High temperature effects on RuBP carboxylase and carbonic anhydrase activity in two tomato cultivars

The effects of temperature on ribulose bisphosphate carboxylase activity were studied in two tomato ( Lycopersicon esculentum Mill.) cultivars which differed in sensitivity to high temperatures. The heat tolerant cultivar, Saladette, had a smaller reduction in photosynthesis and a smaller increase in mesophyll resistance then the sensitive cultivar Roma VF, after 24 h at 35 to 40°C. One hour in vitro treatments at 50°C decreased the activity of ribulose bisphosphate carboxylase extracted from Roma VF by 75%, while Saladette was not affected. Heat stress to the entire plant caused greater inhibition of ribulose bisphosphate carboxylase in the heat sensitive cultivar. Ribulose bisphosphate carboxylase activity in both cultivars decreased with heat treatment but recovered under normal temperatures. Ribulose bisphosphate oxygenase activity decreased similarly in both cultivars under 37/18°C day/night temperatures, which resulted in an apparent change in the relative carboxylase/oxygenase activity of the two cultivars. Carbonic anhydrase activity was slightly greater in Saladette than in Roma VF but no significant decrease in activity was observed in plants exposed to high temperatures.     

Separately and simultaneously induced dark chilling and drought stress effects on photosynthesis, proline accumulation and antioxidant metabolism in soybean

The effects of separately or simultaneously induced dark chilling and drought stress were evaluated in two Glycine max (L.) Merrill cultivars. For the separately induced dark chilling treatment (C), plants were incubated at 8 °C during 9 consecutive dark periods. During the days, plants were kept at normal growth temperatures. For the separately induced drought treatment (D), plants were maintained at normal growth temperatures without irrigation. For the simultaneously induced dark chilling and drought stress treatment (CD), plants were dark chilled without irrigation. All treatments caused similar decreases in pre-dawn leaf water potential, but resulted in distinct physiological and biochemical effects on photosynthesis. In Maple Arrow, where C had the smallest effect on photosynthesis, prolonged CD caused less inhibition of photosynthesis compared to D. Compared to Fiskeby V, the photosynthetic apparatus of Maple Arrow appears to possess superior dark chilling tolerance, a property which probably also conveyed enhanced protection against CD. Proline accumulation was prevented by CD at the ψ_PD where D already resulted in considerable accumulation. The superior capacity for proline accumulation in Maple Arrow would seem to be an important factor in its stress tolerance. Antioxidant activity evoked by CD and D was higher than for C alone. In Fiskeby V, the small increase in ascorbate peroxidase (EC 1.11.1.7) activity, which was in most cases not accompanied by increased gluthatione reductase (EC 1.6.4.2) activity, could impact negatively on its stress tolerance. These results demonstrate large genotypic differences in response to chilling and drought stress, even between soybean cultivars regarded as chilling tolerant.     

Cold-acclimation induced changes in freezing tolerance and translatable RNA content in Citrus grandis and Poncirus trifoliata

Cold-acclimation-induced changes in freezing tolerance and translatable RNA content were compared in seedlings of a relatively cold sensitive citrus species, Citrus grandis L. Osb. cv. Thong Dee (pummelo), and the cold-hardy citrus relative, Poncirus trifoliata L. Raf. cv. Pomeroy (trifoliate orange). Cold acclimation of pummelo (10 days at 15°C followed by 4 weeks at 10°/5°C, day/night) resulted in a decrease in LT₅₀ from −6 to −8°C, while in trifoliate orange (acclimated for 7 weeks at 5°C), the LT₅₀ decreased from −9 to −18°C. Qualitative changes in the in vitro translation profile, revealed by two-dimensional gel electrophoresis, were observed following cold acclimation in both species. An mRNA for a large polypeptide (ca 160 kDa) was detected following cold acclimation of trifoliate orange. A similar change was not observed in pummelo following cold acclimation.     

A comparison of the chilling-stress response in two differentially tolerant cultivars of tomato (Lycopersicon esculentum).

