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In the previous decade, dozens of studies involving thousands of children across several research disciplines have made use of a combined daylong audio-recorder and automated algorithmic analysis called the LENA Ⓡ system, which aims to assess children's language environment. While the system's prevalence in the language acquisition domain is steadily growing, there are only scattered validation efforts on only some of its key characteristics. Here, we assess the LENA Ⓡ system's accuracy across all of its key measures: speaker classification, Child Vocalization Counts (CVC), Conversational Turn Counts (CTC), and Adult Word Counts (AWC). Our assessment is based on manual annotation of clips that have been randomly or periodically sampled out of daylong recordings, collected from (a) populations similar to the system's original training data (North American English-learning children aged 3-36 months), (b) children learning another dialect of English (UK), and (c) slightly older children growing up in a different linguistic and socio-cultural setting (Tsimane' learners in rural Bolivia). We find reasonably high accuracy in some measures (AWC, CVC), with more problematic levels of performance in others (CTC, precision of male adults and other children). Statistical analyses do not support the view that performance is worse for children who are dissimilar from the LENA Ⓡ original training set. Whether LENA Ⓡ results are accurate enough for a given research, educational, or clinical application depends largely on the specifics at hand. We therefore conclude with a set of recommendations to help researchers make this determination for their goals.

Introduction: Metabolomics provides a view of endogenous metabolic patterns not only during plant growth, development and senescence but also in response to genetic events, environment and disease. The effects of the field environment on plant hormone-specific metabolite profiles are largely unknown. Few studies have analyzed useful phenotypes generated by introducing single or multiple gene events alongside the non-engineered wild type control at field scale to determine the robustness of the genetic trait and its modulation in the metabolome as a function of specific agroecosystem environments., Objectives: We evaluated the influence of genetic background (high polyamine lines; low methyl jasmonate line; low ethylene line; and isogenic genotypes carrying double transgenic events) and environments (hairy vetch, rye, plastic black mulch and bare soil mulching systems) on the metabolomic profile of isogenic reverse genetic mutations and selected mulch based cropping systems in tomato fruit. Net photosynthesis and fruit yield were also determined., Methods: NMR spectroscopy was used for quantifying metabolites that are central to primary metabolism. We analyzed both the first moment (means) of metabolic response to genotypes and agroecosystems by traditional univariate/multivariate methods, and the second moment (covariances) of responses by creating networks that depicted changes in correlations of paired metabolites. This particular approach is novel and was necessary because our experimental material yielded highly variable metabolic responses that could not be easily understood using the traditional analytical approaches for first moment statistics., Results: High endogenous spermidine and spermine content exhibited strong effects on amino acids, Krebs cycle intermediates and energy molecules (ADP + ATP) in ripening fruits of plants grown under different agroecosystem environments. The metabolic response to high polyamine genotypes was similar to the response to hairy vetch cover crop mulch; supported by the pattern of changes in correlation between metabolites. Changes in primary metabolites of genotypes mutated for the deficiency of ethylene or methyl jasmonate were unique under all growth conditions and opposite of high polyamine genotype results. The high polyamine trait was found to dominate the low ethylene and low jasmonate mutations under field conditions. For several metabolites low ethylene and low methyl jasmonate genotypes had an inverse relationship. Collectively, these results affirm that interactions between metabolite pathways and growth environments are affected by genotype, and influence the metabolite quality of a crop., Conclusion: This study portrays how metabolite relationships change, both in mean and in correlation, under different genotypic and environmental conditions. Although these networks are surprisingly dynamic, we also find examples of selectively conserved associations.

To examine the impact of projected climate changes on secondary succession, we exposed the same fallow soil with a common seed bank to an in situ gradient of urban to rural macroenvironments that differed in temperature and CO2 concentration ([CO2]). This gradient was established at three locations: Baltimore city center (urban), a city park on the outskirts of Baltimore (suburban), and an organic farm 87 km from the Baltimore city center site (rural). Over a five-year period, the urban site averaged 2.1 degrees C warmer and had a [CO2] that was ~20% higher than at the rural location, indicating that this gradient was a reasonable surrogate for projected changes in those variables for this century. Previous work had demonstrated that other abiotic variables measured across the transect, including tropospheric ozone and nitrogen deposition, did not differ consistently. The first year of exposure resulted in (two- to threefold) greater aboveground biomass in the urban relative to the rural site, but with uniform species composition across sites. Simple regression of abiotic variables indicated that temperature and vapor pressure deficit (VPD) were the best predictors of plant biomass among locations. Stepwise multiple regressions were also performed to analyze the effect of more than one macroenvironmental variable on total plant biomass. The combination of daily CO2 concentration and nighttime temperature explained 87% (P < 0.01) of the variability in total biomass between sites. After five years, the species demography of the plant communities had changed significantly, with a greater ratio of perennials to annuals for the urban relative to the rural location. Greater first-year biomass and litter accumulation at the urban site may have suppressed the subsequent seed germination of annual species, accelerating changes in species composition. If urban macroenvironments reflect future global change conditions, these data suggest a faster rate of secondary succession in a warmer, higher [CO2] world.

