Hybrid physiological models are being increasingly used to assess productivity, carbon sequestration, water and nutrient use and environmental impacts of management decisions. Users include forest managers, politicians, environmental agencies and scientists. However a wider use of these models has been prevented as a result of an incomplete understanding of the mechanisms regulating carbon allocation, nutrient availability in soils and nutrient uptake by trees. On-going innovation in clonal forestry, genetic improvement and vegetation management techniques is also poorly represented in hybrid models. This thesis examines means to represent nutrition and genotype-nutrition interactions in productivity physiological hybrid models. Nutrient limitations and growth differences between genotypes were hypothesized to operate through key physiological processes: photosynthesis, carbon allocation and nutrient internal cycling. In order to accomplish the aims of the study both greenhouse and field experimentation were carried out. In a first experiment, responses of photosynthesis (A) to intercellular CO₂ concentration (Ci) were measured in a fast- and a slow-growing clone of Pinus radiata D. Don cultivated in a greenhouse in a factorial combination of nitrogen and phosphorus supply, and analyzed using the biochemical model of leaf photosynthesis described by Farquhar et al. (1980). There were significant positive linear relationships between the parameters, Vcmax, Jmax, Tp and both foliar nitrogen (Na) and phosphorus (Pa) concentration on an area basis. The study showed that the effects of nitrogen and phosphorus supply on photosynthesis were statistically independent and that the photosynthetic behaviour of the two clones was equivalent. In a similar study, gas exchange and chlorophyll fluorescence were simultaneously measured to determine internal transfer conductance (gm) based on the "constant J method". Transfer conductance may pose significant limitations to photosynthesis which may be differentially affected by nutrition and genotype in Pinus radiata. Values of gm were similar to those of stomatal conductance (gs) and their ratio (gm / gs) was not influenced by nutrient supply or clone being on average (±1 SE) 1.22 ±0.04. Relative mesophyll limitations (LM, 16%) to photosynthesis were marginally greater than those imposed by stomata (LS, 13%), and together smaller than the relative limitations posed to photosynthesis by biochemical processes (LB, 71%). The CO₂ concentration in the intercellular air spaces (Ci) was (±1 SE) 53 ±3 µmol mol-1 lower than in the atmosphere (Ca) while CO₂ concentration in the chloroplasts (Cc) was (±1 SE) 48 ±2 µmol mol-1 less than Ci. Values of LS, LM and LB and CO₂ diffusion gradients posed by gs (Ca-Ci) and gm (Ci-Cc) did not change with nutrient supply or clone. In a third experiment, one-year old Pinus radiata cuttings from four genotypes were cultivated in silica sand with a factorial combination of nitrogen (N0=1.43 and N1=7.14 mM) and phosphorus (P0=0.084 and P1=0.420 mM) supply for 24 months. N supply was enriched with ¹⁵N to 2.5⁰/₀₀ (labelled N) during the first year, then plants transferred to clean sand and cultivated for another year with ¹⁵N at levels close to natural abundance (0.3664899 atom percent ¹⁵N, δ¹⁵N 0.5115 ⁰/₀₀) provided by the source of N in nutrient solution applied during the second year. Recovery of labelled and unlabelled N was used to estimate N remobilization. N remobilization scaled with plant growth, N content and N and P supply. In relative terms, 65% of all stored N was remobilized in the high-nutrient supply regime compared to 42-48% at lower N and P addition rates. Most N remobilization occurred during spring-summer (77%), coincidently with the largest proportion of needle development (80%), indicating that N remobilization was driven by sink-strength. Foliage was by far the main source for internal cycling while roots were the main sink (40%). Clones exhibited differences in N remobilization