Zein accumulation in forage species (<Emphasis Type="Italic">Lotus corniculatus</Emphasis> and <Emphasis Type="Italic">Medicago sativa</Emphasis>) and co-expression of the γ-zein:KDEL and β-zein:KDEL polypeptides in tobacco leaf

Plants of birdsfoot trefoil (Lotus corniculatus), alfalfa (Medicago sativa) and tobacco (Nicotiana tabacum) were transformed by Agrobacterium with binary vectors harbouring genes that code either for wild-type and %-zein:KDEL, or for #-zein:KDEL. The maize seed storage proteins %-zein and #-zein are rich in sulphur amino acids, and our long-term goal is to improve alfalfa and birdsfoot trefoil forage quality. Significant levels of zeins were detected in leaves of birdsfoot trefoil transformants, ranging up to 0.055% and 0.30% of total extractable protein for %-zein and #-zein:KDEL, respectively. In leaves of alfalfa, the %-zein:KDEL expression level was up to 0.026% of the total extractable protein. From a F₁ population of transgenic tobacco, a plant was selected in which the amount of zeins (%-zein:KDEL plus #-zein:KDEL) accounted for 1.1% of the total extractable protein.     

Cryopreservation of alfalfa (Medicago sativa L.) cells by encapsulation-dehydration

Our objective was to establish a cryopreservation protocol for alfalfa (Medicago sativa L.) cells and study the physiological changes occurring in cells during cryopreservation treatment. Cell cultures of Pioneer cvs. 5262 (fall-dormant, winter-hardy) and 5929 (non-dormant, non-hardy) plants initiated regrowth after cryopreservation by encapsulation-dehydration (ED). Pre-treatment of the encapsulated cells for 4 days in B5 medium containing 0.75 M sucrose and dehydration for 4 h in a laminar flow hood were necessary to achieve maximum cell viability after ED and cryopreservation in liquid N₂ (EDN). Viability (measured as triphenyl tetrazolium chloride reduction) of the cv. 5262 cells after cryopreservation was two- to three-fold greater than that of the cv. 5929 cells. Cold acclimation of the cells (10 days at 2°C) improved viability after cryopreservation. The addition of 7.6 µM ABA to the B5 medium enhanced viability in ED but did not improve cell cryopreservability. Cold-acclimated cells had higher protein concentrations, but neither ABA nor cold acclimation influenced protein composition of cold-acclimated cells determined using SDS-PAGE. Encapsulated cells pre-treated for 4 days in B5 medium containing 0.75 M sucrose showed an increased concentration of cell protein and an altered protein composition. Suspension cultures were re-initiated from both ED and EDN treatments by transferring beads sequentially to B5 media containing 0.75, 0.5, 0.25 M sucrose and then to fresh B5 medium. The ED cells resumed rapid growth after two subcultures, whereas EDN cells needed four or five subcultures to resume rapid growth.     

Efficient recovery of transgenic plants through organogenesis and embryogenesis using a cryptic promoter to drive marker gene expression

Our previous studies have shown that tCUP, a cryptic promoter from tobacco, functions in all living plant cell types in a wide range of plant species. This led us to investigate if an enhanced derivative, EntCUP(, could be used to drive the neomycin phosphotransferase II (nptII) gene and select for kanamycin resistance in crop species that regenerate by organogenesis or embryogenesis. Tobacco (leaves), cauliflower (hypocotyls) and alfalfa (leaves, petioles, stems) explants were co-cultivated with Agrobacterium containing either EntCUP(-nptII-nos or 35S-nptII-nos to compare the efficiency of selection for kanamycin resistance. The infected alfalfa explants were placed in somatic embryo induction media, whereas tobacco and cauliflower explants were placed in shoot induction media with kanamycin at concentrations that normally inhibit regeneration. Transgenic plants were recovered from all of the explants with both selectable marker gene constructs. The transformation efficiencies using tCUP(-nptII-nos were comparable to or higher than those using 35S-nptII-nos in all three species tested. This study demonstrated that promoters which are not associated with expressed plant genes can be used as alternatives for the expression of selectable marker genes in a broad range of tissues and species for the generation of transgenic plants.     

