Anna Mensuali-Sodi

Endogenous ethylene requirement for adventitious root induction and growth in tomato cotyledons and lavandin microcuttings in vitro

The role of ethylene in the formation of adventitious roots in vitro was studied in tomato (Lycopersicon esculentum Mill. cv. UC 105) cotyledons and lavandin (Lavandula officinalis Chaix × Lavandula latifolia microshoots. Both systems were able to form roots on hormone-free medium evolving low amounts of ethylene. The addition of 20–50 μM indole-3-acetic acid (IAA) inhibited root formation in tomato cotyledons while increasing ethylene production. Naphthaleneacetic acid (NAA, 3 μM) stimulated root number in lavandin explants and induced a transient rise in ethylene evolution. Enhanced ethylene levels via the endogenous precursors 1-aminocyclopropane-1-carboxylic acid (ACC, 25–50 μM) drastically impaired root regeneration and growth in tomato. In lavandin, 10 μM ACC stimulated ethylene production and significantly inhibited the rooting percentage and root growth. Conversely, ACC enhanced the root number in the presence of NAA only. Severe inhibition of rooting was also caused by ethylene reduction via biosynthetic inhibitors, aminoethoxyvinylglycine (AVG, 5–10 μM) in tomato, and salicylic acid (SA, 100 μM) in lavandin. A strict requirement of endogenous ethylene for adventitious root induction and growth is thus suggested.

Solar UV-B Radiation Influences Carotenoid Accumulation of Tomato Fruit through Both Ethylene-Dependent and - Independent Mechanisms

The effect of UV-B shielding on ethylene production in ripening tomato fruits and the contribution of ethylene and UV-B radiation on carotenoid accumulation and profile during ripening were assessed to get more insight about the interplay between these two regulatory factors. To this aim, rin and nor tomato mutants, unable to produce ripening ethylene, and cv Ailsa Craig were cultivated under control or UV-B depleted conditions until full fruit ripening. The significantly decreased ethylene evolution following UV-B depletion, evident only in Ailsa Craig, suggested the requirement of functional rin and nor genes for UVB-mediated ethylene production. Carotenoid content and profile were found to be controlled by both ethylene and UV-B radiation. This latter influenced carotenoid metabolism either in an ethylene-dependent or -independent way, as indicated by UVB-induced changes also in nor and rin carotenoid content and confirmed by correlation plots between ethylene evolution and carotenoid accumulation performed separately for control and UV-B shielded fruits. In conclusion, natural UV-B radiation influences carotenoid metabolism in a rather complex way, involving ethylene-dependent and -independent mechanisms, which seem to act in an antagonistic way.

Involvement of activated charcoal in the modulation of abiotic and biotic ethylene levels in tissue cultures

Abstract Activated charcoal in the culture medium improved axillary bud proliferation of lavandin and growth of Anemone seedlings, but it negatively influenced root formation. In Anemone, severe medium browning was prevented by activated charcoal (AC). Abiotic ethylene released by the agarized medium was adsorbed by AC, depending upon the culture vessel and the medium volume. Biotic ethylene produced by lavandin nodes, laying directly on the medium surface, was easily adsorbed by AC. On the contrary, Anemone seedlings, 5 cm long, looked healthier in the presence of AC and evolved higher ethylene levels than the control. Therefore, the positive effect of AC on both species cannot be related to ethylene adsorption but probably to the removal of unidentified inhibitory substances from the culture medium.

Leaf ultrastructure, photosynthetic rate and growth of myrtle plantlets under different in vitro culture conditions

The in vitro rooting of myrtle (Myrtus communis L.) plantlets was performed in containers with gas permeable (V) and non-permeable (C) closures characterized by a different number of gas exchanges (1.4 and 0.3 h− 1, respectively). The rooting was induced on Perlite, soaked with half strength Murashige and Skoog (MS) medium with 0.5 mg dm−3 IBA, either with and without 15 g dm−3 of sucrose. During the rooting phase, it was demonstrated that C cultures without sucrose (C−) negatively affect the growth of myrtle plantlets. The net photosynthetic rate and the starch content showed the lowest values in C cultures with and without sucrose (C+ and C−) while chlorophyll a content did not vary among treatments, therefore it could not be considered an indicative parameter to evaluate the autotrophic metabolism in myrtle plantlets. Electron microscopy and image analysis were employed to evaluate the leaf ultrastructure at three sample dates. Plantlet rooted in vented vessels with and without sucrose (V+ and V−) showed chloroplasts with numerous starch inclusions, while several osmiophilic plastoglobules (frequently related with leaf senescence) were found in chloroplast of leaf cells of C− myrtle plantlets.

