Different behaviours in phytoremediation capacity of two heavy metal tolerant poplar clones in relation to iron and other trace elements.

Plant biodiversity and intra-population genetic variability have not yet been properly exploited in the framework of phytoremediation and soil reclamation. For this reason, iron and other metal accumulation capacity of two Cu and Zn tolerant poplar clones, namely AL22 (Populus alba L.) and N12 (Populus nigra L.), was investigated in a pot experiment. Cuttings of the two clones were planted in iron rich soil collected from an urban-industrial area. Concentrations of Cd, Cu, Fe, Pb and Zn were analysed in leaves (at different times), as well as in stems and in roots (at the end of the experiment), both in control plants and in plants grown on a soil whose Fe availability was artificially enhanced. Results showed that Cd and Zn were preferentially accumulated in leaves, whereas Cu, Fe and Pb were mainly accumulated in roots. The main differences in metal accumulation between clones were related to Cd (about tenfold higher concentrations in N12) and Cu (higher concentrations in AL22). Once soil Fe availability was enhanced, the uptake and accumulation of all metals declined, with the exception of Fe at the first sampling time in AL22 leaves. The different behaviour of the two poplar clones suggests that a thoughtful choice should be made for their use in relation to soil heavy metal remediation.

The Effects of Nitrogen Fertilisation on the Anatomical Properties of the Populus alba L. Clone 'Villafranca' Juvenile Wood.

This study investigates the effect of nitrogen fertilisation on the anatomical properties of the juvenile wood of the Populus alba L. clone 'Villafranca' from an experimental trial near the Drava River in Croatia. Nitrogen was applied for two consecutive years, and the immediate and potential post-treatment effects were investigated. The correlation between annual ring width (ARW) and individual wood anatomical properties was also examined. The fertilisation effect was confirmed after the first year of nitrogen application for all wood anatomical properties except the vessel lumen area (VLA). Fibre length (FL) was reduced, and double cell wall thickness (DCWT), ray area (RA), and cell wall area (CWA) increased. In contrast, the vessel lumen diameter (VLD) and vessel lumen area changed inconsistently between treatments. The second year of nitrogen application was determined to be effective for FL only. Due to the insignificant results in the second year of the application of nitrogen, the post-fertilisation effect of nitrogen fertilisation was not confirmed.

Populus alba L., an Autochthonous Species of Spain: A Source for Cellulose Nanofibers by Chemical Pretreatment.

In order to identify new sustainable sources for producing cellulose nanofibers (CNFs), fast-growing poplar (Populus alba L.) wood was evaluated herein. For that purpose, bleached poplar kraft pulp was produced and submitted to TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl radical) mediated oxidation (TEMPO-ox) chemical pretreatment followed by microfluidization. The resulting CNFs were thoroughly characterized, including a rheological study at different pH values. Poplar CNFs showed properties comparable to eucalypt CNFs (reference material for CNFs production), showing high carboxylate content (1048 ± 128 µmol g-1), fibrillation yield (87.3% ± 8.1%), optical transmittance (83% at 700 nm) and thermal stability (up to more than 200 °C). Regarding the rheological study, whereas pH from 4 to 10 did not produce significant changes in rheological behavior, a reduction of pH down to 1 led to an order-of-magnitude increase on the viscoelastic functions. Therefore, poplar CNF shows potential in the pH-sensitive hydrogels application field. Finally, the possible ecotoxicity of poplar CNF was assessed. The decrease in cell viability was very low so that only concentrations causing a 10% cytotoxicity could be calculated for the assay detecting alterations in cell metabolism (10 µg mL-1) and plasma membrane integrity (60 µg mL-1).

Phytodepuration of Nitrate Contaminated Water Using Four Different Tree Species.

Water pollution by excessive amounts of nitrate (NO3-) has become a global issue. Technologies to clean up nitrate-contaminated water bodies include phytoremediation. In this context, this research aimed to evaluate four tree species (Salix alba L., Populus alba L., Corylus avellana L. and Sambucus nigra L.) to remediate nitrate-contaminated waters (100 and 300 mg L-1). Some physiological parameters showed that S. alba L. and P. alba L. increased particularly photosynthetic activity, chlorophyll content, dry weight, and transpired water, following the treatments with the above NO3- concentrations. Furthermore, these species were more efficient than the others studied in the phytodepuration of water contaminated by the two NO3- levels. In particular, within 15 days of treatment, S. alba L. and P. alba L. removed nitrate quantities ranging from 39 to 78%. Differently, C. avellana L. and S. nigra L. did not show particular responses regarding the physiological traits studied. Nonetheless, these species removed up to 30% of nitrate from water. In conclusion, these data provide exciting indications on the chance of using S. alba L. and P. alba L. to populate buffer strips to avoid NO3- environmental dispersion in agricultural areas.

Phosphorus homeostasis in Populus alba L. under excess phosphate conditions, assessed by 31P nuclear magnetic resonance spectroscopy and X-ray microfluorescence.

