Effect of symbiotic N2 fixation on leaf protein contents, protein degradation and nitrogen resorption during leaf senescence in temperate deciduous woody species.

Nitrogen (N) resorption from senescing leaves enables plants to reuse N, making them less dependent on current N uptake from the environment, leading to higher fitness, particularly under low N supply. Species that form a symbiotic association with N2-fixing bacteria have not evolved proficient N resorption, i.e., they retain more N in the senesced leaves than non-N2-fixing species. However, the physiological mechanism underlying the difference is still unknown. Metabolic and structural protein contents in green and senesced leaves, as well as protein degradation during leaf senescence-a critical initial process for subsequent N resorption-were determined in four N2-fixing legumes and in four non-N2-fixers. The metabolic proteins were highly degraded in legumes and to a lesser extent in nonlegumes. Nonetheless, legumes retained more metabolic proteins in their senesced leaves than nonlegumes, because symbiotic N2 fixation improved the metabolic protein content in green leaves. Symbiotic N2 fixation did not change the structural protein content in green leaves. The structural proteins were moderately degraded in nonlegumes, and almost undegraded in legumes, and more structural proteins remained in the senesced leaves of legumes than in those of nonlegumes. The higher metabolic and structural protein contents in the senesced leaves of N2-fixing legumes properly explained the less proficient N resorption. This is an important step in unraveling molecular mechanisms of different N conservation strategies among plant functional types.

Leaf productivity and persistence have been improved during soybean (Glycine max) domestication and evolution.

Artificial and natural selection improved the leaf photosynthetic rate of soybean (Glycine max (L.) Merr. subsp. max). This change may be accompanied by unconscious, undesired changes in other leaf traits, such as decreased leaf persistence, if a finite resource was shared by two or more leaf traits-i.e., if they were traded off. We investigated leaf traits related to productivity (leaf photosynthetic rate, leaf nitrogen content, and stomatal conductance) and those related to persistence (leaf lifespan, leaf mass per unit area, and leaf bulk density) in one wild soybean line and three domesticated soybean lines (a landrace, an old cultivar, and a modern cultivar) in a three year experiment. Some leaf trait values increased while others did not change significantly during domestication and evolution. These results indicate that productivity-related leaf traits and persistence-related leaf traits are not negatively correlated. It was also found that the changes in productivity-related leaf traits and persistence-related leaf traits occurred at different times. Our results indicate that the productivity-related leaf traits and the persistence-related leaf traits have been independently selected for in soybean, and that they were not traded off. Combination of high leaf productivity and high leaf persistence would lead to higher lifetime leaf carbon gain and increased soybean yield.

Species-specific nitrogen resorption proficiency in legumes and nonlegumes.

Nitrogen (N) resorption from senescing leaves enables plants to reuse N, thereby making them less dependent on current N uptake from the environment. Therefore, N resorption is important for survival and fitness, particularly for plants growing under low N supply. We studied N resorption from senescing leaves of 25 legumes and 25 nonlegumes in a temperate region of Japan to test the hypothesis that high N resorption has not evolved in legumes that fix atmospheric N2. The extent of N resorption was quantified by N resorption proficiency that is measured as the level to which leaf N concentration was reduced during senescence, i.e., the lower the senesced leaf N concentration, the lower the N loss through leaf fall and higher the N resorption proficiency. In support of the hypothesis, senesced leaf N concentration was higher in legumes than in nonlegumes, but there was considerable overlap between the groups. The higher senesced leaf N concentration of legumes was associated with a lower proportion of leaf N resorbed during senescence, particularly in species with higher leaf N concentrations. According to a hierarchical partitioning analysis, there was a large contribution of species to the total variance in the senesced leaf N concentration as opposed to a minor contribution of functional group (legume/nonlegume). This study reveals that legumes are not proficient at resorbing N from senescing leaves but that N2-fixation might not be the single most important determinant of N resorption.

Inconsistent intraspecific pattern in leaf life span along nitrogen-supply gradient.

