Photosynthetic and Growth Responses in a Pioneer Tree (Japanese White Birch) and Competitive Perennial Weeds (Eupatorium sp.) Grown Under Different Regimes With Limited Water Supply to Waterlogging.

For a successful natural regeneration of Japanese white birch (Betula platyphylla var. japonica), competitive vegetation should be managed. Here, we clarified how soil water condition modifies the competitiveness of Japanese white birch against perennial weeds, Eupatorium species, based on an ecophysiological approach combining a glasshouse experiment and a field survey. We investigated photosynthetic and growth responses to various water regimes from water deficit to waterlogging (two times-a-week irrigation, three times-a-week irrigation, half waterlogging, and full waterlogging) in pot-grown seedlings of Japanese white birch and the competitive weed Eupatorium makinoi. The ratio of seedling height of Japanese white birch to seedling height of E. makinoi showed a decreasing trend from two times-a-week irrigation to full waterlogging, which suggests a lower competitiveness for light resource in Japanese white birch with increasing soil wetness. The maximum rate of Rubisco carboxylation (Vc,max) based on unit N was lower in waterlogging treatments than in two times- and three times-a-week irrigation in Japanese white birch, whereas E. makinoi showed the opposite response. This suggests that N partitioning into Rubisco and/or Rubisco activation might be suppressed in Japanese white birch but enhanced in E. makinoi under waterlogging. The maximum photochemical efficiency of photosystem II (Fv/Fm) was also lower in seedlings of Japanese white birch grown under waterlogging treatments. We further conducted a field survey on the relationship between Fv/Fm and topographic wetness index (TWI) in seedlings of Japanese white birch and E. glehnii (closely related to E. makinoi) naturally grown in a study site 5 years after canopy tree cutting. Lower Fv/Fm was observed in seedlings of Japanese white birch with increasing TWI, whereas no significant trend was observed in E. glehnii, in agreement with the glasshouse experiment. Thus, keeping soils not always humid might be favorable to photosynthetic performance and growth competitive ability of Japanese white birch against Eupatorium species.

Ethylenediurea (EDU) effects on Japanese larch: an one growing season experiment with simulated regenerating communities and a four growing season application to individual saplings.

Japanese larch (Larix kaempferi (Lamb.) Carr.) and its hybrid are economically important coniferous trees widely grown in the Northern Hemisphere. Ground-level ozone (O3) concentrations have increased since the pre-industrial era, and research projects showed that Japanese larch is susceptible to elevated O3 exposures. Therefore, methodologies are needed to (1) protect Japanese larch against O3 damage and (2) conduct biomonitoring of O3 in Japanese larch forests and, thus, monitor O3 risks to Japanese larch. For the first time, this study evaluates whether the synthetic chemical ethylenediurea (EDU) can protect Japanese larch against O3 damage, in two independent experiments. In the first experiment, seedling communities, simulating natural regeneration, were treated with EDU (0, 100, 200, and 400 mg L-1) and exposed to either ambient or elevated O3 in a growing season. In the second experiment, individually-grown saplings were treated with EDU (0, 200 and 400 mg L-1) and exposed to ambient O3 in two growing seasons and to elevated O3 in the succeeding two growing seasons. The two experiments revealed that EDU concentrations of 200-400 mg L-1 could protect Japanese larch seedling communities and individual saplings against O3-induced inhibition of growth and productivity. However, EDU concentrations ≤ 200 mg L-1 did offer only partial protection when seedling communities were coping with higher level of O3-induced stress, and only 400 mg EDU L-1 fully protected communities under higher stress. Therefore, we conclude that among the concentrations tested the concentration offering maximum protection to Japanese larch plants under high competition and O3-induced stress is that of 400 mg EDU L-1. The results of this study can provide a valuable resource of information for applied forestry in an O3-polluted world.

Constant ratio of Cc to Ci under various CO2 concentrations and light intensities, and during progressive drought, in seedlings of Japanese white birch.

