Eucalyptus Globulus Seedlings Research Articles

USE OF AGROMINERAL AS SUBSTRATE FOR GROWTH OF EUCALYPTUS SEEDLINGS

Success in the production of eucalyptus seedlings directly affects the development of plants in the field. Several factors influence their initial growth, especially the substrate. This study aimed to evaluate the growth of Eucalyptus globulus seedlings and characteristics of the substrate using silicate agromineral (SA) in different proportions (substrate:SA; v/v): T0 – 100:00%, T1 – 90:10%, T2 – 80:20%, and T3 – 50:50%. The treatments were evaluated using a completely randomized design. Measurements of height (H), stem diameter (SD) and calculation of the H/SD ratio of the plants were performed every 3 weeks, totaling 4 measurements. At the end of the experiment (12 weeks), soil fertility analyses, granulometric analysis (sand, silt and clay) and total chemical analysis were carried out. Due to the chemical and mineralogical composition of SA, its use in substrate in a greater proportion (T3) promoted the increase of exchangeable K, Ca and Si contents, which influenced the increase in the sum of bases and cation exchange capacity. The levels of assimilable P and micronutrients Fe, Cu, Zn and Mn were also higher in this treatment. The increase in SA proportions in the substrate composition also favored increase in the sand fraction and reduction in the clay fraction. As for the growth of seedlings, the results were not significant for H and SD as a function of SA doses, leading to the conclusion that the solubilization and slow release of nutrients by SA did not influence the growth of the plants

Elevated CO2 offsets the alteration of foliar chemicals (n-icosane, geranyl acetate, and elixene) induced by elevated O3 in three taxa of O3-tolerant eucalypts

Eucalypts are important forest resources in southwestern China, and may be tolerant to elevated ground-level ozone (O3) concentrations that can negatively affect plant growth. High CO2 may offset O3-induced effects by providing excess carbon to produce secondary metabolites or by inducing stomatal closure. Here, the effects of elevated CO2 and O3 on leaf secondary metabolites and other defense chemicals were studied by exposing seedlings of Eucalyptus globulus, E. grandis, and E. camaldulensis × E. deglupta to a factorial combination of two levels of O3 (< 10 nmol mol−1 and 60 nmol mol−1) and CO2 (ambient: 370 μmol mol−1 and 600 μmol mol−1) in open-top field chambers. GC-profiles of leaf extracts illustrated the effect of elevated O3 and the countering effect of high CO2 on compounds in leaf epicuticular wax and essential oils, i.e., n-icosane, geranyl acetate and elixene, compounds known as a first-line defense against insect herbivores. n-Icosane may be involved in tolerance mechanisms of E. grandis and the hybrid, while geranyl acetate and elixene in the tolerance of E. globulus. Elevated O3 and CO2, singly or in combination, affected only leaf physiology but not biomass of various organs. Elevated CO2 impacted several leaf traits, including stomatal conductance, leaf mass per area, carbon, lignin, n-icosane, geranyl acetate and elixene. Limited effects of elevated O3 on leaf physiology (nitrogen, n-icosane, geranyl acetate, elixene) were commonly offset by elevated CO2. We conclude that E. globulus, E. grandis and the hybrid were tolerant to these O3 and CO2 treatments, and n-icosane, geranyl acetate and elixene may be major players in tolerance mechanisms of the tested species.

Nitrogen loading of Eucalyptus globulus seedlings: nutritional dynamics and influence on morphology and root growth potential

