Costas J. Saitanis

Effects of ozone on chlorophyll and quantum yield of tobacco (Nicotiana tabacum L.) varieties.

Plants of Bel-W3 and of seven commercial tobacco varieties (Nicotiana tabacum L.) were exposed to two relatively low ozone concentrations (90 or 135 ppb) for 20 consecutive days, for 8 h per day. Ozone caused necrotic and chlorotic spots, acceleration of leaf senescence, depression of photosynthetic mechanism, chlorophyll diminution and greater destruction of chl a than of chl b. The higher sensitivity of chl a was also confirmed by exposure of segments of leaves in test tubes to high ozone concentration (>1000 ppb) as well as by bubbling of ozone in extracts of chlorophyll in vitro. The quantum yield (QY) of photosynthesis was positively correlated with the chlorophyll content and negatively correlated with the visible injury and the chl b/a ratio.

Mites associated with stored products in Greece

A faunistic survey of mites was conducted in many product stores during a 6-year study period, 2000–2005, in Greece. A total of 1,073 samples were taken from 34 Greek counties. The survey was carried out on grains (wheat, maize, oat, barley), flour, bran, manufactured agricultural foodstuffs, dried fruits (figs, raisins), residues and dust, stored in varying quantities in five types of storage facilities (stores of agricultural cooperative unions, farm stores, commercial stores, flour mills and silos). Dominance-frequency analysis and redundancy analysis (RDA) were used to reveal the preferences of the collected taxa. Approximately 55% of the samples contained mites and 65 mite taxa were identified, belonging to 15 families in four orders. Six species, namely, Acarus gracilis Hughes, A. immobilis Griffiths, Caloglyphus oudemansi (Zachvatkin), Suidasia medanensis Oudemans, Tyrophagus perniciosus Zachvatkin and Kleemania plumigera (Oudemans), were new to the fauna of Greece. Five species, Tyrophagus similis Volgin, Blattisocius mali (Oudemans), Neoseiulus barkeri (Hughes), Cheyletus cacahuamilpensis Baker and Storchia robustus (Berlese), were recorded for the first time in stored products in Greece. Lepidoglyphus destructor (Schrank), Tyrophagus putrescentiae (Schrank) and Acarus siro L. were dominant or intermediate in all storage facilities examined. Cheyletus malaccensis Oudemans was the most common predatory mite. The highest percentage of infestation (65.3) was recorded in the samples from stores of agricultural cooperative unions. Residue-type materials had the highest degrees and percentages of infestation.

Background ozone monitoring and phytodetection in the greater rural area of Corinth––Greece

Natural background ozone levels were monitored in three places within the greater rural area of Corinth, namely Bogdani Hill, Astronomical Observatory of Krionerion, and Kiato, and compared with ambient ozone monitored in the metropolitan area of Athens. Measurements were made sequentially, for a few weeks at each place, during the summer of 2000. In addition, ozone phytodetection, using tobacco (Nicotiana tabacum L.) plants of the Bel-W3 and Zichnomirodata varieties, was conducted in 12 places (the above included). Moreover, stomatal conductance was measured in the Bel-W3 plants, as well as in leaves of cultivated grape-vines (Vitis vinifera L.) and in needles of Aleppo pine (Pinus halepensis Mill.) trees and compared with the diurnal pattern of ozone concentrations. The 24 and 12 (08:00-20:00) hourly averages of ozone concentrations were high in Athens (37; 51 ppb), at Bogdani Hill (53; 56 ppb) and at the Astronomical Observatory (56; 55 ppb), but relatively low in Kiato (30; 34 ppb). Furthermore, the average daily AOT40 (accumulated exposure over 40 ppb for the daylight hours) (ppbh) was 193 in Athens, 212 at Bogdani Hill, 192 at the Astronomical Observatory and 47 in Kiato. Ozone concentrations exhibited the usual diurnal pattern in Athens (altitude 50 m), where they were maximum during midday and early afternoon hours, as well as at Bogdani Hill (300 m) and in Kiato (5 m) where, however, they were maximal 1-3 h later. At the Astronomical Observatory (altitude 920 m) ozone remained constant during both daylight and night hours. The differences in diurnal patterns are consistent with those in places of different elevation, reported elsewhere. The Bel-W3 plants were injured at all 12 places; Zichnomirodata plants exhibited lower injury and only in some of the places; probable ozone symptoms were also observed on vine plants and pine trees. The greatest injury was observed at the high altitude places of Astronomical Observatory and Mougostos. Stomatal conductance, in all three species, peaked during morning and early midday hours when ozone levels were higher in the high altitude, and lower in the low altitude, places.

