Irrigating Onions and Potatoes with Chromium and Nickel: Its Effects on Catalase and Peroxidase Activities and the Cross-Contamination of Plants

Tropospheric ozone (O₃) has long been documented to cause an injury to plants, but a plants’ protectant, widely applicable in agronomical practice, does not exist. We evaluated the potential antiozonate efficacy of the antitranspirant di-1-p-menthene (Vapor Gard) compared with ethylenediurea (EDU) on Bel-W3 tobacco plants. Plants were treated either with water, or by EDU (10, 100, and 500 mg dm^?3), or by vapor (1, 5, 10, and 50 ml dm^?3) and were exposed either to O₃-enriched (90 ppb) or O₃-free air, for 12 days and 8 h day^?1. EDU when applied at 10 mg dm^?3 did not protect the plants against O₃, but when applied at 100 and 500 mg dm^?3 offered a significant protection to the plants. Vapor, when applied at 1 ml dm^?3 did not protect the plants against O₃, neither by terms of foliar visible injury nor by terms of aboveground biomass. In addition, when applied at 10 and 50 ml dm^?3 caused phytotoxicity to all the plants, which it was expressed as necrotic spots on the leaves’ surface, misshaping of the leaves, or short plants' height_. It is obvious that vapor does not protect Bel-W3 tobacco plants against O₃. The antiozonate role of di-1-p-menthene is species-specific and probably occurs only under short-term exposures.     

Rapid and direct low micromolar NMR method for the simultaneous detection of hydrogen peroxide and phenolics in plant extracts

A rapid and direct low micromolar 1H NMR method for the simultaneous identification and quantification of hydrogen peroxide and phenolic compounds in plant extracts was developed. The method is based on the highly deshielded 1H NMR signal of H?O? at ～10.30 ppm in DMSO-d? and the combined use of picric acid and low temperature, near the freezing point of the solution, in order to achieve the minimum proton exchange rate. Line widths of H?O? below 3.8 Hz were obtained for several Greek oregano extracts which resulted in a detection limit of 0.7 μmol L?1. Application of an array of NMR experiments, including 2D 1H-13C HMBC, spiking of the samples with H?O?, and variable temperature experiments, resulted in the unequivocal assignment of H?O? precluding any confusion with interferences from intrinsic phenolics in the extract.     

The impact of biological pesticides on arbuscular mycorrhizal fungi

Arbuscular mycorrhizal (AM) fungi have a key role for plant nutrition in organic farming systems where crop protection relies on biopesticides. Although these are considered safe, their effects on non-target organisms, such as AM fungi, are not known and should be evaluated. A pot and a field experiment were employed to investigate the impact of biological pesticides (azadirachtin, spinosad, pyrethrum and terpens) on exogenous AM fungal inoculum (pots) and on indigenous AM fungi (field). The synthetic fungicide carbendazim and non-pesticide treated controls with or without mycorrhizal inoculation were also included. Plant growth and root colonization were measured 20 and 40 days post inoculation (dpi) in the pot experiment, or 40 and 90 dpi in the field study. Pesticide effects on the structure of the intraradical AM fungal community were determined via DGGE and cloning. Spinosad, pyrethrum and terpenes did not affect the colonization ability and the structure of the AM fungal community. On the contrary, pot application of azadirachtin resulted in a selective inhibition of the Glomus etunicatum strain of the inoculum. DGGE analysis showed that the field application of azadirachtin induced significant and persistent shifts in the AM fungal community. Carbendazim completely hampered mycorrhizal colonization in pots, compared to its field application which had a transitory effect on the colonization ability and the community structure of indigenous AM fungi. Our study provides first evidence for the effects of biological pesticides on the diversity of AM fungi.     