Achieving successful outplanting of trees on increasingly harsher sites is a global concern. In Chile, for example, new Eucalyptus globulus plantations are being targeted to poorer, low fertility sites where additional stresses often negatively affect seedling performance during the first field season. Nitrogen-loading during the later stages of nursery production can, however, significantly improve seedling physiological performance on harsher sites by increasing plant growth and nutrient storage reserves for subsequent root growth after outplanting. Maximizing these benefits, and applying the concept of loading throughout the entire crop cycle, requires a better understanding of nitrogen (N) dynamics. Thus, for container E. globulus, considered one of the most important plantation species worldwide, we evaluated the effects of increasing N supply (50 to 600 mg N L−1) on seedling morphological traits, biomass production, root growth potential, and foliar N. After 15 weeks of N application, seedlings showed an increase in all growth variables evaluated up to 300 mg N L−1, whereas higher rates allowed luxury consumption. Modeling growth responses revealed, however, an apparent ammonium antagonism with other macronutrient cations suggests that high rates of this N-form may negate the potential benefits of N-loading, or that other modifications to the fertilizer solution may promote additional growth at higher N rates. Foliar N concentration, which was easily and accurately measured with a leaf chlorophyll meter, showed a gradient from basal to apical leaves suggesting a strong translocation of N in E. globulus seedlings. Our results suggest nursery management that supplies 300 mg L−1 of N through the entire growing season could improve seedling nutritional status, morphological attributes, and growth of new roots, attributes that may improve establishment on low fertility sites.

The effect of management operations on the demography of Eucalyptus globulus seedlings

There has been an increased concern about the invasiveness of Eucalyptus globulus Labill. Much research has been devoted to understanding the factors influencing E. globulus recruitment. Standard management operations, performed inside and outside plantations, cause profound disturbances on soil and vegetation, potentially influence E. globulus recruitment, but have been poorly studied. This study, therefore, aims to understand the impacts of standard management operations on seeds and capsules shed prior or after such operations, and, together with weather, their influence on the germination and early survival of E. globulus seedlings. We performed a sowing experiment in Portugal using E. globulus seeds, following a full factorial design with 16 treatments, resulting from three factors: management operations (burning, soil stripping, soil harrowing and control); sowing time (before or after management operations); sowing type (sowing seeds or capsules). Each treatment was replicated 10 times, giving a total of 160 plots (0.5 × 05 m), each sown with 100 seeds or 10 capsules. Seeds and capsules were monitored for approximately one year. We also tested for the presence of seed predation close to the experimental field. Seed sowing resulted in 4.8% germination (379 seedlings), but capsule sowing resulted in only two seedlings. The seed predation experiment, showed that nearly half of the E. globulus seeds went missing, suggesting that predation may contribute to the low germination results. <1.5 months after sowing, 96.3% of the total seedlings had emerged. A Multivariate Regression Tree and a Generalized Linear Mixed Model showed that the number of emerged seedlings was enhanced in plots sown with seeds, and after the management operations, particularly harrowing. Sowing before burning resulted in limited germination. The Cox Model showed that the survival probability was greatest for seeds shed after the soil treatments and in harrowing plots. A Linear Mixed Model showed the negative influence of soil harrowing and the positive effect of crop evapotranspiration on seedling mortality across time. This study shows that E. globulus seeds are able to germinate in distinct soil/vegetation management scenarios, even though some types of management operations, such as soil harrowing, may provide better conditions for germination and survival. Keeping dense competing vegetation is probably the most cost-effective option to minimize unwanted E. globulus recruitment and maximize seedling mortality.

Survival and recovery of Eucalyptus globulus seedlings from severe defoliation

Catastrophic damage that occurs at the early establishment stage of eucalypts, such as decapitation by browsing mammals, may have deleterious effects on survival, growth and tree shape. Such damage may result in a significant loss of productivity from planted forests, as well as represent a major selective force shaping patterns of adaptation in natural forests. An important recovery strategy of many eucalypts is to re-sprout from basal lignotubers, which contain a proliferation of dormant buds as well as stored carbohydrates. We investigated the genetic variation in seedling recovery following decapitation of two Eucalyptus globulus populations that differ in the extent to which they appear to allocate resources to foliar defence as opposed to recovery traits.The two contrasting populations studied were from the continuous eastern distribution of E. globulus on the island of Tasmania. One population, from the wet southern forests, has relatively small lignotubers but high foliar defences; whereas the other from the dry northern forests, has relatively large lignotubers but low foliar defences. Seedlings of 10 families from each population were subjected to artificial decapitation treatments in the nursery at 6months old, planted in a replicated field design and assessed up to 32months after treatment.The two populations differed significantly in their initial response to decapitation, with seedlings from the population with the larger lignotubers and stem base having greater survival and more shoots. Decapitation above the cotyledonary node had a more deleterious effect on survival than decapitation above the first seedling node. The surviving plants exhibited genetic stability not only in constitutive differences, but in the way they changed in response to decapitation. After 20months of growth in the field, there were no lasting treatment effects on important growth traits, with genetic-based population differences overriding the initial treatment effects. These results are consistent with differential resource allocation being part of the adaptive response of E. globulus, and show that early-age decapitation has little long-term impact on the expression of inherent population differences in growth strategies and foliar chemistry.

