Exposure to high temperatures (heat stress) causes reduced yield in tomatoes (Lycopersicon esculentum), mainly by affecting male gametophyte development. Two experiments were conducted where several tomato cultivars were grown under heat stress, in growth chambers (day/night temperatures of 31/25°C) or in greenhouses (day/night temperatures of 32/26°C), or under control (day/night temperatures of 28/22°C) conditions. In heat-sensitive cultivars, heat stress caused a reduction in the number of pollen grains, impaired their viability and germinability, caused reduced fruit set and markedly reduced the numbers of seeds per fruit. In the heat-tolerant cultivars, however, the number and quality of pollen grains, the number of fruits and the number of seeds per fruit were less affected by high temperatures. In all the heat-sensitive cultivars, the heat-stress conditions caused a marked reduction in starch concentration in the developing pollen grains at 3 days before anthesis, and a parallel decrease in the total soluble sugar concentration in the mature pollen, whereas in the four heat-tolerant cultivars tested, starch accumulation at 3 days before anthesis and soluble sugar concentration at anthesis were not affected by heat stress. These results indicate that the carbohydrate content of developing and mature tomato pollen grains may be an important factor in determining pollen quality, and suggest that heat-tolerant cultivars have a mechanism for maintaining the appropriate carbohydrate content under heat stress.

The traditional use of polyols as osmotica in plant culture media is based on the assumption that polyols are not taken up or metabolized by cells. In reality, polyols are significant photosynthetic products and efficiently utilized metabolites in a large number of plants. In addition to these metabolic roles, initial interest in polyols focused primarily on their function as osmoprotectants. This was hypothesized to be due to their ability to act as compatible solutes. More recent research, however, indicates much broader roles for polyols in stress responses based on their significant antioxidant capacity. These include protection against salt and photooxidative stress as well as a potential role in plant pathogen interactions.

Continuous exposure of tomato 'Trust' to high temperatures (day/night temperatures of 32/26 degrees C) markedly reduced the number of pollen grains per flower and decreased viability. The effect of heat stress on pollen viability was associated with alterations in carbohydrate metabolism in various parts of the anther during its development. Under control, favourable temperature conditions (28/22 degrees C), starch accumulated in the pollen grains, where it reached a maximum value 3 d before anthesis; it then diminished towards anthesis. During anther development, the concentration of total soluble sugars gradually increased in the anther walls and in the pollen grains (but not in the locular fluid), reaching a maximum at anthesis. Continuous exposure of the plants to high temperatures (32/26 degrees C) prevented the transient increase in starch concentration and led to decreases in the concentrations of soluble sugars in the anther walls and the pollen grains. In the locular fluid, however, a higher soluble sugar concentration was detected under the high-temperature regime throughout anther development. These results suggest that a major effect of heat stress on pollen development is a decrease in starch concentration 3 d before anthesis, which results in a decreased sugar concentration in the mature pollen grains. These events possibly contribute to the decreased pollen viability in tomato.

The attitude of most conventional plant breeders towards molecular biology ranges from contempt to modest interest. This chapter illustrates, using as an example a plant breeding program on which the authors have been working for the last 7 yr, some general issues in breeding programs that molecular biology needs to address. The example is eventually used to suggest how to bridge the gap between conventional plant breeding and molecular biology. The crop that is used as an example is barley (Hordeum vulgare), a typical crop of marginal, low input, stress environments The chapter also examines a hypothesis of integration between plant breeding and some of the simplest techniques of molecular biology such as the use of molecular markers (RFLP, DNA fingerprinting, RAPDs). If integration proves to be difficult for these techniques, the integration in more advanced areas (transformation, gene isolation, gene transfer, etc.) continues to be a mirage.

Foliar raffinose and sucrose concentrations in eastern white pine (Pinus strobus L.), eastern redcedar (Juniperus virginiana L.), Leyland cypress (×Cupressocyparis leylandii Dallim.), and Virginia pine (Pinus virginiana L.) were measured monthly over 2 years. During cold weather, foliage of white pine and redcedar contained higher concentrations of raffinose and sucrose than did Leyland cypress and Virginia pine. Rafflnose concentrations were highest during winter and were best correlated with the frequency of occurrence of daily minima ≤ 1.7C during the 30 days before sampling. Sucrose concentrations, which also reached maximum levels during the winter, were best correlated with the frequency of occurrence of daily minima ≤ 7.2C in the prior 30 days. Sucrose concentrations were relatively high during fall and spring. Raffinose and sucrose concentrations increased in response to recurring low temperature, with correlations highest for raffinose.