Constant mesophyll conductance (gm), and two-resistance gm model (involved in resistances of cell wall and chloroplast), where gm reaches maximum under higher CO2 concentrations, cannot describe the phenomenon that gm decreases with increasing intercellular CO2 concentration (Ci) under relatively higher CO2 concentrations. Yin et al. (2020) proposed a gm model, according to which the ratio of chloroplastic CO2 concentration (Cc) to Ci is constant in the two-resistance gm model, which can describe the decreasing gm with increasing Ci. In the present study, we investigated the relationship between Cc and Ci in leaves of Japanese white birch by using simultaneous measurements of gas exchange and chlorophyll fluorescence under various CO2 concentrations, light intensities, and during progressive drought. Across the range of ambient CO2 from 50 to 1000 µmol mol-1, and light intensities of 50 to 2000 µmol m-2 s-1, measured under well irrigation, the ratio of Cc to Ci kept constant. During the progressive drought, overestimated Ci due to stomatal patchiness and/or cuticular transpiration was empirically corrected (threshold: stomatal conductance < 0.08 mol H2O m-2 s-1) from the A/Ci response measured under adequate irrigation. The ratio of Cc to Ci during progressive drought (predawn leaf potential reached ≈ - 2 MPa) also remained constant irrespective of soil drying rate in various pot sizes. The present study suggests the involvement of some physiologically regulative mechanisms to keep Cc:Ci ratio constant, which might act on gm in addition to the physical interaction of diffusive resistances in the cell components.

Effects of major vein blockage and aquaporin inhibition on leaf hydraulics and stomatal conductance.

The density and architecture of leaf veins determine the network and efficiency of water transport within laminae and resultant leaf gas exchange and vary widely among plant species. Leaf hydraulic conductance ( Kleaf) can be regulated by vein architecture in conjunction with the water channel protein aquaporin. However, our understanding of how leaf veins and aquaporins affect leaf hydraulics and stomatal conductance ( gs) remains poor. By inducing blockage of the major veins and inhibition of aquaporin activity using HgCl2, we examined the effects of major veins and aquaporins on Kleaf and gs in species with different venation types. A vine species, with thick first-order veins and low vein density, displayed a rapidly declined gs with high leaf water potential in response to vein blockage and a greatly reduced Kleaf and gs in response to aquaporin inhibition, suggesting that leaf aquaporins are involved in isohydric/anisohydric stomatal behaviour. Across species, the decline in Kleaf and gs due to aquaporin inhibition increased linearly with decreasing major vein density, possibly indicating that a trade-off function between vein architecture (apoplastic pathway) and aquaporin activity (cell-to-cell pathway) affects leaf hydraulics.

On the Nonmonotonic, Hormetic Photoprotective Response of Plants to Stress.

Accumulated evidence show that reactive species play a dual role in plants as well, with likely biphasic responses. This suggests that photoprotective mechanisms may also show similar patterns because they are highly related to reactive species. The nonphotochemical quenching (NPQ) is an index of heat dissipation of excitation energy in the antenna system. We present here preliminary evidence from some published studies showing significant biphasic response of NPQ to increasing doses of stress, with U-shaped or inverted U-shaped dose-response relationships, typical of hormesis. This evidence provides a remarkable perspective for designing novel studies where the fate of light energy will be seen through the lens of hormesis.

Springtime photoinhibition constrains regeneration of forest floor seedlings of Abies sachalinensis after a removal of canopy trees during winter.

A clear-cutting of canopy trees during winter often causes severe foliar damage during the following spring in forest floor seedlings of Abies sachalinensis, a typical shade-tolerant evergreen coniferous species. The maximum photochemical efficiency of photosystem II after an overnight dark adaptation showed a temporary decrease immediately before budbreak in 1-year-old shoots of A. sachalinensis seedlings grown under full sunlight in a nursery, suggesting "springtime photoinhibition" related to the phenology of evergreen coniferous species. In the field, a greater rate of canopy tree cutting during winter was associated with more severe photoinhibition in the following spring, immediately before budbreak, which subsequently resulted in a reduction in carbon gain in 1-year-old shoots, and consequently suppressed the growth of current-year shoots. Although photoinhibition under low temperature is a well-known factor to determine the survival rate of tree seedlings during winter in cool regions, the present study additionally proposes that the temporary increase in the susceptibility to photoinhibition in springtime i.e. "springtime photoinhibition" would be a constraint for the regeneration of coniferous seedlings especially when the canopy trees are removed during winter.