Coleomegilla maculata (Coleoptera: Coccinellidae) predation on pea aphids promoted by proximity to dandelions

The impact of a predator on its prey may depend on the presence of other species in the community. In particular, if predators are attracted to areas containing one prey species, another prey species may suffer greater predation if it occurs in the same areas. If the predator is omnivorous, this may occur even if one prey species is an animal and the other is a plant. We investigated the role of local dandelion densities on the impact of the predator Coleomegilla maculata on pea aphids in alfalfa fields. At small spatial scales, increased dandelion densities were associated with high C. maculata densities, presumably because these omnivorous ladybird beetles aggregated to pollen resources. In turn, the high C. maculata densities were associated with low aphid densities, presumably because of increased predation. We used laboratory cages to simulate C. maculata foraging in two adjacent patches of alfalfa, one with dandelions and one without. As in the field, the laboratory experiment showed that C. maculata aggregated to alfalfa interspersed with dandelions, which resulted in increased predation on aphids on alfalfa. This study demonstrates that a pollen-producing plant can indirectly decrease nearby herbivore densities by attracting an omnivorous predator.     

Effects of herbivory, nutrient levels, and introduced algae on the distribution and abundance of the invasive macroalga Dictyosphaeria cavernosa in Kaneohe Bay, Hawaii

Since the 1960s, and possibly earlier, the macroalga Dictyosphaeria cavernosa has overgrown and displaced corals on reef slopes and outer reef flats in Kaneohe Bay, Oahu. This shift in reef community composition is generally attributed to nutrient enrichment resulting from sewage discharge. Following the diversion of most of the sewage effluent in 1977-1978, it was expected that D. cavernosa growth would become nutrient-limited and its abundance would consequently decline, but the alga remains abundant in much of the bay. One explanation for its persistence is that nutrients are once again high enough to support the alga's growth. An alternative explanation is that there has been a reduction in grazing intensity in the bay. In this study we resurveyed the distribution and abundance of D. cavernosa at 120 reef slope sites originally surveyed in 1969. We conducted additional surveys to estimate the biomass of herbivores and the areal coverage of D. cavernosa and other macroalgae on reef slopes and flats. Field experiments were used to determine spatial and temporal patterns of grazing intensity on and growth rates of D. cavernosa and the introduced macroalga Acanthophora spicifera. Laboratory experiments were used to examine preferences among herbivores for some of the most abundant macroalgae on Kaneohe Bay reefs. Twenty years after sewage diversion, D. cavernosa cover on reef slopes has decreased substantially in southern Kaneohe Bay, the site of most of the historical sewage discharge. D. cavernosa cover has changed less in other regions, remaining high in the central bay and low in the north bay. D. cavernosa thalli protected by grazer exclusion cages sustained positive growth rates on reef slopes and flats throughout the bay. Reduced nutrient concentrations may have caused a reduction in D. cavernosa growth rates, and a consequent reduction in D. cavernosa abundance in the south bay shortly after sewage diversion. Measurements of grazing intensity and surveys of herbivorous fish abundance suggest that the continued abundance of D. cavernosa is the result of a reduction in grazing intensity. Reduced grazing intensity on D. cavernosa may in turn be the result of a historical reduction in herbivore biomass or the establishment of several introduced macroalgae on reef flats. The introduced species are preferred by herbivorous fishes over D. cavernosa, as indicated by preference tests. The hypothesis that reduced grazing pressure on D. cavernosa is related to the establishment of introduced species is supported by the observation that D. cavernosa cover is highest on reef slopes where the cover of preferred introduced macroalgae on the adjacent outer reef flat is also high. Conversely, D. cavernosa cover is low or zero on reef slopes where the cover of introduced macroalgae on the adjacent reef flat is low or zero     