A mannitol:mannose 1-oxidoreductase was isolated from celeriac (Apium graveolens var. rapaceum) root tips by fractionation with (NH4)2SO4, followed by chromatography on a Fractogel DEAE column and then concentration with (NH4)2SO4. This newly discovered mannitol dehydrogenase catalyzes the NAD-dependent oxidation of mannitol to mannose, not mannitol to fructose. The sugar product of the enzyme reaction was identified by three independent HPLC systems and by an enzymatically linked system as being mannose and not fructose or glucose. Normal Michaelis--Menten kinetics were exhibited for both mannitol and NAD with Km values of 72 and 0.26 mM, respectively, at pH 9.0. The Vmax was 40.14 mumol/h/mg protein for mannitol synthesis and 0.8 mumol/h/mg protein for mannose synthesis at pH 9.0. In the polyol oxidizing reaction, the enzyme was very specific for mannitol with a low rate of oxidation of sorbitol. In the reverse reaction, the enzyme was specific for mannose. The enzyme was strongly inhibited by NADH and sensitive to alterations of NAD/NADH ratio. The enzyme is of physiological importance in that it is mainly localized in root tips (sink tissue) where it functions to convert mannitol into hexoses which are utilized to support root growth. Product determination and kinetic characterization were carried out on an enzyme preparation with a specific activity (SA) of 30.44 mumol/h/mg protein. Subsequently, the enzyme was further purified to a SA of 201 mumol/h/mg protein using an NAD affinity column. This paper apparently represents the first evidence of the existence of a mannitol:mannose 1-oxidoreductase and also the first evidence of the presence of a mannitol dehydrogenase in vascular plants.

Flooding the roots of greenhouse-grown muskmelon (Cucumis melo L. cv. Noy Yizreel) plants for 4 days reduced sucrose accumulation 36% in the inner mesocarp and 88% in the outer mesocarp of developing fruit. Concentration of the translocated sugars raffinose and stachyose were also lower in fruit on flooded plants than in those from nonflooded plants. In contrast, fruit hexose concentration was similar in both flooded and nonflooded plants. There was no alteration in activities of enzymes associated with sucrose metabolism in the fruit which could explain the decreased sucrose concentration. Four days of root flooding caused no reduction in leaf carbon exchange rate or assimilate export rate, indicating that the reduction in fruit sucrose accumulation was not due to source limitation. Root respiration, measured as CO2 evolution, was approximately 30% lower in anaerobic roots than in aerobic roots. When viewed as carbohydrate consumed, a doubling of glycolytic activity occurred in the anaerobic root mass. Increased demand for carbohydrates by anaerobic roots may lead to a reduction in translocated carbohydrates available for sucrose biosynthesis in the developing fruit.

Two cultivars of greenhouse tomato (Lycopersicon esculentum Mill.) were grown with ambient or 1000 μl CO 2 /liter during Jan.-June 1987 and 1988. In both years, CO 2 -enrichment increased foliar deformation and foliar starch, but during the season, foliar starch levels decreased while deformation increased. `Laura' had more deformation, while `Michigan-Ohio' had higher foliar starch concentration. During an entire season, there was no significant relationship between foliar starch concentration and deformation severity. Foliar C exchange rates in the lower canopy were not affected by severity of deformation. Data from these experiments do not support the hypothesis that excess foliar starch is responsible for foliar deformation at elevated CO 2 .

Yield increases observed among eight genotypes of tomato (Lycopersicon esculentum Mill.) grown at ambient CO2 (about 350) or 1000 microliters per liter CO2 were not due to carbon exchange rate increases. Yield varied among genotypes while carbon exchange rate did not. Yield increases were due to a change in partitioning from root to fruit. Tomatoes grown with CO2 enrichment exhibited nonepinastic foliar deformation similar to nutrient deficiency symptoms. Foliar deformation varied among genotypes, increased throughout the season, and became most severe at elevated CO2. Foliar deformation was positively related to fruit yield. Foliage from the lower canopy was sampled throughout the growing season and analysed for starch, K, P, Ca, Mg, Fe, and Mn concentrations. Foliar K and Mn concentrations were the only elements correlated with deformation severity. Foliar K decreased while deformation increased. In another study, foliage of half the plants of one genotype received foliar applications of 7 millimolar KH2PO4. Untreated foliage showed significantly greater deformation than treated foliage. Reduced foliar K concentration may cause CO2-enhanced foliar deformation. Reduced K may occur following decreased nutrient uptake resulting from reduced root mass due to the change in partitioning from root to fruit.