To examine the impact of climate change on vegetative productivity, we exposed fallow agricultural soil to an in situ temperature and CO2 gradient between urban, suburban and rural areas in 2002. Along the gradient, average daytime CO2 concentration increased by 21% and maximum (daytime) and minimum (nighttime) daily temperatures increased by 1.6 and 3.3 degrees C, respectively in an urban relative to a rural location. Consistent location differences in soil temperature were also ascertained. No other consistent differences in meteorological variables (e.g. wind speed, humidity, PAR, tropospheric ozone) as a function of urbanization were documented. The urban-induced environmental changes that were observed were consistent with most short-term (approximately 50 year) global change scenarios regarding CO2 concentration and air temperature. Productivity, determined as final above-ground biomass, and maximum plant height were positively affected by daytime and soil temperatures as well as enhanced [CO2], increasing 60 and 115% for the suburban and urban sites, respectively, relative to the rural site. While long-term data are needed, these initial results suggest that urban environments may act as a reasonable surrogate for investigating future climatic change in vegetative communities.

Strawberry (Fragaria x ananassa) plants were grown in field plots at the current ambient [CO(2)], and at ambient + 300 and ambient + 600 mumol mol(-1) [CO(2)]. Approximately weekly measurements were made of single leaf gas exchange of upper canopy leaves from early spring through fall of two years, in order to determine the temperature dependence of the stimulation of photosynthesis by elevated [CO(2)], whether growth at elevated [CO(2)] resulted in acclimation of photosynthesis, and whether any photosynthetic acclimation was reduced when fruiting created additional demand for the products of photosynthesis. Stimulation of photosynthetic CO(2) assimilation by short-term increases in [CO(2)] increased strongly with measurement temperature. The stimulation exceeded that predicted from the kinetic characteristics of ribulose-1,5-bisphosphate carboxylase at all temperatures. Acclimation of photosynthesis to growth at elevated [CO(2)] was evident from early spring through summer, including the fruiting period in early summer, with lower rates under standard measurement conditions in plants grown at elevated [CO(2)]. The degree of acclimation increased with growth [CO(2)]. However, there were no significant differences between [CO(2)] treatments in total nitrogen per leaf area, and photosynthetic acclimation was reversed one day after switching the [CO(2)] treatments. Tests showed that acclimation did not result from a limitation of photosynthesis by triose phosphate utilization rate at elevated [CO(2)]. Photosynthetic acclimation was not evident during dry periods in midsummer, when the elevated [CO(2)] treatments conserved soil water and photosynthesis declined more at ambient than at elevated [CO(2)]. Acclimation was also not evident during the fall, when plants were vegetative, despite wet conditions and continued higher leaf starch content at elevated [CO(2)]. Stomatal conductance responded little to short-term changes in [CO(2)] except during drought, and changed in parallel with photosynthetic acclimation through the seasons in response to the long-term [CO(2)] treatments. The data do not support the hypothesis that source-sink balance controls the seasonal occurrence of photosynthetic acclimation to elevated [CO(2)] in this species.

The relative stimulation of photosynthesis by elevated carbon dioxide in C(3) species normally increases strongly with increasing temperature. This results from the kinetic characteristics of Rubisco, and has potentially important implications for responses of vegetation to increasing atmospheric carbon dioxide. It is often assumed that because Rubisco characteristics are conservative, all C(3) species have the same temperature dependence of the response of photosynthesis to elevated carbon dioxide. However, in this field study of Taraxacum officinale, there were no significant differences in the relative stimulation of photosynthesis by elevated carbon dioxide among days with temperatures ranging from 15 to 34 degrees C. Nevertheless, short-term measurements indicated a strong temperature dependence of the stimulation. This suggested that acclimation to temperature caused the lack of variation in the seasonal data. Experiments in controlled environments indicated that complete acclimation of the relative stimulation of photosynthesis by elevated carbon dioxide occurred for growth temperatures of 10 - 25 degrees C. The apparent specificity of Rubisco for carbon dioxide relative to oxygen at 15 degrees C, as assayed in vivo by measurements of the carbon dioxide concentration at which carboxylation equalled oxygenation, also varied with growth temperature. Changes in the apparent specificity of Rubisco accounted for the acclimation of the temperature dependence of the relative stimulation of photosynthesis by elevated carbon dioxide. It is premature to conclude that low temperatures will necessarily reduce the relative stimulation of photosynthesis caused by rising atmospheric carbon dioxide.