PREMISE OF THE STUDY: Leaf life span (LLS) has long been hypothesized to plastically increase with decreasing nitrogen (N) supply from soil to maximize N retention, carbon assimilation, and fitness; however, accumulating evidence shows no consistent trend. The apparent inconsistencies are explained by a recent model that assumes LLS has a hump-shaped quadratic response to the N-supply gradient. The available evidence mostly originates from comparisons of LLS at only two levels of N availability, and the hypothesis remains unanswered. METHODS: We investigated LLS of two asteraceous forbs (Adenocaulon himalaicum and Xanthium canadense) experimentally grown at eight levels of N supply, which covered a range of N supply in their natural habitats. We additionally conducted a literature search to retrieve studies reporting LLS response along an N-supply gradient. KEY RESULTS: The LLS of neither species showed a hump-shaped response along the N-supply gradient. Past studies examining the LLS of an aquatic forb and terrestrial shrubs and trees along the N-supply gradient (more than four levels of N supply) also refuted the hypothesis. CONCLUSIONS: The LLS of a single species exhibited neither an increase nor a hump-shaped response to decreased N supply in a variety of life forms. Comparisons at only a few N levels are misleading with regard to LLS response to N supply.

A meta-analysis of leaf nitrogen distribution within plant canopies.

BACKGROUND AND AIMS: Leaf nitrogen distribution in the plant canopy is an important determinant for canopy photosynthesis. Although the gradient of leaf nitrogen is formed along light gradients in the canopy, its quantitative variations among species and environmental responses remain unknown. Here, we conducted a global meta-analysis of leaf nitrogen distribution in plant canopies. METHODS: We collected data on the nitrogen distribution and environmental variables from 393 plant canopies (100, 241 and 52 canopies for wheat, other herbaceous and woody species, respectively). KEY RESULTS: The trends were clearly different between wheat and other species; the photosynthetic nitrogen distribution coefficient (Kb) was mainly determined by leaf area index (LAI) in wheat, whereas it was correlated with the light extinction coefficient (KL) and LAI in other species. Some other variables were also found to influence Kb We present the best equations for Kb as a function of environmental variables and canopy characteristics. As a more simple function, Kb = 0·5KL can be used for canopies of species other than wheat. Sensitivity analyses using a terrestrial carbon flux model showed that gross primary production tended to be more sensitive to the Kb value especially when nitrogen content of the uppermost leaf was fixed. CONCLUSION: Our results reveal that nitrogen distribution is mainly driven by the vertical light gradient but other factors such as LAI also have significant effects. Our equations contribute to an improvement in the projection of plant productivity and cycling of carbon and nitrogen in terrestrial ecosystems.

Leaf dynamics in growth and reproduction of Xanthium canadense as influenced by stand density.

BACKGROUND AND AIMS: Leaf longevity is controlled by the light gradient in the canopy and also by the nitrogen (N) sink strength in the plant. Stand density may influence leaf dynamics through its effects on light gradient and on plant growth and reproduction. This study tests the hypothesis that the control by the light gradient is manifested more in the vegetative period, whereas the opposite is true when the plant becomes reproductive and develops a strong N sink. METHODS: Stands of Xanthium canadense were established at two densities. Emergence, growth and death of every leaf on the main stem and branches, and plant growth and N uptake were determined from germination to full senescence. Mean residence time and dry mass productivity were calculated per leaf number, leaf area, leaf mass and leaf N (collectively termed 'leaf variables') in order to analyse leaf dynamics and its effect on plant growth. KEY RESULTS: Branching and reproductive activities were higher at low than at high density. Overall there was no significant difference in mean residence time of leaf variables between the two stands. However, early leaf cohorts on the main stem had a longer retention time at low density, whereas later cohorts had a longer retention time at high density. Branch leaves emerged earlier and tended to live longer at low than at high density. Leaf efficiencies, defined as carbon export per unit investment of leaf variables, were higher at low density in all leaf variables except for leaf number. CONCLUSIONS: In the vegetative phase of plant growth, the light gradient strongly controls leaf longevity, whereas later the effects of branching and reproductive activities become stronger and over-rule the effect of light environment. As leaf N supports photosynthesis and also works as an N source for plant development, N use is pivotal in linking leaf dynamics with plant growth and reproduction.

Seasonal change in light partitioning among coexisting species of different functional groups along elevation gradient in subalpine moorlands.