The chilling responses of two differentially cold tolerant cultivars of tomato were monitored through in vivo labelling of polypeptides with [35S]methionine, both during a gradual temperature decrease (2 degrees C/day) and also during a rapid cold shock (4 degrees C). The polypeptides were separated by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and revealed by fluorography. Both cultivars showed changes in the polypeptide profiles resulting from either chilling treatment. During the gradual temperature decrease, there were few differences exhibited between the two cultivars. However, during cold shock both cultivars showed the altered synthesis of several unique polypeptides. Both cultivars showed the appearance of a 35-kDa polypeptide during the gradual temperature decrease and also during the cold shock. The appearance of three high relative mass polypeptides was found in both cultivars only during the gradual temperature decrease. Treatments with cycloheximide and chloramphenicol suggested that cold-shock polypeptides are both nuclear and organelle encoded. The cold-shock response in roots was different from the response in leaves and between cultivars. A comparison of the two cultivars showed a number of differences in polypeptide synthesis which may be related to increased cold tolerance.     

Detection of Two Membrane Polypeptides Induced by Abscisic Acid and Cold Acclimation: Possible Role in Freezing Tolerance

Membrane proteins labeled in vivo from cold-acclimated and ABA-treatedalfalfa seedlings of two cultivars differing in cold-tolerancehave been compared by SDS polyacrylamide gel electrophoresisand fluorography. Results thus obtained indicate that severalqualitative changes occur in the membrane protein-profile specificallyin response to cold acclimation or ABA treatment. While somepolypeptides disappear from the non-acclimated protein patterns,others specifically appear in response to acclimation. Separationby two-dimensional gel electrophoresis and fluorography hasconfirmed the above and has enabled us to detect two proteinsof Mr 42 kDa and 120 kDa that are induced by both acclimationand ABA treatment in the freezing tolerant cultivar. (Received November 30, 1987; Accepted February 22, 1988)     

Soybean (Glycine max) modulation and N2-fixation as affected by exposure to a low root-zone temperature

Low root-zone temperatures (RZTs) are known to reduce soybean N₂-fixation. However, the relative sensitivity of the various stages of symbiosis establishment and function (N₂-fixation) to suboptimal RZTs is unresolved. We conducted experiments to examine the effect of exposure to a RZT of 15°C on nodulation. The control RZT was 25°C. Root temperatures were controlled by circulating cooled water around pots on a growth bench. Soybean seedlings [ Glycine max (L.) Merr. cv. Maple Arrow] were inoculated with 1 ml of a log-phase culture (approximately 10⁻⁸ cells) of Bradyhizobium japonicum strain 532C. They were then (1) maintained continuously at RZTs of 15 or 25°C, transferred to 15 or 25°C from the alternate temperature 7 days after inoculation (DAI), or transferred to 15 or 25°C at 14 DAI, and (2) maintained at 15 or 25°C, or transferred at either 1, 4 or 7 DAI. When seedlings were maintained at a RZT of 25°C nodule primordia (<1 mm) were visible at 7 DAI and N₂-fixation commenced at 14 DAI. Nodule function (N₂-fixation) appeared to be relatively insensitive to low RZTs since exposure of plants to 15°C following the onset of N₂-fixation (14 DAI) resulted in 68% of the N fixed and 78% of the dry weight of the 25°C RZT, although N partitioning to shoot tissues was reduced. In contrast, exposure to the low RZT shortly after inoculation declayed the onset of N₂-fixation for 4 to 6 weeks, primarily by inhibiting the early stages of nodulation. This resulted in fixed N and dry weight levels of 9% and 22% of controls, respectively.     

Growth and photosynthetic response of three soybean cultivars to simultaneous increases in growth temperature and CO2

Three soybean ( Glycine max L. Merr.) cultivars (Maple Glen, Clark and CNS) were exposed to three CO₂ concentrations (370, 555 and 740 μmol mol⁻¹) and three growth temperatures (20/15°, 25/20° and 31/26°C, day/night) to determine intraspecific differences in single leaf/whole plant photosynthesis, growth and partitioning, phenology and final biomass. Based on known carboxylation kinetics, a synergistic effect between temperature and CO₂ on growth and photosynthesis was predicted since elevated CO₂ increases photosynthesis by reducing photorespiration and photorespiration increases with temperature. Increasing CO₂ concentrations resulted in a stimulation of single leaf photosynthesis for 40–60 days after emergence (DAE) at 20/15°C in all cultivars and for Maple Glen and CNS at all temperatures. For Clark, however, the onset of flowering at warmer temperatures coincided with the loss of stimulation in single leaf photosynthesis at elevated CO₂ concentrations. Despite the season-long stimulation of single leaf photosynthesis, elevated CO₂ concentrations did not increase whole plant photosynthesis except at the highest growth temperature in Maple Glen and CNS, and there was no synergistic effect on final biomass. Instead, the stimulatory effect of CO₂ on growth was delayed by higher temperatures. Data from this experiment suggest that: (1) intraspecific variation could be used to select for optimum soybean cultivars with future climate change; and (2) the relationship between temperature and CO₂ concentration may be expressed differently at the leaf and whole plant levels and may not solely reflect known changes in carboxylation kinetics.     

