Studies on lead and cadmium toxicity in Dianthus carthusianorum calamine ecotype cultivated in vitro

• Information on metallophytes during reclamation of land contaminated with heavy metals is sparse. We investigated the response of D. carthusianorum calamine ecotype to Pb and Cd stress. We focused on in vitro selection of tolerant plant material for direct use in chemically degraded areas.
• Shoot cultures were treated with various concentrations of Pb or Cd ions. Plantlet status was estimated as micropropagation efficiency, growth tolerance index (GTI) and through physiological analysis. Moreover, determination of plant Pb, Cd and other elements was performed.
• The application of Pb(NO₃)₂ resulted in stronger growth inhibition than application of CdCl₂. In the presence of Pb ions, a reduction was observed of both, the micropropagation coefficient to 1.1–1.8 and the GTI to 48%. In contrast, Cd ions had a positive influence on tested cultures, expressed as an increase of GTI up to 243% on medium enriched with 1.0 μm CdCl₂. Moreover, photosynthetic pigment content in shoots cultivated on media with CdCl₂ was higher than in control treatment. The adaptation to Cd was associated with decreased accumulation of phenols in the order: 0.0 μm  > 1.0 μm > 3.0 μm  > 5.5 μm CdCl₂. It seems that high tolerance to Cd is related to K uptake, which is involved in antioxidant defence.
• This work presents an innovative approach to the impact of Cd ions on plant growth and suggests a potential biological role of this metal in species from metalliferous areas.

     

Isolation of chlorophylls and carotenoids from freshwater algae using different extraction methods

Usually marine algae are an excellent source of pigments for different commercial sectors. Freshwater macroalgae can be exploited as a good source of biologically active compounds provided an appropriate extraction method is developed. The efficiency of four methods, like microwave‐assisted (MAE), ultrasound‐assisted extraction (UAE), supercritical fluid extraction (SFE) with ethanol as a co‐solvent, as well as conventional Soxhlet extraction were studied in the same conditions (time, solvent and temperature) for the recovery of chlorophylls and carotenoids from three freshwater green algae species: Cladophora glomerata, Cladophora rivularis and Ulva flexuosa. UV‐Vis spectrophotometry was used to determine chlorophyll a, chlorophyll b and total carotenoid content in obtained extracts. The results of this study showed that the advantages of novel extraction techniques (MAE and UAE) include higher yield and, in consequence, lower costs compared to traditional solvent extraction techniques. These methods were much more efficient in freshwater green algae pigment recovery than the classic Soxhlet extraction as well as SFE.     

Microbial and pigment profile of the reddish patch occurring within Tuber magnatum ascomata

Tuber magnatum Pico, the delectable white truffle, is the most prized truffle species. In this study, we examined the reddish pigmentation that frequently occurs in T. magnatum ascomata for the presence of pigment-producing bacteria.The inner part of the reddish-pigmented region of three T. magnatum ascomata collected in North–Central Italy was analysed. This reddish part was used to establish a bacterial culture collection and to extract the total genomic DNA in order to obtain a library of 16S rRNA genes representative of the bacterial community. The molecular approach revealed limited microbial diversity within the reddish-pigmented regions compared to the wider range of bacterial species commonly found at the same maturation stage and season in T. magnatum ascomata. The pigmented regions showed a prevalence of specific bacterial species belonging to α-, β- and γ- Proteobacteria, Actinobacteria and Firmicutes. From the tandem mass spectrometry analysis of the extracted pigment, four compounds were identified: i) bixin, ii) β-carotene, iii) cis-1-glycosyl-apo-8′- lycopene and iv) the fucoxanthin. Carotenoid producing species such as Microbacterium and Chryseobacterium emerged as the most likely cause of the peculiar reddish pigment production. Indeed, our findings suggest that the peculiar reddish pigment might be produced by these bacterial species.     

Marine natural pigments as potential sources for therapeutic applications

In recent years, marine natural pigments have emerged as a powerful alternative in the various fields of food, cosmetic, and pharmaceutical industries because of their excellent biocompatibility, bioavailability, safety, and stability. Marine organisms are recognized as a rich source of natural pigments such as chlorophylls, carotenoids, and phycobiliproteins. Numerous studies have shown that marine natural pigments have considerable medicinal potential and promising applications in human health. In this review, we summarize the marine natural pigments as potential sources for therapeutic applications, including: antioxidant, anticancer, antiangiogenic, anti-obesity, anti-inflammatory activities, drug delivery, photothermal therapy (PTT), photodynamic therapy (PDT), photoacoustic imaging (PAI), and wound healing. Marine natural pigments will offer a better platform for future theranostic applications.     