Observations on population structure and reproductive features of <Emphasis Type="Italic">Laetmonice producta</Emphasis> Grube (Polychaeta,Aphroditidae) in Antarctic waters

The large scale-worm Laetmonice producta Grube 1877 is the most abundant aphroditid polychaete in Antarctic coastal waters. We investigated the demographic structure and some reproductive features of different L. producta populations from high-Antarctic (Weddell Sea) and Antarctic Peninsula (King George Island) shelf bottoms, collected in summer 1996 (ANT-XIII/3, EASIZ I cruise) and autumn 2000 (ANT-XVII/3, EASIZ III cruise). L. producta in the studied geographic areas showed a wide bathymetric range (200-850 m depth), and a different size distribution pattern with depth, characterised by a reduction of large specimens in the deepest stations. The species is gonochoric, with females more abundant in specimens of larger sizes. Eggs at different stages of maturation (ranging from 40 to 320 wm in diameter) were examined in 270 individuals from different stations and size classes. Egg size showed a slightly bimodal trend, with largely overlapping egg cohorts, suggesting a continuous reproduction, and a long-lasting gametogenesis. Significant differences (Kolmogorov-Smirnov test, P<0.05) in egg-size frequency distribution were detected only when data of the two geographic areas were compared (Weddell Sea vs King George Island), and not according to stations within each area, and females' size. The two sets of geographic samples were collected in different seasons and therefore it was not possible to assess if differences observed are due to sampling time or to geographic factors. Mature spermatozoa were recognisable only in autumn male specimens from King George Island, and showed a rounded nucleus and a short conical acrosome. Occurrence of an endosymbiont polychaete, Veneriserva pygoclava meridionalis (new sub-species of Dorvilleidae), was recorded in the coelomic cavity of 163 specimens of L. producta, 125 of which were from the deepest station of the Weddell Sea (stn. 14, 850 m depth). L. producta females with and without the endosymbiont did not show differences in egg-size distribution. The reproductive features of L. producta, together with its large size and slow growth, seem typical of a long-living predator species, and uncoupled from the typical summer environmental conditions and the pulsating system of coastal Antarctic waters.     

Isolation from alfalfa of resistance gene analogues containing nucleotide binding sites

Cloned resistance (R) genes from a broad range of plant species are known to share similarities in DNA sequence and structural motifs. Degenerate oligonucleotide primers designed from conserved regions of the nucleotide binding site (NBS), common to many R genes, were used to amplify the NBS regions from genomic DNA from alfalfa (Medicago sativa L). Sequence comparisons of the amplified fragments indicated that at least 18 families of NBS-containing R genes are present in alfalfa. Comparisons to R genes from other species suggested a polyphyletic origin of these gene families. Using the same degenerate primers, PCR analysis of cDNA prepared from a plant not challenged with a pest or pathogen revealed that many of the NBS-containing gene families were transcribed actively. Amplification of NBS regions from other Medicago species showed the presence of some NBS-containing genes not present in alfalfa. These results indicate that the NBS-containing R genes comprise a large gene family in Medicago, at least some of which are transcribed in healthy plants, and that different Medicago species carry unique NBS genes.     

Evaluating the use of roving diver and transect surveys to assess the coral reef fish assemblage off southeastern Hispaniola

