A mechanistic model for photosynthesis based on the multisubstrate ordered reaction of ribulose 1,5 bisphosphate carboxylase

Abstract. A mechanistic model of photosynthesis is developed based on the characteristics of ribulose 1,5-bisphosphate (RuBP) carboxylase and the assimilation of CO₂ as an ordered reaction with RuBP binding before CO₂. An equation is derived which considers the effects of light (for regeneration of RuBP) and CO₂. Taking values for the maximum turnover of RuBP carboxylase at substrate saturation, the maximum carboxylation efficiency (maximum increase in rate per increase in CO₂ concentration) and the minimum quantum requirement for the C₃ pathway, photosynthesis in the absence of photorespiration is simulated. In the model, at varying concentrations of CO₂, the efficiency of light utilization approaches a maximum value as photon flux density decreases. Similarly, with a given maximum carboxyation capacity, at varying photon flux densities the carboxylation efficiency approaches a constant maximum value (equal to V _max/ K _{m CO2}) as CO₂ is decreased. However, a decrease in the state of activation of RuBP carboxylase under low light results in a lower carboxylation efficiency. Limits on the rate of photosynthesis, as the maximum capacity for regeneration of RuBP is reduced relative to carboxylation potential, or as the maximum capacity of the carboxylase varies, are considered.     

Modelling environmental controls on ecosystem photosynthesis and the carbon isotope composition of ecosystem-respired CO2 in a coastal Douglas-fir forest

We developed and applied an ecosystem-scale model that calculated leaf CO₂ assimilation, stomatal conductance, chloroplast CO₂ concentration and the carbon isotope composition of carbohydrate formed during photosynthesis separately for sunlit and shaded leaves within multiple canopy layers. The ecosystem photosynthesis model was validated by comparison to leaf-level gas exchange measurements and estimates of ecosystem-scale photosynthesis from eddy covariance measurements made in a coastal Douglas-fir forest on Vancouver Island. A good agreement was also observed between modelled and measured δ ¹³C values of ecosystem-respired CO₂ ( δ _R). The modelled δ _R values showed strong responses to variation in photosynthetic photon flux density (PPFD), air temperature, vapour pressure deficit (VPD) and available soil moisture in a manner consistent with leaf-level studies of photosynthetic ¹³C discrimination. Sensitivity tests were conducted to evaluate the effect of (1) changes in the lag between the time of CO₂ fixation and the conversion of organic matter back to CO₂; (2) shifts in the proportion of autotrophic and heterotrophic respiration; (3) isotope fractionation during respiration; and (4) environmentally induced changes in mesophyll conductance, on modelled δ _R values. Our results indicated that δ _R is a good proxy for canopy-level C _c/ C _a and ¹³C discrimination during photosynthetic gas exchange, and therefore has several applications in ecosystem physiology.     

Photoinhibition of photosynthesis in Lemna gibba as induced by the interaction between light and temperature. I. Photosynthesis in vivo

The floating angiosperm Lemna gibba L. was exposed for 2 h to various combinations of photosynthetic photon flux densities and temperature. The extent of photoinhibition of photosynthesis was assayed by measuring the net CO₂ uptake before and after a photoinhibitory treatment, and the time course for photoinhibition was studied. It was found that the maximum quantum yield and the light-saturated rate of CO₂ uptake were affected by the interaction between light and temperature during the photoinhibitory treatment. At a constant photon flux density of 650 μmol m⁻² s⁻¹ the extent of photoinhibition increased with decreasing temperature showing that even a chilling-resistant plant like L. gibba is much more susceptible to photoinhibition at chilling temperatures. About 60% photoinhibition of the quantum yield for CO₂ uptake could be obtained either by a high photon flux density of 1 750 μmol m⁻² s⁻¹ and 25°C or by a moderate photon flux density of 650 μmol m⁻² s⁻¹ and 3°C. The time courses of recovery from 60% photoinhibition produced by either of these two treatments were similar, indicating that the nature of the photoinhibition was intrinsically similar. The extent of photoinhibition was related to the amount of light absorbed in excess to what could be handled by photosynthesis at that temperature. The vital importance of photosynthesis in alleviating photoinhibition is discussed.     