To determine how parameters of a Farquhar-type photosynthesis model varied with measurement temperature and with growth temperature, eight cool and warm climate herbaceous crop and weed species were grown at 15 and 25 degrees C and single leaf carbon dioxide and water vapor exchange rates were measured over the range of 15 - 35 degrees C. Photosynthetic parameters examined were the initial slope of the response of assimilation rate (A) to substomatal carbon dioxide concentration (C(i)), A at high C(i), and stomatal conductance. The first two measurements allow calculation of V(Cmax), the maximum rate of carboxylation of ribulose bisphosphate carboxylase and J(max), the maximum rate of photosynthetic electron transport, of Farquhar-type photosynthesis models. In all species, stomatal conductance increased exponentially with temperature over the whole range of 15 - 35 degrees C, even when A decreased at high measurement temperature. There were larger increases in conductance over this temperature range in the warm climate species (4.3 x) than in the cool climate species (2.5 x). The initial slope of A vs. C(i) exhibited an optimum temperature which ranged from 20 to 30 degrees C. There was a larger increase in the optimum temperature of the initial slope at the warmer growth temperature in the cool climate species than in the warm climate species. The optimum temperature for A at high C(i) ranged from 25 to 30 degrees C among species, but changed little with growth temperature. The absolute values of both the initial slope of A vs. C(i) and A at high C(i) were increased about 10% by growth at the warmer temperature in the warm climate species, and decreased about 20% in the cool climate species. The ratio of J(max) - V(Cmax) normalized to 20 degrees C varied by more than a factor of 2 across species and growth temperatures, but differences in the temperature response of photosynthesis were more related to variation in the temperature dependencies of J(max) and V(Cmax) than to the ratio of their normalized values.

Photosynthetic rates and photosynthate partitioning were studied in three-week-old soybean [Glycine max (L.) Merr. cv. Williams] plants exposed to either ambient (35 Pa) or elevated (70 Pa) CO2 in controlled environment chambers. Ambient CO2-grown plants also were given a single 24 h treatment with 70 Pa CO2 1 d prior to sampling. Photosynthetic rates of ambient CO2-grown plants initially increased 36% when the measurement CO2 was doubled from 35 to 70 Pa. Photosynthetic rates of the third trifoliolate leaf, both after 1 and 21 d of elevated CO2 treatment, were 30 to 45% below those of ambient CO2-grown plants when measured at 35 Pa CO2. These reduced photosynthetic rates were not due to increased stomatal resistance and were observed for 2 to 8 h after plants given 1 d of CO2 enrichment were returned to ambient CO2. Initial and total ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) activities, percent activation, Rubisco protein, soluble protein and leaf chlorophyll content were similar in all CO2 treatments. Quantum yields of photosynthesis, determined at limiting irradiances and at 35 Pa CO2, were 0.049±0.003 and 0.038±0.005 mol CO2 fixed per mol quanta for ambient and elevated CO2-grown plants, respectively (p<0.05). Leaf starch and sucrose levels were greater in plants grown at 70 than at 35 Pa CO2. Starch accumulation rates during the day were greater in ambient CO2-grown plants than in plants exposed to elevated CO2 for either 1 or 21 d. However, the percentage of C partitioned to starch relative to total C fixed was unaffected by 1 d of CO2 enrichment. The above results showed that both photosynthetic and starch accumulation rates of soybean leaflets measured at 35 Pa CO2 were temporarily reduced after 1 and 21 d of CO2 enrichment. The biochemical mechanism affecting these responses was not identified.