Population divergence in the ontogenetic trajectories of foliar terpenes of a Eucalyptus species.

The development of plant secondary metabolites during early life stages can have significant ecological and evolutionary implications for plant-herbivore interactions. Foliar terpenes influence a broad range of ecological interactions, including plant defence, and their expression may be influenced by ontogenetic and genetic factors. This study investigates the role of these factors in the expression of foliar terpene compounds in Eucalyptus globulus seedlings. Seedlings were sourced from ten families each from three genetically distinct populations, representing relatively high and low chemical resistance to mammalian herbivory. Cotyledon-stage seedlings and consecutive leaf pairs of true leaves were harvested separately across an 8-month period, and analysed for eight monoterpene compounds and six sesquiterpene compounds. Foliar terpenes showed a series of dynamic changes with ontogenetic trajectories differing between populations and families, as well as between and within the two major terpene classes. Sesquiterpenes changed rapidly through ontogeny and expressed opposing trajectories between compounds, but showed consistency in pattern between populations. Conversely, changed expression in monoterpene trajectories was population- and compound-specific. The results suggest that adaptive opportunities exist for changing levels of terpene content through ontogeny, and evolution may exploit the ontogenetic patterns of change in these compounds to create a diverse ontogenetic chemical mosaic with which to defend the plant. It is hypothesized that the observed genetically based patterns in terpene ontogenetic trajectories reflect multiple changes in the regulation of genes throughout different terpene biosynthetic pathways.

Germination response of Eucalyptus globulus seeds exposed to low and high temperature stress

Low seedling emergence rates of some seed lots of genetically elite Eucalyptus globulus Labill have been reported by nursery operators and hypothesized to be caused by high temperature stress. This study has confirmed that germination rate was sensitive to exposure to high temperatures. The optimum temperature for the germination rate of normal seedlings in this species was between 24.8°C and 25.5°C. At 25°C, 96% of seeds produced normal seedlings after 21 days. The cardinal temperatures for development of normal Eucalyptus globulus seedlings were between 9.1°C and 10.5°C at low temperature and between 40.3°C and 41.4°C at high temperature. The optimum temperature for maximum attainment of normal seedlings was between 21.2°C and 24.8°C. Temperatures above 30°C generally delayed commencement of germination, reduced total germination percentage, caused a high proportion of seed death and suppressed the development of normal seedlings. Low temperatures also delayed germination but caused less seed mortality. Despite general consistency in responses there was evidence that seed lots exhibit a differential response to temperature, particularly for normal seedling development at the extremes of temperature tested.

Field screening for genetic-based susceptibility to mammalian browsing

Browsing by mammalian herbivores is a major problem in plantation forestry. Seedlings are most vulnerable to browsing during establishment, making protection crucial during this period. Aside from reducing herbivore numbers, browsing can be controlled through the application of tree guards or chemical repellents. These methods are generally short-term options. A promising, potentially longer-term method of reducing browsing damage on plantations is to plant individuals with enhanced natural browse resistance. This requires the development of a rapid, cost-effective means of identifying germplasm with enhanced resistance. Here we present such a screening methodology. We planted Eucalyptus globulus seedlings from 22 different seedlots in randomised blocks along edge rows of six operational plantations. Seedlings were monitored for mammal browsing damage and growth for 2 years. Natural variation between E. globulus seedlots resulted in significant variation in the extent to which they were browsed. Differential browsing was evident just 10 weeks after planting, and caused a significant effect on tree growth after 2 years. Differential browsing was thought to be due to variation in levels of plant secondary metabolites (PSMs). The identification and selective planting of high PSM seedlots in high risk areas could be an effective management tool to reduce browsing in plantation forestry.