Nitrogen allocation in response to partial shading of a plant: Possible mechanisms

The significance of photosynthetic and transpiration rates for the perception by plants of light gradients in leaf canopies has been investigated with regard to nitrogen allocation and re-allocation. A gradient of photon flux density (PFD) over a plant's foliage was simulated by shading one leaf of a pair of primary leaves of bean ( Phaseolus vulgaris L. cv. Rentegever). Photosynthetic rate was manipulated independently of PFD and, to some extent, also of transpiration, by subjecting the leaf to different CO₂ concentrations. Transpiration rate was changed independently of PFD and photosynthetic rate by subjecting the leaf to different vapour pressure differences (VPD). A reduced partial pressure of CO₂ reduced specific leaf mass (SLM) as did a decreased PFD, but did not change leaf N per unit area (N_LA) and light saturated rate of photosynthesis (A_max). A reduced VPD caused several effects consistent with the effect of PFD. It decreased N_LA and A_max and increased the chlorophyll to N ratio in old and young leaves. Furthermore, it decreased the chlorophyll a to b ratio and inhibited leaf growth in young leaves. The transpiration stream is partitioned among the leaves of a plant according to their transpiration rates. The results suggest that relative rates of import of xylem sap into leaves of a plant play an important role in the perception of partial shading of a plant, a situation normally found in dense vegetations. The possible role of cytokinin influx into leaves as controlled by transpiration rate, is discussed.     

Nitrogen allocation in response to partial shading of a plant: Possible mechanisms

The significance of photosynthetic and transpiration rates for the perception by plants of light gradients in leaf canopies has been investigated with regard to nitrogen allocation and re-allocation. A gradient of photon flux density (PFD) over a plant's foliage was simulated by shading one leaf of a pair of primary leaves of bean ( Phaseolus vulgaris L. cv. Rentegever). Photosynthetic rate was manipulated independently of PFD and, to some extent, also of transpiration, by subjecting the leaf to different CO₂ concentrations. Transpiration rate was changed independently of PFD and photosynthetic rate by subjecting the leaf to different vapour pressure differences (VPD). A reduced partial pressure of CO₂ reduced specific leaf mass (SLM) as did a decreased PFD, but did not change leaf N per unit area (N_LA) and light saturated rate of photosynthesis (A_max). A reduced VPD caused several effects consistent with the effect of PFD. It decreased N_LA and A_max and increased the chlorophyll to N ratio in old and young leaves. Furthermore, it decreased the chlorophyll a to b ratio and inhibited leaf growth in young leaves. The transpiration stream is partitioned among the leaves of a plant according to their transpiration rates. The results suggest that relative rates of import of xylem sap into leaves of a plant play an important role in the perception of partial shading of a plant, a situation normally found in dense vegetations. The possible role of cytokinin influx into leaves as controlled by transpiration rate, is discussed.     

Low vapour pressure deficit reduces the beneficial effect of elevated CO2 on growth of N2-fixing alfalfa plants

Plant responses to elevated CO₂ can be modified by many environmental factors, but very little attention has been paid to the interaction between CO₂ and changes in vapour pressure deficit (VPD). Thirty-day-old alfalfa plants ( Medicago sativa L. cv. Aragón), which were inoculated with Sinorhizobium meliloti 102F78 strain, were grown for 1 month in controlled environment chambers at 25/15°C, 14 h photoperiod, and 600 µmol m⁻² s⁻¹ photosynthetic photon flux (PPF), using a factorial combination of CO₂ concentration (400 µmol mol⁻¹ or 700 µmol mol⁻¹) and vapour pressure deficit (0.48 kPa or 1.74 kPa, which corresponded to relative humidities of 85% and 45% at 25°C, respectively). Elevated CO₂ strongly stimulated plant growth under high VPD conditions, but this beneficial effect was not observed under low VPD. Under low VPD, elevated CO₂ also did not enhance plant photosynthesis, and plant water stress was greatest for plants grown at elevated CO₂ and low VPD. Moreover, plants grown under elevated CO₂ and low VPD had a lower leaf soluble protein and photosynthetic activity (photosynthetic rate and carboxylation efficiency) than plants grown under elevated CO₂ and high VPD. Elevated CO₂ significantly increased leaf adaxial and abaxial temperatures. Because the effects of elevated CO₂ were dependent on vapour pressure deficit, VPD needs to be controlled in experiments studying the effect of elevated CO₂ as well as considered in the extrapolations of results to a warmer, high-CO₂ world.     