The relatively little-studied fish fauna off southeastern Hispaniola was rapidly assessed using a combination of visual survey techniques including transects and roving diver surveys. It was found that when combined, both methods provided a more complete overall species assessment than either method was able to provide in isolation. Being able to conduct rapid species assessments is becoming increasingly more important as a conservation tool. Data on species composition, sighting frequency, and abundance of all fishes were collected using both methods. Abundance was recorded in four logarithmic-based categories (roving diver method) while the number of fishes were counted within 40-m² transects (transect method). Both methods were similar in recording the most abundant species, while a greater number of rare species (especially fishery-targeted species) were recorded with the roving diver method. The most abundant groupers were Cephalopholis cruentata and C. fulva. The most abundant parrotfishes were Scarus taeniopterus, Sparisoma aurofrenatum, and Scarus iserti. The most conspicuous differences between fishes off southeastern Hispaniola and elsewhere in the tropical western Atlantic were the low abundance and smaller size of harvested species such as groupers, snappers (Fam. Lutjanidae), and grunts (Fam. Haemulidae). With the roving diver method, more time could be spent surveying (instead of placing transect lines), resulting in a greater number of species being recorded. Additionally, well-trained volunteers can adopt the roving diver method as part of their regular diving program. Transect surveys were able to provide information such as length (biomass) and actual density measures that were not recorded in roving diver surveys. Thus, these methods were complementary and should be used in conjunction when conducting rapid assessments of fish assemblages, especially to detect the effects of overfishing.     

Crystal structure of vestitone reductase from alfalfa (Medicago sativa L.)

Isoflavonoids are commonly found in leguminous plants, where they play important roles in plant defense and have significant health benefits for animals and humans. Vestitone reductase catalyzes a stereospecific NADPH-dependent reduction of (3R)-vestitone in the biosynthesis of the antimicrobial isoflavonoid phytoalexin medicarpin. The crystal structure of alfalfa (Medicago sativa L.) vestitone reductase has been determined at 1.4 A resolution. The structure contains a classic Rossmann fold domain in the N terminus and a small C-terminal domain. Sequence and structural analysis showed that vestitone reductase is a member of the short-chain dehydrogenase/reductase (SDR) superfamily despite the low levels of sequence identity, and the prominent structural differences from other SDR enzymes with known structures. The putative binding sites for the co-factor NADPH and the substrate (3R)-vestitone were defined and located in a large cleft formed between the N and C-terminal domains of enzyme. Potential key residues for enzyme activity were also identified, including the catalytic triad Ser129-Tyr164-Lys168. A molecular docking study showed that (3R)-vestitone, but not the (3S) isomer, forms favored interactions with the co-factor and catalytic triad, thus providing an explanation for the enzyme's strict substrate stereo-specificity.     

Acclimation to sudden increase in light favoring an invasive over native trees in subtropical islands, Japan

Bischofia javanica Blume, an exotic tree, dominates many forest areas of the Bonin Islands in the western Pacific of Japan. The aim of this study was to test the hypothesis that the success of B. javanica (a mid-successional plant species) is related to its high acclimation capacity to sudden light increase due to canopy gap formation. We compared its ecophysiological response to simulated canopy opening with those of native species of different successional status: Trema orientalis Blume, Schima mertensiana (Sieb, et Zucc.) Koidz, Elaeocarpus photiniaefolius Hook.Et Arn. and Ardisia sieboldii Miquel. In all species, transfer of leaves developed in shade (5.3% of full sun) to full sun resulted in a substantial initial reduction in the dark-adapted quantum yield of photosystem II (Fv/Fm). T. orientalis, a pioneer plant species, showed the least reduction (38%), whereas E. photiniaefolius and A. sieboldii, both late-successional plant species, demonstrated large reductions (about 80%). In all four native species, Fv/Fm in shade leaves gradually recovered following transfer, but B. javanica recovered more fully and rapidly than the other species. Unlike Fv/Fm, the chlorophyll content in all species did not recover following the initial decline. This indicates that the recovery of quantum yield (Fv/Fm) was independent of the reduction in chlorophyll. Among all the species, B. javanica showed the highest (1) increase in maximum photosynthetic rate of shade leaves after transfer, (2) production of newly formed sun leaves, and (3) increase in relative growth rate. Ecophysiological characters of B. javanica in simulated canopy openings indicated rapid photosynthetic acclimation in existing shade leaves by minimizing photoinhibition and a rapid deployment of new sun leaves with high photosynthetic capacity. Because its habitats on these Pacific Islands are prone to typhoon disturbance, the successful invasion of B. javanica may lie in the congruence of its acclimation potential and the frequent gap events.     