Muskmelon (Cucumis melo L.) fruit lack a stored starch reserve and therefore depend on translocated photoassimilate from the leaf canopy for sugar accumulation during ripening. The influence of canopy photosynthesis on sucrose' accumulation within muskmelon fruit mesocarp was examined. Canopy photosynthetic activities were estimated in a sweet and a nonsweet genotype. Photosynthetic rate of the nonsweet genotype, on a per-plant basis, was only 56% of that of the sweet genotype. The effect of limiting leaf area of the sweet genotype on carbohydrate concentrations and sucrose metabolizing enzymes within the fruit was evaluated. A 50% reduction of leaf area 8 days before initiation of fruit sucrose accumulation resulted in canopy photosynthesis similar to that of the nonsweet genotype. Reduced photosynthetic activity resulted in slightly lower soluble-carbohydrate concentration in the fruit; however, fruit sucrose concentration was three times higher than that reported previously for the nonsweet genotype. The extent to which `fruit sucrose phosphate synthase (SPS) activity increased during maturation was diminished by leaf removal. Acid invertase activity declined in all fruit in a similar manner irrespective of defoliation. A reduction of leaf area of a sweet genotype reduced sucrose accumulation within the fruit. Lower fruit sucrose concentration was associated with lower concentration of raffinose saccharides and lower SPS activity within the fruit. Additionally, insufficient assimilate supply was judged not to be the factor responsible for low sucrose accumulation in a nonsweet genotype.

ABSTRACTRates and routes of acetic acid penetration into nonheated cherry peppers (Capsicum annuum) were determined by monitoring pH over time at ten interior areas. The pH was determined for fresh red and green peppers, brined peppers, and brined peppers that had been exposed to oxygen prior to brining. The pH measurements, scanning electron microscopy, and dye penetration observations indicated that the primary avenue for acid penetration was through the stem into the placenta. The last area to become acidified (pH < 4.6) was the interior of the fruit wall. Peppers that had been exposed to oxygen were more rapidly acidified (24 hr) as compared to those not exposed to oxygen (150 hr).

The effect of anaerobiosis, imposed during germination of ‘Calypso’ cucumber (Cucumis sativas L.) seeds, was studied. Anaerobic conditions inhibited reserve mobilization from the cotyledons and dry weight gain by the embryonic axis. Within the embryonic axis, lipid degradation was stopped and use of all readily metabolizable carbohydrate reserves was strongly stimulated. By 48 hr of exposure to an anaerobic environment, the axis was nearly depleted of endogenous carbohydrate reserves. Aerobically germinating seeds accumulated a massive concentration of hexose sugars within the axis during the same time period. Thus, growth inhibition within cucumber seeds during anaerobiosis may result in part from carbohydrate deprivation of the embryonic axis.

Abstract The influence of short-term cooling of cucumber ( Cucumis sativus L. ‘Calypso’) roots on leaf expansion and carbohydrate metabolism was studied. Leaf expansion of greenhouse-grown cucumber plants irrigated twice daily with chilled water (5°C) was reduced by about 15% relative to that of plants irrigated with water at ambient greenhouse air temperature, even though the soil was never below 15°C. Leaf carbon exchange rate was not affected by rooting medium temperature. However, photoassimilate export rate was reduced from leaves of plants with cooled roots and starch accumulation in leaves of these plants was increased relative to that in control plants. Galactinol concentrations were significantly higher in leaves of plants that had reduced photoassimilate export rates.

The effects of varied rooting volumes on root growth and source leaf carbohydrate metabolism were studied in greenhouse-grown cucumber (Cucumis sativus L cv Calypso) plants. Plants were grown for 7 weeks in container volumes that ranged from 0.4 to 5.9 liters. Plants grown in the smaller containers exhibited less leaf expansion, lower root and shoot weight, and fewer lateral stems than plants grown in the 5.9 liter containers. Shoot/root ratio was not altered by the container volume, suggesting coordination of root and shoot growth due to rooting volume. Source leaf carbon exchange rates, assimilate export rates, and starch accumulation rates for plants grown in 0.4 liter containers were approximately one-half or less in comparison to those for plants grown in 5.9 liter containers. Starch concentrations per unit leaf area were maintained at high levels in source leaves of plants grown in 0.4 liter containers over the entire day/night cycle. Lower extractable galactinol synthase activities and higher galactinol concentrations occurred in leaves of plants grown in 0.4 liter container volumes. The reduced sink demand, induced by restricted root growth, may have led to increased starch concentrations and to a reduction in stachyose biosynthesis in cucumber source leaves.

