Modelling nitrate influx in young tomato (Lycopersicon esculentum Mill.) plants

The effects of light and NO3^- nutrition on ¹⁵NO3^- influx in roots were investigated in young, 19-d-old, induced tomato plants grown at a constant air and solution temperature of 20C. Nitrate influx was measured by ¹⁵N accumulation for 5 min, on plants exposed to a wide range of exogenous concentrations, from 10 x 10^-3 to 30 mol m^-3. Influx kinetics, fitted to the data following a non-linear procedure, showed multiphasic patterns. The best fits were obtained when three pure and non-additive Michaelis-Menten kinetics were applied, with phase transitions at approximately 0.8 and 4 mol m^-3. In plants grown at 3.0 mol m^-3 NO3^-, the asymptotic maximum influx rate (Imax) of each phase declined during the night until 24 h darkness. At the end of the day period, about a 2-fold enhancement of Imax was observed when plants were pretreated for 3 d with 0.2 instead of 3.0 mol m^-3 NO3^-. The influx rates measured at any given NO3^- concentration and the Imax for any phase showed a negative non-linear correlation with plant nitrate concentration. Furthermore, the results suggest the existence of a set point, approximately 66 mol m^-3 plant nitrate, for which influx is null at any given solution nitrate concentration. A model using modified Michaelis-Menten kinetics is proposed to predict the influx rate as a function of both solution and plant NO3^- concentrations.     

Physiological response and yield of paclobutrazol treated tomato plants (Lycopersicon esculentum Mill.)

Experiments were conducted to study the physiologicaleffect of the plant growth retardant paclobutrazol(PBZ) and its impact on the yield of tomato plants(cv. Precador). Seedlings were treated at the time of prickingout with soil and foliar applications of PBZ atconcentrations of 1.0 and 25.0 mg l^-1respectively. The results established that:-- The reduced height and the increased thickness ofthe young plant stem, as well as the accelerated rootformation are a significant advantage of the PBZtreatment, contributing to the improvement of seedlingquality at planting.-- Soil treatment (1 mg l^-1) and foliar treatment(25 mg l^-1) with PBZ improves the photosyntheticactivity and water balance of tomato cv. Precador.-- PBZ accelerates fruit formation and increases earlyfruit yield.-- The concentrations of the retardant used and themode of its application ensure the production offruits without any residual retardant and harmless tohuman health from a phytosanitary point of view.     

Phosphorus use efficiency in anthocyanin-free tomato (Lycopersicon esculentum Mill.)

An anthocyanin-free tomato plant, H957, and its parental wild type, H883, were hydroponically grown to test for tolerance to a low phosphorus (P) in H957. The tolerance was evaluated by comparing growth and metabolism of H957 vs. H883 at different P concentrations ranging 25–400 μM. Fresh weights were measured weekly. Dry weight, mineral contents, photosynthetic rate, and P utilization ratios of the plants were measured after five weeks of growth in the hydroponic culture. Although the growth of both varieties was severely impaired at 25 μM P, H957 showed a greater fresh weight and dry weight at 50–400 μM P. H957 showed a higher net photosynthetic rate on older leaves while both varieties showed similar photosynthetic rate on young leaves. H957 tissue contains an overall lower P concentration in its tissue than H883. These observations together indicate that the anthocyaninless mutant H957 tolerate to lower P concentration. It does so by utilizing internal P with better efficiency rather than by absorbing external P better.     

Phytochrome-mediated induction of phenylalanine ammonia-lyase in the cotyledons of tomato (Lycopersicon esculentum Mill.) plants

Phenylalanine ammonia-lyase (PAL; EC 4.3.1.5.) induction in cotyledons from 96-h dark-grown Lycopersicon esculentum Mill. was studied in response to continuous light and hourly light pulses (blue, red, far red). The increases of PAL promoted by blue and red pulses are reversed completely by immediately following 758 nm irradiations. The response to continuous red light could be substituted for by hourly 6-min red light pulses. The effect of continuous red treatments is mainly due to a multiple induction effect of phytochrome. In contrast to red light, hourly light pulses with far red and blue, light can only partially substitute for continuous irradiation. The continuous blue response could be due to a combination of a multiple induction response and of a high irradiance response of phytochrome. The continuous far red response, could represent a high irradiance response of phytochrome. Dichromatic irradiations indicate that phytochrome is the photoreceptor controlling the light response (PAL) in tomato seedlings.Abbreviations Norflurazon NF-4-chloro-5-(methylamino)-2-(,,,-trifluoro-m-tolyl)-3 (2H) pyridazinone - PAL phenylalanine ammonia-lyase - phytochrome photoequilibrium P_fr/P_tot - P_fr far-red absorbing form of phytochrome - P_r red absorbing form of phytochrome - P_tot total phytochrome: P_r+P_fr     

Identification and distribution of profilin in tomato (Lycopersicon esculentum Mill.)