How does P affect photosynthesis and metabolite profiles of Eucalyptus globulus?

Phosphorus (P) has multiple effects on plant metabolism, but there are many unresolved questions especially for evergreen trees. For example, we do not know the general effects of P on metabolism, or if P affects photosynthesis via the internal conductance to CO(2) transfer from sub-stomatal cavities to chloroplast or amounts of Rubisco. This study investigates how Pdeficiency affects seedlings of the evergreen tree Eucalyptus globulus grown for 2.5 months with four nutrient solutions differing in P concentration. To determine why photosynthesis was affected by P supply, Rubisco was quantified by capillary electrophoresis, internal conductance was quantified from gas exchange and carbon isotope discrimination, and biochemical parameters of photosynthesis were estimated from A/C(c) responses. Additional insights into the effect of P on metabolism were provided by gas chromatography-mass spectrometry (GC-MS) metabolite profiling. Larger concentrations of P in the nutrient solution led to significantly faster rates of photosynthesis. There was no evidence that stomatal or internal conductances contributed to the effect of P supply on photosynthesis. The increase in photosynthesis with P supply was correlated with V(cmax), and amounts of P, phosphate and fructose 6-phosphate (6-P). Phosphorous supply affected approximately one-third of the 90 aqueous metabolites quantified by GC-MS, but the effect size was generally smaller than reported for experiments on herbaceous species. Phosphorus deficiency decreased concentrations of phosphate, glucose 6-P and fructose 6-P more than it decreased photosynthesis, suggesting faster turnover of smaller pools of phosphate and phosphorylated intermediates. The effect of P supply on most amino acids was small, with the exception of arginine and glutamine, which increased dramatically under P deficiency. P deficiency had small or non-significant effects on carbohydrates and organic acids of the tricarboxylic acid (TCA) cycle. The small effect of P on carbohydrates, organic acids and (most) amino acids likely reflects a functional homeostasis among C metabolism (glycolysis, TCA and pentose P cycles), rates of photosynthesis and growth. The strong functional homeostasis in E. globulus may reflect a conservative, long-term growth and metabolic strategy of evergreen trees.

Differences in Rubisco content and its synthesis in leaves at different positions in Eucalyptus globulus seedlings

The dynamics of ribulose 1.5-bisphosphate carboxylase/oxygenase (Rubisco) content and turnover during leaf development are not well understood in woody plants. Rubisco synthesis, N influx and the mRNA levels of Rubisco-encoding genes were determined as a function of leaf position in 4.5-month-old Eucalyptus globulus seedlings. Rubisco concentration was slightly higher in the top leaves as leaf expansion progressed and was almost maximal in the uppermost fully expanded leaves. Rubisco concentration remained almost constant in the fully expanded leaves at the top and middle positions and then became slightly low at the lowest positions. Rubisco synthesis was active only in the top leaves. These results suggest that Rubisco turnover rate is low in the middle leaves, leading to the maintenance of Rubisco contents, and that Rubisco degradation primarily occurs in the lowest leaves. Changes in the RBCS and rbcL mRNA levels were roughly parallel with Rubisco synthesis, but N influx was more closely correlated with Rubisco synthesis. These results suggest that N influx rather than the transcript abundance of Rubisco-encoding genes is of primary importance in regulating the rate of Rubisco synthesis. Additionally, expression of RBCS multigene family in E. globulus leaves was discussed.

Genotypic variation in morphology and freezing resistance of Eucalyptus globulus seedlings subjected to drought hardening in nursery