Species niches are expected to differ between different functional groups and between species with different functional traits. However, it is still unclear how functional traits contribute to niche separation between species coexisting in a community and between sites along environmental gradients. We studied seasonal changes in light partitioning among coexisting species belonging to different functional groups in moorland plant communities at different altitudes. We estimated the lifetime light absorption per unit invested leaf biomass (ΦLleafmass) as a measure of the benefit/cost ratio of light acquisition. Evergreen species absorbed more light in spring, whereas deciduous species absorbed more light in summer. A similar tradeoff was also found between short and tall species within each functional group. As a result, evergreen and shorter species had comparable ΦLleafmass values to those of deciduous and taller species. Evergreen species had higher ΦLleafmass at higher altitudes relative to deciduous species, suggesting that evergreen habit is more advantageous for the lifetime light interception at higher altitudes. Our results demonstrate that phenological tradeoffs for light partitioning can contribute to the coexistence of species with different functional traits. Our results also reveal that the most advantageous traits differ depending on environment.

Mean residence time of leaf number, area, mass, and nitrogen in canopy photosynthesis.

Mean residence time (MRT) of plant nitrogen (N), which is an indicator of the expected length of time N newly taken up is retained before being lost, is an important component in plant nitrogen use. Here we extend the concept MRT to cover such variables as leaf number, leaf area, leaf dry mass, and nitrogen in the canopy. MRT was calculated from leaf duration (i.e., time integral of standing amount) divided by the total production of leaf variables. We determined MRT in a Xanthium canadense stand established with high or low N availability. The MRT of leaf number may imply longevity of leaves in the canopy. We found that the MRT of leaf area and dry mass were shorter than that of leaf number, while the MRT of leaf N was longer. The relatively longer MRT of leaf N was due to N resorption before leaf shedding. The MRT of all variables was longer at low N availability. Leaf productivity is the rate of canopy photosynthesis per unit amount of leaf variables, and multiplication of leaf productivity by MRT gives the leaf photosynthetic efficiency (canopy photosynthesis per unit production of leaf variables). The photosynthetic efficiency of leaf number implies the lifetime carbon gain of a leaf in the canopy. The analysis of plant-level N use efficiency by evaluating the N productivity and MRT is a well-established approach. Extension of these concepts to leaf number, area, mass, and N in the canopy will clarify the underlying logic in the study of leaf life span, leaf area development, and dry mass and N use in canopy photosynthesis.

Demand and supply of N in seed production of soybean (Glycine max) at different N fertilization levels after flowering.

Nitrogen (N) has been suggested as a determinant of seed production especially in species with high seed N content. Assuming that seed yield was determined as the balance between N demand and supply for seed production, we studied the effect of N fertilization after flowering on soybean (Glycine max L. Merr.) yield. Seed N concentration was nearly constant irrespective of N fertilization, indicating that seed production was proportional to the amount of N available for seed growth. N demand for seed production was analyzed as the product of seed number, the rate of N filling in individual seeds, and the length of the reproductive period. N fertilization increased seed number and the reproductive period, but did not influence the N filling rate. Seed number was positively correlated with dry mass productivity after flowering. Three N sources were distinguished: mineral N uptake, symbiotic N(2) fixation and N remobilization from vegetative body. N fertilization increased N uptake and N remobilization, but lowered N(2) fixation. We concluded that N availability in the reproductive period determined seed yield directly through increasing N supply for seed growth and indirectly through increasing seed N demand with enhanced plant dry mass productivity.

Supra-optimal leaf area index of a temperate liana Pueraria lobata for competition with Solidago altissima at the expense of canopy photosynthesis.

Leaf area index (LAI) measured for the actual plant canopy is higher than the LAI that maximizes canopy photosynthesis (referred to as optimal LAI) because each individual can increase its light interception by unilaterally producing more leaf area. The LAI of an invasive woody vine Pueraria lobata (kudzu) is one of the highest among plant species, sometimes attaining nearly 10 m2 m-2. The high LAI casts heavy shade over neighboring plants, making their survival difficult. Interesting to note is that the high LAI also increases self-shading, thereby decreasing its own photosynthesis processes. In the present study, the influences of the high LAI on light interception and canopy photosynthesis, as well as on the inter-specific competition was investigated on a roadside P. lobata vegetation in Japan. With the aid of a canopy photosynthesis model and a sensitivity analysis, it was revealed that the actual LAI was 2.2-3.0 times higher than the optimal LAI for maximizing canopy photosynthesis. In the following year, a field experiment was conducted where a nearly optimal LAI was maintained throughout the growth period by regularly clipping the leaves of P. lobata. Ultimately, the field results revealed that even with a nearly optimal LAI, P. lobata was outcompeted by a competing alien weed, Solidago altissima (tall goldenrod). These results indicate that the supra-optimal leaf area, rather than maximum canopy carbon gain, makes P. lobata the dominating species in light-competing environments.

