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T. Casey Garvey and John D. Hewitt

A study using Lycopersicon cheesmanii Riley LA 1449 (typicum), a low soluble solids content (SSC) accession, and L. cheesmanii f. minor LA 528 (minor), a high SSC accession, was undertaken to characterize the accumulation of starch, sugar, and total SSC. Fruit of each accession was sampled throughout development to identify differences in SSC, starch accumulation, and sugar distribution. Osmetric analysis indicated that the minor race had higher SSC content throughout the ontogeny of fruit development than the typicum. Typicum contained more starch than minor, and both accessions showed a rapid decline in percent starch as the fruit ripened. Sucrose remained low throughout all stages of fruit development for both accessions. Glucose increased in the minor and declined in the typicum. Fructose increased in both accessions. Total reducing sugar content at the full ripe stage was higher in minor than the typicum.

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Takashi Nishizawa, Sayuri Nagasawa, Yuko Mori, Yuko Kondo, Yuka Sasaki, Jorge B. Retamales and Arturo Lavin

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Kenji Kobashi, Hiroshi Gemma and Shuichi Iwahori

A water stress treatment was imposed on peach trees [Prunus persica (L.) Batsch `Kansuke Hakuto' (Peach Group)] to elucidate the relationship among sugar accumulation, sugar metabolism, and abscisic acid (ABA) in fruit under water stress. Treatment was carried out on peach trees grown in containers from 8 July 1996 [80 days after full bloom (DAFB)] for 16 days, to achieve a predawn water potential of -0.8 to -1.1 MPa compared to that of -0.4 to -0.6 MPa in control trees. Levels of sorbitol, sucrose, and total sugars, as well as the activity of sorbitol oxidase increased in fruit of water stressed trees under moderate water stress (-0.8 MPa), whereas under severe water stress (-1.1 MPa), no difference between the waterstressed trees and the controls was observed. Water stress also induced an increase in ABA in the fruit. These initial results indicated that water-stress-induced ABA accelerated sugar accumulation in peaches by activating sorbitol metabolism.

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T.M. Mills, M.H. Behboudian, P.Y. Tan and B.E. Clothier

Five-year-old `Braeburn' apple trees (Malus domestica Borkh.) on MM.106 rootstock were studied to determine plant and fruit quality responses to reduced plant water status late in the season. Trees were irrigated or not irrigated. Those not irrigated developed reduced xylem water potential and stomatal conductance from 110 and 132 days after full bloom (DAFB), respectively. However, they showed no reduction in photosynthetic rates. Fruit were harvested at stage 1 (S1), starting 167 DAFB, or stage 2 (S2), starting 180 DAFB. At S1, fruit had higher soluble solids concentrations, enhanced red skin pigmentation, and a tendency for higher sorbitol concentrations. Total soluble sugar concentrations at final harvest showed no difference between treatments, but fruit from the nonirrigated trees showed earlier sugar accumulation during the season. Such fruit also had reduced Ca+2 concentrations at S1 and S2 relative to those on plants that were irrigated. No incidence of any disorder was found in fruit from either treatment at harvest or after 12 weeks of 0C storage.

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Motoko Iida, Nancy A. Bantog, Kunio Yamada, Katsuhiro Shiratake and Shohei Yamaki

The regulation of NAD+-dependent sorbitol dehydrogenase (NAD-SDH, EC 1.1.1.14) by sugar was investigated by using sliced tissues of japanese pear (Pyrus serotina Nakai cv. Kousui) fruit in order to determine its role in the mechanism of sugar accumulation in fruit tissue. The results of the activities and steady-state levels of the protein and mRNA indicate that NAD-SDH in japanese pear fruit is among the sugar-inducible genes. By preincubating the sliced tissues for 16 hours in a medium without sugar, NAD-SDH activity declined and reached a stable level that was maintained for up to 40 hours. The washing procedure also reduced the sugar concentration in the apoplast and cytosol of the sliced tissues to low concentrations and enabled them to be manipulated by exogenous applications of carbohydrate solutions. Incubation of tissues in 50 or 100 mm sorbitol for 8 hours led to enhanced expression of the NAD-SDH gene as determined by increased mRNA and protein levels and enhanced enzyme activity. The presence of 100 mm glucose, sucrose, or mannitol also gave significant stimulation on the levels of activity, protein, and mRNA of NAD-SDH compared with those of control tissues bathed in media in which the osmotic potential had been adjusted to that of the sugar solutions by adding polyethylene glycol. However, fructose was ineffective in stimulating NAD-SDH activities and the level of the protein was not enhanced but the level of mRNA was increased. Therefore, it is suggested that NAD-SDH gene transcription is enhanced by each sugar investigated, and fructose appears to be unique as it also influences NAD-SDH at a post-transcriptional level.

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John R. Stommel and Kathleen G. Haynes

Fruit of the cultivated tomato (Lycopersicon esculentum Mill.) store predominantly glucose and fructose whereas fruit of the wild species L. hirsutum Humb. & Bonpl. characteristically accumulate sucrose. Reducing sugar and sucrose concentrations were measured in mature fruit of parental, F1, F2, and backcross (BC1) populations derived from an initial cross of L. esculentum `Floradade' × L. hirsutum PI 390514. Generational means analysis demonstrated that additive effects were equal to dominance effects for percentage of reducing sugar. It was determined that a single major gene, dominant for a high percentage of reducing sugar, regulates the percentage of reducing sugar in tomatoes. We propose that this gene be designated sucr. Only additive effects were demonstrated to be important for glucose: fructose ratios. Using L. hirsutum as a donor parent for increasing total soluble solids concentration in the cultivated tomato is discussed.