Rosmarinic acid content in basil plants grown in vitro and in hydroponics

The accumulation of selected caffeic acid derivatives (CADs), in particular rosmarinic acid (RA), was investigated in different tissues (leaves, roots and plantlet shoots) of sweet basil (Ocimum basilicum L.) plants grown either in vitro or in hydroponic culture (floating system) under greenhouse conditions. Two cultivars with green leaves (Genovese and Superbo) and one with purple leaves (Dark Opal) were tested. The content of CADs in HCl-methanol extracts was determined by HPLC. LC-MS and LC-MS-MS were used to confirm the identification of the metabolites of interest. Apart from rosmarinic acid (RA) and a methylated form of this substance, no other CADs were detected at significant level in any of the analyzed samples. The content of RA ranged approximately from 4 to 63 mg/g DW, depending on the growing system. The highest RA content was found during the in vitro multiplication, in the acclimatized plants and in the roots of hydroponically-grown seedlings at full bloom. In vitro, 6-benzyladenine reduced the accumulation of RA in purple-leaf Dark Opal cultivar, but an opposite effect of this growth regulator was observed in the green-leaf genotypes. Our findings suggest the possibility to scale-up RA production by means of in vitro or hydroponic culture of sweet basil.

Effect of thidiazuron and gibberellic acid on leaf yellowing of cut stock flowers

Plant hormones such as cytokinins and gibberellins are able to inhibit leaf yellowing in different species of cut flowers and potted plants. These hormones can be used alone or in combination among them for preserving chlorophyll in floriculture items. In the present study thidiazuron was tested alone or combined with GA3 for delaying leaf yellowing of cut stock flowers during vase life. Cut flowers were placed in a controlled environment and treated for 24 hours with the following solutions: distilled water (control) or solutions containing 5, 10 µM thidiazuron (TDZ), 0.5 mM gibberellic acid (GA3), or a combination of 0.5 mM GA3 with 5 µM TDZ. The effect of treatments was evaluated by measuring chlorophyll content, ethylene production, leaf gas exchanges and chlorophyll a fluorescence. Results showed that TDZ was able to delay leaf yellowing in light during whole experimental period (30 days). The effect of TDZ on dark stored flowers was less effective, and also delayed chlorophyll losses for 10–12 days. TDZ and GA3 combination did not show any synergistic nor beneficial effect. Gas exchange values such as net photosynthesis, vapour pressure deficit, stomatal conductance and water use efficiency were higher in the TDZ only treatment.

Aroma characterisation and UV elicitation of purple basil from different plant tissue cultures

Exposure to stressful environmental conditions can induce severe metabolic variations in basil (Ocimum basilicum) aroma. The aromatic profiles of Dark Opal and Red Rubim varieties (in vivo plants, in vitro shoots, callus, and suspension cultures) were investigated for the first time. The established calli represented the most interesting miniaturised aromatic plant systems, as they were able to emit many typical basil volatiles with very low amounts of phenylpropanoids (1-2%). The hydrocarbon monoterpenes and oxygenated volatiles emitted from calli of both varieties were greatly and conversely affected by UV-C and UV-B, in comparison with the non-irradiated samples. As calli of both varieties still maintained very low levels of phenylpropanoids even after UV elicitation, they might be regarded not only as efficient in vitro plant models to study volatile compounds under UV stress conditions, but also as safe aromatic biomass in comparison with in vivo basil plants.

Novel Prunus rootstock somaclonal variants with divergent ability to tolerate waterlogging.