capacity, but these differences were completely explained by the size of the N pool before remobilization took place, indicating that N remobilization performance was similar among clones. In a fourth study, four clones were cultivated in silica sand with a factorial combination of nitrogen and phosphorus supply for ten months, and patterns of carbon allocation examined using a carbon balance approach. Gross-primary productivity (GPP) scaled mainly with nitrogen but also with phosphorus supply. The fraction of GPP (GPP = ANPP + APR + TBCA) allocated to above-ground components (ANPP) increased with N and P supply at the expense of total-below ground C allocation (TBCA) with no apparent effect on the fraction of GPP partitioned to above-ground plant respiration (APR). Carbon use efficiency (NPP:GPP) scaled with nutrient supply, being 0.42 in the low-nutrient supply regime compared to 0.51 in the high-nutrient supply regime, suggesting that in poor fertility environments a larger proportion of the C budget is respired compared to the net productivity. Fast-growing clones allocated about 2-4% more carbon to above-ground components (ANPP) at the expense of carbon allocated below-ground (TBCA) with no effect on carbon respired above-ground (APR), indicating that faster-growing genotypes allocate more carbon to leaf area which may compound and increase overall GPP over time. The field component of this thesis was conducted in a subset of locations where ENSIS (formerly New Zealand Forest Research Institute) had established trials to test the influence of species, soil disturbance and plant nutrition on sustainability indicators. Plots were small in size (3 m × 3 m) with trees spaced at 0.5 m × 0.5 m (40 000 trees ha-1) with nine measurement trees surrounded by a two-row buffer. All sites were planted in winter 2001 and harvested in spring 2005. The aim of this pilot study was to examine patterns of carbon allocation during the fourth year after planting in control and fertilized mini-plots of Pinus radiata in five sites with contrasting climate and soil conditions in the South Island of New Zealand. The study showed that the fraction of gross-primary productivity allocated belowground increased as the soil C:N ratio increased. However, these results should be interpreted with caution due to the unusual nature of the trial and the reduced number of sites studied. Two existing physiological models were selected for the discussion in this thesis (3-PG, Landsberg and Waring 1997; canopy net carbon exchange model, Whitehead et al. 2002). Potential improvements for the nutritional component of 3-PG comprise: accounting for reductions in carbon use efficiency (NPP:GPP) in poor-fertility environments, adding a preliminary fertility modifier (FN, 0-1) driven by soil C : N ratio and soil N, adding a preliminary relationship between carbon allocation to roots and the soil C : N ratio and representing faster-growing genotypes by increasing their leaf area but not their photosynthetic performance. The canopy net carbon exchange model (NCE) combines the coupled model of leaf photosynthesis - stomatal conductance described by Leuning (1995) with canopy structure and a water balance model to scale carbon assimilation from leaves to canopies. Potential improvements to account for nutrient deficiencies in the leaf model by Leuning (1995), comprise using nutrient ratios to discriminate nitrogen (Na/Pa < 23 mol mol-1) from phosphorus deficiencies (Na/Pa > 23 mol mol-1), adding relationships between photosynthetic model parameters Vcmax and Jmax to Pa, and correcting the estimation of photosynthetic parameters Vcmax and Jmax by accounting for transfer conductance (gm). The canopy net carbon exchange model may be also modified to account for carbon-use efficiency, carbon allocation to roots and genotype in a similar form to that proposed for 3-PG. The results previously outlined provide a preliminary framework to represent tree and soil nutrition in physiological hybrid productivity models.