Physiological Responses of <i>Dendrobium officinale</i> under Exposure to Cold Stress with Two Cultivars

This study aimed to explore the cold tolerance of two cultivars of Dendrobium officinale (MG1, MG2) grown in different regions of China. Under -2°C incubation, cultivar MG1 remained active after 3 d, and continued to grow after returning to room temperature. However, MG2 could only maintain its activity after 2 d treatment at −2°C, and the seedlings died with the low temperature treatment time. Investigation of the characteristics of the plants grown in the south (Hangzhou) or north (Zhengzhou) of China indicated that the leaves of MG1 also had reduced stomatal density, the highest thickness, and a compact microstructure. The contents of proline and soluble sugars were higher in MG1 than those in MG2. The cultivar MG1 had higher SOD enzyme activity than MG2, while CAT and POD activities in samples from Zhengzhou were higher than those from Hangzhou. The contents of polysaccharides and alkaloids in stems of in MG1 were higher than those in MG2, while the content of flavonoids in the Zhengzhou samples was higher than that in the Hangzhou samples. In addition, plant heights, stem diameters, and chlorophyll content were higher in MG1. Overall, MG1 had better cold resistance than MG2. MG1 is a cold tolerant cultivar with thick leaves and reduced stomatal density, higher contents of soluble sugars, proline, CAT, POD, polysaccharides, flavonoids and alkaloids, which together make it more adaptable to low temperatures. Thus, the cultivar MG1, with its demonstrated cold tolerance, can accordingly be grown on a large scale in cold regions, thereby expanding the available planting area for this important traditional medicinal plant to meet the increasing commercial demand for it.     

Cold-induced accumulation of protein in the leaves of spring and winter barley cultivars

Electrophoretic pattern and quantitative changes in soluble proteins were determined in the leaves of spring and winter cultivars of barley (Hordeum vulgare L., cv. Makouei and cv. Reyhan, respectively) exposed to 4 degrees C for 14 d. Seedlings were grown in a controlled growth chamber for 2 weeks at a constant air temperature of 20 degrees C and then transferred to constant 4 degrees C for 14 d followed by returning to 20 degrees C (cold treatment), or they were maintained throughout at 20 degrees C during the experimental period of 40 d (control treatment). Plants were sampled every 48 h for leaf fresh weight measurements. Total leaf soluble proteins were extracted and their concentration was either determined by a colorimetric method, or size-fractionated on SDS-PAGE. Low temperature-induced increases in protein amount occurred over the second week of exposure to cold treatment irrespective of cultivar: the winter cultivar was 2 d prior in this response. The protein patterns and their density showed differences between-cultivars and between-temperature treatments. A new cold-induced polypeptide was recognized in the leaves of winter barley cultivar on day 22 (8 d at 4 degrees C) compared to the control. This polypeptide was produced earlier over the first 48 h of low temperature in the winter cultivar compared with the spring one, recognizing in the leaves of cold-treated seedling until day 26. This more rapid response to a low temperature by the winter barley cultivar indicates a more sensitive response compared with the spring barley, probably cold-shock protein is a component of this cold-induced response.     

Effects of long-term chilling on ultrastructure and antioxidant activity in leaves of two cucumber cultivars under low light

Cucumber ( Cucumis sativus L.) cv. Xintaimici (a chilling-resistant cultivar) and cv. Jinyan no. 4 (a chilling-sensitive cultivar) were subjected to two temperatures (15/15 and 25/18°C) under low light (100 μmol m⁻² s⁻¹) to understand the relationship between ultrastructural changes and the antioxidant abilities caused by low temperature (15/15°C). We also aimed to find indicators for chilling resistance that could be used on a routine basis in breeding programs of greenhouse crops. At the 15/15°C treatment, the membranes of chloroplast, mitochondrion, ER and plasma were not significantly changed in Xintaimici, whereas they were seriously affected in Jinyan no. 4. This result was consistent with the changes of malonaldehyde in chilling-stressed cucumber leaves. The antioxidant activities were changed under low temperature according to cultivar-expected resistance, relating in part to the described ultrastructural changes. The activities of superoxide dismutase (EC 1.15.1.1) and guaiacol peroxidase (EC 1.11.1.7) increased in chilling-stressed leaves of both cultivars, but the two enzymes were not responsible for the difference between cucumber cultivars. At 15/15°C, contents of GSH and activities of glutathione reductase (GR, EC 1.6.4.2) increased more in leaves of Xintaimici than in those of Jinyan no. 4, while catalase (CAT, EC 1.11.1.6) activities decreased less. GSH, GR and CAT were affected by low temperature and cultivars and correlated with the difference in ultrastructure between chilling-stressed cucumber cultivars. We propose that the three antioxidants might be therefore used as biochemical indicators to screen chilling-resistant cucumber cultivars.     