The effects of photosynthetic periods and light intensity on cucumber (Cucumis sativus L.) carbon exchange rates and photoassimilate partitioning were determined in relation to the activities of galactinol synthase and sucrose-phosphate synthase. Carbon assimilation and partitioning appeared to be controlled by different mechanisms. Carbon exchange rates were influenced by total photon flux density, but were nearly constant over the entire photoperiod for given photoperiod lengths. Length of the photosynthetic periods did influence photoassimilate partitioning. Assimilate export rate was decreased by more than 60% during the latter part of the short photoperiod treatment. This decrease in export rate was associated with a sharp increase in leaf starch acccumulation rate. Results were consistent with the hypothesis that starch accumulation occurs at the expense of export under short photoperiods. Galactinol synthase activities did not appear to influence the partitioning of photoassimilates between starch and transport carbohydrates. Sucrose phosphate synthase activities correlated highly with sugar formation rates (sucrose, raffinose, stachyose + assimilate export rate, r = 0.93, alpha = 0.007). Cucumber leaf sucrose phosphate synthase fluctuated diurnally in a similar pattern to that observed in vegetative soybean plants.

Abstract Length and width measurements of leaf lamina from six fresh-market and nine pickling cucumber (Cucumis sativus L.) cultivars were used in the development of leaf area prediction models. Models using the single independent variable of length (L) were selected from regression analysis using length (L), width (W), and measured leaf areas (A) of 188 ‘Calypso’ cucumber leaves. Tests for equality of regressions (α= 0.05) indicated that a single model accurately predicted leaf area for greenhouse-grown ‘Calypso’ cucumbers (grown in a soil medium) as well as 15 field-grown cultivars. A separate prediction model was required for ‘Calypso’ cucumber plants grown in hydroponic sand culture. Inclusion of leaf width in a multiple regression model provided accurate leaf area prediction for all environments, culture systems, and cultivars tested.

Seasonal changes in soluble carbohydrates of Fraser fir [Abies fraseri (Pursh) Poir.] needles were monitored in Fall 1984, Spring 1985, and Fall 1985 through Spring 1986. Raffinose concentration increased in the fall and decreased in the spring. There was a 23-fold increase in raffinose concentration from Aug. 1985 to Jan. 1986. Sucrose concentration varied from fall to spring with the lowest concentration occurring in February. Postharvest needle abscission from harvested branches held 6 weeks without water was inversely correlated with raffinose concentration at the time of harvest. Diurnal fluctuations in soluble carbohydrates were monitored on 12 July and 26 Oct. 1985. Raffinose concentration fluctuated slightly on both dates with a decrease during the dark period. On 12 July, sucrose increased during the day and decreased at night, whereas hexoses decreased in the day and increased at night. No significant diurnal changes in sucrose or hexose were evident on 26 Oct. Controlled-environment studies at 24° (day)/18°C (night), 18°/12°, and 12°/6° showed that most of the raffinose accumulation was due to low temperature; the remainder to short days. Postharvest needle loss was lowest in plants with high needle raffinose concentrations resulting from the 12°/6° temperature. Storage without water resulted in significant postharvest needle loss for shoots from plants preconditioned with 24°/18° and 18°/12°, but not for those exposed to 12°/76°. Compared to long days, plants preconditioned with short days lost fewer needles following harvest.

Galactinol synthase (UDP-galactose:inositol galactosyltransferase) is the first unique enzyme in the biosynthetic pathway of raffinose saccharides. Its role as a regulator of carbon partitioning between sucrose and raffinose saccharides in developing soybean (Glycine max L. Merrill) seeds was examined. Galactinol synthase activity and concentrations of sucrose, stachyose, and raffinose were compared during seed development between two genotypes that were high and two genotypes that were low in mature seed raffinose saccharide concentration. In all genotypes, sucrose concentration increased as seed development progressed, but in both low raffinose saccharide genotypes, greater increases in sucrose concentration were observed late in seed development. Sucrose to stachyose ratios in mature seeds were 2.3-fold greater in low raffinose saccharide genotypes than in the high raffinose saccharide genotypes. During seed development, higher levels of galactinol synthase activity were observed in the high raffinose saccharide genotypes than in the low raffinose saccharide genotypes. A common linear relationship for all four soybean genotypes was shown to exist between galactinol formed estimated from galactinol synthase activity data and the concentration of galactose present in raffinose saccharides. Results of this study implied that galactinol synthase is an important regulator of carbon partitioning between sucrose and raffinose saccharides in developing soybean seeds.