Effects of elevated CO2 concentration on seed production in C3 annual plants.

The response of seed production to CO(2) concentration ([CO(2)]) is known to vary considerably among C(3) annual species. Here we analyse the interspecific variation in CO(2) responses of seed production per plant with particular attention to nitrogen use. Provided that seed production is limited by nitrogen availability, an increase in seed mass per plant results from increase in seed nitrogen per plant and/or from decrease in seed nitrogen concentration ([N]). Meta-analysis reveals that the increase in seed mass per plant under elevated [CO(2)] is mainly due to increase in seed nitrogen per plant rather than seed [N] dilution. Nitrogen-fixing legumes enhanced nitrogen acquisition more than non-nitrogen-fixers, resulting in a large increase in seed mass per plant. In Poaceae, an increase in seed mass per plant was also caused by a decrease in seed [N]. Greater carbon allocation to albumen (endosperm and/or perisperm) than the embryo may account for [N] reduction in grass seeds. These differences in CO(2) response of seed production among functional groups may affect their fitness, leading to changes in species composition in the future high-[CO(2)] ecosystem.

Light interception in species with different functional groups coexisting in moorland plant communities.

Competition for light is one of the most essential mechanisms affecting species composition. It has been suggested that similar light acquisition efficiency (Φ(mass), absorbed photon flux per unit aboveground mass) may contribute to species coexistence in multi-species communities. On the other hand, it is known that traits related with light acquisition vary among functional groups. We studied whether Φ(mass) was similar among species with different functional groups coexisting in moorland communities. We conducted stratified clipping in midsummer when the stand biomass reached a maximum. Light partitioning among species was estimated using a model accounting for both direct and diffuse light. Evergreen species were found to have a significantly lower Φ(mass) than deciduous species, which resulted from their lower absorbed photon flux per unit leaf area and lower specific leaf area. Shrubs had a smaller leaf mass fraction, but their Φ(mass) was not lower than that of herbs because they had a higher leaf position due to the presence of wintering stems. Species with vertical leaves had a higher Φ(mass) than those with horizontal leaves despite vertical leaves being a decided disadvantage in terms of light absorption. This higher Φ(mass) was achieved by a greater leaf height in species with vertical leaves. Our results clearly demonstrate that light acquisition efficiency was different among the functional groups. However, the trend observed is not necessarily the same as that expected based on prior knowledge, suggesting that disadvantages in some traits for light acquisition efficiency are partly compensated for by other traits.

Does leaf shedding increase the whole-plant carbon gain despite some nitrogen being lost with shedding?

When old leaves are shed, part of the nitrogen in the leaf is retranslocated to new leaves. This retranslocation will increase the whole-plant carbon gain when daily C gain : leaf N ratio (daily photosynthetic N-use efficiency, NUE) in the old leaf, expressed as a fraction of NUE in the new leaf, becomes lower than the fraction of leaf N that is resorbed before shedding (R(N)). We examined whether plants shed their leaves to increase the whole-plant C gain in accord with this criterion in a dense stand of an annual herb, Xanthium canadense, grown under high (HN) and low (LN) nitrogen availability. The NUE of a leaf at shedding expressed as a fraction of NUE in a new leaf was nearly equal to the R(N) in the LN stand, but significantly lower than the R(N) in the HN stand. Thus shedding of old leaves occurred as expected in the LN stand, whereas in the HN stand, shedding occurred later than expected. Sensitivity analyses showed that the decline in NUE of a leaf resulted primarily from a reduction in irradiance in the HN stand. On the other hand, it resulted from a reduction in irradiance and also in light-saturated photosynthesis : leaf N content ratio (potential photosynthetic NUE) in the LN stand.

Leaf shedding increases the photosynthetic rate of the canopy in N2-fixing and non-N2-fixing woody species.