Genetic improvement of a number of crops including soybean (Glycine max L. [Merr]) has been associated with stay-green. Research on stay green genes has focused primarily on genes involved with photosynthesis and chlorophyll degradation. The current study explores the impact of a group of developmental genes, known as the E gene series, on the rate of soybean leaf senescence. The objective of this experiment was to determine the role of E-genes in the control of leaf senescence in soybean. The experiment was conducted in a split-plot design with three replications. The main plots were two photoperiods imposed following R1; i) natural day length (Amb) and ii) incandescent day length extension of 3 hours (Amb+3). The split plots were five E-gene near-isogenic lines (NILs), planted on different dates to obtain synchronous flowering. Phenology, photosynthesis, normalized difference vegetative index (NDVI) and fluorescence measurements were taken including, dark adapted photosynthetic efficiency (Fv/Fm), electron transport rate (ETR), and leaf chlorophyll concentration (SPAD). Leaf tissues were also analyzed for gene expression patterns among Harosoy isolines. Yield parameters like dry matter accumulation, harvest index and grain yields were recorded. The leaf net photosynthesis was more closely related to ETR than to SPAD values, suggesting that visual observation of stay-green may not be as effective in evaluating functional senescence as measurement of ETR. Cultivars with the dominant E1 allele maintained functional photosynthesis for longer, such that full senescence was delayed by 10-15 days in these cultivars. This phenomenon was observed under both photoperiod treatments and irrespective of the genetic background (Clark and Harosoy) in which the alleles appeared. Maintenance of functional photosynthesis by the E1 dominant allele can be attributed to maintenance of high ETR, and Fv/Fm, as well as delayed decline in leaf chlorophyll concentrations. Expression of senescence related genes were delayed in the isoline which had delayed leaf senescence phenotype. Consistent with the effect on leaf senescence, the dominant alleles also reduced the rate of phenological development, such that R5 occurred later in genotypes with dominant alleles and under the Amb+3 treatment. Cultivars with the dominant E1 allele under extended photoperiod treatment accumulated more biomass and had decreased apparent harvest index which caused no change in grain yields. The dominant E allele may delay leaf senescence directly or indirectly, through its delay of reproductive development.

Leaf senescence is the final stage of leaf development, during which the leaf relocates most of itsvaluable nutrients to developing or storing parts of the plant. As this process progresses, leaves losetheir green color and their capacity to perform photosynthesis. Shade and darkness are well-knownas factors inducing leaf senescence and it has been proposed that senescence can be initiated byreductions in photosynthesis, photomorphogenesis and transpiration. However, despite the fact thatthe signaling mechanisms regulating each of these processes have been extensively described,particularly in seedlings, their contribution to the initiation of senescence in mature leaves stillremains unclear. Furthermore, the use of different experimental systems to study shade-inducedleaf senescence has yielded several divergent results, which altogether complicate the overallunderstanding of leaf senescence. To address this, darkened plants and individually darkened leaves, which show different rates of leafsenescence, were studied. Comparing the transcriptome and metabolome of these two darktreatmentsrevealed that they differed distinctly with regard to their metabolic strategies. Wholedarkened plants were severely carbohydrate-starved, accumulated amino acids and slowed downtheir metabolism. In contrast, individually darkened leaves showed continued active metabolismcoupled to senescence-associated degradation and relocation of amino acids. This knowledge was used to set up a new system to study how shade affects leaf senescence in themodel plant Arabidopsis thaliana. Use of this system revealed that different senescence-associatedhallmarks appeared in response to different intensities of shade. Some of these hallmarks werefurther shown to be part of both leaf senescence and photosynthetic acclimation to low light. Finally, using this system on phytochrome mutants revealed that loss of phytochrome A increasedthe loss of chlorophyll under shade, without increasing the expression of senescence-associatedgenes. Together, these findings suggest that shade-induced leaf senescence, which is generally perceived asa single process, is actually an intricate network of different processes that work together tomaintain an optimal distribution of nutrients within the plant.