Eucalyptus globulus Labill is one of the most planted species in Chile, because of its fast growth and superior pulp qualities. Nevertheless, the incidence of drought and frost damage immediately after planting is frequent. The purpose of this work was to study the effect of drought hardening on frost resistance and on variations in morphological traits that may increase drought resistance at nursery phase in four genotypes of E. globulus Labill. Drought hardening treatments consisted in induced water stress by watering restriction, until pre-dawn stem xylem water potentials (Ψ pd ) reached -0.2, -1.8 and -2.6 MPa. Two water stress-rewatering cycles were applied during 54 days of hardening. Plant and root biomasses were affected by the interaction of drought hardening and genotypes. The rest of morphological and alometrical traits were affected independently by drought or genotype. Plant height, leaf area, specific leaf area (SLA), stem, and leaf biomasses decreased with drought hardening, while collar diameter was not affected. Genotypes responded differentially to drought hardening in plant height, leaf area, SLA, and stem, and leaf biomasses. Ice nucleation temperature (INT), and freezing temperatures (FRT), and 50% freezing damage index of leaves (LT 50 ) were affected by the interaction between drought hardening and genotypes. EG-13, EG-23 and EG-22 genotypes became freezing tolerant with drought hardening (-2.6 MPa). Additionally, EG-14 genotype increased its freezing resistance at -1.8 MPa. Therefore, freezing resistance levels and mechanism depend on genotype and drought hardening treatment. The success in tree breeding by genetic selection should be facilitated by improved understanding of the physiology of stress resistance development and survival during water supply limitations. The knowledge of morphological and freezing resistance dependency on the interaction between genotype and drought hardening may be useful nursery management information to improve plantation success.

Do artificial and natural defoliation have similar effects on physiology of Eucalyptus globulus Labill. seedlings?

• Artificial defoliation is often used to simulate defoliation by herbivory and is usually considered a good indication of a plant’s response to a given type of damage. However, the findings of studies directly comparing the two defoliation types are inconsistent. • Here, the short term effects of artificial and insect defoliation by larvae of Paropsisterna agricola on growth, biomass allocation and photosynthetic capacity of Eucalyptus globulus seedlings were compared in a glasshouse experiment. The artificial defoliation was carried out to closely resemble the spatial patterns observed for insect defoliation. • Height and diameter increments were reduced as a result of insect defoliation, whereas artificial defoliation had no significant effect on height. Increased photosynthetic capacity was observed in response to both treatments, but the magnitude of this increase was larger in insect- than in artificially-defoliated seedlings. Significant reductions in foliar carbohydrate content and total biomass were noticeable in artificially-defoliated seedlings. Although the foliar carbohydrate levels also decreased across the crown zones following insect defoliation treatment, seedlings allocated a large amount of their biomass in the branches of the damaged zone. • Despite our best endeavours to simulate insect defoliation in the artificial treatment, the latter may not reflect accurately the full strength of the effects. However, artificial and insect defoliation were similar in their direction of the responses they caused in E. globulus seedlings.

Plasticity in maximum stomatal conductance constrained by negative correlation between stomatal size and density: an analysis using Eucalyptus globulus

ABSTRACTMaximum stomatal conductance to water vapour and CO2 (gwmax, gcmax, respectively), which are set at the time of leaf maturity, are determined predominantly by stomatal size (S) and density (D). In theory, many combinations of S and D yield the same gwmax and gcmax, so there is no inherent correlation between S and D, or between S, D and maximum stomatal conductance. However, using basic equations for gas diffusion through stomata of different sizes, we show that a negative correlation between S and D offers several advantages, including plasticity in gwmax and gcmax with minimal change in epidermal area allocation to stomata. Examination of the relationship between S and D in Eucalyptus globulus seedlings and coppice shoots growing in the field under high and low rainfall revealed a strong negative relationship between S and D, whereby S decreased with increasing D according to a negative power function. The results provide evidence that plasticity in maximum stomatal conductance may be constrained by a negative S versus D relationship, with higher maximum stomatal conductance characterized by smaller S and higher D, and a tendency to minimize change in epidermal space allocation to stomata as S and D vary.

Plasticity in maximum stomatal conductance constrained by negative correlation between stomatal size and density: An analysis usingEucalyptus globulus

Maximum stomatal conductance to water vapour and CO2 (gwmax, gcmax, respectively), which are set at the time of leaf maturity, are determined predominantly by stomatal size (S) and density (D). In theory, many combinations of S and D yield the same gwmax and gcmax, so there is no inherent correlation between S and D, or between S, D and maximum stomatal conductance. However, using basic equations for gas diffusion through stomata of different sizes, we show that a negative correlation between S and D offers several advantages, including plasticity in gwmax and gcmax with minimal change in epidermal area allocation to stomata. Examination of the relationship between S and D in Eucalyptus globulus seedlings and coppice shoots growing in the field under high and low rainfall revealed a strong negative relationship between S and D, whereby S decreased with increasing D according to a negative power function. The results provide evidence that plasticity in maximum stomatal conductance may be constrained by a negative S versus D relationship, with higher maximum stomatal conductance characterized by smaller S and higher D, and a tendency to minimize change in epidermal space allocation to stomata as S and D vary.

