Agostino Sorgonà

Nitrate uptake along the maize primary root: an integrated physiological and molecular approach

The spatial variability of the nitrate (NO₃-) uptake along the maize primary root axis was investigated at physiological and molecular levels. Net NO₃- uptake rate (NNUR) and its kinetic parameters, together with the gene expression of a high-affinity NO₃- transporter (NRT2.1), were evaluated. The activity and the expression of plasma membrane H⁺ -ATPase (PM H⁺ -ATPase), key enzyme in plant nutrition, were also analysed. The NNUR showed a heterogeneous spatial pattern along the root, where the regions closer to the root tip early exhibited higher capacity to absorb NO₃- than the basal regions, because of a higher maximum NNUR and faster induction of the inducible high-affinity transport system (iHATS), the presence of the high-affinity transport system (HATS) also at external NO₃- concentrations >100 µm and an improved NO₃- transport because of lower K(m) values. ZmNRT2.1 transcript abundances were not spatially correlated with NNUR, suggesting that post-translational effects or NAR2 protein co-expression could be involved. By contrast, PM H⁺ -ATPase displayed a similar spatial-temporal pattern as that of nitrate uptake, resulting in higher activity in the root tip than in the basal regions. Increased activities of the enzyme after nitrate supply resulted in enhanced expression of MAH3 and MAH4, PM H⁺ -ATPase subfamily II genes, while MAH1 was not expressed.

Coumarin Differentially Affects the Morphology of Different Root Types of Maize Seedlings

The effects of coumarin on the length, diameter, and branching density of different root types in maize seedlings (Zea mays L. cv. Cecilia) were investigated. The maize root system represents a useful model for morphological studies, as it consists of radicle, seminal, and nodal roots whose origin and development are quite different. Maize seedlings were grown in a hydroponic culture for 6 days, and then coumarin (at concentrations of 0, 25, 100, and 400μM) was added to the nutrient solution. Coumarin inhibited root length, but effects differed depending on the root type. C1/2 values, representing the coumarin concentration causing a 50% inhibition of the root length, were calculated by nonlinear regression. Six, 1, and 0.25 mM coumarin were sufficient to reduce the radicle, seminal, and nodal root lengths by 50%, respectively. At the highest coumarin concentration, the subapical root zone showed swelling. The degree recorded by average diameter was higher in nodal roots than in seminal and radicle roots. Furthermore, coumarin decreased the number of lateral roots and branching density more in the seminal than in the radicle roots. These results suggest the following order of sensitivity to coumarin: nodal > seminal > radicle roots. The observed spatial effects of coumarin could be ecologically significant, since taprooted species could benefit at the disadvantage of fibrous-rooted species and could modify community composition.

Coumarin inhibits the growth of carrot (Daucus carota L. cv. Saint Valery) cells in suspension culture

We used a carrot (Daucus carota L. cv. Saint Valery) cell suspension culture as a simplified model system to study the effects of the allelochemical compound coumarin (1,2 benzopyrone) on cell growth and utilisation of exogenous nitrate, ammonium and carbohydrates. Exposure to micromolar levels of coumarin caused severe inhibition of cell growth starting from the second day of culture onwards. At the same time, the presence of 50 mumol/L coumarin caused accumulation of free amino acids and of ammonium in the cultured cells, and stimulated their glutamine synthetase, glutamate dehydrogenase, glucose-6-phosphate dehydrogenase and phosphoenolpyruvate carboxylase activities. Malate dehydrogenase, on the other hand, was inhibited under the same conditions. These effects were interpreted in terms of the stimulation of protein catabolism and/or interference with protein biosynthesis induced by coumarin. This could have led to a series of compensatory changes in the activities of enzymes linking nitrogen and carbon metabolism. Because coumarin seemed to abolish the exponential phase and to accelerate the onset of the stationary phase of cell growth, we hypothesise that such allelochemical compounds may act in nature as an inhibitor of the cell cycle and/or as a senescence-promoting substance.

Cell wall immobilisation and antioxidant status of Xanthoria parietina thalli exposed to cadmium

Total and cell wall-bound cadmium and the major antioxidants were measured in thalli of the lichen Xanthoria parietina (L.) Th. Fr. exposed to two Cd concentrations, namely 4.5 or 9.0 μm, in liquid medium during exposure periods of either 24 or 48 h. Total Cd in the thalli was within the range of previous field measurements and was proportional to the exposure concentration, but less than proportional with respect to exposure duration. More than half of the total Cd was immobilised by the cell wall. The adopted conditions of Cd stress caused: (i) no changes in dry weight and protein concentration; (ii) an increase in the level of ascorbic acid and a decrease in that of reduced glutathione, as well as an increase in guaiacol peroxidase activity; (iii) no changes or moderate decreases in the activities of superoxide dismutase, catalase, dehydroascorbate-, NADPH-dependent glutathione disulfide-, and monodehydroascorbate reductases and of ascorbate peroxidase; (iv) an increase of the level of thiobarbituric acid-reactive substances, assumed to reflect malondialdehyde formation arising from membrane lipid peroxidation. Thus, X. parietina might withstand realistic levels of Cd stress by: (1) intercepting the heavy metal at cell wall level, (2) the intervention of antioxidant metabolites, and (3) a moderate increase in guaiacol peroxidase activity.

