The intergenic spacer region of the rDNA in Haplopappus gracilis (Nutt.) Gray

In this paper we provide further information on the genome organization of Haplopappus gracilis, one of the six angiosperms showing the lowest chromosome number, i.e. 2n=4, by determining the nucleotide sequence of the Intergenic Spacer region of the rRNA genes and its cytological localization on metaphase chromosomes. DNA sequence analysis reveals the occurring of a product of 4,382 bp in length, characterized by the presence of four blocks of different repeated sequences. Our analysis also evidenced putative promoter regions with three Transcription Initiation Sites for Polymerase I, as previously reported in Artemisia absinthium, belonging to the same Asteraceae family. A fluorescent in situ hybridization with the Intergenic Spacer probe indicates the presence of rDNA genes only in the satellited chromosomes of H. gracilis; besides differences in the signal intensity between homologous chromosomes were frequently observed, so suggesting, for these chromosome sites, the presence of a variable number of rDNA gene copies, even if a divergent chromatin organization in corresponding regions can not be ruled out

Characterization of normal and "albino" phenotypes in Erythrina crista-galli

We present here a characterization of Erythrina crista-galli L. (syn: Erythrina lamifolia Jacq.) seedlings, obtained from a plant from the Botanical Garden of Pisa University. This plant produces seeds that, during germination, have shown two different seedling phenotypes: normal (NT, 75%) and "albino" types (AT, 25%). Albino seedlings survive only 8–9 weeks and their growth is dramatically reduced when compared with wild type seedlings. Biochemical investigations have shown that albino seedlings completely lack chlorophyll and carotenoids and also soluble sugar levels are lower than in the normal type. We have also conducted sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) experiments and silver staining analysis on different protein extracts from shoots and leaves of both phenotypes, and demonstrated strong differences in protein patterns. The almost total absence of putative small and large RuBisCo bands in albino seedlings should be emphasized. We have also microspectrophotometrica..

TiO2 nanoparticles may alleviate cadmium toxicity in co-treatment experiments on the model hydrophyte Azolla filiculoides

The hydrophyte Azolla filiculoides can be a useful model to assess if TiO2 NPs may in some way alleviate the Cd injuries and improve the ability of the plant to cope with this metal. With this mechanistic hypothesis, after a pre-treatment with TiO2 NPs, A. filiculoides plants were transferred to cadmium-contaminated water with or without TiO2 nanoparticles. After 5 days of treatment, cadmium uptake, morpho-anatomical, and physiological aspects were studied in plants. The continuous presence of TiO2 nanoparticles, though not increasing the uptake of cadmium in comparison with a priming treatment, induced a higher translocation of this heavy metal to the aerial portion. Despite the translocation factor was always well below 1, cadmium contents in the fronds, generally greater than 100 ppm, ranked A. filiculoides as a good cadmium accumulator. Higher cadmium contents in leaves did not induce damages to the photosynthetic machinery, probably thanks to a compartmentalization strategy aimed at confining most of this pollutant to less metabolically active peripheral cells. The permanence of NPs in growth medium ensured a better efficiency of the antioxidant apparatus (proline and glutathione peroxidase and catalase activities) and induced a decrease in H2O2 content, but did not suppress TBARS level

Karyological and molecular characterisation of subgenus Vicia (Fabaceae)

In the present report, we have analysed the subgenus Vicia by karyological and molecular approaches with the aim to clarify the relationships among Vicia species included in this subgenus by previously evidenced morphological investigations. Multivariate analysis using several karyomorphological parameters in addition to symmetry indices has allowed the construction of a dendrogram of linkage distances very useful to compare and to include in a phylogenetic tree obtained by internal transcribed spacer (ITS) rDNA sequences. Moreover, a separate analysis was performed combining our molecular data on ITS sequences with those reported in the literature for the section Vicilla. Our analyses partly confirm the monophyletic status of the various sections in which the subgenus Vicia has been divided, however questioning, in some cases, the real need to maintain all the nine sections so far accepted and the placement of some individual species in the two subgenera Vicia and Vicilla

Chromatin organisation and computer aided karyotyping of Triticum durum Desf. cv. Timilia

