Ioannis Therios

Melatonin promotes adventitious root regeneration in in vitro shoot tip explants of the commercial sweet cherry rootstocks CAB‐6P (Prunus cerasus L.), Gisela 6 (P. cerasus × P. canescens), and MxM 60 (P. avium × P. mahaleb).

Abstract:  The objectives of this study were to test the effects of melatonin (N‐acetyl‐5‐methoxytryptamine), a natural compound of edible plants on the rooting of certain commercial sweet cherry rootstocks. Shoot tip explants from previous in vitro cultures of the cherry rootstocks CAB‐6P (Prunus cerasus L.), Gisela 6 (P. cerasus × P. canescens), and M × M 60 (P. avium × P. mahaleb) were included in the experiment. The effect of indole‐3‐acetic acid (IAA) and indole‐3‐butyric acid (IBA) alone or in combination with melatonin was tested concerning their rooting potential. Seven concentrations of melatonin (0, 0.05, 0.1, 0.5, 1, 5, and 10 μm) alone or in combination with 5.71 μm of IAA or 4.92 μm of IBA were tested. For each rootstock, 21 treatments were included. The explants were grown in glass tubes containing 10 mL of substrate. The parameters measured include rooting percentage, number of roots per rooted explant, root length, and callus formation. The data presented in this study show that melatonin has a rooting promoting effect at a low concentration but a growth inhibitory effect at high concentrations. In the absence of auxin, 1 μm melatonin had auxinic response concerning the number and length of roots, but 10 μm melatonin was inhibitory to rooting in all the tested rootstocks. The final conclusion of this experiment is that exogenously applied melatonin acted as a rooting promoter and its action was similar to that of IAA.

NUTRITIONAL STATUS, GROWTH, CO2 ASSIMILATION, AND LEAF ANATOMICAL RESPONSES IN TWO KIWIFRUIT SPECIES UNDER BORON TOXICITY

ABSTRACT Two Actinidia species [Actinidia deliciosa (A.Chev.) C.F. Liang et A.R. Ferguson var. deliciosa cv. Hayward] and (Actinidia arguta Planch.) were grown in a sand-perlite mixture (1 : 1) in a growth room and irrigated with Hoaglands nutrient solutions containing 20, 50, 100, 200, and 500 μM boron (B). Growth, leaf ion concentration, photosynthetic rate and B induced changes in leaf anatomy were investigated. The greatest shoot length of the two species was recorded with 50 μM B. Boron concentration was highest in the leaf margin, intermediate in the remaining leaf blade and minimum in petioles. Boron toxicity induced a decrease of photosynthetic rate (Pn), of the volume of mesophyll cells, an increase of the volume of intercellular spaces and cell damage. Inclusion of 500 μM of B in the nutrient solution decreased calcium (Ca) and manganese (Mn) concentrations in kiwifruit leaves but did not affect the concentration of other mineral nutrients. Finally, the two species did not differ significantly in leaf B accumulation and sensitivity to B.

Response of two citrus genotypes to six boron concentrations: concentration and distribution of nutrients, total absorption, and nutrient use efficiency

A greenhouse experiment was performed to study the effects of boron (B) on growth, nutrient concentration and distribution, nutrient use efficiency, and total nutrient absorption of 2 citrus genotypes. The experimental layout was a 6 × 2 factorial, with 6 B concentrations (0.05, 0.25, 0.50, 1.00, 2.00, and 5.00 mg/L) on 2 genotypes: the sour orange (Citrus aurantium L.) and the Swingle citrumelo (C. paradisi Macf. × Poncirus trifoliata L.). The plants were grown for 3 months in a B-free sand : perlite (1 : 1) medium that was irrigated with 6 half-strength Hoaglands nutrient solutions. Increasing B supply in the nutrient solution increased the B concentration linearly in all parts of the plant in the following order: basal leaves > top leaves > bark > root > stems > wood. There was no consistent effect of B supply on the concentration of other fundamental elements (P, K, Ca, Mg, Mn, Zn, Fe). Furthermore, none of the tested B concentrations significantly affected the total plant content and consequently the absorption of any other element. A concentration of 1.00 mg B/L or higher resulted in less B absorption by the Swingle citrumelo than by the sour orange. Furthermore, the Swingle citrumelo has the ability to retain more B in its stems and roots than the sour orange, thus preventing B transport to leaves. Finally, B and Mn use efficiency in both genotypes correlated significantly and negatively with the B supply.