Canopy nitrogen distribution is optimized to prevent photoinhibition throughout the canopy during sun flecks.

As photoinhibition primarily reduces the photosynthetic light use efficiency at low light, sunfleck-induced photoinhibition might result in a fatal loss of carbon gain in the shade leaves within a canopy with barely positive carbon balance. We hypothesized that shade leaves at the lower canopy might retain a certain amount of leaf nitrogen (NL) to maintain energy consumption via electron transport, which contributes to circumventing photoinhibition during sunflecks to keep efficient utilization of low light during the rest period of daytime. We investigated excess energy production, a potential measure of susceptibility to photoinhibition, as a function of NL distribution within a Japanese oak canopy. Optimal NL distribution, which maximizes canopy carbon gain, may lead to a higher risk of photoinhibition in shade leaves during sunflecks. Conversely, uniform NL distribution would cause a higher risk of photoinhibition in sun leaves under the direct sunlight. Actual NL distribution equalized the risk of photoinhibition throughout the canopy indicated by the constant excess energy production at the highest light intensities that the leaves received. Such a homeostatic adjustment as a whole canopy concerning photoinhibition would be a key factor to explain why actual NL distribution does not maximize canopy carbon gain.

A practical approach to estimate diffusional leakages of leaf chamber of open gas exchange systems using intact leaves.

We propose a novel approach to determine the coefficient of CO2 diffusional leakage (k_CO2 ) in the chamber of a portable open gas exchange system (Li-6400, Li-Cor Inc., NE, USA) using intact leaves. Assuming that dark respiration rate (Rn ) is constant at various CO2 concentrations, at least in a short term, k_CO2 should be the value that makes the leak-corrected Rn constant. We used 2 representative tree species native to northern Japan, Japanese white birch (Betula platyphylla var. japonica) and Japanese oak (Quercus mongolica var. crispula), to determine k_CO2 , by measuring Rn at various CO2 concentrations. Irrespective of the species, k_CO2 estimated by the present approach was well correlated with the diffusional leak coefficient (k_flow ) estimated by the generally used approach that utilizes CO2 concentrations inside and outside the chamber based on thermally killed leaves at various flow rates. Furthermore, the present approach does not require the ambient CO2 concentration outside the chamber (Ca ) if Ca in the laboratory is stable, which provides a feasible way to correct the photosynthetic rate, taking diffusional leakage into account.

Overwintering evergreen oaks reverse typical relationships between leaf traits in a species spectrum.

The leaf economics spectrum has given us a fundamental understanding of the species variations in leaf variables. Across plant species, tight correlations among leaf mass per area (LMA), mass-based nitrogen (N m) and photosynthetic rate (A m) and leaf lifespan have been well known as trade-offs in leaf carbon economy. However, the regional or biome-level correlations may not be necessary to correspond with the global-scale analysis. Here, we show that almost all leaf variables in overwintering evergreen oaks in Japan were relatively well included within the evergreen-broadleaved trees in worldwide temperate forests, but N m was more consistent with that in deciduous broadleaved trees. Contrary to the universal correlations, the correlation between A m and N m among the evergreen oaks was negative and the correlation between A m and LMA disappeared. The unique performance was due to specific nitrogen allocation within leaves, i.e. the evergreen oaks with later leaf maturation had lower N m but higher nitrogen allocation to photosynthetic enzymes within leaves, to enhance carbon gain against the delayed leaf maturation and the shortened photosynthetic period due to cold winters. Our data demonstrate that correlations between leaf variables in a local scale are occasionally different from averaged global-scale datasets, because of the constraints in each biome.