This study investigated the basis of the negative relationship between leaf size and photosynthetic rate per unit of area among five cultivars of soybeans. Exposure of developing mainstem leaves to light, and sizes and light saturated photosynthesis rates of those leaves at maturity were compared in cultivars grown in field plots for two years at Beltsville, Maryland, USA. Plants were grown both in stands at 2.5 cm by 1 m spacing and as isolated plants. While cultivar differences in leaf size were large and consistent in both planting arrangements, significant cultivar differences in light saturated photosynthetic rates were found only in plants grown in stands. Similarly, leaf size was significantly correlated with specific leaf weight only for plants grown in stands. The mainstem apex and developing mainstem leaves experienced more severe shading in large-leaved cultivars than in small-leaved cultivars when plants were grown in stands. Thus, cultivar differences in photosynthetic capacity were probably a consequence of differences in the exposure of developing leaves to light.

Daytime rates of net photosynthesis of upper canopy leaflets of soybeans were compared on 17 days for leaflets exposed to air at the ambient humidity and at a higher humidity. Leaflets at the higher humidity had higher rates of net photosynthesis on 16 of the 17 days. The daily total of net photosynthesis of leaflets at the higher humidity was on average 1.32 times that for leaflets at ambient humidity. A strong limitation of net photosynthesis by ambient humidity was found throughout the growing season.

Photosynthetic characteristics at high measurement irradiance were analyzed for single leaves of two C3 and one C4 species grown under twenty one combinations of irradiance level, irradiance duration, and air temperature in order to test the idea that photosynthetic characteristies developed by leaves in different environments are controlled by the daily amount of photosynthesis. Photosynthetic rates per unit area and mesophyll conductances at 25°C and air levels of CO2 and O2, and parameters for two photosynthesis models were used to characterize the photosynthetic properties of the leaves. Leaves with highest values of the photosynthetic parameters for each species were often developed in environments with irradiance levels below saturation for photosynthesis, and with only 12 hours of irradiance per day. Lower air temperature during growth increased the photosynthetic characteristics for a given irradiance regime. Photosynthetic characteristics had higher correlation coefficients with daily photosynthesis of mature leaves divided by 24-hour leaf elongation rates of young leaves, than with daily photosynthesis alone, indicating that photosynthetic characteristics may be related to a balance between photosynthesis and leaf expansion.

Net photosynthetic rates and mesophyll conductances at 25 °C at light saturation and air levels of carbon dioxide and oxygen were measured on recently fully expanded leaflets of second trifoliolate leaves of soybeans (Glycine max cv. Kent). Plants were grown outdoors in pots at Beltsville, Maryland with 14 planting times from May through August, 1983. Air temperature and humidity, and photosynthetically active radiation (PAR) were measured for the expansion periods of the second trifoliolate leaves. Rates of net photosynthesis ranged from 24 to 33 μmol m(-2) s(-1), and mesophyll conductances from 0.24 to 0.35 cm s(-1) for the different planting dates. Mean 24-h air temperatures ranged from 20.6 to 29.0 °C, and mean daily PAR ranged from 29.4 to 58.4 mol m(-2) d(-1) for the leaf expansion periods. There was a positive relationship between photosynthetic characteristics and PAR during leaf expansion, and a negative relationship between photosynthetic characteristics and leaf expansion rates, with 96% of the variation in photosynthetic characteristics accounted for by these two variables. Leaf expansion rates were highly correlated with air temperature.

Responses to humidity of net photosynthesis and leaf conductance of single attached leaves were examined in populations of herbs from wet soil sites in Beltsville, Maryland and Davis, California, USA. Plants were grown in controlled environments under three conditions which differed in the magnitude of the day-night temperature difference and in daytime air saturation deficit. No population differences in response were found in Abutilon theophrasti. In Amaranthus hybridus stomatal conductance and net photosynthesis were more reduced by increasing leaf to air water vapor pressure difference (VPD) in the population from Beltsville, but only for the growth condition with a constant 25°C temperature. In Chenopodium album, stomatal conductance was more sensitive to VPD in the population from Davis, but only for the growth condition with 28/22°C day/night temperatures. Population differences in the sensitivity to VPD of leaf conductance were associated with differences in leaf area to root weight ratio. The relative reduction of net photosynthesis as VPD increased was greater than, equal to, or less than the relative decrease in substomatal carbon dioxide partial pressure. The pattern depended on species, and on growth condition. From these results one can not conclude that environmental humidity has been a strong selective force in determining sensitivity to humidity of stomatal conductance.