When light energy input exceeds the capacity for photosynthesis the plant need to dissipate the excess energy and this is done through non-photo-chemical quenching (NPQ). Photochemical quenching (photosynthesis), NPQ and fluorescence are three alternative faiths of excited chlorophylls. PsbS associates to photosystem II and is involved in NPQ. The results presented in this thesis were generated on Arabidopsis plants and mainly based on wildtype Col-0 together with a mutant deficient in PsbS (npq4) and a transgene overexpressing PsbS (oePsbS). We connect light and herbivore stress and show that the level of PsbS influences the food preference of both a specialist (Plutella) and a generalist (Spodoptera) herbivore as well as oviposition of Plutella. Level of PsbS also affects both metabolomics and transcriptomics of the plant; up-regulation of genes in the jasmonic acid (JA) -pathway and amount of JA has been found in the npq4 plants after herbivory. Since many experiments were performed in field we have also characterized the field plant and how it differs from the commonly used lab plant. We have also studied the natural variation of NPQ in Arabidopsis plants both in the field and the lab. The results show surprisingly no correlation. / Överskottsenergi kan vara skadligt för en växts membran och fotosynteskomplex. Vid överskott av solenergi blir fotosystemen mättade och växten behöver därför ett sätt för att göra sig av med all överskottsenergi, detta kallas för ”icke-fotokemisk quenching” (NPQ). Fotokemisk quenching (fotosyntes), NPQ och fluoresens är tre alternativa vägar för exalterade klorofyller. PsbS är involverad i NPQ och associerar med fotosystem II. De resultat som presenteras i denna avhandling kommer från studier av modellväxten Arabidopsis thaliana (Backtrav), i huvudsak gjorda på vildtypen i jämförelse med en mutant som saknar PsbS (npq4) och en transgen som överuttrycker PsbS (oePsbS). Vi har försökt att undersöka kopplingen mellan ljus- och herbivoristress och visar här att mängden PsbS påverkar både en specialist (Plutella) och en generalist (Spodoptera) insekt vid val av föda, samt Plutella även vid äggläggning. Växternas nivå av PsbS visade sig även påverka metabolomet och transkriptomet, och vi fann en uppreglering av gener i biosyntesen för jasmonat samt mer av själva hormonet jasmonat i npq4 växter efter herbivori. Eftersom vi har gjort många av experimenten ute i fält har vi även karakteriserat en typisk Arabidopsis växt i fält samt hur denna skiljer sig från den vanligt använda lab-växten. Dessutom har vi även undersökt naturlig variation av NPQ av Arabidopsis både i fält och på lab och resultaten visar, till vår förvåning, att det inte går att finna någon korrelation mellan dessa.

The C3 photosynthetic pathway uses light energy to reduce CO2 to carbohydrates and other organic compounds and is a central component of biological metabolism. In C3 photosynthesis, CO2 assimilation is catalyzed by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), which reacts with both CO2 and O2. While competitive inhibition of CO2 assimilation by oxygen is suppressed at high CO2 concentrations, O2 inhibition is substantial when CO2 concentration is low and O2 concentration is high; this inhibition is amplified by high temperature and aridity (Sage 2004). Atmospheric CO2 concentration dropped below saturating levels 25-30 million years ago (Tipple & Pagani 2007), and the C4 photosynthetic pathway is hypothesized to have first evolved in warm, low latitude environments around this time (Christin et al. 2008a). The primary feature of C4 photosynthesis is suppression of O2 inhibition through concentration of CO2 around Rubisco. This pathway is estimated to have evolved almost 50 times across 19 angiosperm families (Muhaidat et al. 2007), a remarkable example of evolutionary convergence. In several C4 lineages, there are species with photosynthetic traits that are intermediate between the C3 and C4 states, known as C3-C4 intermediates. In two eudicot genera, Flaveria (Asteraceae) and Alternanthera (Amaranthaceae), there is evidence that these species represented an intermediate state in the evolution of the C4 pathway (McKown et al. 2005; Sanchez-del Pino 2009). The purpose of this thesis is to ascertain the specific benefits to plant carbon balance and resource-use efficiencies of the C3-C4 pathway relative to C3 species, particularly at low CO2 concentrations and high temperatures, factors which are thought to have been important in selecting for C3-C4 traits (Ehleringer et al. 1991). This will provide information on the particular advantages of the C3-C4 pathway in warm, often arid environments and how these advantages may have been important in advancing the initial stages of C4 evolution in eudicots. This thesis addresses the physiological intermediacy of previously uncharacterized C3-C4 species of Heliotropium (Boraginaceae); the water- and nitrogen-use efficiencies of C3-C4 species of Flaveria; and the photosynthetic performance and acclimation of C3, C4 and C3-C4 species of Heliotropium, Flaveria and Alternanthera grown at low and current ambient CO2 levels and high temperature.