Effect of leaf toughness on the susceptibility of the leaf-cutting ant Atta sexdens to attacks of a phorid parasitoid

Summary: Because the size of Atta spp. along foraging trails is partly determined by the characteristics of the plants harvested, and considering that parasitic phorid flies are attracted mostly to large individuals, we hypothesized that plant toughness affects the susceptibility of Atta spp. to these parasitoids. To test this hypothesis, we evaluated parasitism rates of the phorid Neodohrniphora sp. and its effect on Atta sexdens (L.) foragers in a laboratory colony. We manipulated forager size by alternating tough (Anthocephalus chinensis, Rubiaceae) and tender (Rosa chinensis, Rosaceae) plants given to the colony. Ants foraging on tough leaves were larger than ants foraging on soft leaves, and there was a significant reduction in forager size for both plants when the colony was exposed to Neodohrniphora sp. However, there were no relative differences on forager size between the two plants after the introduction of the parasitoid. The lack of response of Neodohrniphora sp. to the increase in ant size when the colony was given tough leaves may be attributed to the unusually large number of suitable hosts in a laboratory colony. However, large foragers are much less abundant in the field, in which case shifts in the size of the workforce triggered by different substrates could affect the incidence of parasitism.     

Competition and intraguild predation among three sympatric carnivores

We examined the relative roles of dominance in agonistic interactions and energetic constraints related to body size in determining local abundances of coyotes (Canis latrans, 8-20 kg), gray foxes (Urocyon cinereoargenteus, 3-5 kg) and bobcats (Felis rufus, 5-15 kg) at three study sites (hereafter referred to as NP, CP, and SP) in the Santa Monica Mountains of California. We hypothesized that the largest and behaviorally dominant species, the coyote, would exploit a wider range of resources (i.e., a higher number of habitat and/or food types) and, consequently, would occur in higher density than the other two carnivores. We evaluated our hypotheses by quantifying their diets, food overlap, habitat-specific abundances, as well as their overall relative abundance at the three study sites. We identified behavioral dominance of coyotes over foxes and bobcats in Santa Monica because 7 of 12 recorded gray fox deaths and 2 of 5 recorded bobcat deaths were due to coyote predation, and no coyotes died as a result of their interactions with bobcats or foxes. Coyotes and bobcats were present in a variety of habitats types (8 out of 9), including both open and brushy habitats, whereas gray foxes were chiefly restricted to brushy habitats. There was a negative relationship between the abundances of coyotes and gray foxes (P=0.020) across habitats, suggesting that foxes avoided habitats of high coyote predation risk. Coyote abundance was low in NP, high in CP, and intermediate in SP. Bobcat abundance changed little across study sites, and gray foxes were very abundant in NP, absent in CP, and scarce in SP; this suggests a negative relationship between coyote and fox abundances across study sites, as well. Bobcats were solely carnivorous, relying on small mammals (lagomorphs and rodents) throughout the year and at all three sites. Coyotes and gray foxes also relied on small mammals year-round at all sites, though they also ate significant amounts of fruit. Though there were strong overall interspecific differences in food habits of carnivores (P<0.0001), average seasonal food overlaps were high due to the importance of small mammals in all carnivore diets [bobcat-gray fox: 0.79ǂ.09 (SD), n=4; bobcat-coyote: 0.69ǂ.16, n=6; coyote-gray fox: 0.52ǂ.05, n=4]. As hypothesized, coyotes used more food types and more habitat types than did bobcats and gray foxes and, overall, coyotes were the most abundant of the three species and ranged more widely than did gray foxes. We propose that coyotes limit the number and distribution of gray foxes in Santa Monica Mountains, and that those two carnivores exemplified a case in which the relationship between their body size and local abundance is governed by competitive dominance of the largest species rather than by energetic equivalences. However, in the case of the intermediate-sized bobcat no such a pattern emerged, likely due to rarity or inconsistency of agonistic interactions and/or behavioral avoidance of encounters by subordinate species.     