Instrumental recording and biomonitoring of ambient ozone in the Greek countryside.

Among eight commercial Greek varieties of tobacco (Nicotiana tabacum L.) tested for their ozone-sensitivity levels, the Zichnomirodata (KK6/5) variety was found to be the most sensitive, although less sensitive than the well-known super-sensitive Bel-W3. Besides qualitative differences in the appearance of macroscopic symptoms these two varieties can be used simultaneously as a reliable pair of ozone bioindicators. The occurrence of ozone in the Greek countryside was surveyed by biomonitoring in 14 rural regions over the country and by a simultaneous biomonitoring and instrumental recording of ozone concentrations at a single remote side (Pournaria, Arcadia). Phytotoxic symptoms were observed mainly on the leaves of Bel-W3 and occasionally on those of Zichnomirodata varieties, suggesting that ozone levels were high enough to affect at least sensitive species. The instrumental monitoring (during a total period of 912 h) revealed maximum hourly O3 concentration 62 ppb, while the thresholds of 30, 40 and 50 ppb were exceeded for 40%, 20% and 6% of the recording period, respectively. The accumulated exposure over 40 ppb (AOT40) for the daylight hours over the 38 monitored days was 680 ppb h.

High doses of ethylene diurea (EDU) are not toxic to willow and act as nitrogen fertilizer

UNLABELLED Ethylene diurea (EDU) is synthetic chemical which protects plants against damage caused by ground level O3 and is used experimentally as a biomonitoring tool at doses usually ranging from 200 to 400mgL(-1) a.i. Although several studies have investigated the protective action of EDU, this mechanism remains unclear. Important uncertainties in EDU action are whether EDU acts as a source of nitrogen (N) to plants and whether high doses are phytotoxic. In order to answer these questions, we conducted an open-field experiment where potted willow (Salix sachalinensis Fr. Schm) plants were exposed to ambient O3 conditions and treated with 0, 800 or 1600mgL(-1) EDU as a soil drench, every nine days, for about 2.5months. We examined approximately 50 response variables. Based on N content in different plant organs, we found that (a) all EDU was transferred to the leaves and (b) high doses of EDU increased the leaf N content. However, EDU did not affect the C content and distribution within the plant body. Still, even at the highest dose, EDU was not toxic to this fast-growing species (however such a high dose should not be applied in uncontrolled environments); and there was no EDU persistence in the soil, as indicated by soil N content. Notably, our soil was free from organic matter and N-poor. KEY MESSAGE EDU per se does not cause toxicity to willow plants when applied as drench to a soil with no organic matter, rather, high EDU doses may act as nitrogen fertilizer in a nitrogen-poor soil.