主催：JST/CREST，Vrije University, ALTERRA, IBPC

Thesis (Ph. D.)--University of Hawaii at Manoa, 1996. / Includes bibliographical references. / Microfiche. / xxi, 302 leaves, bound ill 29 cm

Obligate shade plants possess adaptations that enable them to photosynthesise in the low light environment of the forest floor. Adaptations that facilitate light scavenging may compromise capacity for high rates of photosynthesis. This study compares the responses of obligate shade and facultative shade plant species upon exposure to elevated light. The obligate shade plants were two commercially grown medicinal herb species of ginseng, Panax ginseng C.A. Meyer and Panax quinquefolius L.; and goldenseal - Hydrastis canadensis L. Comparison was made to Arabidopsis thaliana and Pisum sativum L. as facultative shade species. Panax ginseng (Korean ginseng) and Panax quinquefolius (American ginseng) are obligate shade plants found in broadleaf forests of Eastern Asia and North America, respectively. Studies on these plants have shown optimal growth at light intensities between 200-300 [mu]mol photons. m⁻�. s⁻�, or 10-15% of full sunlight, and at intensities greater than 500 [mu]mol photons. m⁻�. s⁻� characteristic photoinbibitory symptoms develop. An atypical response to methyl viologen in photosynthetic electron transport assays was observed in ginseng in both isolated thylakoid membranes and whole leaves. No correlation was found between detectable superoxide dismutase activity and altered methyl viologen reactions. In a mutagenesis study using the model cyanobacterium Synechocystis sp. PCC 6803, a unique amino acid residue in the terminal electron acceptor PsaC, found only in ginseng, was changed and found to have no effect on methyl viologen reactions. Electron transfer to methyl viologen was examined in both isolated thylakoid membranes and whole leaves using chlorophyll a fluorescence and the apparent ability for methyl viologen to act as an electron acceptor was observed to differ between ginseng species. Obligate shade species were observed to possess alternate pools of photosystem II centres that potentially provide a mechanism to maximise photosynthetic gain under low light and during short periods of increased illumination. In experiments designed to identify physiological processes that contribute to increased susceptibility to photoinhibition in obligate shade plants, responses were observed and characterised following a moderate increase in illumination (140 to 400 [mu]mol photons. m⁻� . s⁻�) using chlorophyll a fluorescence induction curve analysis. The obligate shade species exhibited varied responses to elevated light and showed increased susceptibility, to photoinhibition. Photoprotective non-photochemical dissipative capacity was quantified and found to be comparable between all species studied.

Although it has long been recognized that physiological acclimation of photosynthesis and respiration can occur in plants exposed to changing environmental conditions (e.g. light, temperature or stress), the extent of acclimation in different tissues (i.e. pre-existing and new foliage) however, has not received much attention until recently. Furthermore, few studies have investigated the extent of photosynthetic and respiratory acclimation under natural conditions, where air temperatures vary diurnally and seasonally. In this study, the effects of variations in temperature on respiratory CO2 loss and photosynthetic carbon assimilation were examined under both controlled and natural environments. The purpose of the investigations described in this thesis was to identify the effects acclimation would have on two key metabolic processes in plants exposed to temperature change, with emphasis also placed on the role of nutrition (nitrogen) and respiratory enzymatic characteristics on the potential for acclimation in two contrasting tree species, Pinus radiata and Populus deltoides. Controlled-environment studies (Chapter 2 and 3) established that rates of foliar respiration are sensitive to short-term changes in temperature (increasing exponentially with temperature) but in the longer-term (days to weeks), foliar respiration acclimates to temperature change. As a result, rates of dark respiration measured at any given temperature are higher in cold-acclimated and lower in warm-acclimated plants than would be predicted from an instantaneous response. Acclimation in new foliage (formed under the new temperature environment) was found to result in respiratory homeostasis (i.e. constant rates of foliar respiration following long-term changes in temperature, when respiration is measured at the prevailing growth temperature). Available evidence suggests that substantial adjustments in foliar respiration tend to be developmentally dependent. This may in part explain why respiratory homeostasis was only observed in new but not in pre-existing tissues. Step changes in temperature (cold and warm transfers) resulted in significant changes in photosynthetic capacity. However, in stark contrast to the findings of respiration, there was little evidence for photosynthetic acclimation to temperature change. The results obtained from field