Long-Term Leaf Production Response to Elevated Atmospheric Carbon Dioxide and Tropospheric Ozone

Elevated concentrations of atmospheric CO₂ and tropospheric O₃ will profoundly influence future forest productivity, but our understanding of these influences over the long-term is poor. Leaves are key indicators of productivity and we measured the mass, area, and nitrogen concentration of leaves collected in litter traps from 2002 to 2008 in three young northern temperate forest communities exposed to elevated CO₂ and/or elevated O₃ since 1998. On average, the overall effect of elevated CO₂ (+CO₂ and +CO₂+O₃ versus ambient and +O₃) was to increase leaf mass by 36% whereas the overall effect of elevated O₃ was to decrease leaf mass by 13%, with similar effects on stand leaf area. However, there were important CO₂ × O₃ × year interactions wherein some treatment effects on leaf mass changed dramatically relative to ambient from 2002 to 2008. For example, stimulation by the +CO₂ treatment decreased (from +52 to +25%), whereas the deleterious effects of the +O₃ treatment increased (from −5 to −18%). In comparison, leaf mass in the +CO₂+O₃ treatment was similar to ambient throughout the study. Forest composition influenced these responses: effects of the +O₃ treatment on community-level leaf mass ranged from +2 to −19%. These findings are evidence that community composition, stand development processes, CO₂, and O₃ strongly interact. Changes in leaf nitrogen concentration were inconsistent, but leaf nitrogen mass (g m⁻²) was increased by elevated CO₂ (+30%) and reduced by elevated O₃ (−16%), consistent with observations that nitrogen cycling is accelerated by elevated CO₂ but retarded by elevated O₃.     

General Metabolic Changes in Leaf Lipids in Response to Ozone

Of eight species of higher plants examined, lettuce was themost resistant to ozone. Lettuce leaves showed no visible changeswhile in other species visible injuries were apparent afterplants had been exposed to 0.5 µl liter^–1 ozonein the light for 6 h and then kept in the light for a further20 h. Analysis of leaf lipids immediately after 4–6 hof treatment with ozone revealed decreases in levels of monogalactosyldiacylglycerol(MGDG) and increases in levels of triacylglycerol (TG) in allplants including lettuce, but the extent of the changes variedamong the plant species. The fatty acids esterified to TG inozone-treated leaves were mainly     

Response of Forest Trees to Increased Atmospheric CO 2

The CO ₂ fertilization hypothesis stipulates that rising atmospheric CO ₂ has a positive effect on tree growth due to increasing availability of carbon. The objective of this paper is to compare the recent literature related to both field CO ₂ -enriched experiments with trees and empirical dendrochronological studies detecting CO ₂ fertilization effects in tree-rings. This will allow evaluation of tree growth responses to atmospheric CO ₂ enrichment by combining evidence from both ecophysiology and tree-ring research. Based on considerable experimental evidence of direct CO ₂ fertilization effect (increased photosynthesis, water use efficiency, and above- and belowground biomass), and predications from the interactions of enriched CO ₂ with temperature, nitrogen and drought, we propose that warm, moderately drought-stressed ecosystems with an ample nitrogen supply might be the most CO ₂ responsive ecosystems. Empirical tree-ring studies took the following three viewpoints on detecting CO ₂ fertilization effect in tree-rings: 1) finding evidence of CO ₂ fertilization effect in tree-rings, 2) attributing growth enhancement to favorable climate rather than atmospheric CO ₂ enrichment, and 3) considering that tree growth enhancement might be caused by synergistic effects of several factors such as favorable climate change, CO ₂ fertilization, and anthropogenic atmospheric deposition (e.g., nitrogen). At temperature-limiting sites such as high elevations, nonfindings of CO ₂ fertilization evidence could be ascribed to the following possibilities: 1) cold temperatures, a short season of cambial division, and nitrogen deficiency that preclude a direct CO ₂ response, 2) old trees past half of their maximum life expectancy and consequently only a small increase in biomass increment due to CO ₂ fertilization effect might be diminished, 3) the elimination of age/size-related trends by statistical detrending of tree-ring series that might remove some long-term CO ₂ -related trends in tree-rings, and 4) carbon partitioning and growth within a plant that is species-specific. Our review supports the atmospheric CO ₂ fertilization effect hypothesis, at least in trees growing in semi-arid or arid conditions because the drought-stressed trees could benefit from increased water use efficiency to enhance growth.     