Quantitative bioluminometric method for DNA-based species/varietal identification in food authenticity assessment

A method is reported for species quantification by exploiting single-nucleotide polymorphisms (SNPs). These single-base changes in DNA are particularly useful because they enable discrimination of closely related species and/or varieties. As a model, quantitative authentication studies were performed on coffee. These involved the determination of the percentage of Arabica and Robusta species based on a SNP in the chloroplastic trnL(UAA)-trnF(GAA) intraspacer region. Following polymerase chain reaction (PCR), the Robusta-specific and Arabica-specific fragments were subjected to 15 min extension reactions by DNA polymerase using species-specific primers carrying oligo(dA) tags. Biotin was incorporated into the extended strands. The products were captured in streptavidin-coated microtiter wells and quantified by using oligo(dT)-conjugated photoprotein aequorin. Aequorin was measured within 3 s via its characteristic flash-type bioluminescent reaction that was triggered by the addition of Ca(2+). Because of the close resemblance between the two DNA fragments, during PCR one species serves as an internal standard for the other. The percentage of the total luminescence signal obtained from a certain species was linearly related to the percent content of the sample with respect to this species. The method is accurate and reproducible. The microtiter well-based assay configuration allows high sample throughput and facilitates greatly the automation.     

Bioactive Lipids of Marine Microalga Chlorococcum sp. SABC 012504 with Anti-Inflammatory and Anti-Thrombotic Activities

Microalgae are at the start of the food chain, and many are known producers of a significant amount of lipids with essential fatty acids. However, the bioactivity of microalgal lipids for anti-inflammatory and antithrombotic activities have rarely been investigated. Therefore, for a sustainable source of the above bioactive lipids, the present study was undertaken. The total lipids of microalga Chlorococcum sp., isolated from the Irish coast, were fractionated into neutral-, glyco-, and phospho-lipids, and were tested in vitro for their anti-inflammatory and antithrombotic activities. All tested lipid fractions showed strong anti-platelet-activating factor (PAF) and antithrombin activities in human platelets (half maximal inhibitory concentration (IC₅₀) values ranging ~25–200 μg of lipid) with the highest activities in glyco- and phospho-lipid fractions. The structural analysis of the bioactive lipid fraction-2 revealed the presence of specific sulfoquinovosyl diacylglycerols (SQDG) bioactive molecules and the HexCer-t36:2 (t18:1/18:1 and 18:2/18:0) cerebrosides with a phytosphingosine (4-hydrosphinganine) base, while fraction-3 contained bioactive phosphatidylcholine (PC) and phosphatidylethanolamine (PE) molecules. These novel bioactive lipids of Chlorococcum sp. with putative health benefits may indicate that marine microalgae can be a sustainable alternative source for bioactive lipids production for food supplements and nutraceutical applications. However, further studies are required towards the commercial technology pathways development and biosafety analysis for the use of the microalga.     

Immunohistochemical expression of dogTERT in canine testicular tumours in relation to PCNA,ki67 and p53 expression.

The objective of the study was to determine the immunohistochemical expression of canine TERT in canine testicular tumours comparing two different antibodies for TERT, and to correlate them with well established markers specific to dividing cells such as PCNA and ki67, and with expression of the p53 tumour suppressor gene. The study included 36 cases of canine testicular tumours, which were categorized as 12 Sertoli Cell Tumours (SCT), 20 seminomas, 3 interstitial cell tumours and 1 mixed germ cell-sex cord stromal tumour (MT). Two antibodies for hTERT were examined; a highly specific TERT antibody, RCK-hTERT, was evaluated for the first time. Immunodetection of RCK-hTERT was observed in 31% of tumours examined (6/20 Seminomas, 4/12 SCT, 1/3 interstitial cell tumour and 0/1 mixed germ cell-sex cord stromal tumour), while the NCL-hTERT in 67% of them (15/20 Seminomas, 6/12 SCT, 3/3 interstitial cell tumour and 0/1 ΜΤ). PCNA immunoreactivity was detected in all cases. Regarding ki67, 3 SCT, 12 seminomas and all interstitial cell tumours showed clear immunoreaction. p53 immunoreactivity was detected in 6 SCT, 15 seminomas and all interstitial cell tumours. The immunohistochemical expression of both TERT antibodies are discussed and compared in order to clarify their potential usefulness in canine testicular malignancies in relation to the expression of well known cell cycle markers. Our results indicate that TERT and PCNA are useful proliferation markers but not helpful to evaluate prognosis. Instead of that ki67 and p53 could be used for predicting aggressiveness in this group of tumours     

