Leaf water relations determine the trade-off between ozone resistance and stomatal functionality in urban tree species.

Urban trees possess different capacities to mitigate ozone (O3) pollution through stomatal uptake. Stomatal closure protects trees from oxidative damage but limits their growth. To date, it is unclear how plant hydraulic function affect stomatal behaviour and determine O3 resistance. We assessed gas exchange and hydraulic traits in three subtropical urban tree species, Celtis sinensis, Quercus acutissima, and Q. nuttallii, under nonfiltered ambient air (NF) and elevated O3 (NF60). NF60 decreased photosynthetic rate (An) and stomatal conductance (gs) only in Q. acutissima and Q. nuttallii. Maintained An in C. sinensis suggested high O3 resistance and was attributed to higher leaf capacitance at the full turgor. However, this species exhibited a reduced stomatal sensitivity to vapour pressure deficit and an increased minimal gs under NF60. Such stomatal dysfunction did not decrease intrinsic water use efficiency (WUE) due to a tight coupling of An and gs. Conversely, Q. acutissima and Q. nuttallii showed maintained stomatal sensitivity and increased WUE, primarily correlated with gs and leaf water relations, including relative water content and osmotic potential at turgor loss point. Our findings highlight a trade-off between O3 resistance and stomatal functionality, with efficient stomatal control reducing the risk of hydraulic failure under combined stresses.

Pinofuranoxins A and B, Bioactive Trisubstituted Furanones Produced by the Invasive Pathogen Diplodia sapinea.

Two new bioactive trisubstituted furanones, named pinofuranoxins A and B (1 and 2), were isolated from Diplodia sapinea, a worldwide conifer pathogen causing severe disease. Pinofuranoxins A and B were characterized essentially by NMR and HRESIMS spectra, and their relative and absolute configurations were assigned by NOESY experiments and computational analyses of electronic circular dichroism spectra. They induced necrotic lesions on Hedera helix L., Phaseolus vulgaris L., and Quercus ilex L. Compound 1 completely inhibited the growth of Athelia rolfsii and Phytophthora cambivora, while 2 showed antioomycetes activity against P. cambivora. In the Artemia salina assay both toxins showed activity inducing larval mortality.

How Quercus ilex L. saplings face combined salt and ozone stress: a transcriptome analysis.

BACKGROUND: Similar to other urban trees, holm oaks (Quercus ilex L.) provide a physiological, ecological and social service in the urban environment, since they remove atmospheric pollution. However, the urban environment has several abiotic factors that negatively influence plant life, which are further exacerbated due to climate change, especially in the Mediterranean area. Among these abiotic factors, increased uptake of Na + and Cl - usually occurs in trees in the urban ecosystem; moreover, an excess of the tropospheric ozone concentration in Mediterranean cities further affects plant growth and survival. Here, we produced and annotated a de novo leaf transcriptome of Q. ilex as well as transcripts over- or under-expressed after a single episode of O3 (80 nl l-1, 5 h), a salt treatment (150 mM for 15 days) or a combination of these treatments, mimicking a situation that plants commonly face, especially in urban environments. RESULTS: Salinity dramatically changed the profile of expressed transcripts, while the short O3 pulse had less effect on the transcript profile. However, the short O3 pulse had a very strong effect in inducing over- or under-expression of some genes in plants coping with soil salinity. Many differentially regulated genes were related to stress sensing and signalling, cell wall remodelling, ROS sensing and scavenging, photosynthesis and to sugar and lipid metabolism. Most differentially expressed transcripts revealed here are in accordance with a previous report on Q. ilex at the physiological and biochemical levels, even though the expression profiles were overall more striking than those found at the biochemical and physiological levels. CONCLUSIONS: We produced for the first time a reference transcriptome for Q. ilex, and performed gene expression analysis for this species when subjected to salt, ozone and a combination of the two. The comparison of gene expression between the combined salt + ozone treatment and salt or ozone alone showed that even though many differentially expressed genes overlap all treatments, combined stress triggered a unique response in terms of gene expression modification. The obtained results represent a useful tool for studies aiming to investigate the effects of environmental stresses in urban-adapted tree species.