Metabolic changes associated with cold-acclimation in contrasting cultivars of barley

Cereal plants become more resistant to freezing when first exposed to a period of cold-acclimation. Many physiological and molecular changes have been shown to occur at low temperatures, but the role and the contribution of each to frost resistance is still poorly understood. Two cultivars of barley ( Hordeum vulgare L.), the winter barley Onice and the spring barley Gitane, were acclimated under controlled conditions under an 8-h photoperiod at 4°C (light) and 2°C (dark) for 21 days. Changes in free proline, ABA, water-soluble carbohydrates and free fatty acids were measured to assess their involvement in cold-acclimation and to explain the different frost-resistant capacities of the two cultivars. Exposure of barley plants to low temperature resulted in an equal increase in proline in both cultivars. During the first days of cold acclimation, ABA levels showed a peak in the frost-resistant cultivar, lasting about 24 h, followed by a decrease. The water soluble carbohydrates reached their highest content after 3 days of hardening, although after 14 to 21 days of acclimation the carbohydrate content was similar to that of unhardened plants. The frost-resistant Onice had a much higher free fatty acid content than the frost-sensitive Gitane. Furthermore in Onice 86% of free farty acids was represented by unsaturated molecular species. Inolenic acid alone being 71%. In contrast, in the frost-sensitive cultivar only 31% of free fatty acids was unsaturated and a large amount of 9-oxo-nonanoic acid, a product present in the linolenic acid cascade, was also detected.
The ABA content after 2 days of hardening and the free fatty acid composition were clearly different between the two cultivars and may explain, at least in part, the different frost-resistant capacities of Onice and Gitane.     

Sucrose and fructan metabolism of different wheat cultivars at chilling temperatures

Four wheat ( Triticum aestivum L.) varieties cultivated in different climates from subtropics to North Patagonia were used to study sucrose and fructan metabolism in plants when submitted to a cold period. Higher levels of sugars were found in the more cold tolerant cultivars. Sucrose synthase (EC 2.4.1.13) and sucrose phosphate synthase (EC 2.4.1.14) activities showed a 2–3 fold increase when plants were grown at 4°C for 10 days. The more cold-tolerant wheat cultivars also showed the higher levels of enzyme activities. These metabolical changes were not due to anatomical or morphological differences produced during growth at 4°C     

The effects of low root-zone temperature stress on two soybean (Glycine max) genotypes when combined with Bradyrhizobium strains of varying geographic origin

In areas with short growing seasons, poor early vegetative growth of soybean (Glycine max [L.] Merr.) is often attributed to the restrictive effect of cool soil conditions on nodulation and N₂-fixation by this subtropical grain legume. However, there are few studies regarding potential genetic variability of soybean and Bradyrhizobium japonicum genotypes for nodulation at cool root-zone temperatures (RZT). Experiments were conducted to (1) test for a threshold temperature for low RZT inhibition of soybean nodulation and (2) ascertain whether this threshold temperature response depends mainly on the micro- or macrosymbiont. In experiment 1 soybean seedlings (Glycine max [L.] Merr. cv. Maple Arrow) were inoculated with 1 ml of a log phase culture of B. japonicum strain 532C, H8 or H15 (the latter two strains were isolated from cold soils of Hokkaido, northern Japan) and maintained at either 16, 17.5, 19 or 25°C RZT. In experiment 2 seedlings of cv. Maple Arrow and a cold-tolerant Evans isoline were combined with strain 532C and two Hokkaido strains (H5, H30) at both 19 and 25°C RZT. Results indicated that N₂-fixation at 44 days after inoculation was substantially reduced (30–40%) by RZT as high as 19°C, due to development of less nodule mass and to a delay in the onset of N₂-fixation and a small decrease in the number of nodules formed. However, the number of nodules formed was sharply reduced and the time required for the first appearance of nodules was significantly delayed below an RZT of 17.5°C. Differences between cultivars for nodulation and N accumulation were apparent at 25°C, but were abolished by growth at 19°C, indicating that, in spite of differences in growth potential between the cultivars under optimum RZT, both cultivars were equally limited by low RZT. Differences between B. japonicum strains were consistent across temperatures and were largely attributable to higher rates of specific nodule activity recorded for strain 532C, which seemed well adapted to low RZT. These results suggest that the host plant mediates the sensitivity of N₂-fixation under low RZT and that inoculation with B. japonicum strains from cold environments is unlikely to enhance soybean N₂-fixation under cool soil conditions.     