Stomatal responses to humidity in air and helox

Abstract. Stomatal responses to humidity were studied in several species using normal air and a helium: oxygen mixture (79:21 v/v, with CO₂ and water vapour added), which we termed 'helox'. Since water vapour diffuses 2.33 times faster in helox than in air, it was possible to vary the water-vapour concentration difference between the leaf and the air at the leaf surface independently of the transpiration rate and vice versa. The CO₂ concentration at the evaporating surfaces ( c_i ), leaf temperature and photon flux density were kept constant throughout the experiments. The results of these experiments were consistent with a mechanism for Stomatal responses to humidity that is based on the rate of water loss from the leaf. Stomata apparently did not directly sense and respond to either the water vapour concentration at the leaf surface or the difference in water vapour concentration between the leaf interior and the leaf surface. In addition, stomatal responses that caused reductions in transpiration rate at low humidities were accompanied by decreases in photosynthesis at constant c_i , suggesting heterogeneous (patchy) stomatal closure.     

The significance of differences in the mechanisms of photosynthetic acclimation to light, nitrogen and CO2 for return on investment in leaves

1. We report changes in photosynthetic capacity of leaves developed in varying photon flux density (PFD), nitrogen supply and CO₂ concentration. We determined the relative effect of these environmental factors on photosynthetic capacity per unit leaf volume as well as the volume of tissue per unit leaf area. We calculated resource-use efficiencies from the photosynthetic capacities and measurements of leaf dry mass, carbohydrates and nitrogen content.
2. There were clear differences between the mechanisms of photosynthetic acclimation to PFD, nitrogen supply and CO₂. PFD primarily affected volume of tissue per unit area whereas nitrogen supply primarily affected photosynthetic capacity per unit volume. CO₂ concentration affected both of these parameters and interacted strongly with the PFD and nitrogen treatments.
3. Photosynthetic capacity per unit carbon invested in leaves increased in the low PFD, high nitrogen and low CO₂ treatments. Photosynthetic capacity per unit nitrogen was significantly affected only by nitrogen supply.
4. The responses to low PFD and low nitrogen appear to function to increase the efficiency of utilization of the limiting resource. However, the responses to elevated CO₂ in the high PFD and high nitrogen treatments suggest that high CO₂ can result in a situation where growth is not limited by either carbon or nitrogen supply. Limitation of growth at elevated CO₂ appears to result from internal plant factors that limit utilization of carbohydrates at sinks and/or transport of carbohydrates to sinks.     

Does photosynthetic acclimation to elevated CO2 increase photosynthetic nitrogen-use efficiency? A study of three native UK grassland species in open-top chambers

1. The photosynthetic response to elevated CO₂ and nutrient stress was investigated in Agrostis capillaris, Lolium perenne and Trifolium repens grown in an open-top chamber facility for 2 years under two nutrient regimes. Acclimation was evaluated by measuring the response of light-saturated photosynthesis to changes in the substomatal CO₂ concentration.
2. Growth at elevated CO₂ resulted in reductions in apparent Rubisco activity in vivo in all three species, which were associated with reductions of total leaf nitrogen content on a unit area basis for A. capillaris and L. perenne . Despite this acclimation, photosynthesis was significantly higher at elevated CO₂ for T. repens and A. capillaris , the latter exhibiting the greatest increase of carbon uptake at the lowest nutrient supply.
3. The photosynthetic nitrogen-use efficiency (the rate of carbon assimilation per unit leaf nitrogen) increased at elevated CO₂, not purely owing to higher values of photosynthesis at elevated CO₂, but also as a result of lower leaf nitrogen contents.
4. Contrary to most previous studies, this investigation indicates that elevated CO₂ can stimulate photosynthesis under a severely limited nutrient supply. Changes in photosynthetic nitrogen-use efficiency may be a critical determinant of competition within low nutrient ecosystems and low input agricultural systems.     