Role of corticular photosynthesis following defoliation in Eucalyptus globulus

Defoliation can reduce net fixation of atmospheric CO(2) by the canopy, but increase the intensity and duration of photosynthetically active radiation on stems. Stem CO(2) flux and leaf gas exchange in young Eucalyptus globulus seedlings were measured to assess the impact of defoliation on these processes and to determine the potential contribution of re-fixation by photosynthetic inner bark in offsetting the effects of defoliation in a woody species. Pot and field trials examined how artificial defoliation of the canopy affected the photosynthetic characteristics of main stems of young Eucalyptus globulus seedlings. Defoliated potted seedlings were characterized by transient increases in foliar photosynthetic rates and concomitant decreases in stem CO(2) fluxes (both in the dark and light). Defoliated field-grown seedlings showed similar stem CO(2) flux responses, but of reduced magnitude. Despite demonstrating increased re-fixation capability, defoliated potted-seedlings had slowed stem growth. The green stem of seedlings exhibited largely shade-adapted characteristics. Defoliation reduced stem chlorophyll a/b ratio and increased carotenoid concentration. An increased capacity to re-fix internally respired CO(2) (up to 96%) suggested that stem re-fixation represents a previously unexplored mechanism to minimize the impact of foliar loss by maximizing the contribution of all photosynthetic tissues, particularly for young seedlings.

Effects of leaf position, surface, and entry sites on Quambalaria eucalypti infection in eucalypt

The objectives of this study were to determine the effects of leaf position, surface, and the need for entry sites, such as wounds, other pathogen lesions or natural openings, to promote infection of Quambalaria eucalypti in Eucalyptus spp. We carried out a series of artificial inoculations in wounded and unwounded leaves from seedlings of Eucalyptus globulus and two hybrid clones (UFV-100 - E. grandis x E. urophylla and UFV-101 - E. saligna x E. maidenii). E. globulus was more susceptible to infection than clones UFV-100 and UFV-101. Artificial wounds greatly favored pathogen penetration. However, old lesions caused by Cylindrocladium pteridis and Puccinia psidii did not provide infection sites for Q. eucalypti. Differences in fungal spore germination and leaf surface colonization were not observed between expanding and fully expanded leaves. Q. eucalypti penetrated through the wounded leaves and we didn't observe direct penetration thought the epidermis. Stomata neither attracted nor repelled germ tube and somatic hyphae. Young expanding leaves were more susceptible to Q. eucalypti infection than fully expanded leaves located in the upper third of the plant. Infection took place preferentially on the abaxial leaf surface.

Weeds in Eucalyptus globulus subsp. maidenii (F. Muell) establishment: effects of competition on sapling growth and survivorship

The effect of herbaceous vegetation on growth and survival was assessed in planted eucalyptus saplings grown under four levels of weed cover. Seedlings of Eucalyptus globulus subsp. maidenii were planted with 0 (W0), 25 (W25), 50 (W50) and 100% (W100) weed cover. Weed species composition and aboveground biomass was determined. Soil water content was evaluated by the gravimetric method. Seedling leaf area, diameter and height were evaluated at planting and during the 3 months following establishment. Tree height, diameter and stem volume was estimated at 12, 24 and 36 months. First year tree survival was recorded. The ratio of cumulative stem growth under W100: cumulative stem growth under W0 was used to measure competitive performance. Regression analysis was used to determine competition thresholds. Weeds seriously threaten the growth of E. globulus subsp. maidenii. Early negative effects of competition on growth were evident as from the second month after establishment, and both seedlings and 1-year-old saplings were more affected than 2- and 3-year-old ones. A “minimum-response threshold” was determined near 500 kg/ha (corresponding to W25 cover). No clear effect was found between sapling survival and weed biomass.