Impacts of ethylenediurea (EDU) soil drench and foliar spray in Salix sachalinensis protection against O3-induced injury

It is widely accepted that elevated levels of surface ozone (O3) negatively affect plants. Ethylenediurea (EDU) is a synthetic substance which effectively protects plants against O3-caused phytotoxicity. Among other questions, the one still open is: which EDU application method is more appropriate for treating fast-growing tree species. The main aims of this study were: (i) to test if chronic exposure of Salix sachalinensis plants to 200-400mgEDUL-1, the usually applied range for protection against O3 phytotoxicity, is beneficial to plants; (ii) to evaluate the effects of chronic exposure to elevated O3 on S. sachalinensis; (iii) to assess the efficacy of two methods (i.e. soil drench and foliar spray) of EDU application to plants; (iv) to investigate the appropriate concentration of EDU to protect against elevated O3-induced damage in S. sachalinensis; and (v) to compare the two methods of EDU application in terms of effectiveness and EDU consumption. Current-year cuttings grown in infertile soil free from organic matter were exposed either to low ambient O3 (AOZ, 10-h≈28.3nmolmol-1) or to elevated O3 (EOZ, 10-h≈65.8nmolmol-1) levels during daylight hours. Over the growing season, plants were treated every nine days with 200mL soil drench of 0, 200 or 400mgEDUL-1 or with foliar spray of 0, 200 or 400mgEDUL-1 (in two separate experiments). We found that EDU per se had no effects on plants exposed to AOZ. EOZ practically significantly injured S. sachalinensis plants, and the impact was indifferent between the experiments. EDU did not protect plants against EOZ impact when applied as soil drench but it did protect them when applied as 200-400mgL-1 foliar spray. We conclude that EDU may be more effective against O3 phytotoxicity to fast-growing species when applied as a spray than when applied as a drench. Keymessage: Soil-drenched EDU was ineffective in protecting willow plants against O3-induced injury, whereas foliar-sprayed EDU was effective even at the concentration of 200mgL-1.

Morphology and life cycle of Marchalina hellenica (Gennadius) (Hemiptera: Margarodidae) on pine (Parnis Mt.) and fir (Helmos Mt.) forests of Greece

Abstract Marchalina hellenica (Gennadius) (Hemiptera: Margarodidae) is a scale insect, endemic in pine (Pinus halepensis) forests of Greece and other Mediterranean countries, which plays a major role in the production of honeydew honey. We investigated the morphological and the biological characteristics of M. hellenica in the pine forests of Mt. Parnis in comparison with those in the high-altitude fir (Abies cephalonica) forest of Mt. Helmos where it has been recently established, after anthropogenic intervention. Morphologically, the final body size of the 1st instar and the adult stages of M. hellenica in the fir forest were equal to those in the pine forest. Biologically, in the fir forest, the insect exhibited a long 1st instar’s period, which was the stage of its overwintering. In the pine forest, the 1st instar period was short and the insect overwinters in the stage of 2nd instar. The number of eggs per female in the fir forest was quite low (25–145) compared with the number of eggs in the pine forest (200–300). Concerning the qualitative parameters of the resulting honeydew-honey, statistically significant differences were found in Diastase and HMF (p < 0.0001). No difference was found in sucrose, fructose + glucose and water content, neither to electrical conductivity and total acidity. All values were within the EU limits (EU Directive 2001).

Application and further characterization of the snap bean S156/R123 ozone biomonitoring system in relation to ambient air temperature

Increased mixing ratios of ground-level ozone (O3) threaten individual plants, plant communities and ecosystems. In this sense, O3 biomonitoring is of great interest. The O3-sensitive S156 and the O3-tolerant R123 genotypes of snap bean (Phaseolus vulgaris L.) have been proposed as a potential tool for active biomonitoring of ambient O3. In the present study, an O3 biomonitoring was conducted, with the S156/R123 tool, along with a monitoring of O3 and other environmental conditions in an urban area in Athens, Greece, during the growing seasons of 2012 and 2013. Plant yield was evaluated to assess the effectiveness of AOT40 in interpreting O3-induced phytotoxicity. Across the two genotypes, an approximately two times lower total number of pods - and consequently lower bulk mass of seeds - was found in 2012 than in 2013, although there was no significant difference in the final AOT40 between the two years. No significant differences were observed in the stomatal density or conductance between the two genotypes, whereas it was estimated that, in both genotypes, the abaxial leaf surface contributes 2.7 fold to O3 intake in comparison to the adaxial one. By testing the role of ambient air temperature in outdoor plant environment chambers (OPECs), it was found that increased temperature limits mature pod formation and complicates interpretation of O3 impacts in terms of S156/R123 yields ratios. This is the first study providing evidence for a hormetic response of plants to ambient air temperature. This study also points out the complexity of using yield as a measure of O3 impact across different environments with the snap bean system, whereas visible foliar injury is more consistently related to O3 effects.

