Antonio Lupini

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.

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.

Root Architecture Plasticity of Citrus Rootstocks in Response to Nitrate Availability

ABSTRACT Under nutrient heterogeneous soil conditions, the ability of plants to compete for resource acquisition was closely related to the plasticity of root architecture, intended as a change of topology and geometry. Root architecture variations in response to different nitrate supplies of co-generic citrus rootstocks, ‘Rough Lemon’ (Citrus jambhiri Lush), ‘Sweet Orange’ (Citrus sinensis (L) Osbeck), ‘Cleopatra Mandarin’ (Citrus reshni Hort ex Tan.), and ‘Sour Orange’ (Citrus aurantium L.) are reported here. Root architecture was gauged by an image-specific analysis system (WinRHIZO). Based on the topological index, root topology of citrus rootstocks varied in response to nitrate treatments. In terms of root geometry, external-external and, to a lesser degree, internal-internal links of all rootstocks changed in response to nitrate supply. Among the rootstocks, ‘Cleopatra Mandarin’, both in terms of root topology (high TI) and root geometry (high length of exterior and interior link), could be considered less competitive owing to resource acquisition compared to the other rootstocks.

Allelopatic Potential of Dittrichia viscosa (L.) W. Greuter Mediated by VOCs: A Physiological and Metabolomic Approach

Dittrichia viscosa (L.) W. Greuter is a pioneer species belonging to the Compositae family. It is widespread in the Mediterranean basin, where it is considered invasive. It is a source of secondary metabolites, playing an important ecological role. D. viscosa plant extracts showed a phytotoxic activity on several physiological processes of different species. In the current study, the allelopathic potential of D. viscosa VOCs, released by its foliage, was evaluated on seed germination and root growth of lettuce. The VOCs effect was also studied on lettuce adult plants in microcosm systems, which better mimicked the open field conditions. D. viscosa VOCs inhibited both seed germination and root growth of lettuce. The VOCs composition revealed a large presence of terpenoids, responsible of the effects observed. Moreover, D. viscosa VOCs caused an alteration on plant water status accompanied by oxidative damages and photoinhibition on lettuce adult plants.

High-throughput 18K SNP array to assess genetic variability of the main grapevine cultivars from Sicily

The viticulture of Sicily, for its vocation, is one of the most important and ancient forms in Italy. Autochthonous grapevine cultivars, many of which known throughout the world, have always been cultivated in the island from many centuries. With the aim to preserve this large grapevine diversity, previous studies have already started to assess the genetic variability among the Sicilian cultivars by using morphological and microsatellite markers. In this study, simple sequence repeat (SSR) were utilized to verify the true-to-typeness of a large clone collection (101) belonging to 21 biotypes of the most 10 cultivated Sicilian cultivars. Afterwards, 42 Organization Internationale de la Vigne et du Vin (OIV) descriptors and a high-throughput single nucleotide polymorphism (SNP) genotyping array (Vitis18kSNP) were applied to assess genetic variability among cultivars and biotypes of the same cultivar. Ampelographic traits and high-throughput SNP genotyping platforms provided an accuracy estimation of genetic diversity in the Sicilian germplasm, showing the relationships among cultivars by cluster and multivariate analyses. The large SNP panel defined sub-clusters unable to discern among biotypes, previously classified by ampelographic analysis, belonging to each cultivar. These results suggested that a very large number of SNP did not cover the genome regions harboring few morphological traits. Genetic structure of the collection revealed a clear optimum number of groups for K = 3, clustering in the same group a significant portion of family-related genotypes. Parentage analysis highlighted significant relationships among Sicilian grape cultivars and Sangiovese, as already reported, but also the first evidences of the relationships between Nero d’Avola and both Inzolia and Catarratto. Finally, a small panel of highly informative markers (12 SNPs) allowed us to isolate a private profile for each Sicilian cultivar, providing a new tool for cultivar identification.

Long- and short-term effects of boron excess to root form and function in two tomato genotypes

Boron (B) is an essential plant nutrient, but when present in excess it is toxic. Morphological measurements were made to assess the impact of B toxicity on the growth of two different tomato hybrids, Losna and Ikram. Contrasting long and short-term B responses in these tomato hybrids, were observed. Losna showed less toxicity symptoms, maintaining higher growth and showing much less B content in both root and shoot tissues compared to Ikram. Root morphological differences did not explain the tolerance between the two hybrids. Under excess B supply, a significant inhibition on net nitrate uptake rate was observed in Ikram, but not in Losna. This effect may be explained by a decrease of nitrate transporter transcripts in Ikram, which was not measured in Losna. There was a different pattern of B transporter expression in two tomatoes and this can explain the contrasting tolerance observed. Indeed, Losna may be able to exclude or efflux B resulting in less accumulation in the shoot. Particularly, SlBOR4 expression showed significant differences between the tomato hybrids, with higher expression in Losna explaining the improved B-tolerance.

Phytotoxic Potential and Biological Activity of Three Synthetic Coumarin Derivatives as New Natural-Like Herbicides

Coumarin is a natural compound well known for its phytotoxic potential. In the search for new herbicidal compounds to manage weeds, three synthetic derivatives bearing the coumarin scaffold (1–3), synthesized by a carbonylative organometallic approach, were in vitro assayed on germination and root growth of two noxious weeds, Amaranthus retroflexus and Echinochloa crus-galli. Moreover, the synthetic coumarins 1–3 were also in vitro assayed on seedlings growth of the model species Arabidopsis thaliana to identify the possible physiological targets. All molecules strongly affected seed germination and root growth of both weeds. Interestingly, the effects of synthetic coumarins on weed germination were higher than template natural coumarin, pointing out ED50 values ranging from 50–115 µM. Moreover, all synthetic coumarins showed a strong phytotoxic potential on both Arabidopsis shoot and root growth, causing a strong reduction in shoot fresh weight (ED50 values ≤ 60 µM), accompanied by leaf development and a decrease in pigment content. Furthermore, they caused a strong alteration in root growth (ED50 values ≤ 170 µM) and morphology with evident alterations in root tip anatomy. Taken together, our results highlight the promising potential herbicidal activity of these compounds.