Profilin is a G-actin monomer-binding protein which has been shown to participate in actin-based tipgrowth of animal cells. The abundance of profilin in pollen and its occurrence in several vegetable foods raises the question of the role of profilin in plants. First, its distribution throughout various parts of the plant needs to be determined. This paper describes observations on the presence of profilin in the tomato plant (Lycopersicon esculentum Mill.). The distribution of profilin in flower buds, stems, leaves, roots, and fruits of tomato was determined by immunoblotting and by tissue printing, showing that profilin is present in most if not all parts of the tomato plant.We gratefully acknowledge the help provided by Dr. A.T. Jagendorf and the donation of tomato seeds and maize pollen by N. Eanetta and Dr. M. Smith, respectively. The use of Dr. R. Wayne's SZH ILLD dissecting microscope is gratefully acknowledged. This work was aided by helpful discussions with C.S. Combs, Dr. C.A. Conley, and Dr. J. Andersland. This work was supported by a Hatch grant and NRI Competitive Grants Program/USDA 94-37304-1046 to MVP. This material is based upon work supported under a National Science Foundation Graduate Research Fellowship to DWD.     

Purification and characterization of a glucokinase from young tomato (Lycopersicon esculentum L. Mill.) fruit

In order to clearly establish the properties of the enzymes responsible for hexose phosphorylation we have undertaken the separation and characterization of these enzymes present in tomato fruit (Martinez-Barajas and Randall 1996). This report describes the partial purification and characterization of glucokinase (EC. 2.7.1.1) from young green tomato fruit. The procedure yielded a 360-fold enrichment of glucokinase. Tomato fruit glucokinase is a monomer with a molecular mass of 53 kDa. Glucokinase activity was optimal between pH 7.5 and 8.5, preferred ATP as the phosphate donor (K _m = 0.223 mM) and exhibited low activity with GTP or UTP. The tomato fruit glucokinase showed highest affinity for glucose (K _m = 65 μM). Activity observed with glucose was 4-fold greater than with mannose and 50-fold greater than with fructose. The tomato fruit glucokinase was sensitive to product inhibition by ADP (K _i = 36 μM). Little inhibition was observed with glucose 6-phosphate (up to 15 mM) at pH 8.0; however, at pH 7.0 glucokinase activity was inhibited 30–50% by physiological concentrations of glucose 6-phosphate. Received: 4 October 1997 / Accepted: 10 January 1998     

Isolation and characterization of nitrate reductase-deficient mutants in tomato (Lycopersicon esculentum Mill.).

Summary Five nitrate reductase-deficient mutants of tomato were isolated from an M2 population after ethyl-methanesulphonate (EMS) seed treatment by means of selection for chlorate resistance. All mutations were monogenic and recessive and complementation analysis revealed that they were non-allelic. Biochemical and molecular characterization of these mutants showed that four of them are cofactor mutants while one is an apoenzyme mutant.     

The histogenesis of somaclones from tomato (Lycopersicon esculentum Mill.) cotyledons

Summary A histological study ofin vitro cultured cotyledonary expiants of tomato (Lycopersicon esculentum) was performed in order to determine the site (differentiated tissue or developing callus) and the mode of plant regeneration.Results have shown that callus develops at the excision sites of cotyledonary expiants and that shoots are formed exclusively within the unorganized callus: excision areas are the only morphogenetic sites and the proximal excision is the preferred site for plant regeneration.Shoots differentiate by organogenesis within the superficial region of the callus. Few neocambial cells cooperate in the neoformation. Origin from a single cell is highly unlikely since rarely observed single activated cells never developed into shoots.Regenerated plants may be chimeras if invitro culture induces genetic diversity in the initial cells.Abbreviations IAA Indole-3-acetic acid - c callus - d vegetative dome - s shoot - ad adaxial - ab abaxial - t tracheid - p parenchyma - S sieve tube     