Modification of the culture medium to produce aluminum toxicity in cell suspensions of coffee (Coffea arabica L.)

Coffee (Coffea arabica) plants are usually grown in soils containing high levels of organic materials. Under these conditions, aluminum (Al) is toxic because of the acidic nature of the soils. Al is the most abundant metal found in the earth's crust and occurs in a number of different forms in soil. In acid soils, Al toxicity is a global problem that limits crop productivity. A major problem in obtaining cellular lines displaying Al tolerance in culture is the composition of the medium. In the experiments presented here, we modified the composition of the culture medium for a C. arabica cell line to produce Al toxicity. Murashige-Skoog media was used, complete (MS) and half ionic strength (MSHIS), at either pH 5.8 or 4.3. We found that MSHIS and pH 4.3 provided the optimal conditions to obtain Al toxicity as measured by the ability to grow in a range of Al concentrations (25-1,000 µM). The lethal dose (LD₅₀) under these conditions was 25 µM. The concentrations of free Al in the culture medium were corroborated by the fluorescent compound Morin. Al was found to enter the cell after 30 min, and the signal was then retained for up to 2 h.     

Effects of high nitrogen load on growth, photosynthesis and nutrient status of Cryptomeria japonica and Pinus densiflora seedlings

To evaluate the sensitivity of Japanese cedar (Cryptomeria japonica D. Don) and Japanese red pine (Pinus densiflora Sieb. et Zucc.) to high N deposition, 1-year-old seedlings were grown in brown forest soil treated with N as NH₄NO₃ at 0, 25, 50, 100 and 300 mg l^-1 fresh soil volume, equivalent to 0, 28, 57, 113 and 340 kg N ha^-1. Net photosynthetic rate and whole-plant dry mass of C. japonica seedlings were increased by the N treatment, whilst those of P. densiflora seedlings were significantly reduced by the highest N treatment. The reduction in the net photosynthesis of P. densiflora seedlings was mainly due to a depression of carboxylation efficiency accompanied by a decrease in concentration and activity of Rubisco in the needles. In P. densiflora seedlings, needle concentrations of P and Mg were decreased, and the concentrations of N and Mn were increased by the highest N treatment. The reductions in needle protein concentration and Rubisco activity were negatively correlated with the ratios of N/P and Mn/Mg in the needles. These results suggest that nutrient imbalances of these elements may be induced in P. densiflora seedlings grown under high N deposition. We conclude that P. densiflora is more sensitive to high N deposition than C. japonica, and that the relatively high atmospheric N deposition to Japanese forest ecosystems may adversely affect the health of N-sensitive tree species such as P. densiflora.     

Influence of groundwater depth on the seasonal sources of water accessed by <Emphasis Type="Italic">Banksia</Emphasis> tree species on a shallow,sandy coastal aquifer