Photoprotection of evergreen and drought-deciduous tree leaves to overcome the dry season in monsoonal tropical dry forests in Thailand.

In tropical dry forests, uppermost-canopy leaves of evergreen trees possess the ability to use water more conservatively compared with drought-deciduous trees, which may result from significant differences in the photoprotective mechanisms between functional types. We examined the seasonal variations in leaf gas exchange, chlorophyll fluorescence and the amounts of photosynthetic pigments within lamina of the uppermost-canopy leaves of three drought-deciduous trees (Vitex peduncularis Wall., Xylia xylocarpa (Roxb.) W. Theob., Shorea siamensis Miq.), a semi-deciduous tree (Irvingia malayana Miq.) and two evergreen trees (Hopea ferrea Lanessan and Syzygium cumini (L.) Skeels) in Thailand. Area-based maximum carbon assimilation rates (Amax) decreased during the dry season, except in S. siamensis. The electron transport rate (ETR) remained unchanged in deciduous trees, but decreased during the dry season in evergreen and semi-deciduous trees. In the principal component analysis, the first axis (Axis 1) accounted for 44.3% of the total variation and distinguished deciduous from evergreen trees. Along Axis 1, evergreen trees were characterized by a high Stern-Volmer non-photochemical quenching coefficient (NPQ), high xanthophyll cycle pigments/chlorophyll and a high de-epoxidation state of the xanthophyll cycle, whereas the deciduous trees were characterized by a high ETR, a high quantum yield of PSII (ΦPSII = (Fm(') -F)/Fm(')) and a high mass-based Amax under high-light conditions. These findings indicate that drought-deciduous trees showing less conservative water use tend to dissipate a large proportion of electron flow through photosynthesis or alternative pathways. In contrast, the evergreens showed more conservative water use, reduced Amax and ETR and enhanced NPQ and xanthophyll cycle pigments/chlorophyll during the dry season, indicating that down-regulated photosynthesis with enhanced thermal dissipation of excess light energy played an important role in photoprotection. Trees with different water uses and leaf lifespans appear to employ different photoprotective mechanisms to overcome the unfavorable dry-season drought. Our data may suggest that future changes in precipitation will strongly impinge on forest structure and functions.

Seasonal variations of gas exchange and water relations in deciduous and evergreen trees in monsoonal dry forests of Thailand.

This study compared leaf gas exchange, leaf hydraulic conductance, twig hydraulic conductivity and leaf osmotic potential at full turgor between two drought-deciduous trees, Vitex peduncularis Wall. and Xylia xylocarpa (Roxb.) W. Theob., and two evergreen trees, Hopea ferrea Lanessan and Syzygium cumini (L.) Skeels, at the uppermost canopies in tropical dry forests in Thailand. The aims were to examine (i) whether leaf and twig hydraulic properties differ in relation to leaf phenology and (ii) whether xylem cavitation is a determinant of leaf shedding during the dry season. The variations in almost all hydraulic traits were more dependent on species than on leaf phenology. Evergreen Hopea exhibited the lowest leaf-area-specific twig hydraulic conductivity (leaf-area-specific K(twig)), lamina hydraulic conductance (K(lamina)) and leaf osmotic potential at full turgor (Ψ(o)) among species, whereas evergreen Syzygium exhibited the highest leaf-area-specific K(twig), K(lamina) and Ψ(o). Deciduous Xylia had the highest sapwood-area-specific K(twig), along with the lowest Huber value (sapwood area/leaf area). More negative osmotic Ψ(o) and leaf osmotic adjustment during the dry season were found in deciduous Vitex and evergreen Hopea, accompanied by low sapwood-area-specific K(twig). Regarding seasonal changes in hydraulics, no remarkable decrease in K(lamina) and K(twig) was found during the dry season in any species. Results suggest that leaf shedding during the dry season is not always associated with extensive xylem cavitation.