Plant species differ in early seedling growth and tissue nutrient responses to arbuscular and ectomycorrhizal fungi.

Experiments with plant species that can host both arbuscular mycorrhizal fungi (AMF) and ectomycorrhizal fungi (EMF) are important to separating the roles of fungal type and plant species and understanding the influence of the types of symbioses on plant growth and nutrient acquisition. We examined the effects of mycorrhizal fungal type on the growth and tissue nutrient content of two tree species (Eucalyptus grandis and Quercus costaricensis) grown under four nutrient treatments (combinations of low versus high nitrogen (N) and phosphorus (P) with different N:P ratios) in the greenhouse. Trees were inoculated with unidentified field mixtures of AMF or EMF species cultivated on root fragments of AMF- or EMF-specific bait plants. In E. grandis, inoculation with both AMF and EMF positively affected belowground plant dry weight and negatively affected aboveground dry weight, while only inoculation with AMF increased tissue nutrient content. Conversely, Q. costaricensis dry weight and nutrient content did not differ significantly among inoculation treatments, potentially due to its dependence on cotyledon reserves for growth. Mineral nutrition of both tree species differed with the ratio of N to P applied while growth did not. Our results demonstrate that both tree species' characteristics and the soil nutrient environment can affect how AMF and EMF interact with their host plants. This research highlights the importance of mycorrhizal fungal-tree-soil interactions during early seedling growth and suggests that differences between AMF and EMF associations may be crucial to understanding forest ecosystem functioning.

Growth duration is a better predictor of stem increment than carbon supply in a Mediterranean oak forest: implications for assessing forest productivity under climate change.

Understanding whether tree growth is limited by carbon gain (source limitation) or by the direct effect of environmental factors such as water deficit or temperature (sink limitation) is crucial for improving projections of the effects of climate change on forest productivity. We studied the relationships between tree basal area (BA) variations, eddy covariance carbon fluxes, predawn water potential (Ψpd ) and temperature at different timescales using an 8-yr dataset and a rainfall exclusion experiment in a Quercus ilex Mediterranean coppice. At the daily timescale, during periods of low temperature (< 5°C) and high water deficit (< -1.1 MPa), gross primary productivity and net ecosystem productivity remained positive whereas the stem increment was nil. Thus, stem increment appeared limited by drought and temperature rather than by carbon input. Annual growth was accurately predicted by the duration of BA increment during spring (Δtt0-t1 ). The onset of growth (t0 ) was related to winter temperatures and the summer interruption of growth (t1 ) to a threshold Ψpd value of -1.1 MPa. We suggest that using environmental drivers (i.e. drought and temperature) to predict stem growth phenology can contribute to an improvement in vegetation models and may change the current projections of Mediterranean forest productivity under climate change scenarios.

Sweets for the foe - effects of nonstructural carbohydrates on the susceptibility of Quercus robur against Phytophthora quercina.

The root-rot pathogen Phytophthora quercina is a key determinant of oak decline in Europe. The susceptibility of pedunculate oak (Quercus robur) to this pathogen has been hypothesized to depend on the carbon availability in roots as an essential resource for defense. Microcuttings of Q. robur undergo an alternating rhythm of root and shoot growth. Inoculation of mycorrhizal (Piloderma croceum) and nonmycorrhizal oak roots with P. quercina was performed during both growth phases, that is, root flush (RF) and shoot flush (SF). Photosynthetic and morphological responses as well as concentrations of nonstructural carbohydrates (NSC) were analyzed. Infection success was quantified by the presence of pathogen DNA in roots. Concentrations of NSC in roots depended on the alternating root/shoot growth rhythm, being high and low during RF and SF, respectively. Infection success was high during RF and low during SF, resulting in a significantly positive correlation between pathogen DNA and NSC concentration in roots, contrary to the hypothesis. The alternating growth of roots and shoots plays a crucial role for the susceptibility of lateral roots to the pathogen. NSC availability in oak roots has to be considered as a benchmark for susceptibility rather than resistance against P. quercina.