The Response of Plastidome and Chondriome in Leaf Chlorenchyma to Prolonged Atmospheric Pollution

The quantitative changes in the major organelles, chloroplasts and mitochondria, were followed in order to evaluate plant cell responses to prolonged atmospheric pollution. Chlorenchyma cells were compared in needle and leaves of evergreen (Pinus sylvestris, Arctostaphylos uva-ursi, and Vaccinium vitis-idaea), deciduous (Betula pubescens, V. myrtillus, and V. uliginosum), and herbaceous (Cornus suecica, Potentilla erecta, and Solidago lapponica) plants growing at distances of 65 to 70 km (an undisturbed habitat) and 8 to 10 km (a heavily damaged habitat) from the Severonikel industrial complex in the town of Monchegorsk on the Kola peninsula. Chlorenchyma cells with the structure undamaged by atmospheric pollination were used for comparison. In undamaged (depressed) chlorenchyma cells of needles and leaves affected by heavy pollination, the morphometric analysis showed that the chloroplast density did not change, while the mitochondrial density increased in most species under study. The considerable increase in the number of mitochondria in the depressed chlorenchyma cells of needles and leaves in evergreen plant species seems to constitute the mechanism for plant compensation under prolonged stress conditions.     

Compositional Characterization and Pyrolysis of Loblolly Pine and Douglas-fir Bark

Two potential biofuel resources, Douglas-fir and Loblolly pine bark, were subjected to extensive chemical and compositional analysis. The barks were initially extracted with dichloromethane, and the resulting extracted compounds were characterized by gas chromatography coupled with mass spectrometric analysis. Characterization of the major bark biocomponents indicated that Douglas-fir and Loblolly pine bark contained 22.5 and 13.2 % tannins, 44.2 and 43.5 % lignin, 16.5 and 23.1 % cellulose, and 7.6 and 14.1 % hemicellulose, respectively. Of particular interest is the high content of tannins and lignin, which make these barks excellent potential precursors for bio-oils and/or other value-added chemicals. ¹³C nuclear magnetic resonance (NMR) was used to characterize the chemical structure of the lignin and tannins. These samples were also analyzed by ³¹P NMR after phosphitylation of the hydroxyl groups in lignin and tannins. The NMR spectral data indicated that the lignin in both barks contained p-hydroxyphenyl (h) and guaiacyl (g) of lignin monomers with an h/g ratio of 10:90 and 22:78 for Douglas-fir and Loblolly pine bark, respectively. Gel permeation chromatography was used to analyze the molecular weight distributions of extracted tannins, isolated cellulose, and ball-milled lignin. The pyrolysis of Douglas-fir and pine bark at 500°C in a tubular reactor generated 48.2 and 45.2 % of total oil, of which the light oil contents are 14.1 and 20.7 % and heavy oil are 34.1 and 24.4 %. Similarly, fast pyrolysis at 375°C yielded 56.1 and 49.8 % of total oil for Douglas-fir and pine bark, respectively.     

不同施肥条件下夏玉米光谱特征与叶绿素含量和LAI的相关性

在不同施肥方式下研究了长期试验地夏玉米在4个典型生育期的叶绿素相对含量(SPAD)、叶面积指数(LAI)和冠层光谱特征,系统分析了单波段以及由可见光和近红外波段组成的归一化差异植被指数(NDVI)与叶片叶绿素含量和LAI的相关性.结果表明:在同一个生育期,NK、PK处理和CK的光谱曲线特征相似,NPK、NP、OM、1/2OM1/2NPK处理的光谱特征曲线相似.除460nm外,其它单波段反射率与叶绿素含量和LAI都显著相关,尤其在510~1100nm波段相关性最为密切;不同波段组合的NDVI与这2个指标也显著相关,且相关系数大于单波段.从拔节期到乳熟期,绝大部分NDVI都可以有效地拟合叶绿素含量和LAI的变化,选用NDVI(560,950)和NDVI(660,760)建立拟合模型(R2>0.60),并以NDVI(560,950)拟合效果最佳(R2>0.80).可见,不同施肥方式条件下长期试验地夏玉米冠层光谱特征与叶绿素含量、LAI都具有良好的相关性,可以为夏玉米肥料管理提供科学依据.     

The Physiological Response of Phyllostachys edulis under the Elevated Atmospheric Ozone

In order to provide theoretical evidences for bamboo cultivation under the background of climate change, the method of open top chambers (OTCs) was employed to approach the effects of elevated ozone on photosynthetic pigments content, lipid peroxidation and anti oxidation enzymes in the leaves of Phyllostachys edulis. The result indicated that with the increasing of ozone concentration, the content of the chlorophyll, carotenoid and SOD activity decreased, while the content of souble protein, relative electrical conductivity, contents of MDA, O2- and the activity of POD increased.Correlation analyses showed that there was significant negative correlation between O2- content and SOD activity as well as photosynthetic pigment,whereas significant positive correlation between O2- and relative electeical conductivity, MDA, POD as well as soluble protein. It was summarried that, on one hand, long time exposure to high ozone concentration exposure, accelerated the aging of leaves, intensified the damage to membrane lipid, and destroyed the structure of cell membrane and the function of anti oxidation system as well. It affected the regular growth of P.edulis. This research was conducted to provide theoretical evidence for bamboo cultivation during this time of possible climate change.     