Acquisition and allocation of carbon and nitrogen by Danthonia richardsonii in response to restricted nitrogen supply and CO2 enrichment

Dry weight (DW) and nitrogen (N) accumulation and allocation were measured in isolated plants of Danthonia richardsonii (Wallaby Grass) for 37 d following seed imbibition. Plants were grown at ≈ 365 or 735 μ L L^–1 CO₂ with N supply of 0·05, 0·2 or 0·5 mg N plant^–1 d^–1. Elevated CO₂ increased DW accumulation by 28% (low-N) to 103% (high-N), following an initial stimulation of relative growth rate. Net assimilation rate and leaf nitrogen productivity were increased by elevated CO₂, while N concentration was reduced. N uptake per unit root surface area was unaffected by CO₂ enrichment. The ratio of leaf area to root surface area was decreased by CO₂ enrichment. Allometric analysis revealed a decrease in the shoot-N to root-N ratio at elevated CO₂, while the shoot-DW to root-DW ratio was unchanged. Allometric analysis showed leaf area was reduced, while root surface area was unchanged by elevated CO₂, indicating a down-regulation of total plant capacity for carbon gain rather than a stimulation of mineral nutrient acquisition capacity. Overall, growth in elevated CO₂ resulted in changes in plant morphology and nitrogen use, other than those associated simply with changing plant size and non-structural carbohydrate content.     

Elevated CO2 and ozone reduce nitrogen acquisition by Pinus halepensis from its mycorrhizal symbiont

The effects of 700 μmol mol⁻¹ CO₂ and 200 nmol mol⁻¹ ozone on photosynthesis in Pinus halepensis seedlings and on N translocation from its mycorrhizal symbiont, Paxillus involutus, were studied under nutrient-poor conditions. After 79 days of exposure, ozone reduced and elevated CO₂ increased net assimilation rate. However, the effect was dependent on daily accumulated exposure. No statistically significant differences in total plant mass accumulation were observed, although ozone-treated plants tended to be smaller. Changes in atmospheric gas concentrations induced changes in allocation of resources: under elevated ozone, shoots showed high priority over roots and had significantly elevated N concentrations. As a result of different shoot N concentration and net carbon assimilation rates, photosynthetic N use efficiency was significantly increased under elevated CO₂ and decreased under ozone. The differences in photosynthesis were mirrored in the growth of the fungus in symbiosis with the pine seedlings. However, exposure to CO₂ and ozone both reduced the symbiosis-mediated N uptake. The results suggest an increased carbon cost of symbiosis-mediated N uptake under elevated CO₂, while under ozone, plant N acquisition is preferentially shifted towards increased root uptake.     

An experimental test of the eddy correlation technique over a Mediterranean macchia canopy

Abstract. Flux densities of water vapour and carbon dioxide were measured for a Mediterranean macchia canopy. Results show good agreement between the measured available energy and the sum of latent sensible and heat flux densities determined with the eddy correlation technique. Joint evaluation of the Bowen ratio, aerodynamic resistance, canopy resistance and the 'omega factor' suggests that the macchia canopy is intermediate in aerodynamic roughness between coniferous and deciduous canopies. Maximum daytime carbon flux densities ranged from -14 to -22(μnol m⁻² s⁻¹ on a ground area basis. The ratio of transpiration to assimilation (E/A) was a function of incident photo-synthetic photon flux density below about 400 μmol m⁻²s⁻¹ and above it was fairly constant at 272 mol mol⁻¹ (H₂O/CO₂). The relationship between carbon influx and canopy conductance was linear. Results show promising applications of the eddy correlation technique for evaluating physiological features of canopies, treated as unitary functional systems.     

Investigations on Net CO2 Assimilation, Transpiration and Root Growth of Fagus sylvatica Infested with Four Different Phytophthora Species

Abstract: In this contribution, we compare the influence of four different Phytophthora species on root development, net CO₂ assimilation and transpiration of beech seedlings and saplings. Some few days after inoculation, photosynthesis and transpiration of seedlings infected with either P. citricola or P. cambivora were strongly reduced. In parallel, about 60 % of their root systems was destroyed compared to control plants. Three weeks after infection, all seedlings were dead, showing severe wilt symptoms on leaves. Remarkably, P. syringae and P. undulata infected seedlings and older beech plants did not differ from controls regarding photosynthesis and transpiration, although the root systems were damaged. However, a significant influence on net CO₂ assimilation and transpiration of P. citricola infected beech saplings was visible after bud break in the following year. Some days before plants started to wilt, photosynthesis and transpiration were reduced to almost zero. Water use efficiency data (WUE) clearly indicated that infected plants suffered from severe drought.     