Characterization of Microbial Mats from a Desert Wadi Ecosystem in the Sultanate of Oman

Desert wadis are widespread in the Arabian Peninsula and play a vital role in the ecology of the region; nevertheless, these ecosystems are among the least studied. Various types of microbial mats are predominant in wadis, but information on their bacterial diversity and spatial distribution is very scarce. We investigated bacterial diversity, pigments and lipid composition of ten mats located at the down-, mid- and upstream of a desert wadi in Oman. Direct microscopy revealed the existence of different unicellular and filamentous cyanobacteria, with the dominance of the heterocystous genera Calothrix and Scytonema. The majority of MiSeq 16S rRNA sequences (44-76%) were affiliated to Cyanobacteria and Proteobacteria. While Alphaproteobacteria was the most dominant proteobacterial class (10 to 48% of total sequences), Gamma- and Deltaproteobacteria were subdominant. Cluster analysis showed that the mats’ bacterial communities at the different locations along the wadi were different and shared less than 60% of their operational taxonomic units (OTUs). Chlorophyll a and scytonemin were the most predominant pigments in all mats. Different saturated, branched and mono- and poly-unsaturated fatty acids were detected in all mats, with C₁₆ and C₁₈ compounds as most dominant. The detected pigments and fatty acids indicate a major role of cyanobacteria in the wadi mats and the adaptation of microorganisms therein to the harsh wadi environment. Detection of diadinoxanthin and fucoxanthin confirmed the presence of diatoms. We conclude that microbial mats are important elements in wadi ecosystems and exist in a great variety of structure and community composition.     

The transiently pubescent young leaves of plane (Platanus orientalis) are deficient in photodissipative capacity

We have analyzed reflectance changes and carotenoid composition of young and mature leaves of Platanus orientalis L. in order to test the hypothesis that the transient occurrence of highly absorptive and reflective leaf hairs of young leaves (M. Ntefidou and Y. Manetas 1996, Aust. J. Plant Physiol. 23: 535–538) may be correlated to a weakly developed photodissipative capacity in the chloroplast. Compared to mature leaves, young leaves showed negligible reflectance changes at 530 nm upon sudden illumination, possibly indicating a limited production of zeaxanthin. In addition, actual pigment analysis confirmed lower pools of xanthophyll cycle components and reduced capacity for violaxanthin photoconversion in young leaves. Accordingly, the epoxidation state at saturating photon fluence rates was particularly high. A notable feature of xanthophyll cycle interconversions in young leaves was the inability to drive the system to complete de-epoxidation, as antheraxanthin in the light was always higher than zeaxanthin. Among the rest of the carotenoids, the levels of β-carotene were particularly low. Moreover, most of the photosynthetic pigments were considerably bleached when young leaves were exposed to high light. The above results strongly suggest that young leaves possess a limited photodissipative capacity and therefore, the presence of leaf hairs affords protection against excess light. When the leaf has matured and presumably the concentrations of photoprotective compounds are adequate, the loss of hairs is not of consequence. In fact, their presence on mature leaves may reduce the photosynthetically active radiation to non-saturating levels for photosynthesis.     

Ultraviolet-B radiation effects on the Mediterranean ruderal Dittrichia viscosa

Young seedlings of Dittrichia viscosa L. (syn. Inula viscosa (L.) Aiton) (Asteraceae) were extensively treated with artificial rain in order to remove the water soluble component of their epicuticular UV-B absorbing compounds. As a result, 75% of the epicuticular absorbing capacity at 300 nm was lost. The seedlings were subsequently grown in a naturaly lit glasshouse for 80 days under 0.06, 6.41 and 10.14 kJ m^-2 day ^-1 biologicaly effective UV-B radiation doses. The initial, pre-rain values of the water soluble, epicuticular UV-B absorbing potential was restored in about three weeks. During this transient period the plants were exposed to the enhanced UV-B radiation doses with part of their UV-B radiation screen removed. Although a trend for increased accumulation of epicuticular UV-B absorbing capacity was observed with increasing UV-B radiation doses, the allelopathic potential of the epicuticular material remained unchanged. Internal (cellular) UV-B absorbing compounds and chlorophylls were unaffected, but total carotenoids were increased, indicating a possible protective role against UV-B radiation damage. Leaf, stem and root dry mass were the same under all treatments but UV-B radiation caused a reduction in the dry mass invested per unit leaf area with a concomitant increase in leaf area. The importance of this UV-B radiation induced selective allocation of photosynthate to the production of assimilative surfaces is discussed.     

