QTL analysis of horticultural traits differentiating the cultivated tomato from the closely related species Lycopersicon pimpinellifolium

Molecular markers were used to map and characterize quantitative trait loci (QTLs) for several characters of agronomic and biological importance in an interspecific backcross of tomato. The parents of the cross were an elite processing inbred Lycopersicon esculentum cv M82-1-7 and the closely related red-fruited wild species L. pimpinellifolium (LA1589). A total of 257 BC₁ plants were grown under field conditions in Ithaca, New York and scored for 19 quantitative traits. A genetic linkage map was constructed for the same population using 115 RFLP, 3 RAPD and 2 morphological markers that spanned 1,279 cM of the tomato genome with an average interval length of 10.7 cM. A minimum of 54 putatively significant QTLs (P<0.001; LOD> 2.4) were detected for all characters with a range of 1–7 QTLs detected per character. Of the total 54 QTLs 11% had alleles with effects opposite to those predicted by the parental phenotypes. The percentage of phenotypic variation associated with single QTLs ranged from 4% to 47%. Multilocus analysis showed that the cumulative action of all QTLs detected for each trait accounted for 12–59% of the phenotypic variation. The difference in fruit weight was controlled largely by a single major QTL (fw2.2). Digenic epistasis was not evident. Several regions of the genome (including the region near sp on chromosome 6) showed effects on more than one trait. Implications for variety improvement and inferences about the domestication of the cultivated tomato are discussed.     

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.     

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.     

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     

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.     

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     

Purification and characterization of fructokinase from developing tomato (Lycopersicon esculentum Mill.) fruits

A procedure is described which allows the purification of fructokinase (EC 2.7.1.4) from young tomato fruit. The procedure yielded a 400-fold purification and two isoenzymes designated fructokinase I and II (FKI and FKII) were separated by anion-exchange chromatography. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) the molecular mass was estimated to be 35 kDa. Gel filtration on Sepharose-12 indicated that for both fructokinases the functional form is a dimer. Two dimensional isoelectric focusing/SDS-PAGE combined with immunoblotting showed that FKI has two components with isoelectric points (pIs) of 6.42 and 6.55, while four components with pIs from 6.07 to 6.55 were detected for FKII. A mixture of both fructokinases showed that the components of FKI match the more alkaline components of FKII. The activity of both fructokinases increased with increasing pH to around 8.0 and equal activity was observed from 8.0 to 9.5. Both fructokinases were specific for fructose with K _m values for fructose of 0.131 and 0.201 mM for FKI and FKII, respectively. At high concentrations (> 0.5 mM), fructose was also a strong inhibitor with inhibition constants (K _i) of 1.82 and 1.39 mM for FKI and FKII, respectively. The preferred phosphate donor for both isoforms was ATP, and K _m values of 0.11 and 0.15 mM were observed for FKI and FKII. At low concentrations (0.05–0.2 mM), fructose exhibited noncompetitive inhibition with respect to ATP for both fructokinases. This inhibition pattern changed to uncompetitive when higher fructose concentrations (0.5–10 mM) were used. These data indicated that substrate addition is ordered, with ATP adding first. Inhibition by ADP was also affected by the fructose concentrations. At 0.5 mM fructose, FKI showed non-competitive inhibition by ADP with respect to ATP and this inhibition changed to uncompetitive when 3 mM fructose was used. The isoform FKII showed a competitive inhibition pattern for ADP at 0.5 mM fructose which also changed to uncompetitive when 3 mM fructose was used. The features of the regulation of both fructokinases suggest that this enzyme might have a relevant role in carbon metabolism during tomato fruit development.     

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.     

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.     

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.     

Effect of salinity on tomato (Lycopersicon esculentum Mill.) during seed germination stage

A study was conducted using ten genetically diverse genotypes along with their 45F₁ (generated by diallel mating) under normal and salt stress conditions. Although, tomato (Lycopersicon esculentum Mill.) is moderately sensitive to salinity but more attention to salinity is yet to be required in the production of tomato. In present study, germination rate, speed of germination, dry weight ratio and Na⁺/K⁺ ratio in root and shoot, were the parameters assayed on three salinity levels; control, 1.0 % NaCl and 3.0 % NaCl with Hoagland’s solution. Increasing salt stress negatively affected growth and development of tomato. When salt concentration increased, germination of tomato seed was reduced and the time needed to complete germination lengthened, root/shoot dry weight ratio was higher and Na⁺ content increased but K⁺ content decreased. Among the varieties, Sel-7 followed by Arka Vikas and crosses involving them as a parent were found to be the more tolerant genotypes in the present study on the basis of studied parameters.     

Allotriploid somatic hybrids of diploid tomato (Lycopersicon esculentum Mill.) and monoploid potato (Solanum tuberosum L.)