Competitive interactions of fifty barley cultivars with Avena sterilis and Asperugo procumbens

A 2-year study was conducted in northern Greece to assess the ability of 29 six-row and 21 two-row barley (Hordeum vulgare L.) cultivars to compete with winter wild oat [Avena sterilis spp. ludoviciana (Durieu) Gill & Magne] and German-madwort (Asperugo procumbens L.). The fresh weight of both weeds grown in competition with six of the most competitive six-row barley cultivars (Bulk Resist. Desc-Sutter, Lignee 640, Paico's, Mantnan's-EH 165, Pistacho and Malebo) was reduced by 61–75% compared with one of the least competitive six-row cultivars [Gloria's-Copal's (30B-4Y-20M)]. The corresponding fresh weight reduction of both weeds grown in competition with three of the most competitive two-row barley cultivars (Goldmarker, Shanon and Helena) ranged from 59 to 76% compared with one of the least competitive two-row cultivars (Guanajuato). The ability to withstand competition (AWC) from winter wild oat and German-madwort of the six-row and two-row cultivars ranged from 33.7 to 78.3% and 26.7 to 69.2%, respectively. Representative yield losses ranged from 24 to 68% and 30 to 75%, respectively. The corresponding ability to compete (AC) indices ranged from 22.5 to 64.8% and 21.1 to 59.1%. The results of this study indicated that the barley cultivars Ligne 640 and Pistacho (2B-1Y-1B) had high AWC from winter wild oat and German-madwort and provided high grain yield in both weedy and weed-free conditions. Consequently, they could successfully be used in Mediterranean organic (no herbicide use) and conventional agricultural systems with reduced herbicide use. Barley cultivars with high AC and ability to produce high grain yield under weedy conditions [with Bulk Resist. Desc-Sutter, Arupo's and Gloria's-Copal's (30B-4Y-23M) among the most efficient] could also successfully be used in organic fields infected by the two weeds.     

Nutritional quality of greenhouse lettuce at harvest and after storage in relation to N application and cultivation season

The effect of five levels of nitrogen fertilization on the growth and nutritional quality of Cos lettuce (Lactuca sativa L. cv. Parris Island) at harvest and after storage was studied during autumn and winter in South-West Greece. Plants were cultivated hydroponically in a greenhouse and the nitrate, chlorophyll and ascorbic acid (vitamin C) concentrations within the plant tissues were measured at harvest and following storage at 5 or 10 °C for 10 days. Nitrate accumulated in the leaves with increasing amounts of N within the nutrient solution and was higher in the winter than in the autumn. At the lowest N level (20 mg L⁻¹), the inner leaves accumulated more nitrate than the outer leaves, whereas at higher N levels (140, 200 or 260 mg L⁻¹) nitrate accumulation was higher in the outer leaves. Overall, the highest nitrate concentrations were detected in the petiole and the proximal end of the leaf, but at the lowest N application rate (20 mg L⁻¹) nitrate accumulated in the distal region of the leaf too. Although the nitrate concentrations within the leaves did not change significantly during 10 days storage at 5 or 10 °C, the chlorophyll and vitamin C concentrations decreased. Chlorophyll loss was higher in lettuce that was grown under low N levels and was higher at 10 °C than at 5 °C, but was reduced by enclosure of the lettuce in polyethylene film. It is concluded that the optimum N application rate for Cos lettuce grown hydroponically under cover during autumn and winter in South-West Greece, and in other areas with a similar climate, is 200 mg N L⁻¹ because at this N rate yield is satisfactory and leaf nitrate concentrations are below the maximum acceptable level for human consumption. Nutritional value (vitamin C concentration) and market quality (chlorophyll content) are highest at harvest and decrease during storage, but quality in terms of nitrate concentration does not change.