Volatile Constituents and Antioxidant Activity of Peel, Flowers and Leaf Oils of Citrus aurantium L. Growing in Greece

The volatile constituents of the essential oils of the peel, flower (neroli) and leaves (petitgrain) of bitter orange (Citrus aurantium L.) growing in Greece were studied by GC-MS. The analytical procedures enabled the quantitative determination of 31 components. More specifically, the components of the essential oils identified were: twelve in the peel, twenty-six in the flowers, and twenty and sixteen in old and young leaves, respectively. The major constituents of the different parts of Citrus aurantium L. essential oils were: β-pinene (0.62%–19.08%), limonene (0.53%–94.67%), trans-β-ocimene (3.11%–6.06%), linalool (0.76%–58.21%), and α-terpineol (0.13%–12.89%). The DPPH test demonstrated that the essential oils in the old leaves had the maximum antioxidant activity, followed by the flowers, young leaves and the peel in that order. This study updates the data in the literature on the essential oils of bitter orange, and provides information on the composition of the oils for a further evaluation of this product.

Melatonin enhances root regeneration, photosynthetic pigments, biomass, total carbohydrates and proline content in the cherry rootstock PHL-C (Prunus avium × Prunus cerasus)

The present study, investigates the effects of melatonin (0, 0.05, 0.1, 0.5, 1, 5 and 10 μM) on the morphogenic and biochemical responses in the cherry rootstock PHL-C (Prunus avium L. × Prunus cerasus L.), from shoot tip explants. The incorporation of melatonin (0-10 μM) in the Murashige and Skoog (MS) medium, greatly influenced rooting either positively or negatively. Melatonin, irrespective of its concentration, had a negative effect concerning the number of roots. However, application of 0.5 μM melatonin significantly increased the root length; while 1 μM melatonin increased the root length by 2.5 times, and the fresh weight of the roots by 4 times, in comparison to the control. Although 0.05 μM melatonin increased rooting by 11.11%, 5 μM melatonin had a significant reduction on the number, the fresh weight of roots, and the rooting percentage. Melatonin concentration of 0.1 μM resulted in the greatest chlorophyll (a + b) content, and 5-10 μM reduced the chlorophyll concentration by 2 times, compared to the control. The high melatonin concentrations (5 and 10 μM), increased the levels of proline and carbohydrates in leaves by 3-4 times. In the roots, 0.5 μM of melatonin concentration increased the carbohydrate levels by 1.5 times, while 0.05, 0.1 and 1 μM melatonin concentration significantly reduced the proline content.

Changes in Peroxidases and Catalase Activity During in vitro Rooting

Enzyme changes in non-rooted (treated with Fe-EDTA) and rooted (treated with Fe-EDDHA) stems of rootstock GF-677 (Prunus amygdalus×P. persica) during adventitious root formation in vitro have been recorded. The first roots appeared approximately after 12 d on the rooting medium. By contrast to non-rooted stems, rooted stems showed a maximum of soluble peroxidase activity on the 9th day, of ionically bound peroxidase to cell wall on the 6th and 12th day and of catalase on the 6th and the 15th day. A time course study of changes of soluble peroxidases isoenzymes showed that there was a band visible only in the rooted stems and also a new band appeared three days before the emergence of roots.

Melatonin combined with ascorbic acid provides salt adaptation in Citrus aurantium L. seedlings.

Ascorbic acid (AsA) and melatonin (Mel) are known molecules participating in stress resistance, however, their combined role in counteracting the impact of salinity in plants is still unknown. In this work the effect of exogenous application of 0.50 mΜ AsA, 1 μΜ Mel and their combination (AsA + Mel) on various stress responses in leaves and roots of Citrus aurantium L. seedlings grown under 100 mΜ NaCl for 30 days was investigated. Application of AsA, Mel or AsA + Mel to saline solution decreased NaCl-induced electrolyte leakage and lipid peroxidation and prevented NaCl-associated toxicity symptoms and pigments degradation. Also, leaves exposed to combined AsA + Mel treatment displayed lower Cl(-) accumulation. Treatments with AsA and/or Mel modulated differently carbohydrates, proline, phenols, glutathione and the total antioxidant power of tissues as well as the activities of SOD, APX, POD, GR and PPO compared to NaCl alone treatment. Exposure of leaves and roots to chemical treatments and especially to combined AsA and Mel application was able to regulate CaMIPS, CaSLAH1 and CaMYB73 expression, indicating that sugar metabolism, ion homeostasis and transcription regulation were triggered by AsA and Mel. These results provide evidence that the activation of the metabolic pathways associated with combined AsA and Mel application are linked with salt adaptation in citrus plants.