Theory and practice of a portable photosynthesis instrument

Abstract. Theory and practice of non-steady-state portable photosynthesis instruments (LI-6000 and 6200, LI-COR Inc., Nebraska, U.S.A.) are presented. Mass balance equations for the time dependence of H₂O and CO₂ mol fractions within the leaf chamber were used to describe instrument function. Measurements for each run were fitted to an exponential function to estimate average rates of CO₂ assimilation and transpiration during the measurement period. Stomatal conductances and intercellular CO₂ mol fractions were also computed. Linear data analysis used in the LI-6200 produced similar results for assimilation rates, stomatal conductances and intercellular CO₂ concentrations compared to a more rigorous nonlinear analysis, provided humidity within the chamber was kept constant during the measurement period. Instrument performance for CO₂ fluxes was confirmed by injecting pure CO₂ at steady rates from a microsyringe into the chamber. Miniature evaporimeters were designed to check H₂O flux measurements. Significant discrepancies were observed between LI-6200 estimates of H₂O fluxes and direct measurement and errors were attributed to adsorption desorption of water vapour on chamber walls or to leaks. The leaf chamber should be stored at humidities and temperatures similar to those during measurement conditions for maximum reliability of results.     

An apparatus for pulse and pulse-chase experiments with 14CO2 on attached leaves with known, steady-state rates of photosynthesis

Abstract. An apparatus is described to carry out pulse and pulse-chase experiments with ¹⁴CO₂ on intact, attached leaves with known, steady-state rates of photosynthesis under defined conditions of temperature, vapour pressure deficit and photon flux density. Data are presented which show that the pattern of distribution of ¹⁴C between compounds in extracts of such leaves is a true reflection of the pathways of photosynthetic carbon metabolism in the leaf during steady-state photosynthesis.     

Leaf Gas Exchange in Canopy Species of a Venezuelan Cloud Forest

Tropical cloud forests are considered humid ecosystems with frequent cloud cover down to the ground surface. However, seasonal variation in precipitation may induce short-term water stress. For canopy leaves, this water stress may also be a consequence of large atmospheric vapor pressure deficits. The objective of this work was to study five canopy cloud forest species to determine if there are restrictions to leaf gas exchange as a consequence of seasonality in precipitation and to daily water deficit due to air evaporative demand mainly during maximum incoming radiation hours. Seasonal daily courses of microclimatic variables (air temperature, relative humidity, photosynthetic photon flux density) and plant responses (leaf water potential, stomatal conductance, CO₂ assimilation rates, leaf nitrogen concentration) were measured at 2400 m asl in Monterrey, an intermontane valley of the Venezuelan Andes. A gradient in terms of responses to water stress conditions was observed between the species, with Clusia multiflora (a 46% reduction in stomatal conductance between seasons) as the most affected and Miconia resimoides (increased stomatal conductance) responding more favorably to slight water stress conditions. If we consider the limitations of water stress and/or light conditions on CO₂ assimilation we may arrange the species into those in which water stress conditions have a greater impact on leaf carbon gain, those where light conditions are determinant and one in which both water stress and light conditions may affect leaf carbon assimilation.     

Photosynthesis and Optimal Light Microhabitats for a Rare Cactus, Mammillaria gaumeri, in Two Tropical Ecosystems

Mammillaria gaumeri , an endemic cactus from Yucatan, is threatened due to human-mediated habitat fragmentation but knowledge regarding factors that would inform management plans is limited, including information on optimal microhabitats and growth. We studied seasonal patterns of microclimate, net CO₂ uptake and growth of adult individuals during the dry, rainy and northwind (late fall to winter) seasons in separate populations located in a coastal dune and in a dry forest. To provide an assay of primary productivity, we measured the production of new axillary buds (areoles) from the apex. We found that patterns of net CO₂ uptake in the three seasons were consistent with obligate crassulacean acid metabolism. The reduction in soil water potential and the increase in water vapor pressure deficit led to a reduction in 24-h net CO₂ uptake from 18.0 to 1.3 mmol CO₂/m², after 30 d of drought. Dry-weight gain was maximal during the rainy season and there was no growth during the dry season, thus seasonal patterns of net CO₂ uptake and growth for M. gaumeri reflected soil and atmospheric water contents. Annual dry-weight gain increased linearly with plant size and with total daily photosynthetic photon flux density (PPFD) up to 80 percent of ambient. Maximal growth was in exposed microhabitats receiving 60–80 percent of total ambient PPFD. We hypothesize that low-magnitude disturbances, resulting in more exposed microhabitats, might be beneficial for growth and conservation of adult individuals of this rare species.     