Water vapor was added differentially to the gas streams entering the cells of three makes of differential infrared carbon dioxide analysers. Analyser deflections were compared with those expected from dilution of the carbon dioxide by the additional gas. Tests were made at 0, 365, and 730 cm(3) m(-3) concentrations of carbon dioxide, and with the dewpoint in one cell of the analysers held constant at 15, 20, or 25°C. None of the analysers always responded in the ways predicted from dilution. The results showed that errors of a few cm(3) m(-3) could occur in estimates of carbon dioxide differentials using the theoretical correction for dilution. Furthermore the amount of error varied with the carbon dioxide range, the difference in water content, and in some cases the dewpoint range.

Photosynthetic characteristics at high measurement irradiance were analyzed for single leaves of two C3 and one C4 species grown under twenty one combinations of irradiance level, irradiance duration, and air temperature in order to test the idea that photosynthetic characteristics developed by leaves in different environments are controlled by the daily amount of photosynthesis. Photosynthetic rates per unit area and mesophyll conductances at 25°C and air levels of CO2 and O2, and parameters for two photosynthesis models were used to characterize the photosynthetic properties of the leaves. Leaves with highest values of the photosynthetic parameters for each species were often developed in environments with irradiance levels below saturation for photosynthesis, and with only 12 hours of iradiance per day. Lower air temperature during growth increased the photosynthetic characteristics for a given irradiance regime. Photosynthetic characteristics had higher correlation coefficients with daily photosynthesis of mature leaves divided by 24-hour leaf elongation rates of young leaves, than with daily photosynthesis alone, indicating that photosynthetic characteristics may be related to a balance between photosynthesis and leaf expansion.

Large underestimates of the limitation to photosynthesis imposed by stomata can occur because of an error in the standard method of calculating average substomatal pressures of carbon dioxide when heterogeneity of those pressures occurs across a leaf surface. Most gas exchange data supposedly indicating nonstomatal inhibition of photosynthesis by water stress could have this error. However, if no stomatal closure occurs, any reduction in photosynthesis must be due to nonstomatal inhibition of photosynthesis. Net carbon dioxide exchange rates and conductances to water vapor were measured under field conditions in upper canopy leaves of tomato plants during two summers in Beltsville, Maryland, USA. Comparisons were made near midday at high irradiance between leaflets in air with the ambient water vapor content and in air with a higher water content. The higher water content, which lowered the leaf to air water vapor pressure difference (VPD), was imposed either one half hour or several hours before measurements of gas exchange. In both seasons, and irrespective of the timing of the imposition of different VPDs, net photosynthesis increased 60% after decreasing the VPD from 3 to 1 kPa. There were no differences in leaf conductance between leaves at different VPDs, thus transpiration rates were threefold higher at 3 than at 1 kPa VPD. It is concluded that nonstomatal inhibition of photosynthesis did occur in these leaves at high transpiration rate.

Light-saturated photosynthetic rates at air levels of carbon dioxide were measured about weekly in upper canopy leaves of two soybean cultivars grown at stand densities of 40 and 100 plants per square meter. Early in the season, when leaf area indices differed between stand densities, plants of both cultivars grown at high stand density had photosynthetic rates which averaged 23% lower than plants at low stand density. Later in the season, when there were no differences in leaf area index between stand densities, there were no differences in photosynthetic rates in the cultivar Kent, but rate differences of about 14% persisted in the cultivar Williams. In Williams mainstem leaves emerged into full sunlight later in their development at high than at low stand density. In both cultivars the oldest fully exposed leaves were photosynthetically immature for much of the season, as higher rates could be achieved by lower leaves which were shaded in situ. The results identify shading of young developing leaves and photosynthetic immaturity of fully exposed leaves as factors limiting canopy photosynthesis in soybeans, and indicate cultivar differences in how much high stand density reduces photosynthetic capacity.

Net photosynthetic rates and mesophyll conductances were measured under standardized conditions for leaves of two C3 and one C4 annual species grown at temperatures of 20 to 32°C. Plants were grown with varying day and night temperatures, and also at constant temperatures equal to all the day and night temperatures used. Plants were grown with 8, 12, and 16 hours of light per day. This design allowed determination of whether photosynthetic characteristics were best correlated with day, night, mean, or time-weighted mean temperatures, The results showed that for Glycine max (L.) Merr. (C3) night temperature was most important in determining photosynthetic characteristics, while in Helianthus annuus L. (C3) and Amaranthus hypochondriacus L. (C4) the time-weighted mean temperature was most important. The results for all species were consistent with the hypothesis that development of photosynthetic characteristics is related to a balance between the rate of leaf expansion and the rate of photosynthesis under the growth conditions.