Influence of photoperiod on low temperature acclimation for cold-hardiness in Drosophila auraria

ABSTRACT. Imagines of Drosophila auraria Peng, a reproductive diapause species, developed cold-hardiness at low temperatures to a greater extent when exposed to a diapause-inducing photoperiod (LD10:14 h) than when exposed to a diapause-preventing photoperiod (LD 16:8h). Imagines kept at 18°C, which was the temperature at which they were reared to eclosion, did not survive a test exposure to -5°C for 8 days regardless of age or photoperiod. When transferred to 10 or 5°C, either from eclosion or from 8 days after eclosion, the survival rate, on testing, rose with time since transfer and rose faster and higher with a photoperiod of LD 10:14h than with LD16:8h. Flies transferred to 15°C only showed improved ability to survive the test if they were kept in LD 10:14h. When cultured at 18°C to the age of 8 days after eclosion, diapause was terminated in about 30% of females even at LD 10:14h. In these post-diapause females the ability to develop cold-hardiness at lower temperatures was somewhat less than in the diapausing females, but apparently greater than in the non-diapause females. These results suggest that the physiological mechanism which promotes cold-hardiness under a diapause-inducing photoperiod is not directly linked to the process causing reproductive diapause.
In Sapporo, flies from a natural population became tolerant to cold in October when they entered diapause and daily mean temperature fell below 15°C and the light/dark cycle fell below LD 12:12h.     

The state of water in acclimating vegetative buds from Malus and Amelanchier and its relationship to winter hardiness

The relationship between freezable water and cold hardiness during acclimation was studied using vegetative buds from several apple ( Malus domestica Borkh) cultivars and from one saskatoonberry ( Amelanchier alnifolia Nutt. cv. Smoky) cultivar. According to leakage data and visual assessments of cortical browning, vegetative buds of all cultivars were most tolerant to subfreezing temperatures in January. The hardy condition was also associated with maximum tolerance to desiccation. Qualitative features of freezing exotherms (number of peaks and temperature of the transition) were not correlated with the hardy condition in the tissues. However, the amount of unfrozen water, determined by quantifying the energy of the exotherms, increased with increasing hardiness. In buds that survived exposure to −45°C, freezing reduced the intracellular water content, but only to levels above the critical moisture content for desiccation damage. In buds that did not survive exposure to −45°C, freezing reduced the water content to levels equal to or less than the critical moisture content for desiccation damage. These observations suggest that the freezing of water in nonhardy tissue dried the tissue to moisture levels at which severe dehydration damage occurred. It appears that acclimation of vegetative apple buds involves at least two processes: (1) an increase in tolerance to dehydration and (2) an increase in the level of unfreezable water.     

Induction ofent-kaurene biosynthesis by low temperature in dwarf peas

Germinating pea (Pisum sativum L.) seeds of two dwarf cultivars, “Progress No. 9” and “Green Arrow”, and two tall cultivars, “Alaska” and “Alderman”, were treated with low temperature (3–5°C) for 14 days and then transferred to normal growing conditions (19–21°C for 16 h/14.5–16.5°C for 8 h) for an additional 10 days. Biosynthesis of [¹⁴C]ent-kaurene from [¹⁴C]2-mevalonic acid (2-MVA) was assayed in cell-free enzyme extracts prepared from shoot tips 10 days after cold treatment and was compared with activity in enzyme extracts prepared from noncold-treated, 10-day-old control plants. Shoot lengths of cold-treated plants were measured throughout a 35-day period and compared with shoot lengths of plants grown without cold treatment for 25–35 days. Low temperature induced a five-to 10-fold enhancement ofent-kaurene, hence potentially gibberellin (GA), biosynthesis in seedlings of the two dwarf cultivars but not in the tall cultivars. However, the lack of an increase in growth rate in the cold-treated dwarfs indicated that endogenous GA biosynthesis remained blocked at some point beyondent-kaurene in the biosynthetic pathway. Since the late-flowering “Alderman” cultivar did not exhibit enhanced biosynthesis ofent-kaurene, it appears that if vernalization in late-flowering cultivars of peas is correlated with enhanced GA biosynthesis, it is not the early part of the biosynthetic pathway which is affected.     