Influence of leaf age and light environment on the gas exchange of lupins and wheat

Rates of net photosynthesis (A), transpiration (E) and leaf conductance to water vapour transfer (gH₂O) were measured on leaves of Lupinus angustifolius L. cv. Ritson's and L. cosentinii Guss. cv. Eregulla throughout development and on flag leaves of wheat ( Triticum aestivum L. cvs Gutha, Gamenya and Warigal) after full expansion. Plants were grown in large containers of soil, in a naturally-lit, temperature controlled glasshouse. Throughout most of their life, lupin leaves had higher photosynthetic rates and leaf conductances than found for wheat. During leaf ageing in lupins, photosynthesis and conductance changed proportionately such that leaf intercellular CO₂ concentration was maintained relatively constant at about 200 ppm. Under continuously cloudy conditions, leaf conductance at midday of lupins and wheat was higher than at similar photon flux densities at other times of day on cloudless days. On cloudy days the relationship between gH₂O and photon flux density in lupins was very different from that derived from diurnal measurements on clear days. The potentially low water use efficiency under cloud, evident as decreases in the A/gH₂O ratio, was rarely realised in practise due to a reduction in leaf-to-air water vapour concentration difference on cloudy days. The possible reasons for the high conductance on cloudy days are discussed.     

Do tomatoes on the plant behave as climacteric fruits?

I considered the possibility that changes in fruit photosynthesis obscure the occurrence of the climacteric rise in respiration in tomato fruits attached to the plant. Internal CO₂ and ethylene concentrations in tomatoes ( Lycopersicon esculentum Mill. cv. OH 7814) were analyzed after direct sampling through polyethylene tubes implanted in the external pericarp. Fruits which were shaded with aluminium foil contained up to 60 ml 1⁻¹ CO₂, until the internal ethylene concentration exceeded 1 μl l⁻¹, when CO₂ concentration declined to below 40 ml l⁻¹; the CO₂ concentration in fruits exposed to light only occasionally exceeded 40 ml 1⁻¹. The internal CO₂ concentration of detached fruits first declined and then increased along with ethylene concentration, as expected for the climacteric. Detached green fruits under continuous low photosynthetic photon flux density (100 μmol m⁻² s⁻¹) contained almost no internal CO₂ and produced no CO₂. Changes in photosynthesis and an associated CO₂-generating system in green fruits are thought to obscure the climacteric rise in tomato fruits developing on the plant.     

Stomatal and photosynthetic responses to shade in sorghum, soybean and eastern gamagrass

We studied photosynthetic and stomatal responses of grain sorghum ( Sorghum bicolor [L.] Moench cv. Pioneer 8500), soybean ( Glycine max L. cv. Flyer) and eastern gamagrass ( Tripsacum dactyloides L.) during experimental sun and shade periods simulating summer cloud cover. Leaf gas exchange measurements of field plants showed that short-term (5 min) shading of leaves to 300–400 μmol m⁻² s⁻¹ photosynthetic photon flux density reduced photosynthesis, leaf temperature, stomatal conductance, transpiration and water use efficiency and increased intercellular CO₂ partial pressure. In all species, photosynthetic recovery was delayed when leaves were reilluminated, apparently by stomatal closure. The strongest stomatal response was in soybean. Photosynthetic recovery was studied further with soybeans grown indoors (maximum photosynthetic photon flux density 1 200 μmol m⁻² s⁻¹). Plants grown indoors had responses to shade similar to those of field plants, except for brief nonstomatal limitation immediately after reillumination. These responses indicated the importance of the light environment during leaf development on assimilation responses to variable light, and suggested different limitations on carbon assimilation in different parts of the soybean canopy. Photosynthetic oxygen evolution recovered immediately upon reillumination, indicating that the light reactions did not limit soybean photosynthetic recovery. While shade periods caused stomatal closure and reduced carbon gain and water loss in all species, the consequences for carbon gain/water loss were greatest in soybean. The occurrence of stomatal closure in all three species may arise from their shared phenologies and herbaceous growth forms.