In Mediterranean ecosystems vegetation overlying shallow, transient aquifers is often dominated by woody phreatophytes, trees and shrubs that have been shown to be dependent on groundwater for their water requirements. Natural and anthropogenic alterations of groundwater tables (abstraction) are of clear importance to phreatophytic vegetation as reduction of water tables may sever these plants from their natural water sources. Seasonal water sources were determined for species growing on a coastal dune system that overlies a shallow sandy aquifer in south-western Australia. The plants studied grew over groundwater that ranged in depth from 2.5 to 30 m. The naturally occurring stable isotope of hydrogen (deuterium, '²H) was used to distinguish potential water sources. Isotopic ratios from vascular water of the dominant species of the study area (Banksia ilicifolia R. Br. and Banksia attenuata R. Br. trees) were compared with those of potential sources of precipitation, soil moisture and groundwater. A relatively shallow-rooted perennial shrub, Hibbertia hypericoides Benth., was also included as an isotopic reference. The results suggest that both B. attenuata and B. ilicifolia are phreatophytic as they derived some of their water from groundwater throughout the dry-wet cycle, with the exception of B. attenuata at the site of greatest depth to groundwater (30 m) which did not use groundwater. A high proportion (>50%) of groundwater use was not maintained throughout all seasons. With the onset of the hot Mediterranean summer, progressive drying of the surface soils resulted in increased use of groundwater and deep soil moisture. During the wet winter plants used proportionately more water from the upper layers of the soil profile. The degree to which groundwater was utilised by the study species was dependent on the proximity of groundwater, availability of moisture in shallower horizons of the soil profile, root system distribution and maximum root depth.     

Interactive effects of ultraviolet radiation and salinity on the ecophysiology of two Arctic red algae from shallow waters

In a comparative ecophysiological study, the abundant red macroalgae Devaleraea ramentacea (L.) Guiry and Palmaria palmata (L.) O. Kuntze from shallow waters of the Arctic Kongsfjord (Spitsbergen) were exposed to hyposaline and hypersaline media, in combination with and without artificial UV radiation, to evaluate the interactive effects of both environmental parameters on optimum quantum yield of photosynthesis, as well as on the physiological capability to synthesise and accumulate photoprotective mycosporine-like amino acids (MAAs). While D. ramentacea exhibited euryhaline features and acclimated well to the UV radiation applied, P. palmata can be characterised as a stenohaline plant because of its high mortality even under mild hyposaline conditions (15 PSU). In addition, the latter species showed a limited ability to acclimate to changing PAR/UV radiation, pointing to a relatively low physiological plasticity. Both species synthesised and accumulated MAAs after UV treatment. However, only in D. ramentacea was a correlation between increasing MAA concentration and decreasing photosynthetic sensitivity under UV observed. All ecophysiological data from the laboratory correlate well with field observations, where both red-algal species co-exist in the same shallow-water habitat of the Kongsfjord. However, while P. palmata becomes more often greenish, sometimes slightly bleached over the summer months, D. ramentacea appears much more healthy under the prevailing environmental conditions.     

The effects of desiccation on seeds of Acer saccharinum and Aesculus pavia: recalcitrance in temperate tree seeds

This study was undertaken to determine how the results from lipid, moisture, and differential scanning calorimetry analyses conducted on silver maple (Aceraceae: Acer saccharinum L.) and red buckeye (Hippocastanaceae: Aesculus pavia L.) compared with those obtained from previous studies on white and water oaks (Fagaceae: Quercus alba and Q. nigra), and the tropical zone species American muskwood (Meliaceae: Guarea guidonia) and carapa (Meliaceae: Carapa guianensis). Seeds were air-dried at room temperature for 9-11 days. At intervals, germination was tested, moisture determined, and lipids extracted. It was found that, like the other recalcitrant seeds, (1) viability was greatly reduced or lost after 11 days of drying, (2) percentage changes in individual fatty acids were not related to seed viability, and (3) results from the differential scanning calorimetry studies revealed a strong relationship between enthalpy/onset data from the embryo and cotyledon tissues and loss of viability. Also, silver maple seeds experienced a 50% reduction in viability by day 5 of drying and retained an axis moisture content over 25% throughout the experiment. However, unlike the other recalcitrant seeds surveyed, both silver maple and red buckeye had a significant reduction in the total amount (mg/g) of cotyledon lipids as the experiment progressed. However, no decrease in the unsaturated/saturated fatty acid ratio was found, so we conclude that in these species lipid peroxidation is not a marker of declining seed viability. Also, red buckeye seeds did not lose 50% viability until after day 8 of the experiment, and axis moisture content fell well below 20% as the seeds dried.