Allotriploid somatic hybrids were obtained from fusions between protoplasts of diploid tomato and monohaploid potato. The selection of fusion products was carried out in two different ways: (1) The fusion of nitrate reductase-deficient tomato with potato gave rise only to hybrid calli if selection was performed on media lacking ammonium. Parental microcalli were rarely obtained and did not regenerate. (2) The fusion of cytoplasmic albino tomato with potato gave rise to albino and green hybrid calli and plants. Allotriploids were identified from the two somatic hybrid populations by counting chloroplast numbers in leaf guard cells and by flow cytometry of leaf tissue. Although some pollen fertility of allotriploids and pollen-tube growth of tomato, potato andLycopersicon pennellii into the allotriploid style were observed, no progeny could be obtained. The relevance of allotriploid somatic hybrids in facilitating limited gene transfer from potato to tomato is discussed.     

Effects of different nitrogen forms on the growth and cytokinin content in xylem sap of tomato (Lycopersicon esculentum Mill.) seedlings

In order to investigate the effects of homogeneous and localized supply of different nitrogen forms (nitrate, NO₃ ^? vs ammonium, NH₄ ⁺) on the growth of tomato seedlings, root morphology and six cytokinin (CTK) fractions in xylem sap were analyzed. Whole roots were supplied with different ratios of NO₃ ^? to NH₄ ⁺ (100:0, as 100-0NA; 75:25, as 75-25NA; 50:50, as 50-50NA) under homogeneous supply. In split-root experiments, three treatments were compared: a sole NO₃ ^? supply (N|N), a spatially separated supply of NO₃ ^? and NH₄ ⁺ (N|A), and a spatially separated supply of NO₃ ^? and a mixture of NO₃ ^? and NH₄ ⁺ nutrition at a ratio of 75:25 (N|AN). All concentrations of total N were set at 5 mM. The results showed that (1) homogeneous 75% NO₃ ^? plus 25% NH₄ ⁺ supply to the whole root zone led to maximum shoot and root dry matter (DM), root surface area (RS) and root volume (RV). The spatially separated supply of NO₃ ^? and NH₄ ⁺ (N|A) resulted in a contrasting effect on root morphology: in comparison to N|N, root DM in the NO₃ ^?-containing pot was increased by 50% whereas it was depressed by 50% in the NH₄ ⁺-containing pot. The 75% NO₃ ^? plus 25% NH₄ ⁺ supply in the split-root experiment led to no significant effects either on shoot DM and root DM, or on RS and RV when compared to N|N. (2) The presence of NH₄ ⁺ in the external medium led to a significantly reduced total xylem-CTK concentration, and a close negative correlation was found between xylem NH₄ ⁺ and total CTK concentration irrespective of culture mode. A relatively high level of zeatin riboside (ZR) was maintained both in 75-25NA and N|A treatments. It was concluded that, in addition to the percentage of NH₄ ⁺ to NO₃ ^? in the nutrient solution, whether NH₄ ⁺ was supplied to the whole root system or to only part of the root system was also an important factor affecting plant growth. The fact that the 75-25NA and N|A treatments resulted in optimal growth of tomato seedlings might be attributed to the higher ZR concentration in xylem.     

ACC-deaminase and/or nitrogen-fixing rhizobacteria and growth response of tomato (Lycopersicon pimpinellfolium Mill.)

The study aimed to identify and select important plant growth-promoting rhizobacteria (PGPR) and examine the response of tomato growth upon inoculation. Inoculation with rhizobacterial isolates increased all the measured physical, chemical, and enzymatic growth parameters compared to control. However, the TAN1 isolate had the highest effect, and significantly (P < 0.05) increased the root length (8.25-fold), root fresh (8.36-fold) and dry (12.6-fold) weight, shoot length (6.92-fold), shoot fresh (7.18-fold) and dry (6.90-fold) weight, number of leaves (11.0-fold), chlorophyll a (6.25-fold), chlorophyll b (10.7-fold), carotenoid contents (8.80-fold), seedlings fresh (9.0-fold) and dry (8.71-fold) weight, plant macronutrient uptake, i.e. N (7.7- and 8.9-fold), P (10.5- and 11.4-fold), K (7.8- and 8.8-fold), Ca (12.7- and 8.2-fold), and Mg (12.6- and 9-fold) in shoot and root, plant micronutrient uptake, i.e. Zn (6.6-, 10.2-), Cu (9.3-, and 10.3-fold), Fe (7.7- and 10.7-fold), and Mn (4.7- and 5.7-fold) in shoot and root and plant antioxidant enzymes, i.e. glutathione S-transferase (10.7-fold), peroxidase (8.1-fold), and catalase (10.5-fold). Our results concluded that inoculation of agricultural crops with rhizobacteria is a very useful approach to increase the plant growth. The rhizobacteria having both 1-aminocyclopropane-1-carboxylate (ACC) deaminase and nitrogen-fixing activity are more effective than rhizobacteria possessing either ACC-deaminase or nitrogen-fixing activity alone for growth promotion of crops.