毛竹对大气臭氧浓度倍增的生理响应

为了给气候变化背景下的竹林培育应对策略提供理论依据,以毛竹 （Phyllostachys edulis） 为材料,运用开顶式气室 （OTCs） 模拟当前环境背景大气O3浓度（NF,40~45nL·L-1）的倍增1倍（TR1,92~106nL·L-1）和倍增2倍（TR2, 142~160nL·L-1）情景,分析叶片光合色素、脂质过氧化及抗氧化酶等生理指标的变化规律。结果表明：大气O3浓度倍增处理90d时,毛竹叶片Chl、Car含量和SOD活性显著降低,叶片相对电导率和可溶性蛋白、MDA、O2-含量及POD活性显著升高。叶片O2-含量与SOD活性、光合色素含量呈极显著负相关,而与叶片相对电导率、POD活性和MDA、可溶性蛋白含量呈显著正相关。研究表明大气O3浓度倍增会导致毛竹叶片老化加快,光合色素降解或合成受阻,膜脂过氧化,膜结构和氧化系统破坏,影响毛竹的正常生长。本文从抗性生理方面揭示了毛竹对大气臭氧胁迫的耐受性,为竹子应对气候变化研究提供了参考。     

Episodic Growth and Relative Shoot: Root Balance in Loblolly Pine Seedlings

Leaf, root and stem systems of loblolly pine seedlings are characterizedby a seasonal periodicity in growth, during which they alternatein spurts of activity. Despite this periodicity, the allometriccoefficient describing the ratio of the relative growth ratesof leaf to root remains constant for at least the first twoyears of development. In part, constancy results from the inabilityof variation in the allocation of growth increment during briefperiods to change a pre-existing structure accumulated overthe life of the seedling. In addition, alternating periods ofleaf, root and stem growth may represent the action of feedbackmechanisms which operate to maintain an adaptive balance betweenorgan systems. Pinus taeda L., loblolly pine, allometric coefficient, homeostatic control of growth     

Germination Promotion of Loblolly Pine and Baldcypress Seeds by Stratification and Chemical Treatments

The effects of 4 chemicals on the germination promotion of stratified and unstratified seeds of loblolly pine (Pinus taeda) and baldcypress (Taxodium distichum) were studied. The chemicals used were gibberellic acid, kinetin, potassium nitrate and thiourea, each at 3 different concentrations. Stratification promoted the germination of both seed species. Certain concentrations of gibberellic acid, potassium nitrate and thiourea improved the germination of unstratified loblolly pine and baldcypress seeds while kinetin had no stimulatory effect. All 4 chemicals at specific concentrations promoted the germination of loblolly pine seeds stratified for a short period of time. Considering both speed and completeness of germination, best results were obtained when 21-day stratified seeds were treated with either gibberellic acid (100 mg/1) or kinetin (10 mg/1). In baldcypress, on the other hand, none of these chemicals had any stimulatory effect on the germination of stratified seeds. Germination of both species of seeds was either partially or completely inhibited by the highest concentration of thiourea (30,000 mg/1) used.     

Complexities in Understanding Ecosystem Response to Ozone

Ecological risk assessment of O₃ impact requires consideration of many factors that, perhaps, are not of concern in human health risk assessment. The episodic nature of O₃ exposure, functional complexity of species assemblages, and the broad spatial and temporal scales characteristic of natural ecosystems make ecological risk assessment extremely difficult. The majority of exposure studies using plants have examined the sensitivity of individual species, growing under controlled conditions. Research has shown that individuals growing in plant mixtures may not respond the same way to O₃ as when growing alone. In addition, other naturally occurring stresses can modify plant response to O₃. Understanding the effect of O₃ on natural systems and protecting vegetation resources represent significant scientific and regulatory challenges. Here we review several factors that need to be considered when evaluating ecosystem response to O₃. Then we briefly present two examples of controlled seedling studies that were conducted to better understand mechanisms of tree response to O₃. In the first example controlled exposure studies revealed responses in tree roots that led to hypothesis testing in the field in ponderosa pine ecosystems. Field experiments have confirmed a similar response in root biomass and carbohydrates across a natural O₃ gradient in S. California, suggesting at least a partial role for O₃ in the response. The second example illustrates the difficulty of understanding mechanistic interactions to O₃ stress even in controlled chamber studies. The second example also illustrates the difficulty of using chamber studies to understand responses in the field. While our knowledge of vegetation response to O₃ is extensive and compelling, important questions remain about how to quantify these effects in the field, assess their magnitude, and establish a suitable standard that is protective of ecosystems.     