Foliar antioxidant status of adult Mediterranean oak species (Quercus ilex L. and Q. pubescens Willd.) exposed to permanent CO2-enrichment and to seasonal water stress

Foliar antioxidants were measured in adult individuals of holm oak (Quercus ilex L.) and white oak (Q. pubescens Willd.) growing in the field either within the vicinity of natural CO2 springs or at a nearby control site under ambient CO2, which had been previously exposed to either daily irrigation or no irrigation. In oak trees permanently exposed to elevated CO2 the activities of antioxidant enzymes tended to be lower and the ascorbate pool was larger and more in reduced form, suggesting an attenuation of the oxidative risk in the CO2-enriched trees. In the enriched individuals of both species. the imposition of water shortage significantly increased the size of the glutathione pool and the total superoxide dismutase activity in a species-specific manner. Moreover, water-stressed trees exposed to elevated CO2 tended to have higher catalase and ascorbate peroxidase activities than water-stressed control trees. Such changes may reflect the need for an enhanced compensatory effort when trees acclimated to elevated CO2 are exposed to oxidative stress-promoting environmental factors, such as water shortage.

Allelochemical effects on net nitrate uptake and plasma membrane H+-ATPase activity in maize seedlings

Seven-day-old maize seedlings grown in a nitrogen-free hydroponic culture were exposed for 48 h to 0, 100 and 300 μM trans-cinnamic, p-coumaric, ferulic, caffeic acids, umbelliferone and 200 μM KNO3. Net nitrate uptake was affected by trans-cinnamic, ferulic and p-coumaric acids in a concentration-dependent manner, and trans-cinnamic acid appeared to be the strongest inhibitor. Conversely, at low concentrations, caffeic acid stimulated net nitrate uptake while umbelliferone did not influence it. After 24 h of treatment, plasma membrane H+-ATPase activity significantly decreased in a concentration-dependent manner in response to trans-cinnamic, ferulic and p-coumaric acids, while umbelliferone and caffeic acid had no effect on H+-ATPase activity.

Allelopathic potential of Artemisia arborescens: Isolation, identification and quantification of phytotoxic compounds through fractionation-guided bioassays

The aerial part of Artemisia arborescens L. (Asteraceae) was extracted with water and methanol, and both extracts were fractionated using n-hexane, chloroform, ethyl acetate and n-butanol. The potential phytotoxicity of both crude extracts and their fractions were assayed in vitro on seed germination and root growth of lettuce (Lactuca sativa L.), a sensitive species largely employed in the allelopathy studies. The inhibitory activities were analysed by dose–response curves and the ED 50 were estimated. Crude extracts strongly inhibited both germination and root growth processes. The fraction-bioassay indicated the following hierarchy of phytotoxicity for both physiological processes: ethyl acetate ≥ n-hexane > chloroform ≥ n-butanol. On the n-hexane fraction, GC–MS analyses were carried out to characterise and quantify some of the potential allelochemicals. Twenty-one compounds were identified and three of them, camphor, trans-caryophyllene and pulegone were quantified.

Short-term effects of coumarin along the maize primary root axis

The short-term effects of coumarin on three different maize primary root zones, transition zone (TZ, 3 mm), and two non-growing zones (NGZ1 and NGZ2 at 20 and 50 mm, respectively), were studied in order to investigate the effect of the allelochemical on maize root elongation rate (RER). The RER, plasma membrane (pm) H+-ATPase activity, quantitative pH changes and cell membrane potentials were evaluated. The results showed that coumarin caused at the TZ 1) an increased RER; 2) an enhancement of pm H+-ATPase activity and proton extrusion; and 3) a transient depolarization followed by a hyperpolarization of cell membrane potential. These observations were not evident in the NGZ1 and NGZ2 of the maize root. Coumarin-treatment in the NGZ1 did not change RER, but caused a membrane depolarization, while the NGZ2 was mostly insensitive to the allelochemical. These data suggested that the primary maize root was sensitive to coumarin within a 20 mm section from the root tip, but the more distal NGZ2 was not involved in coumarin-elicited physiological responses.

Phytotoxic activity of foliar volatiles and essential oils of Calamintha nepeta (L.) Savi

Foliar volatiles and essential oils of Calamintha nepeta (L.) Savi, a Mediterranean plant species belonging to the Labiatae family, were investigated for their phytotoxic activities on seed germination and root growth of crops (Lactuca sativa L. and Raphanus sativus L.) and weed species (Lolium perenne L. and Amaranthus retroflexus L.). Foliar volatiles of C. nepeta (L.) Savi strongly inhibited both germination and root growth of lettuce, and its essential oils, especially at 125, 250 and 500 μL/L, inhibited both processes in lettuce, radish and A. retroflexus L. species, while displaying a little effect on L. perenne L. By GC–MS, 28 chemicals were identified: 17 monoterpenes, 8 sesquiterpenes, 1 diterpene and 2 miscellaneous. Pulegone was the main constituent of the C. nepeta (L.) Savi essential oils. The terpenic components of essentials oils were probably responsible for the phytotoxic activities.

Compost from Fresh Orange Waste: A Suitable Substrate for Nursery and Field Crops?

Composting represents a valuable strategy for recycling orange processing waste for use as soil conditioner, provided compost maturity is duly evaluated. Following a 5-month aerobic bioconversion, orange waste reached an acceptable degree of maturity in terms of humification parameters, absence of phytotoxicity (determined on test plants according to ISO methods) and low content of simple phenolic compounds. A greenhouse experiment with two nursery crops (tomato and zucchini) showed that, depending on the characteristics of the growing substrate (vermiculite or perlite), orange compost addition selectively induced pH and electrical conductivity (EC) increases, which in turn would affect plant growth responses. In field crops shoot mass weight was increased after compost addition. Results suggest that at field scale, orange compost may be used for organic fertilization; while in nursery crops it can be mixed with commercial potting substrates selected in relation to plant sensitivity. In horticultural crops, for its greater ability to buffer pH and EC changes, vermiculite could represent the preferred complementary substrate compared to perlite.