Alleviation of Zn toxicity by low water availability.

Heavy metal contamination and drought are expected to increase in large areas worldwide. However, their combined effect on plant performance has been scantly analyzed. This study examines the effect of Zn supply at different water availabilities on morpho-physiological traits of Quercus suber L. in order to analyze the combined effects of both stresses. Seedlings were treated with four levels of zinc from 3 to 150 µM and exposed to low watering (LW) or high watering (HW) frequency in hydroponic culture, using a growth chamber. Under both watering regimes, Zn concentration in leaves and roots increased with Zn increment in nutrient solution. Nevertheless, at the highest Zn doses, Zn tissue concentrations were almost twice in HW than in LW seedlings. Functional traits as leaf photosynthetic rate and root hydraulic conductivity, and morphological traits as root length and root biomass decreased significantly in response to Zn supply. Auxin levels increased with Zn concentrations, suggesting the involvement of this phytohormone in the seedling response to this element. LW seedlings exposed to 150 µM Zn showed higher root length and root biomass than HW seedlings exposed to the same Zn dose. Our results suggest that low water availability could mitigate Zn toxicity by limiting internal accumulation. Morphological traits involved in the response to both stresses probably contributed to this response.

Effects of three fire-suppressant foams on the germination and physiological responses of plants.

Suppressant foams used to fight forest fires may leave residual effects on surviving biota that managers need to consider prior to using them. We examined how three fire-suppressant foams (FSFs) (Forexpan S, Phos-Chek-WD881, and Silv-ex) affected seed germination and physiological responses of three plant species. Exposure to FSFs, whether in diluted concentrations or those typical in the field, reduced final germination percentages of seeds grown in petri dishes and within growth chambers. However, the FSFs did not cause total germination failure in any treatment. Inhibition of germination increased with longer exposure times, but only to diluted FSF solutions. Unlike in the laboratory experiments, none of the three FSFs affected seedling emergence when tested in field conditions. Further, we found no evidence of long-term phytotoxic effects on antioxidant enzyme activity nor chlorophyll content of the plant saplings. Therefore, although the three FSFs showed evidence of phytotoxicity to plants in laboratory tests, their actual impact on terrestrial ecosystems may be minimal. We suggest that the benefits of using these FSFs to protect plants in threatened forest ecosystems outweigh their minor risks.

Physiological response and molecular mechanism of Quercus variabilis under cadmium stress.

Heavy metal pollution is a global environmental problem, and Quercus variabilis has a stronger tolerance to Cd stress than do other species. We aimed to explore the physiological response and molecular mechanisms of Q. variabilis to Cd stress. In this study, the antioxidant enzyme activities of leaves were determined, while the photosynthetic parameters of leaves were measured using Handy PEA, and ion fluxes and DEGs in the roots were investigated using noninvasive microtest technology (NMT) and RNA sequencing techniques, respectively. Cd stress at different concentrations and for different durations affected the uptake patterns of Cd2+ and H+ by Q. variabilis and affected the photosynthetic efficiency of leaves. Moreover, there was a positive relationship between antioxidant enzyme (CAT and POD) activity and Cd concentration. Transcriptome analysis revealed that many genes, including genes related to the cell wall, glutathione metabolism, ion uptake and transport, were significantly upregulated in response to cadmium stress in Q. variabilis roots. WGCNA showed that these DEGs could be divided into eight modules. The turquoise and blue modules exhibited the strongest correlations, and the most significantly enriched pathways were the phytohormone signaling pathway and the phenylpropanoid biosynthesis pathway, respectively. These findings suggest that Q. variabilis can bolster plant tolerance by modulating signal transduction and increasing the synthesis of compounds, such as lignin, under Cd stress. In summary, Q. variabilis can adapt to Cd stress by increasing the activity of antioxidant enzymes, and regulating the fluxes of Cd2+ and H+ ions and the expression of Cd stress-related genes.