Effects of Boron on Growth, and Chlorophyll and Mineral Contents of Shoots of the Apple Rootstock MM 106 cultured in vitro

The in vitro cultures of apple rootstock MM 106 produced the highest fresh mass (FM) when 0.1 mM B was included in the culture medium. By increasing B concentration of the culture medium from 0.1 to 6.0 mM, FM and contents of B, P, Ca, and Mg in explants increased, whereas K, Fe, Mn, and Zn contents decreased. SPAD units of leaves characterizing chlorophyll contents declined as B concentration of the culture medium increased from 0.1 to 6.0 mM.

Comparative Transcriptome Analysis of Two Olive Cultivars in Response to NaCl-Stress

Background Olive (Olea europaea L.) cultivation is rapidly expanding and low quality saline water is often used for irrigation. The molecular basis of salt tolerance in olive, though, has not yet been investigated at a system level. In this study a comparative transcriptomics approach was used as a tool to unravel gene regulatory networks underlying salinity response in olive trees by simulating as much as possible olive growing conditions in the field. Specifically, we investigated the genotype-dependent differences in the transcriptome response of two olive cultivars, a salt-tolerant and a salt-sensitive one. Methodology/Principal Findings A 135-day long salinity experiment was conducted using one-year old trees exposed to NaCl stress for 90 days followed by 45 days of post-stress period during the summer. A cDNA library made of olive seedling mRNAs was sequenced and an olive microarray was constructed. Total RNA was extracted from root samples after 15, 45 and 90 days of NaCl-treatment as well as after 15 and 45 days of post-treatment period and used for microarray hybridizations. SAM analysis between the NaCl-stress and the post-stress time course resulted in the identification of 209 and 36 differentially expressed transcripts in the salt–tolerant and salt–sensitive cultivar, respectively. Hierarchical clustering revealed two major, distinct clusters for each cultivar. Despite the limited number of probe sets, transcriptional regulatory networks were constructed for both cultivars while several hierarchically-clustered interacting transcription factor regulators such as JERF and bZIP homologues were identified. Conclusions/Significance A systems biology approach was used and differentially expressed transcripts as well as regulatory interactions were identified. The comparison of the interactions among transcription factors in olive with those reported for Arabidopsis might indicate similarities in the response of a tree species with Arabidopsis at the transcriptional level under salinity stress.

Growth, Nutritional Status, Chlorophyll Content, and Antioxidant Responses of the Apple Rootstock MM 111 Shoots Cultured Under High Boron Concentrations In Vitro

ABSTRACT The in vitro response of the apple rootstock MM 111 to increasing concentrations of boron (B) (0.1, 0.5, 1, 3, and 6 mM) in MS medium is reported. The in vitro cultures of MM 111 shoots produced the highest fresh mass when 0.1 mM B was included in the medium. By increasing B concentration of the culture medium from 0.1 to 6 mM, B, phosphorus (P), calcium (Ca), and magnesium (Mg) concentrations of explants increased, whereas potassium (K), iron (Fe), manganese (Mn), and zinc (Zn) concentrations decreased. Chlorophyll content (SPAD units) of leaves declined as B concentration of the culture medium increased from 0.1 to 6 mM. The highest peroxidase (POD) activity in leaves was recorded in the presence of 6 mM B in the medium. By increasing B concentration of the medium from 0.1 to 3 mM, catalase (CAT) activity increased in leaves. Superoxide dismutase (SOD) activity in leaves and stems increased as B concentration of the medium increased. The non-enzymatic antioxidant power activity of leaves (FRAP values) increased gradually as B concentration of the medium increased.