studies (Chapter 4) show that in the long-term over a full year, dark respiration rates in both tree species were insensitive to temperature but photosynthesis retained its sensitivity, increasing with increasing temperature. Respiration in both species showed a significant downregulation during spring and summer and increases in respiratory capacity were observed in autumn and winter. Thermal acclimation of respiration was associated with a change in the concentration of soluble sugars. Hence, acclimation of dark respiration under a naturally changing environment is characterized by changes in the temperature sensitivity and apparent capacity of the respiratory apparatus. The results from controlled and natural-environment studies were used to drive a leaflevel model (which accounted for dark respiratory acclimation) with the aim of forecasting the overall impact of responses of photosynthesis and respiration in the long term (Chapter 5). Modellers utilise the temperature responses of photosynthesis and respiration to parameterize carbon exchange models but often ignore acclimation and use only instantaneous responses to drive such models. The studies here have shown that this can result in erroneous estimates of carbon exchange as strong respiratory acclimation occurs over longer periods of temperature change. For example, it was found here that the failure to factor for dark respiratory acclimation resulted in the underestimation of carbon losses by foliar respiration during cooler months and an overestimation during warmer months - such discrepancies are likely to have an important impact on determinations of the carbon economy of forests and ecosystems. The overall results substantiate the conclusion that understanding the effect of variations in temperature on rates of carbon loss by plant respiration is a prerequisite for predicting estimates of atmospheric CO2 release in a changing global environment. It has been shown here that within a moderate range of temperatures, rate of carbon uptake by photosynthesis exceeds the rate of carbon loss by plant respiration in response to warming as a result of strong respiratory acclimation to temperature change. This has strong implications for models which fail to account for acclimation of respiration. At present, respiration is assumed to increase with increasing temperatures. This erroneous assumption supports conclusions linking warming to the reinforcement of the greenhouse effect.

Marine phytoplankton ecophysiological state and primary production measurements have typically been controversial due to potential impacts of measurement techniques. Advances in chl-a fluorescence techniques have provided a means for rapid, non-invasive measurement of electron transport through photosystem 2 (PSII) in dilute phytoplankton suspensions. While studies on higher plants have outlined a close relationship between PSII electron transport and carbon fixation, results from studies on microalgae reveal significant variations in the relationship. Three species of phytoplankton representing three major taxonomic groups of the marine phytoplankton were used in this study: (1) Chaetoceros muelleri CS176 Lemmermann (Bacillariophyta), (2) Isochrysis galbana CS177 Parke (Haptophyta) and, (3) Nannochloropsis oculata CS179 (Droop) Hibberd (Ochrophyta, eustigmatophyte). Each species was cultured in semicontinuous culture and primary production was estimated using oxygen evolution and carbon fixation techniques and compared against predictions based on chl-a fluorescence measurements. It was found that predicted values of primary production both under-estimated and overestimated actual carbon fixation measured via radioisotope (14C) techniques. This variation was primarily explained by probable errors in the assumed values for PSII density. The relationship between oxygen evolution or carbon fixation with chl-a fluorescence-derived measures was commonly linear below the light saturation parameter, with a departure from linearity occurring at higher irradiances. This departure from linearity was greatest in cultures adapted to low light conditions. At higher light intensities alternative electron pathways such as the Mehler reaction and/or chlororespiration are likely to be more active in low light-adapted cultures, leading to this greater non-linearity. Chl-a fluorescence measurements were also found to be a useful in characterising ecophysiology using photosynthesis-versus irradiance curves. However, an important caveat on this is the measurement of PSII density (çPSII) rather than use of an assumed value as changes in çPSII can have a profound impact on light curve parameters. A field study in Fremantle Harbour found a healthy (negligible nutrient starvation), diatom dominated, phytoplankton community. Results suggest that phytoplankton are able to begin boosting photosynthetic capability just prior to morning twilight. Waters in the harbour were well mixed via tidal motion and substantial midday photoinhibition was not observed. Data suggest levels of primary production at the mouth of the harbour are similar to those of coastal waters in the plume of the Ocean Reef wastewater outfall.