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Zhifang Gao, Sastry Jayanty, Randolph Beaudry and Wayne Loescher

In apple (Malus ×domestica Borkh.), where sorbitol is a primary photosynthetic product that is translocated throughout the plant, accumulation of sorbitol in sink cells appears to require an active carrier-mediated membrane transport step. Recent progress in isolation and characterization of genes for sorbitol transporters in sour cherry (Prunus cerasus L.) and mannitol transporters in celery (Apium graveolens L.) suggested that similar transporters may be present in apple tissues. A defect in these transporters could also explain the occurrence of the fruit disorder watercore, characterized by the accumulation of fluids and sorbitol in the apoplasmic free space. Our objectives therefore included isolation and characterization of genes for sorbitol transporters in apple tissues and comparisons of expression of transporter genes, especially in various sink tissues including watercored and non-watercored fruit tissues. We have isolated and characterized two sorbitol transporter genes, MdSOT1 and MdSOT2. Sequence analyses indicated that these are members of the major facilitator transporter superfamily that gives rise to highly hydrophobic integral membrane proteins. Heterologous expression and measurement of sorbitol uptake in yeast indicated that these are specific and with high affinities for sorbitol, with Kms for sorbitol of 1.0 and 7.8 mm for MdSOT1 and MdSOT2, respectively. Sorbitol transporter expression was evident in all sink tissues tested with the exception of watercore-affected fruit tissues. Sorbitol accumulation in apple sink tissues thus involves an apoplasmic active membrane transport step and watercore results from a defect in that process.

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Md. Shahidul Islam, S. Khan and T. Matsui

Seasonal fluctuations of carbohydrate levels and compositions and the activities of related enzymes of three cultivated tomato (Lycopersicon esculentum Mill. cv. Lady First, Momotaro, and Minicarol) cultivars were examined at 45-days interval with seven different sowing in the relatively warm climate of Japan. Fruits picked on early winter to spring seasons had higher sugar concentrations compared to hot season. Fructose and glucose in nearly equal amounts were the predominant sugar in all the seasons. Sucrose was present in trace quantities, but cherry cultivar Minicarol accumulated higher levels than the other two large-fruited types.

Acid invertase (EC 3.2.1.26) was highest at red stage during December to April, while fruit matured during May to August had lowest activity. The activity levels of soluble invertase were predominant compared to cell wall-bound fraction. The sucrose synthase (EC 2.4.1.13) showed highest activity in rapidly growing fruits followed by a very low activity with fruit maturation. Sucrose synthase showed the higher activity during November to February, and almost low activity during all the experimental periods. The sucrose phosphate synthase (EC 2.4.1.14) also showed higher activity during October to February, but the activity levels did not change drastically throughout the fruit development. The results substantiate the conclusion that, in all the planting seasons, acid invertase is a principal enzyme in the process of tomato fruit ripening and during early stage of tomato fruit development, sucrose synthase is the dominant enzyme, which, in turn, plays a part in regulating the translocation of sucrose into the fruit.

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Mohammed Dafiri and André Gosselin

Tomato plants were submitted to three photosynthetic photon fluxes (PPF) of 50, 100, and 150 μmol. m-2s-1 and cluster-pruned according to different scenarios. The highest PPF combined with severe cluster pruning produced the highest yield and the best fruit quality. The highest PPF increased growth, photosynthesis and leaf sugar content. Severe cluster pruning increased the average fruit weight, leaf sucrose and glucose content, but reduced photosynthesis. Data will be discussed in relation to crop management and efficiency.

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Azusa Sato, Hiroshi Okubo and Kazuyuki Saitou

The aim of this study was to investigate physiological and biochemical mechanisms of shoot elongation after cold period in hyacinth (Hyacinthus orientalis L. cv. Delft Blue). Hyacinth shoot rapidly elongated during hydro-culture period in cooled bulbs, but not in non-cooled bulbs. Alpha-amylase (EC 3.2.1.1.) is a key enzyme involved in starch hydrolysis. Alpha-amylase activity increased during the cold storage period and was low during rapid shoot elongation in hyacinth. In the non-cooled bulbs, its activity remained at the similar level. Sucrose content increased during the cold storage period in the shoot, but not in the scales. We, for the first time, isolated cDNA for cold-responsive alpha-amylase gene (HoAmy1A, accession No. AB198975) from hyacinth, and presented that HoAmy1A expression increased in the scale during the cold storage period, but the level was very low during shoot elongation. We also found that promoter region of HoAmy1A contained CArG element, which is related to the response to low temperature. In tulip (Tulipa genesriana L.), the most studied bulbous plant, dramatic increase in alpha-amylase activity and translocation of sugars from the scales to shoot occurred during the growth stage, following cold treatment (Komiyama et al., 1997; Lambrechts et al., 1994). Our results suggest that there are two types (tulip and hyacinth types) of sprouting mechanisms in bulbous plants.