Differential effects of elevated ultraviolet-B radiation on plant species of a dune grassland ecosystem

In a greenhouse study, plants of three monocotyledonous and five dicotyledonous species, which occur in a Dutch dune grassland, were exposed to four levels of ultraviolet-B (UV-B) radiation. UV-B levels simulated up to 30% reduction of the stratospheric ozone column during summertime in The Netherlands. Six of the plant species studied in the greenhouse were also exposed to enhanced UV-B irradiance in an experimental field study. In the field experiment plants either received the ambient UV-B irradiance (control) or an enhanced UV-B level simulating 15–20% ozone depletion during summertime in The Netherlands. The purpose of both experiments was to determine the response of the plant species to UV-B radiation and to compare results obtained in the greenhouse with results of the field experiment. Large intraspecific differences in UV-B sensitivity were observed in the greenhouse study. Total dry matter accumulation of monocotyledons was increased, while dry matter accumulation of dicotyledons remained unaffected or decreased. The increase in biomass production of monocotyledons at elevated UV-B was not related to the rate of photosynthesis but to alterations in leaf orientation. In the greenhouse study, UV-B radiation also affected morphological characteristics. Shoot height or maximum leaf length of five out of eight species was reduced. In the field study only one species showed a significantly decreased maximum leaf length at enhanced UV-B. Possible reasons for this discrepancy are discussed. The absorbance of methanolic leaf extracts also differed between species. UV absorbance of field-grown plants was higher than greenhouse-grown plants. In the greenhouse study, the highest UV-B level increased UV-B absorbance of some species. In the field study however, this stimulation of UV absorbance was not observed. In general, results obtained in the greenhouse study were similar to results obtained in the field study. Difficulties in extrapolating results of UV-B experiments conducted in the greenhouse to the field situation are discussed.     

Impact of Atmospheric Sulfur Deposition on Sulfur Metabolism in Plants: H2S as Sulfur Source for Sulfur Deprived Brassica oleracea L.

Brassica oleracea L. was rather insensitive to atmospheric H₂S: growth was only negatively affected at ≥0.4 μl I^?1. Shoots formed a sink for H₂S and the uptake rate showed saturation kinetics with respect to the atmospheric concentration. The H₂S uptake rate was high in comparison with other species, which may reflect the high sulfur need of Brassica. The net uptake of sulfate by roots of hydroponically grown plants was substantially reduced after one week of exposure to 0.25 μl l^?1 H₂S, indicating that plants switched in part from sulfate to H₂S as sulfur source for plant growth. Plants were sulfur deficient after two weeks of sulfur deprivation, illustrated by reduced growth, which was more pronounced for shoots than for roots, and in enhanced shoot dry matter content. The latter could for the greater part be attributed to enhanced levels of soluble sugars and starch. Sulfur deficiency was further characterized by a low pigment content, extremely low levels of sulfate and water-soluble non-protein thiols, and by enhanced levels of nitrate and free amino acids, particularly in the shoots. Furthermore, sulfur deficient plants contained a lower total lipid content in shoots, whereas its content in roots was unaffected. The level of sulfolipids was decreased in both roots and shoots. When sulfur deprived plants were exposed to 0.25 μl I^?1 H₂S for one week, all sulfur deficiency symptoms were abolished and growth was restored. Furthermore, plants were able to grow with 0.4 μl I^?1 H₂S as the sole sulfur source. Water-soluble non-protein thiol content was enhanced in both shoots and roots of H₂S exposed plants, irrespective of the sulfate supply to the roots, whereas plants grown with H₂S as sole sulfur source contained very low sulfate levels. The interaction between atmospheric and pedospheric sulfur nutrition in plants is discussed.     

Effects of lifelong [CO2] enrichment on carboxylation and light utilization of Quercus pubescens Willd. examined with gas exchange,biochemistry and optical techniques

Lifelong exposure to elevated concentrations of atmospheric CO₂ may enhance carbon assimilation of trees with unlimited rooting volume and consequently may reduce requirements for photoprotective pigments. In early summer the effects of elevated [CO₂] on carboxylation and light utilization of mature Quercus pubescens trees growing under chronic [CO₂] enrichment at two CO₂ springs and control sites in Italy were examined. Net photosynthesis was enhanced by 36 to 77%. There was no evidence of photosynthetic downregulation early in the growing season when sink demand presumably was greatest. Specifically, maximum assimilation at saturating [CO₂], electron transport capacity, and Rubisco content, activity and carboxylation capacity were not significantly different in trees growing at the CO₂ springs and their respective control sites. Foliar biochemical content, leaf reflectance index of chlorophyll pigments (NDVI), and photochemical efficiency of PSII (ΔF/F_m′) also were not significantly affected by [CO₂] enrichment except that starch content and ΔF/F_m′ tended to be higher at one spring (42 and 15%, respectively). Contrary to expectation, prolonged elevation of [CO₂] did not reduce xanthophyll cycle pigment pools or alter mid‐day values of leaf reflectance index of xanthophyll cycle pigments (PRI), despite the enhancement of carbon assimilation. However, both these pigments and PRI were well correlated with electron transport capacity.