Effects of drought on mesophyll conductance and photosynthetic limitations at different tree canopy layers.

In recent years, many studies have focused on the limiting role of mesophyll conductance (gm ) to photosynthesis (An ) under water stress, but no studies have examined the effect of drought on gm through the forest canopy. We investigated limitations to An on leaves at different heights in a mixed adult stand of sessile oak (Quercus petraea) and beech (Fagus sylvatica) trees during a moderately dry summer. Moderate drought decreased An of top and lowest beech canopy leaves much more than in leaves located in the mid canopy; whereas in oak, An of the lower canopy was decreased more than in sunlit leaves. The decrease of An was probably not due to leaf-level biochemistry given that VCmax was generally unaffected by drought. The reduction in An was instead associated with reduction in stomatal and mesophyll conductances. Drought-induced increases in stomatal limitations were largest in leaves from the top canopy, whereas drought-induced increases in mesophyll limitations were largest in leaves from the lowest canopy. Sensitivity analysis highlighted the need to decompose the canopy into different leaf layers and to incorporate the limitation imposed by gm when assessing the impact of drought on the gas exchange of tree canopies.

The effect of induced heat waves on Pinus taeda and Quercus rubra seedlings in ambient and elevated CO2 atmospheres.

Here, we investigated the effect of different heat-wave intensities applied at two atmospheric CO2 concentrations ([CO2]) on seedlings of two tree species, loblolly pine (Pinus taeda) and northern red oak (Quercus rubra). Seedlings were assigned to treatment combinations of two levels of [CO2] (380 or 700 µmol mol(-1)) and four levels of air temperature (ambient, ambient +3°C, or 7-d heat waves consisting of a biweekly +6°C heat wave, or a monthly +12°C heat wave). Treatments were maintained throughout the growing season, thus receiving equal heat sums. We measured gas exchange and fluorescence parameters before, during and after a mid-summer heat wave. The +12°C heat wave, significantly reduced net photosynthesis (Anet) in both species and [CO2] treatments but this effect was diminished in elevated [CO2]. The decrease in Anet was accompanied by a decrease in Fv'/Fm' in P. taeda and ΦPSII in Q. rubra. Our findings suggest that, if soil moisture is adequate, trees will experience negative effects in photosynthetic performance only with the occurrence of extreme heat waves. As elevated [CO2] diminished these negative effects, the future climate may not be as detrimental to plant communities as previously assumed.

Cyclobotryoxide, a phytotoxic metabolite produced by the plurivorous pathogen Neofusicoccum australe.

Two isolates of Neofusicoccum australe belonging to ITS haplotypes H4 and H1 and associated with grapevine cordon dieback and branch dieback of Phoenicean juniper, respectively, have been shown to produce in vitro structurally different secondary metabolites. From the strain BOT48 of N. australe (haplotype H4) a new cyclohexenone oxide, namely, cyclobotryoxide, was isolated together with 3-methylcatechol and tyrosol. Cyclobotryoxide was characterized as (1S,5R,6S)-5-hydroxy-3-methoxy-4-methyl-7-oxabicyclo[4.1.0]hept-3-en-2-one by spectroscopic, optical, and chemical methods. The strain BL24 (haplotype H1) produced tyrosol along with botryosphaerone D and (3S,4S)-3,4,8-trihydroxy-6-methoxy-3,4-dihydro-1(2H)-naphthalenone. The metabolites obtained from both strains were tested at four concentrations on leaves of grapevine cv. Cannonau, holm oak, and cork oak by the leaf puncture assay. Cyclobotryoxide proved to be the most phytotoxic compound. Tyrosol and cyclobotryoxide were also tested on detached grapevine leaves at concentrations of 0.25 and 0.5 mg/mL. Only cyclobotryoxide was found to be active in this bioassay.