Discrimination between seedlings of Eucalyptus globulus, E. nitens and their F1 hybrid using near-infrared reflectance spectroscopy and foliar oil content

Summary Identification of plant hybrids produced from closely related species can be difficult using morphological characteristics alone, particularly when identifying young seedlings. In this study, we compared the performance of three calibration models developed to discriminate between seedlings of Eucalyptus globulus, E. nitens and their first-generation hybrid using either foliar oil chemistry or near-infrared reflectance spectral data from fresh, whole leaves. Both oil and near-infrared reflectance spectroscopy (NIRS) models were developed using partial least-squares discriminant analysis and showed high classification accuracy, all correctly classifying more than 91% of samples in cross-validation. Additionally, we developed a larger, “global” and independently validated NIRS model specifically to discriminate between E. globulus and F1 hybrid seedlings of different ages. This model correctly classified 98.1% of samples in cross-validation and 95.1% of samples from an independent test set. These results show that NIRS analysis of fresh, whole leaves can be used as a rapid and accurate alternative to chemical analysis for the purpose of hybrid identification.

Novel mannose‐sequestration technique reveals variation in subcellular orthophosphate pools do not explain the effects of phosphorus nutrition on photosynthesis in Eucalyptus globulus seedlings

Although only a small proportion of plant phosphorus (P) is used for photosynthesis, the relationships between P and photosynthesis can be strong. It was hypothesized, in this study, that variation in the allocation of orthophosphate (Pi) between active (cytoplasmic) and nonactive (vacuolar) pools would underpin differences in rates of photosynthesis in 4-month-old Eucalyptus globulus seedlings grown with a varying P supply. Photosynthetic biochemistry was assessed by the response of net photosynthesis to increasing intercellular [CO2]. Cytoplasmic Pi was sequestered as mannose 6-phosphate. Total P and the proportion of P as Pi were positively related to P supply. The ratios of active : stored Pi (10-24%) varied little over the range of treatments. Active Pi was positively related to P supply, as was photosynthesis (7 micromol CO2 m(-2) s(-1) with 0 mM P vs. 16 micromol CO2 m(-2) s(-1) with 0.32 mM P). Positive relationships between P supply and photosynthesis were explained best by leaf P content, not by active pools of Pi. The distribution of Pi between the vacuole and the cytoplasm had little impact on the photosynthetic phosphorus-use efficiency (PPUE), and reductions in cytoplasmic Pi had little effect on photosynthesis. Hence, PPUE is an unsuitable guide for assessing plant responses to increasingly unavailable P in the environment.

Increased photosynthesis following partial defoliation of field-grown Eucalyptus globulus seedlings is not caused by increased leaf nitrogen

Increased photosynthetic rates following partial defoliation may arise from changes in leaf biochemistry, water relations or nutrient status. Twelve-month-old field-grown Eucalyptus globulus Labill. seedlings were pruned from below to reduce the green crown depth by 50 (D50) or 70% (D70). Photosynthetic responses to light and CO2 concentration were examined before and one, three and five weeks after partial defoliation. One week after defoliation, photosynthetic rates were greater in seedlings in the D50 (21 micromol m(-2) s(-1)) and D70 (23 micromol m(-2) s(-1)) treatments than in control seedlings (15 micromol m(-2) s(-1)); however, there was little difference in photosynthetic rates between partially defoliated seedlings and control seedlings after 5 weeks. An analysis of the sensitivity of photosynthesis to biochemical parameters revealed that the transient increase in photosynthetic rate in response to partial defoliation was largely a function of the maximum rate of carboxylation (85-87%) and the maximum rate of RuBP regeneration (55-60%) rather than stomatal conductance (12-13%). Nitrogen increased in leaves following partial defoliation (increases of 0.6 and 1.2 g m(-2) for D50 and D70, respectively), but was accumulated in a non-photosynthetic form (i.e., there was no increase in nitrogen concentration of Rubisco or chlorophyll). Increased photosynthetic rates immediately following partial defoliation were primarily a result of increased activity rather than amount of photosynthetic machinery. There was no evidence that phosphorus was responsible for the increase in photosynthetic rates after partial defoliation.