Association Mapping of Total Carotenoids in Diverse Soybean Genotypes Based on Leaf Extracts and High-Throughput Canopy Spectral Reflectance Measurements

Carotenoids are organic pigments that are produced predominantly by photosynthetic organisms and provide antioxidant activity to a wide variety of plants, animals, bacteria, and fungi. The carotenoid biosynthetic pathway is highly conserved in plants and occurs mostly in chromoplasts and chloroplasts. Leaf carotenoids play important photoprotective roles and targeted selection for leaf carotenoids may offer avenues to improve abiotic stress tolerance. A collection of 332 soybean [Glycine max (L.) Merr.] genotypes was grown in two years and total leaf carotenoid content was determined using three different methods. The first method was based on extraction and spectrophotometric determination of carotenoid content (eCaro) in leaf tissue, whereas the other two methods were derived from high-throughput canopy spectral reflectance measurements using wavelet transformed reflectance spectra (tCaro) and a spectral reflectance index (iCaro). An association mapping approach was employed using 31,253 single nucleotide polymorphisms (SNPs) to identify SNPs associated with total carotenoid content using a mixed linear model based on data from two growing seasons. A total of 28 SNPs showed a significant association with total carotenoid content in at least one of the three approaches. These 28 SNPs likely tagged 14 putative loci for carotenoid content. Six putative loci were identified using eCaro, five loci with tCaro, and nine loci with iCaro. Three of these putative loci were detected by all three carotenoid determination methods. All but four putative loci were located near a known carotenoid-related gene. These results showed that carotenoid markers can be identified in soybean using extract-based as well as by high-throughput canopy spectral reflectance-based approaches, demonstrating the utility of field-based canopy spectral reflectance phenotypes for association mapping.     

Importance of Seasonality for the Response of a Mesic Temperate Grassland to Increased Precipitation Variability and Warming

Timing of precipitation events within the growing season and the non-uniformity of warming might be decisive for alterations in productivity and community composition, with consequences for ecosystem functioning. The responses of aboveground production, community composition, functional group and species evenness to altered intra-annual precipitation variability and their interactions with winter or summer warming were examined in European, mesic temperate grassland. Increased precipitation variability with an induced spring drought resulted in a 17% reduction in ANPP, and late drought reduced ANPP by 18% compared to regular rainfall patterns throughout the entire growing season. Winter warming increased ANPP by 12%, whereas summer warming showed no significant effect on biomass but decreased species richness. The effects of increased precipitation variability and warming on ANPP were independent of each other. Forbs benefited from high precipitation variability with spring drought events, likely due to reduced competitive pressure by decreasing, water stressed grasses. Increased precipitation variability coinciding with higher summer temperatures led to reduced species evenness and likely promoted the establishment of specialists and drought-tolerant species. Seasonality of climatic factors, here early versus late drought events in the high precipitation variability treatments, was important in driving shifts in community composition but not for decreases in ANPP. Non-uniform warming, here winter versus summer, affected the direction of response of both community composition and ANPP. Variability of resources is affecting ecosystem processes and species interactions. Recognition of seasonality and non-uniformity of climatic factors will improve predictions of plant performance and biotic interactions in response to climate change.     

植物激素和抗生素调节的转基因火炬松愈伤组织的生长和分化

包含质粒载体pBI121的农杆菌菌株LBA4404被用于转化火炬松的成熟合子胚。质粒载体pBI121含有胭脂碱合成酶基因的启动子驱动的新霉素磷酸转移酶基因（npt Ⅱ)和花椰菜花叶病毒的35S启动子驱动的GUS基因。器官发生的转基因愈伤组织和转基因的再生植株已经获得,并经GUS组织化学染色、聚合酶链式反应和Southern杂交分析证实。植物激素（BA/IBA）和抗生素对转基因愈伤组织的生长和分化的影响被研究。500mg/L羧苄青霉素和2mg/LBA、0.5mg/L IBA的组合导致转基因愈伤组织的生长增加54.2%,分化增加45.7%。500mg/L Claforan和2mg/L BA、0.5mg/L IBA的组合导致转基因愈伤组织的生长增加40.8%,分化增加38.7%。高浓度的植物激素和抗生素降低了转基因愈伤组织的生长和分化。实验结果有助于建立一个高效的农杆菌介导的火炬松遗传转化系统,也有助于未来针叶树的遗传转化研究。     