Air pollution from gas refinery through contamination with various elements disrupts semiarid Zagros oak (Quercus brantii Lindl.) forests, Iran.

Soils and oak trees (Quercus brantii Lindl.) in Zagros forests are suffering from the air pollution caused by the Ilam Gas Refinery. Thus, for the first time, we investigated the contamination level of sulfur and trace elements in these ecosystems. Sampling of soil and tree leaves was carried out in different seasons of 2019 and at different distances from the gas refinery. The results showed that soils and leaves at the various distances compared with control distance (10,000 m) were more affected by the gas refinery. Distance from the pollution source and physicochemical properties of soils were the main factors affecting contamination of soil elements contents. The soils with pollution load indices (PLI) of 4.54 were in the highly polluted category. Sulfur was at highly polluted category in soils and were highly enriched in trees. The trees mainly absorbed studied elements via their aerial organs. Our findings indicated that oak trees with the highest value of metal accumulation index are influence tools for monitoring various elements in the polluted air produced by the gas refinery. It is recommended that the ecosystem components near the refinery be studied to accurately evaluate disorders in the food chain.

Plant-soil distribution of potentially toxic elements in response to elevated atmospheric CO2.

The distribution of contaminant elements within ecosystems is an environmental concern because of these elements' potential toxicity to animals and plants and their ability to hinder microbial ecosystem services. As with nutrients, contaminants are cycled within and through ecosystems. Elevated atmospheric CO2 generally increases plant productivity and alters nutrient element cycling, but whether CO2 causes similar effects on the cycling of contaminant elements is unknown. Here we show that 11 years of experimental CO2 enrichment in a sandy soil with low organic matter content causes plants to accumulate contaminants in plant biomass, with declines in the extractable contaminant element pools in surface soils. These results indicate that CO2 alters the distribution of contaminant elements in ecosystems, with plant element accumulation and declining soil availability both likely explained by the CO2 stimulation of plant biomass. Our results highlight the interdependence of element cycles and the importance of taking a broad view of the periodic table when the effects of global environmental change on ecosystem biogeochemistry are considered.

Effects of light spectra and 15N pulses on growth, leaf morphology, physiology, and internal nitrogen cycling in Quercus variabilis Blume seedlings.

Light spectra of sunlight transmittance can generate an interactive effect with deposited nitrogen (N) on regenerated plants across varied shading conditions. Total N content in understory plants can be accounted for by both exogeneous and endogenous sources of derived N, but knowledge about the response of inner N cycling to interactive light and N input effects is unclear. We conducted a bioassay on Chinese cork oak (Quercus variabilis Blume) seedlings subjected to five-month N pulsing with 15NH4Cl (10.39 atom %) at 120 mg 15N plant-1 under the blue (48.5% blue, 33.7% green, and 17.8% red), red (14.6% blue, 71.7% red, 13.7% green), and green (17.4% blue, 26.2% red, 56.4% green) lighting-spectra. Half of the seedlings were fed twice a week using a 250 ppm N solution with micro-nutrients, while the other half just received distilled water. Two factors showed no interaction and neither affected growth and morphology. Compared to the red-light spectrum, that in blue light increased chlorophyll and soluble protein contents and glutamine synthetase (GS) activity, root N concentration, and N derived from the pulses. The green-light spectrum induced more biomass allocation to roots and a higher percentage of N derived from internal reserves compared to the red-light spectrum. The 15N pulses reduced the reliance on N remobilization from acorns but strengthened shoot biomass, chlorophyll content, GS activity, and N concentration. In conclusion, light spectrum imposed an independent force from external N pulse to modify the proportion of N derived from internal sources in total N content in juvenile Q. variabilis.

Effects of nitrogen additions on mesophyll and stomatal conductance in Manchurian ash and Mongolian oak.