It has long been hypothesized that timing of leaf shedding is critical for plant fitness but there is little experimental evidence to support the hypothesis. According to an optimality theory, shedding of old leaves increases canopy photosynthesis despite some nitrogen (N) being lost as litterfall, when the ratio of daily photosynthesis to leaf N (N-use efficiency, ε) in old leaves, expressed as a fraction of ε in new leaves, becomes lower than the fraction of leaf N that is resorbed before shedding (RN). This was shown to be true for N-poor plants but not for N-rich plants in a pot experiment; however, the use of planting pots imposes a variety of physical, chemical and biological constraints that could change the experimental results. Here we conducted a 3-year field survey in a cool temperate deciduous forest to examine whether Alnus sieboldiana Matsum. (N2-fixing) and Carpinus tschonoskii Maxim. (non-N2-fixing) shed their leaves to increase canopy photosynthesis in accord with the above criterion. These species often grow sympatrically and were chosen as representatives of N-rich and N-poor plants, respectively. Overall, daily photosynthesis decreased with leaf age, accompanied by small changes in leaf N, resulting in a decrease in ε. In both species, ε of leaves at shedding expressed as a fraction of ε in new leaves was nearly equal to RN in all years, implying that the old leaves were shed to increase canopy photosynthesis. Our results, together with those of previous field surveys, suggested that the timing of leaf shedding is explained by N use in maximizing canopy photosynthesis across broad groups of species.

Crossover from localized to itinerant states in hydrocarbon Mott insulators.

Crossover from an itinerant state to an isolated electronic state in electron-doped polycyclic aromatic hydrocarbon (PAH) was studied for the two smallest zigzag-type molecules of naphthalene (NN) and anthracene (AN) by focusing on their 1 : 1 stoichiometry, A1(NN) and A1(AN), with alkali metals (A = K and Rb). The competition between on-site Coulombic repulsion energy (U) and bandwidth (W) was argued in terms of their magnetic and electrical properties upon lattice expansion, when A varies from K, with a smaller ionic radius, to Rb, with a larger ionic radius. The temperature-dependence of magnetic susceptibility shows a pronounced hump associated with antiferromagnetic (AFM) interactions for Rb1(NN), being similar to those of K1(NN) and K1(AN) in the earlier report. On the other hand, Rb1(AN) intriguingly exhibits paramagnetic susceptibility, observed in a nearly localized electron system, being apart from an highly correlated Mott insulating state. Crystal structural analyses of the X-ray diffraction profiles show a small difference in lattice parameters of the ab plane among K1(NN), K1(AN), and Rb1(NN), whereas Rb1(AN) exhibits a significantly larger value than those of the others, being indicative of greatly modified interaction energies. The different magnetic properties observed in Rb1(AN) are interpreted from its modified intermolecular distance. The possibility of an emergent metallic state of K1(AN) under high pressure has also been described, referring to electrical transport measured under high pressure.

Changes in leaf area, nitrogen content and canopy photosynthesis in soybean exposed to an ozone concentration gradient.

Influences of ozone (O3) on light-saturated rates of photosynthesis in crop leaves have been well documented. To increase our understanding of O3 effects on individual- or stand level productivity, a mechanistic understanding of factors determining canopy photosynthesis is necessary. We used a canopy model to scale photosynthesis from leaf to canopy, and analyzed the importance of canopy structural and leaf ecophysiological characteristics in determining canopy photosynthesis in soybean stands exposed to 9 concentrations of [O3] (37-116 ppb; 9-h mean). Light intensity and N content peaked in upper canopy layers, and sharply decreased through the lower canopy. Plant leaf area decreased with increasing [O3] allowing for greater light intensity to reach lower canopy levels. At the leaf level, light-saturated photosynthesis decreased and dark respiration increased with increasing [O3]. These data were used to calculate daily net canopy photosynthesis (Pc). Pc decreased with increasing [O3] with an average decrease of 10% for an increase in [O3] of 10 ppb, and which was similar to changes in above-ground dry mass production of the stands. Absolute daily net photosynthesis of lower layers was very low and thus the decrease in photosynthesis in the lower canopy caused by elevated [O3] had only minor significance for total canopy photosynthesis. Sensitivity analyses revealed that the decrease in Pc was associated with changes in leaf ecophysiology but not with decrease in leaf area. The soybean stands were very crowded, the leaves were highly mutually shaded, and sufficient light for positive carbon balance did not penetrate to lower canopy leaves, even under elevated [O3].

