivity in both soluble and cell wall fractions. The inductionof activity in the soluble fraction was suppressed by the antisensegene, while that in the cell wall fraction was unaffected. Thesefindings suggest that mRNA for some other invertase, in particular,the mRNA for a cell wall-bound invertase, was present in leaves. ¹Present address: Plant Breeding and Genetics Research Laboratory,Japan Tobacco Inc., 700 Higashibara, Toyoda, Iwata, Shizuoka,438 Japan. ²Present address: National Institute of Agrobiological Resources,Kannondai, Tsukuba, Ibaraki, 305 Japan.     

Overexpression of sucrose phosphate synthase increases sucrose unloading in transformed tomato fruit

Sucrose unloading and sink activity were examined in tomato plants (Lycopersicon esculentum) overexpression sucrose phosphate synthase (SPS; EC 2.3.1.14). Like the leaves, the fruit of the transformed tomato plants had elevated (2.4-fold) SPS activity. SPS over-expression in tomato fruit did not significantly change acid invertase, and only slightly reduced ADPglc ppase activity, but enhanced sucrose synthase activity by 27%. More importantly, the amount of sucrose unloaded into the fruit was considerably increased. Using [³H]- (fructosyl)-sucrose in in vitro unloading experiments with harvested 20-d-old fruit, 70% more sucrose was unloaded into the transformed fruits compared to the untransformed controls. Furthermore, the turnover of the sucrose unloaded into the fruit of transformed plants was 60% higher than that observed in the untransformed controls. Taken together, these results demonstrate that SPS overexpression increases the sink strength of transformed tomato fruit.