The response of plant CO2 diffusion conductances (mesophyll and stomatal conductances, gm and gsc) to soil drought has been widely studied, but few studies have investigated the effects of soil nitrogen addition levels on gm and gsc. In this study, we investigated the responses of gm and gsc of Manchurian ash and Mongolian oak to four soil nitrogen addition levels (control, low nitrogen, medium nitrogen and high nitrogen) and the changes in leaf anatomy and associated enzyme activities (aquaporin (AQP) and carbonic anhydrase (CA)). Both gm and gsc increased with the soil nitrogen addition levels for both species, but then decreased under the high nitrogen addition level, which primarily resulted from the enlargements in leaf and mesophyll cell thicknesses, mesophyll surface area exposed to intercellular space per unit leaf area and stomatal opening status with soil nitrogen addition. Additionally, the improvements in leaf N content and AQP and CA activities also significantly promoted gm and gsc increases. The addition of moderate levels of soil nitrogen had notably positive effects on CO2 diffusion conductance in leaf anatomy and physiology in Manchurian ash and Mongolian oak, but these positive effects were weakened with the addition of high levels of soil nitrogen.

Effect of elicitors on holm oak somatic embryo development and efficacy inducing tolerance to Phytophthora cinnamomi.

Holm oak trees (Quercus ilex L.) mortality is increasing worryingly in the Mediterranean area in the last years. To a large degree this mortality is caused by the oomycete Phytophthora spp., which is responsible for forest decline and dieback in evergreen oak forest areas of the southwestern Iberian Peninsula. This study is based on the possibility of applying chemical elicitors or filtered oomycete extracts to holm oak somatic embryos (SE) in order to induce epigenetic memory, priming, that may increase tolerance to the pathogen in future infections. To this end, we first examined the effect of priming treatments on SE development and its oxidative stress state, to avoid elicitors that may cause damage to embryogenic tissues. Both, the sterile oomycete extracts and the chemical elicitor methyl jasmonate (MeJA) did not produce any detrimental effect on SE growth and development, unlike the elicitors benzothiadiazole (BTH) and p-aminobenzoic acid (PABA) that reduced the relative weight gain and resulted in necrotic and deformed SE when were applied at high concentrations (25 µM BTH or 50 µM PABA) in accordance with their high malondialdehyde content. No significant differences among elicitation treatments were found in dual culture bioassays, although those SEs elicited with 50 µM MeJA increased H2O2 production after challenged against active oomycete indicating the activation of stress response. Since this elicitation treatment did not produce any adverse effect in the embryogenic process we suggest that could be used in further priming experiments to produce holm oak plants adapted to biotic stress.

[Protection by some plant methanol extracts of cherry tomatoes (Solanum lycopersicum var. Cerasiforme) from fungic infection by Alternaria alternata]. / Utilisation d'extraits méthanoliques de plantes pour la protection des cultures de tomates-cerises (Solanum lycopersicum var. cerasiforme) contre l'infection fongique par Alternaria alternata.

Cherry tomato is very susceptible to fungal infections that can cause considerable damage in crops and during storage. Alternaria infection is one of the most common and dangerous alterations for this fruit. They are caused by Alternaria alternata or some other species belonging to the same genus. In this work, we tested the antifungal activity of methanol extracts from five plants harvested in the region of Jijel (Algeria) on A. alternata. The activity was first tested in vitro and then on greenhouse cherry tomato plants: extracts were applied to healthy plants before infection in order to test their preventive action, and after infection to determine whether they are able to knock out Alternaria. Results showed that Rosmarinus officinalis and Lavandula angustifolia extracts were the most active in vitro on A. alternata. Microscopic observations of the mold indicated that these extracts inhibited the dictyospores production. The antifungal activity tested on the plants grown in greenhouse revealed that R. officinalis extract still was the most active. Extracts of L. angustifolia and Punica granatum did not protect the plants from Alternaria infection, but provided a total cure at the end of the treatment. Extracts from Quercus suber and Eucalyptus globulus were the least active. They did not bestow any protection nor complete healing of the plants. Dictyospores counting on fruits at the end of the treatment confirmed the results obtained for the greenhouse crops.