Role of extensin peroxidase in tomato (Lycopersicon esculentum Mill.) seedling growth

 It is proposed that inhibition of extensin peroxidase activity leads to a less rigid cell wall and thus promotes cell expansion and plant growth. A low-molecular-weight inhibitor derived from the cell walls of suspension-cultured tomato cells was found to completely inhibit extensin peroxidase-mediated extensin cross-linking in vitro at a concentration of 260 μg/ml. The inhibitor had no effect upon guaiacol oxidation catalyzed by extensin peroxidase or horseradish peroxidase. We have demonstrated that the light-irradiated inhibition of plant growth may be partially offset by inhibition of endogenous extensin peroxidase activity. Overall plant growth was enhanced by up to 15% in the presence of inhibitor relative to control plants. Inhibitor-treated and illuminated tomato hypocotyls grew up to 15% taller than untreated controls. The inhibitor had no effect upon etiolated plants over a 15-d period, suggesting that only low levels of peroxidase-mediated cross-linking can be found in the cell walls of etiolated plants. SDS-PAGE/Western blots of ionically bound protein from both etiolated and illuminated hypocotyls identified a doublet at 57/58.5 kDa which is immuno-reactive with antibodies raised to tomato extensin peroxidase. Levels of the 58.5-kDa protein, determined by SDS-PAGE, were at least threefold higher in illuminated tomato hypocotyls than in etiolated hypocotyls. Three fold higher levels of extensin peroxidase, elevated in-vitro extensin cross-linking activity and 15% higher levels of cross-linked, non-extractable extensin were observed in illuminated tomato hypocotyls compared with etiolated tomato hypocotyls. This suggests that white-light inhibition of tomato hypocotyl growth appears to be mediated, at least partially, by deposition of cell wall extensin, a process regulated by M_r-58,500 extensin peroxidase. Our results indicate that the contribution of peroxidase-mediated extensin deposition to plant cell wall architecture may have an important role in plant growth. Received: 22 July 1999 / Accepted: 11 October 1999     

Characterization of the subtilase gene family in tomato (Lycopersicon esculentum Mill.)

The gene family of subtilisin-like serine proteases (subtilases, SBTs) in tomato (Lycopersicon esculentum Mill.) comprises at least 15 members, 12 of which have been characterized in this study. Sequence comparison revealed that tomato subtilases fall into 5 distinct subfamilies. Single genes were shown to exist for LeSBT1, LeSBT2 and tmp, while 5 and 6 genes were found in the LeSBT3/4 and P69 subfamilies, respectively. With the exception of tmp, tomato subtilase genes were found to lack introns. Expression of subtilase genes was confirmed at the mRNA level by northern blot analysis and/or by primer extension experiments. For each of the 5 subtilase subfamilies, a distinctive pattern of expression was observed in tomato organs. At least one of the subtilases was found to be expressed in each organ analysed. Structural features evident from deduced amino acid sequences are discussed with reference to the related mammalian proprotein convertases.     

Diurnal control of the drought-inducible putative histone H1 gene in tomato (Lycopersicon esculentum Mill. L.)

The mRNA of genes le20, lcyP2, lhcll, and asr1 was quantified in leaves and roots of tomato plants (Lycopersicon esculentum Mill. L.) during three day/night cycles and 48 h constant illumination. lcyp2 and lhcll are known to exhibit diurnal or circadian rhythms in leaf tissue while asr1 has shown no evidence of diurnal fluctuations. Previously reported diurnal fluctuations of le20 mRNA in leaves could have been due to either changes in plant water status and abscisic acid concentration (le20 is a drought- and ABA-inducible gene) or changes in climate variables. Plants were grown hydroponically and at constant temperature (20.6 0.5C) and humidity (66 1%) such that no changes in plant water status or tissue ABA concentration were detectable. le20 and lcyP2 and lhcll mRNAs all fluctuated diurnally in leaf tissue and all reached a maximum during the light period. Surprisingly, le20 and lcyPs mRNA showed diurnal cycles in root tissue. There was no evidence for diurnal trends in asr1 mRNA, but levels increased steadily during constant light in both leaves and roots. le20, lcyP2 and lhcll mRNA showed only one cycle during 48h illumination and while carbon assimilation remained high and constant during this period, stomatal conductance decreased after 6 h light and then remained low. Photosystem II efficiency decreased during illumination, recovered during dark periods and showed a weak rhythm during constant light. It was concluded that le20 and lcyP2 have a diurnal component controlling their expression in leaves and roots, responding to light/dark cycles independently of water status or ABA concentration.Keywords: Tomato, histone H1, le20, lcyP2, diurnal trends.      