The phosphates (Pi) are nowadays recognized as pollutants. We studied the effect of Pi (0.625-12.500 mM KH2PO4) in the culture medium on in vitro grown 2-month-old Populus alba trees. The levels of sugar phosphates and vacuolar and cytoplasmic Pi in cell compartments of roots and stems were determined using 31P NMR, while tissue-specific micro- and macroelements mapping on stem cross-sections were performed using synchrotron-based X-ray microfluorescence. Plants grown on 0.625 mM Pi (MS/2 medium) showed a survival rate of 70%. With the increase in Pi concentrations up to 6.250 mM, plant growth and survival increased, without changes in total P content per mass or in the levels of cytoplasmic and vacuolar phosphates, in both stems and roots, while the levels of Fe, Cu, Zn, Ca and Mn in stems increased. Further increase in Pi to 9.375 and 12.500 mM in the medium resulted in inhibited growth comparable with plants grown on MS/2, with the increase in total P content per mass up to 50%, in both stems and roots, but with no changes in cytoplasmic and vacuolar phosphates; 12.500 mM Pi affected even plant survival (70%) and thus might be considered as mildly toxic. 31P NMR results indicate that the high tolerance of P. alba to increased Pi could result from its ability to maintain an intracellular P homeostasis, despite P accumulation up to 50%, in both stems and roots, indicating P. alba as a promising wood species for dendroremediation.

Cadmium impact, accumulation and detection in poplar callus cells.

Effect of cadmium cations and their interaction with silicon cations was determined in poplar calli and expressed as changes in callus growth, cell viability and cadmium cation accumulation. Cell viability throughout culture versus cadmium cation accumulation in cells is discussed. At the same time, the study sought appropriate methods for cadmium cation detection in callus cells and also in experiments with low plant material (e.g. protoplasts). Cadmium cations were determined by atomic absorption spectroscopy and using fluorescence microscopy with a specific cadmium cation fluorescent dye. The detection of cadmium cations in callus cells by the latter method appears suitable because the callus cells are surrounded by primary cell walls without auto-fluorescence and these values fit well with atomic absorption spectroscopy quantification. However, the visualisation method has some limits discussed below.

Anthocyanins of the anthers as chemotaxonomic markers in the genus Populus L.. Differentiation between Populus nigra, Populus alba and Populus tremula.

Three main species of Popululs L. (Salicaceae) have been reported to occur in the Iberian Peninsula: Populus nigra L., Populus alba L. and Populus tremula L. The degree of pilosity of the bracts of the male catkins is a key character for their differentiation. The anthers of these poplar species possess anthocyanins that provide them a red colouration. Since these poplars are wind-pollinated and, consequently, do not need to attract pollinators, anthocyanins in the anthers might be acting as photoprotectors, shielding pollen grains from excessive sunlight. In order to verify this hypothesis, the first objective of this study was to establish if there is any relationship between the degree of pilosity of the bracts (related to the physical shading of the pollen grains) and the levels and types of anthocyanins in the anthers of these three species. This study also aimed to check the usefulness of the anthocyanins of the anthers as chemotaxonomic markers, through the study of the differences in the anthocyanin composition between these poplar species. Anthocyanins were identified from the data supplied by HPLC-DAD-MS(n) analyses. Seventeen different compounds, including mono-, di- and triglycosides and anthocyanin-derived pigments (F-A(+) dimers) have been identified. Cyanidin 3-O-glucoside was the major compound in all the samples (>60% of the total content), which may be in accordance with the photoprotective role proposed for them. However, qualitative and quantitative differences were detected among samples. Cyanidin and delphinidin 3-O-sambubiosides have been detected only in the anthers of P. tremula as well as cyanidin 3-O-(2″-O-xyloxyl)rutinoside, making them valuable chemotaxonomic markers for this species. Hierarchical Cluster and Principal Components Analyses (HCA and PCA) carried out with the anthocyanin percent composition data have allowed a separation of the samples that is in accordance with the initial classification of the samples made from the morphological characters of the specimens. Furthermore, these analyses have revealed intraspecific differences among samples that point out to different clones or varieties of a same species.

Cryopreservation of white poplar (Populus alba L.) by vitrification of in vitro-grown shoot tips.

Shoot tips from in vitro-grown, cold-hardened stock plants of white poplar (Populus alba L.) were successfully cryopreserved at -196  °C by one-step vitrification. After preculturing at 5  °C for 2 days on hormone-free MS medium containing different sucrose concentrations, and loading for 20 min with 2 M glycerol and 0.4 M sucrose, shoot tips were treated with the PVS2 vitrification solution and plunged directly into liquid nitrogen. Best survival rate (90%) was obtained when shoot tips were precultured on 0.09 M sucrose, hormone-free MS medium, vitrified by exposure to PVS2 solution for 60 min at 0  °C and, following cryopreservation, rewarmed at 40  °C and washed in 1.2 M sucrose solution for 20 min. Regrowth was improved by plating shoot tips on a gelled MS medium containing 1.5 µM N6-benzyladenine plus 0.5 µM gibberellic acid, while shoot rooting was achieved on MS medium containing 3 µM indole-3-butyric acid. Following this procedure, almost 60% rooted shoots were obtained from cryopreserved shoot tips.

Transformation of elite white poplar (Populus alba L.) cv. ' Villafranca' and evaluation of herbicide resistance.

Transgenic white poplar plants (Populus alba L.) expressing the nptII gene and the bar gene from Streptomyces hygroscopicus have been produced using Agrobacterium tumefaciens-mediated gene transfer. Eleven kanamycin-resistant plant lines were obtained with a transformation frequency of 7%. Successful genetic transformation was confirmed by Southern and northern analyses. The level of resistance to the commercial preparation of phosphinothricin (Basta; Roussel-Hoechst Agrovet) was evaluated by in vitro and in vivo assays. Using in vitro selective conditions for phosphinothricin, only plantlets from four kanamycin-resistant independent lines remained green and continued to grow and root. After transfer to the growth chamber, all selected transgenic lines were shown to be completely resistant to the herbicide Basta with doses equivalent to 6 l ha-1 (normal field dosage) and were tolerant at concentration of 12 l ha-1. This is the first report describing the genetic transformation of a P. alba clonal cultivar of commercial interest with a gene of agronomic value.