TITLE: Utilisation d'extraits méthanoliques de plantes pour la protection des cultures de tomates-cerises (Solanum lycopersicum var. cerasiforme) contre l'infection fongique par Alternaria alternata. ABSTRACT: La tomate-cerise est un fruit très sujet aux infections fongiques qui peuvent causer des dégâts considérables dans les cultures et lors de la conservation. Les alternarioses comptent parmi les altérations les plus répandues et dangereuses pour ce fruit. Elles sont causées par Alternaria alternata ou d'autres espèces appartenant au même genre. Dans ce travail, nous avons testé l'activité antifongique d'extraits méthanoliques de cinq plantes récoltées dans la région de Jijel (Algérie) sur A. alternata. L'activité a d'abord été testée in vitro, puis sur des plants de tomates-cerises cultivés sous serre : les extraits ont été appliqués sur des plants sains, avant l'infection, afin de tester leur action préventive, et après l'infection pour déterminer s'ils sont capables de traiter l'alternariose. Les résultats ont montré que les extraits de Rosmarinus officinalis et Lavandula angustifolia étaient les plus actifs in vitro sur A. alternata. L'observation microscopique de la moisissure a indiqué que ces extraits agissaient en inhibant sa production de dictyospores. L'activité antifongique testée sur les plants cultivés sous serre a révélé que l'extrait de R. officinalis était toujours le plus actif. Venaient ensuite les extraits de L. angustifolia et Punica granatum qui n'ont pas permis la protection des plants contre l'alternariose, mais qui ont néanmoins donné une guérison totale à la fin du traitement. Les extraits de Quercus suber et Eucalyptus globulus étaient les moins actifs. Ils n'ont permis ni la prévention, ni la guérison complète des plants. Le comptage des dictyospores réalisé sur les fruits à la fin du traitement a confirmé les résultats obtenus pour les cultures sous serre.

Genetic transformation of cork oak (Quercus suber L.) for herbicide resistance.

The bar gene was introduced into the cork oak genome. Cork oak embryogenic masses were transformed using the Agrobacterium strain AGL1 which carried the plasmid pBINUbiBar. This vector harbours the genes, nptII and bar, the latter under control of the maize ubiquitin promoter. The transgenic embryogenic lines were cryopreserved. Varying activities of phosphinothricin acetyl transferase were detected among the lines, which carried 1-4 copies of the insert. Molecular and biochemical assays confirmed the stability and expression of the transgenes 3 months after thawing the cultures. These results demonstrate genetic engineering of herbicide tolerance in Quercus spp.

Image analysis of epicuticular damage to foliage caused by dry deposition of the air pollutant nitric acid.

Nitric acid vapor is produced by the same photochemical processes that produce ozone. In the laboratory, concentrated nitric acid is a strong acid and a powerful oxidant. In the environment, where the concentrations are much lower, it is an innocuous source of plant nitrogen. As an air pollutant, which mode of action does dry deposition of nitric acid follow? We investigated the effects of dry deposition of nitric acid on the foliage of four tree species native to the western United States. A novel controlled environment, fumigation system enabled a four-week exposure at concentrations consistent with ambient diurnal patterns. Scanning electron microscopy and automated image analysis revealed changes in the epicuticular wax layer during fumigation. Exposure to nitric acid resulted in a reproducible suite of damage symptoms that increased with increasing dose. Each tree species tested exhibited a unique set of damage features, including cracks, lesions, and conformation changes to epicuticular crystallite structures. Dry deposition of atmospheric nitric acid caused substantial perturbation to the epicuticular surface of all four tree species investigated, consistent with the chemical oxidation of epicuticular waxes. Automated image analysis eliminated many biases that can trouble microscopy studies. Trade names and commercial enterprises or products are mentioned solely for information. No endorsements by the U.S. Department of Agriculture are implied.