Changes in Endogenous Growth Kegulators in Loblolly Pine Seeds During the Process of Stratification and Germination

The seeds of loblolly pine (Pirns taeda L.) were cold-stratified for 0, 14, 28, 42, and 56 days. Endogenous growth regulators were extracted from these seeds, and also from the germinating and the 28-day warm stratified seeds. Partially purified extracts were separated on chromatographic paper. The chromatograms were cut into 10 equal segments, and these were tested for biological activity using three different btoassays. The results indicated that the unstratified seeds and those stratified up to a period of 28 days contained very little or no growth promoter (GA-like substances), and a relatively high concentration of an inhibitor (presumably abscisic acid), Following 42-day stratification, the promoter concentration gradually increased while the inhibitor level fell almost to zero. A high level of promoter but no inhibitor was detected in germinating seeds. No auxin-like activity was noted in the unstratified seeds. This activity slowly increased up to a period of 28 days and remained at this level for the subsequent stratification periods. However, the activity greatly increased in the germinating seeds. Very little changes in the levels of growth regulators were noted in warm-stratified seeds as compared to the unstrati-fied controls.     

Purification and Characterization of Catalase from Loblolly Pine (Pinus taeda L.) Megagametophytes

Catalase (EC 1.11.1.6) was purified to near homogeneity from isolated megagametophytes of germinated loblolly pine (Pinus taeda L.) seeds, and monospecific antibodies were elicited in rabbits. Following a procedure that involved acetone extraction, (NH4)2SO4 fractionation, and four chromatographic steps (i.e. DE-52 cellulose, Superdex-200, hydroxylapatite, and phenyl-Sepharose CL-4B), catalase was purified about 140-fold to a final specific activity of 2215 mmol min-1 mg-1 of protein. Cotton isocitrate lyase antibodies were used, and protein immunoblots revealed that the resolution on hydroxylapatite and phenyl-Sepharose allowed for the complete separation of catalase from contaminating isocitrate lyase. The molecular masses of the native enzyme and its subunit are 235 and 59 kD, respectively, indicating that the pine holoenzyme is a homotetramer. Loblolly pine catalase exists as multiple isoforms. When megagametophytes taken 7 d after imbibition at 30[deg]C were extracted, subjected to nondenaturing isoelectric focusing, and stained for catalase activity, at least four catalase isoforms were observed, including one dominant form with an isoelectric point of 6.87. Purified pine catalase is not a glycoprotein and has a ratio of absorbance at 208 nm to absorbance at 405 nm of 1.5. When probed with loblolly pine catalase antibodies, protein blots of cell-free extracts from megagametophytes of mature, stratified, and germinated loblolly pine seeds, the megagametophyte glyoxysomal fraction, and purified loblolly pine catalase all revealed one immunoreactive 59-kD polypeptide. This indicates that no detectable change in the enzyme's monomeric molecular mass occurs during seed stratification and germination, early seedling growth, and purification.     

Effect of Nitrogen Fertilization and Growth Suppression on Pitch Canker Development on Loblolly Pine Seedlings

One-yr-old loblolly pine seedlings of two half-sib families, grown in sand, were fertilized three times per week with nutrient solution containing 20 μg/ml (low) or 80 μg/ml (high) nitrogen. Nitrogen concentration in the nutrient solution was either constant throughout the experiment, or interehanged after the inoculation of stems or shoots with Fusarium subglutinans, 55 days after initiation of fertilization. Growth was suppressed by a weekly excision of shoots branching from the stem apex. Either high nitrogen nutrition or shoot excision generally enhanced canker elongation on stem inoculated plants; the combination of both was extremely conducive for disease development. With intact plants of family 8–68, interchange of pre-inoculation low nitrogen nutrition with high nitrogen after inoculation enhanced canker elongation and rate of wilt. Nitrogen content varied in wood, bark and needles, as well as with time intervals, but was consistently in accordance with nitrogen level in the nutrient solution. In shoot excised plants, nitrogen content was higher than in the respective treatment without shoot excision. The higher nitrogen nutrient accelerated disease development on inoculated shoots, compared to low nitrogen, on both pine families. With respective treatments, stem cankers were larger and rates of shoots exhibiting lesions or wilt were higher on plants of family 8–68 than on 8–61. It is postulated that the disease enhancing effect associated with higher nitrogen content in stem tissues results from an increased nitrogen availability to the pathogen.