Plants require access to free water for nutrient uptake, but excess water surrounding the roots can be injurious or even lethal because it blocks the transfer of free oxygen between the soil and the atmosphere. Genetic improvement efforts in this study were focused on the increased tolerance in roots to waterlogging. Among a pool of clones generated in vitro from leaf explants of rootstock Mr.S.2/5 of Prunus cerasifera L., the S.4 clone was flood tolerant whereas the S.1 clone was sensitive. The S.4 clone formed adventitious roots on exposure to flooding. Moreover, the chlorophyll content and mitochondrial activity in the leaf and root, soluble sugar content, alcohol dehydrogenase activity and ethylene content were different between the clones. The sorbitol transporter gene (SOT1) was up-regulated during hypoxia, the alcohol dehydrogenase genes (ADH1 and ADH3) were up-regulated in the leaves and down-regulated in the roots of the S.4 clone during hypoxia, and the 1-aminocyclopropane-1-oxidase gene (ACO1) was up-regulated in the leaves and roots of the S.4 clone during hypoxia and down-regulated in the wild-type roots. In addition, in the S.4 root, hypoxia induced significant down-regulation of a glycosyltransferase-like gene (GTL), which has a yet-undefined role. Although the relevant variation in the S.4 genome has yet to be determined, genetic alteration clearly conferred a flooding-tolerant phenotype. The isolation of novel somaclonals with the same genomic background but with divergent tolerance to flooding may offer new insights in the elucidation of the genetic machinery of resistance to flooding and aid in the selection of new Prunus rootstocks to be used in various adverse environments.

Effects of ethylene and cytokinins on vase life of cut Eucalyptus parvifolia Cambage branches

The role of ethylene and cytokinins was investigated during postharvestsenescence of cut Eucalyptus parvifolia Cambage branches.The effect of endogenous and exogenous ethylene on the vase life of the cutbranches was studied using 2 mM 1-aminocyclopropane-1-carboxylicacid (ACC) as a continuous treatment or “pulse” treatment for 48h with 20, 40 and 80 μl l−1ethylene. Both endogenous and exogenous ethylene reduced the vase life of thebranches; however, the effect of the endogenous hormone was stronger than theexogenous applications. Ethylene biosynthesis was inhibited by pulse treatmentfor 24 h using 1 mM AOA or 2 mMCoCl2. The latter treatment significantly extended the vase life ofthe branches by delaying senescence. The effect of cytokinins was evaluated onthe vase life of cut E. parvifolia branches by pulsetreatment for 24 h with 10, 50 and 100 mM thidiazuron(TDZ) or 85, 130 and 260 mM N6-benzyladenine (BA). The resultsobtained showed that no response was observed following pulse treatment with BAwhile, although TDZ had little effect on vase life, it was a good inhibitor ofchlorophyll degradation.

Effects of Promoters and Inhibitors of Ethylene and ABA on Flower Senescence of Hibiscus rosa-sinensis L.

Hibiscus rosa-sinensis L. flowers (cv La France) senesce and die over a 12-h period after opening. The aim of this study was to examine the physiological mechanisms regulating the senescence process of ephemeral hibiscus flowers. Different flower stages and floral organs were used to determine whether any interaction existed during flower senescence between endogenous abscisic acid (ABA) and the predisposition of the tissue to ethylene synthesis. This was carried out on whole flowers treated with promoters and inhibitors of ethylene and ABA synthesis or a combination of them. Treatments with 1-aminocyclopropane-1-carboxylic acid (ACC), a precursor of ethylene biosynthesis, enhanced flower senescence, whereas amino-oxyacetic acid (AOA) and fluridone, an ethylene and an ABA inhibitor, respectively, extended flower longevity. These effects were more significant when applied before anthesis. Ethylene evolution was substantially reduced in all organs from open and senescent flowers treated with fluridone and AOA. Similarly, endogenous ABA accumulation was negatively affected by AOA and fluridone treatments. Application of fluridone plus ACC reduced ethylene evolution and increased ABA content in a tissue-specific manner but did not overcome the inhibitor effect on flower longevity. AOA plus fluridone treatment slightly accelerated flower longevity compared to AOA-treated flowers. Application of ABA alone promoted senescence, suppressed ethylene production, and, when applied with fluridone, countered the fluridone-induced increase in flower longevity. Taken together, these results suggest that the senescence of hibiscus flowers is an endogenously regulated ethylene- and ABA-dependent process.