Reproductive yield of individuals competing for light in a dense stand of an annual, Xanthium canadense.

In a dense stand, individuals compete with each other for resources, especially for light. Light availability decreases with increasing depth in the canopy, thus light competition becoming stronger with time in the vegetative phase. In the reproductive phase, on the other hand, leaves start senescing, and the light environment, particularly of smaller individuals, will be improved. To study the effect of change in light climate on reproduction of individuals, we established an experimental stand of an annual, Xanthium canadense, and assessed temporal changes in whole plant photosynthesis through the reproductive phase with particular reference to light availability of individuals. At flowering, 83% of individuals were still alive, but only 27% survived to set seeds. Most of the individuals that died in the reproductive phase were smaller than those that produced seeds. Individuals that died at the early stage of the reproductive phase had a lower leaf to stem mass ratio, suggesting that the fate of individuals was determined partly by the pattern of biomass allocation in this period. At the early stage of the reproductive phase, leaf area index (LAI) of the stand was high and larger individuals had higher whole plant photosynthesis than smaller individuals. Although light availability at later stages was improved with reduction in LAI, whole plant photosynthesis was very low in all individuals due to a lower light use efficiency, which was caused by a decrease in photosynthetic N use efficiency. We conclude that light competition was still strong at the early stage of the reproductive phase and that later improvement of light availability did not ameliorate the photosynthesis of smaller individuals.

Leaf lifespan and lifetime carbon balance of individual leaves in a stand of an annual herb, Xanthium canadense.

Leaf lifespan in response to resource availability has been documented in many studies, but it still remains uncertain what determines the timing of leaf shedding. Here, we evaluate the lifetime carbon (C) balance of a leaf in a canopy as influenced by nitrogen (N) availability. Stands of Xanthium canadense were established with high-nitrogen (HN) and low-nitrogen (LN) treatments and temporal changes of C gain of individual leaves were investigated with a canopy photosynthesis model. Daily C gain of a leaf was maximal early in its development and subsequently declined. Daily C gain at shedding was nearly zero in HN, while it was still positive in LN. Sensitivity analyses showed that the decline in the daily C gain resulted primarily from the reduction in light level in HN and by the reduction in leaf N in LN. Smaller leaf size in LN than in HN led to higher light levels in the canopy, which helped leaves of the LN stand maintain for a longer period. These results suggest that the mechanism by which leaf lifespan is determined changes depending on the availability of the resource that is most limiting to plant growth.

Dynamics of leaf area and nitrogen in the canopy of an annual herb, Xanthium canadense.

We studied leaf area and nitrogen dynamics in the canopy of stands of an annual herb Xanthium canadense, grown at a high (HN)- and a low-nitorgen (LN) availability. Standing leaf area increased continuously through the vegetative growth period in the LN stand, or leveled off in the later stage in the HN stand. When scaled against standing leaf area, both production and loss rates of leaf area increased but with different patterns: the production rate was retarded, while the loss rate was accelerated, implying an upper limit of standing leaf area of the canopy. The rate of leaf-area production was higher in the HN than in the LN stand, which was caused by the higher rate of leaf production per standing leaf area as well as the greater standing leaf area in the HN stand. Although the rate of leaf-area loss was higher in the HN than in the LN stand, it was not significantly different between the two stands when compared at a common standing leaf area, suggesting involvement of light climate in determination of the leaf-loss rate. On the other hand, the rate of leaf-area loss was positively correlated with nitrogen demand for leaf area development across the two stands, suggesting that leaf loss was caused by retranslocation of nitrogen for construction of new leaves. A simple simulation model of leaf and nitrogen dynamics in the canopy showed that, at steady state, where the rate of leaf-area loss becomes equal to the production rate, the standing leaf area was still greater in the HN than in the LN stand. Similarly, when the uptake and loss of nitrogen are equilibrated, the standing nitrogen was greater in the HN than in the LN stand. These results suggest that leaf-area production is strongly controlled by nitrogen availability, while both nitrogen and light climate determine leaf-loss rates in the canopy.