A Comparison of the Effect of Salt on Polypeptides and Translatable mRNAs in Roots of a Salt-Tolerant and a Salt-Sensitive Cultivar of Barley

The effect of salt stress on polypeptide and mRNA levels in roots of two barley (Hordeum vulgare L.) cultivars differing in salt tolerance (cv CM 72, tolerant; cv Prato, sensitive) was analyzed using two-dimensional polyacrylamide gel electrophoresis. Preliminary experiments indicated that germination of Prato was inhibited significantly in the presence of NaCl, but growth of the surviving Prato seedlings was not substantially different from that of CM 72. Fluorographs of two-dimensional gels containing in vivo labeled polypeptides or in vitro translation products were computer analyzed to identify and quantitate changes that resulted when plants were grown in the presence of 200 millimolar NaCl for 6 days. The patterns of in vivo labeled polypeptides and in vitro products of CM 72 and Prato were qualitatively the same. Salt caused quantitative changes in numerous polypeptides and translatable mRNAs, but, overall, the changes were relatively small. Salt did not induce the synthesis of unique polypeptides or translatable mRNAs and did not cause any to disappear. Because of the similarities of the two cultivars with respect to growth and polypeptide patterns and the slight changes in polypeptide and translation product levels caused by salt, specific polypeptides or translatable mRNAs that are related to salt tolerance in barley could not be identified.     

Regulation of miR319 during cold stress in sugarcane

MicroRNAs (miRNAs) are part of a novel mechanism of gene regulation that is active in plants under abiotic stress conditions. In the present study, 12 miRNAs were analysed to identify miRNAs differentially expressed in sugarcane subjected to cold stress (4 °C). The expression of miRNAs assayed by stem–loop RT‐PCR showed that miR319 is up‐regulated in sugarcane plantlets exposed to 4 °C for 24 h. The induction of miR319 expression during cold stress was observed in both roots and shoots. Sugarcane miR319 was also regulated by treatment with abscisic acid. Putative targets of this miRNA were identified and their expression levels were decreased in sugarcane plantlets exposed to cold. The cleavage sites of two targets were mapped using a 5′ RACE PCR assay confirming the regulation of these genes by miR319. When sugarcane cultivars contrasting in cold tolerance were subjected to 4 °C, we observed up‐regulation of miR319 and down‐regulation of the targets in both varieties; however, the changes in expression were delayed in the cold‐tolerant cultivar. These results suggest that differences in timing and levels of the expression of miR319 and its targets could be tested as markers for selection of cold‐tolerant sugarcane cultivars.     

Ureide degradation pathways in intact soybean leaves

Ureides dramatically accumulate in shoots of N(2)-fixing soybean (Glycine max L. Merr.) under water deficit and this accumulation is higher in cultivars that have N(2) fixation that is sensitive to water deficit. One possible explanation is that ureide accumulation is associated with a feedback inhibition of nitrogenase activity. A critical factor involved in ureide accumulation is likely to be the rate of ureide degradation in the leaves. There exists, however, a controversy concerning the pathway of allantoic acid degradation in soybean. Allantoate amidinohydrolase was reported to be the pathway of degradation in studies using the cultivar Maple Arrow and allantoate amidohydrolase was the pathway that existed in the cultivar Williams. This investigation was undertaken to resolve the existence of these two pathways. An in situ technique was developed to examine the response of ureide degradation in leaf tissue to various treatments. In addition, the response of ureide accumulation and N(2) fixation activity was measured for intact plants in response to treatments that differentially influenced the two degradation pathways. The results from these studies confirmed that Maple Arrow and Williams degraded allantoic acid by different pathways as originally reported. The existence of two degradation pathways within the soybean germplasm opens the possibility of modifying ureide degradation to minimize the influence of soil water deficits on N(2) fixation activity.