Seasonal variations in water relations in current-year leaves of evergreen trees with delayed greening.

We investigated seasonal patterns of water relations in current-year leaves of three evergreen broad-leaved trees (Ilex pedunculosa Miq., Ligustrum japonicum Thunb., and Eurya japonica Thunb.) with delayed greening in a warm-temperate forest in Japan. We used the pressure-volume method to: (1) assess the extent to which seasonal variation in leaf water relations is attributable to leaf development processes in delayed greening leaves versus seasonal variation in environmental variables; and (2) investigate variation in leaf water relations during the transition from the sapling to the adult tree stage. Leaf mass per unit leaf area was generally lowest just after completion of leaf expansion in May (late spring), and increased gradually throughout the year. Osmotic potential at full turgor (Psi(o) (ft)) and leaf water potential at the turgor loss point (Psi(w) (tlp)) were highest in May, and lowest in midwinter in all species. In response to decreasing air temperature, Psi(o) (ft) dropped at the rate of 0.037 MPa degrees C(-1). Dry-mass-based water content of leaves and the symplastic water fraction of total leaf water content gradually decreased throughout the year in all species. These results indicate that reductions in the symplastic water fraction during leaf development contributed to the passive concentration of solutes in cells and the resulting drop in winter Psi(o) (ft). The ratio of solutes to water volume increased in winter in current-year leaves of L. japonicum and E. japonica, indicating that osmotic adjustment (active accumulation of solutes) also contributed to the drop in winter in Psi(o) (ft). Bulk modulus of elasticity in cell walls fluctuated seasonally, but no general trend was found across species. Over the growing season, Psi(o) (ft) and Psi(w) (tlp) were lower in adult trees than in saplings especially in the case of I. pedunculosa, suggesting that adult-tree leaves are more drought and cold tolerant than sapling leaves. The ontogenetic increase in the stress resistance of I. pedunculosa may be related to its characteristic life form because I. pedunculosa grows taller than the other species studied.

Coordination of crown structure, leaf plasticity and carbon gain within the crowns of three winter-deciduous mature trees.

We examined the vertical profiles of leaf characteristics within the crowns of two late-successional (Fagus crenata Blume and Fagus japonica Maxim.) and one early-successional tree species (Betula grossa Sieb. et Zucc.) in a Japanese forest. We also assessed the contributions of the leaves in each crown layer to whole-crown instantaneous carbon gain at midday. Carbon gain was estimated from the relationship between electron transport and photosynthetic rates. We hypothesized that more irradiance can penetrate into the middle of the crown if the upper crown layers have steep leaf inclination angles. We found that such a crown has a high whole-crown carbon gain, even if leaf traits do not change greatly with decreasing crown height. Leaf area indices (LAIs) of the two Fagus trees (5.26-5.52) were higher than the LAI of the B. grossa tree (4.50) and the leaves of the F. crenata tree were more concentrated in the top crown layers than were leaves of the other trees. Whole-crown carbon gain per unit ground area (micromol m(-2) ground s(-1)) at midday on fine days in summer was 16.3 for F. crenata, 11.0 for F. japonica, and 20.4 for B. grossa. In all study trees, leaf dry mass (LMA) and leaf nitrogen content (N) per unit area decreased with decreasing height in the crown, but leaf N per unit mass increased. Variations (plasticity) between the uppermost and lowermost crown layers in LMA, leaf N, the ratio of chlorophyll to N and the ratio of chlorophyll a to b were smaller for F. japonica and B. grossa than for F. crenata. The light extinction coefficients in the crowns were lower for the F. japonica and B. grossa trees than for the F. crenata tree. The leaf carbon isotope ratio (delta(13)C) was higher for F. japonica and B. grossa than for F. crenata, especially in the mid-crown. These results suggest that, in crowns with low leaf plasticity but steep leaf inclination angles, such as those of F. japonica and B. grossa trees, irradiance can penetrate into the middle of the crowns, thereby enhancing whole-crown carbon gain.