Volatile Compounds in Thymus sect. Teucrioides (Lamiaceae): Intraspecific and Interspecific Diversity, Chemotaxonomic Significance and Exploitation Potential

Thymus sect. Teucrioides comprises three species, namely, T. hartvigii, T. leucospermus, and T. teucrioides, distributed in Greece and Albania. The volatile constituents of all species of the section were obtained by hydrodistillation and investigated by GC‐FID and GC/MS analyses. Twenty populations were sampled and a total of 103 compounds were identified, representing 98.0–99.9% of the oil compositions. The oils were mainly characterized by high contents of monoterpene hydrocarbons (42.7–92.4%), with the exception of three oils for which oxygenated monoterpenes were the dominating constituents, viz., that of T. hartvigii ssp. macrocalyx, with linalool as main compound (89.2±0.5%), and those of T. hartvigii ssp. hartvigii and of one population of T. teucrioides ssp. candilicus, containing thymol as major component (46.4±3.1 and 38.2±3.9%, resp.). The most common compound in the oils of the 20 populations of the section was p‐cymene. Considerable variation was detected within and among populations, and seven chemotypes were distinguished, i.e., p‐cymene, linalool, p‐cymene/thymol, p‐cymene/γ‐terpinene, p‐cymene/borneol, p‐cymene/γ‐terpinene/borneol, and p‐cymene/linalool chemotypes. Different chemotypes may exist in the same population. Multivariate statistical analyses enabled the segregation of the oils within Thymus sect. Teucrioides into two groups, one consisting of the three subspecies of T. teucrioides and the second comprising the species T. hartvigii and T. leucospermus. A linalool‐rich chemotype, unique within the section, distinguished the oil of T. hartvigii ssp. macrocalyx from all other oils. The high oil content of p‐cymene and the preference for serpentine substrates render T. teucrioides species promising for future exploitation.

High doses of ethylenediurea (EDU) as soil drenches did not increase leaf N content or cause phytotoxicity in willow grown in fertile soil

Ground-level ozone (O3) levels are nowadays elevated in wide regions of the Earth, causing significant effects on plants that finally lead to suppressed productivity and yield losses. Ethylenediurea (EDU) is a chemical compound which is widely used in research projects as phytoprotectant against O3 injury. The EDU mode of action remains still unclear, while there are indications that EDU may contribute to plants with nitrogen (N) when the soil is poor in N and the plants have relatively small leaf area. To reveal whether the N content of EDU acts as a fertilizer to plants when the soil is not poor in N and the plants have relatively large total plant leaf area, willow plants (Salix sachalinensis Fr. Schm) were exposed to low ambient O3 levels and treated ten times (9-day interval) with 200mL soil drench containing 0, 800 or 1600mg EDU L-1. Fertilizer was added to a nutrient-poor soil, and the plants had an average plant leaf area of 9.1m2 at the beginning of EDU treatments. Indications for EDU-induced hormesis in maximum electron transport rate (Jmax) and ratio of intercellular to ambient CO2 concentration (Ci:Ca) were observed at the end of the experiment. No other EDU-induced effects on leaf greenness and N content, maximum quantum yield of photosystem II (Fv/Fm), gas exchange, growth and matter production suggest that EDU did not act as N fertilizer and did not cause toxicity under these experimental conditions.