New cases of somatic conversion (paramutation) in tomato (Lycopersicon esculentum Mill.)

Summary The genetic behaviour of three chlorophyll variegated F₁ tomato plants, derived from irradiated gametophytes, was analyzed over several generations of selfing and backcrossing. The results suggest that irradiation has put genes, different in all three mutants, into a labile state, n*, remaining so after fertilization. This state had the power of converting the associated wild allele N into a deficient form.Somatic conversion was soon followed, in Plant C₁₁ always and nearly always in Plant C₁₂, by stabilization of both alleles in a conversion-inactive recessive state, genetically similar, and stable except in special conditions.In the other type, found seldom in C₁₂, always in C₆, the n* state was permanent and transmissible. Conversion occurred with a certain frequency determined by developmental and genotypic influences, and the converted allele also acquired conversion power, so that gametes from an N n* plant were of three kinds: N, n* and n*. This process corresponds to paramutation (Brink 1958).Results were compared and contrasted with other published data.Contribution nr. 748 of the Biology Division of Euratom.     

Characterization of a cDNA encoding 5-aminolevulinic acid dehydratase in tomato (Lycopersicon esculentum Mill.)

Summary A full-length cDNA clone encoding tomato (Lycopersicon esculentum Mill.) 5-aminolevulinic acid dehydratase (ALAD) was isolated and characterized. The primary structure predicts a 430-amino acid precursor which comprises a 41.7 kDa, 388-amino acid mature protein and a 47-amino acid transit sequence. The tomato primary sequence shows extensive homology to those of pea and spinach. Southern analysis indicated that 1 to 2 copies of the ALAD gene are present in the tomato genome. Northern blot analysis shows differential expression in various tomato organs, and constitutive developmental expression in tomato fruits.Abbreviations ALA 5-aminolevulinic acid - ALAD 5-aminolevulinic acid dehydratase (EC 4.2.1.24)     

Translocation of copper and other micronutrients in tomato plants (Lycopersicon esculentum Mill.): nicotianamine-stimulated copper transport in the xylem

The influence of the endogenous micronutrient chelator, nicotianamine(NA), and of Cu nutrition on the distribution of Cu, Fe, Mn,Zn, and NA was investigated in eight different shoot organs,roots, and in xylem exudates of the NA-containing tomato wildtype Lycopersicon esculentum Mill. cv. Bonner Beste and itsNA-less mutant chloronerva. Contrary to the other heavy metals, copper transport in thexylem was inefficient in the mutant and was enhanced by an applicationof NA to the roots or leaves in proportion to the applied NAconcentration. Also, with NA application, the Cu concentrationin mutant roots decreased significantly, and increased in theshoot. Fe and Mn transport in the xylem was greater in the mutantthan in the wild type, and was decreased in the mutant by theapplication of NA to the leaves. Zn transport in the xylem wasthe same in both genotypes and was unaffected by NA application.After application of NA to leaves and roots of the mutant itwas possible to detect NA in the xylem exudate (up to 2nmolNA(g^–1 root FWh^–1). High Cu supply (3 µM) resulted in higher Cu and Mn concentrationsin all organs of the wild type as compared to mutant organs,but Fe concentrations were not influenced. Under high Cu supply(3µM) the NA concentrations of roots and the three youngestleaves of the wild type were higher than under normal Cu supply(0.3 µM). The highest concentrations were found in theshoot apex under both Cu conditions (up to 361 nmol NAg^–1FW). It is concluded from our experiments and from the high stabilityconstant of the NA-Cu-complex (log K= 18.6) that NA is involvedin Cu translocation whereas for the translocation of Fe, Mn,and Zn, NA is not essential. Key words: Copper transport, micronutrients, mobilization, nicotianamine, xylem