Abstract The tribe Triticeae includes three cereal genera Secale, Hordeum and Triticum and because of their economical and agronomical importance, the relationships among species on the tribe have been extensively investigated. The wild wheat relatives are an important source of genetic variation for cultivated species and wheat is an important crop of the mediterranean region. Banding pattern of metaphase chromosomes and nuclear DNA content in root meristematic cells of an old sicilian landrace "Timilia" were determined. Microdensitometric evaluation of nuclear absorption at different thresholds of optical density indicates the organization of chromatin in the interphase nuclei. Chromosome morphometric data, karyotype simmetry, the TF% values and Syi indices were determined. The results are compared with the data of other durum wheat varieties as Capeiti and Simeto

Cadmium uptake, localization and stress-induced morphogenic response in the fern Pteris vittata.

Cadmium uptake, tissue localization and structural changes induced at cellular level are essential to understand Cd tolerance in plants. In this study we have exposed plants of Pteris vittata to different concentrations of CdCl2 (0, 30, 60, 100 μM) to evaluate the tolerance of the fern to cadmium. Cadmium content determination and its histochemical localization showed that P. vittata not only takes up, but also transports and accumulates cadmium in the aboveground tissues, delocalizing it mainly in the less bioactive tissues of the frond, the trichomes and the scales. Cadmium tolerance in P. vittata was strictly related to morphogenic response induced by the metal itself in the root system. Adaptive response regarded changes of the root apex size, the developmental pattern of root hairs, the differentiation of xylem elements and endodermal suberin lamellae. All the considered parameters suggest that, in our experimental conditions, 60 μM of Cd may represent the highest concentration that P. vittata can tolerate; indeed this Cd level even improves the absorbance features of the root and allows good transport and accumulation of the metal in the fronds. The results of this study can provide useful information for phytoremediation strategies of soils contaminated by Cd, exploiting the established ability of P. vittata to transport, delocalize in the aboveground biomass and accumulate polluting metals

Chromosome numbers for the Italian flora: 3

In this contribution new chromosome data obtained from material collected in Italy are presented. It includes four chromosome counts for the following genera: Bellevalia (Asparagaceae), Genista (Fabaceae), Linaria (Plantaginaceae), and Teucrium (Lamiaceae)

Mineral nutrients in soil and pea plants after exposition to TiO2 nanoparticles through a biosolid-amended soil

In addition to the benefits derived from nanotechnology, there is also concern about the potential risks of engineered nanoparticles (ENPs) when released into the environment. Their possible accumulation and effects in agricultural soils and Nanonnovation 2018 Conference & Exibition crops are closely linked to food and agriculture safety. Particular attention has been focused on the reuse of biosolids from wastewater treatment plants that are considered a cost-effective practice for the improvement of nutrients and organic matter in agricultural soils and, but also a sink of contaminants such as nanoparticles (NPs). TiO2NPs have a global production of about 10.000 tons/year and it are among the most extensively used ENPs. Moreover, dissimilar or inconclusive results have been reported concerning the impact of TiO2 NPs on the soil-crop system, thus more information regarding their behavior are necessary. This study aimed to evaluate the potential effects of TiO2 NPs (anatase and rutile) and larger particles (bulk) on the availability of soil nutrients and on the nutritional status of Pisum sativum plants, simulating low (80 mg/kg) and high load of TiO2 (800 mg/kg ) in a biosolid-amended soil. Treated soils were analyzed for N, P, K, Ca, Mg, Mn, Fe, Cu, Zn, soil microbial community, and plants grown in laboratory for 30d were analyzed for growth, pigments and mineral nutrition. Results showed that the treatment with TiO2 at macro- and nano-scale significantly reduced the availability of Mn, Fe and P in soils, this last more evident for the NPs treatments. Indeed, the soil bacterial diversity was reduced when the mixture of anatase and rutile were spiked in the biosolid-amended soil at high concentration. Moreover, the pea plants from treated soils showed an imbalance in the mineral nutrition, with reduction in the plant tissues of Mn and K and increase of N. This study pose a reflection on the use of biosolid, which could act as a vehicle for the spread and accumulation of ENPs in agro-ecosystems

DNA methylation patterns of Brachypodium distachyon chromosomes and their alteration by 5-azacytidine treatment

Sequential immunolocalisation of 5-methylcytosine (5-MeC) and fluorescence in situ hybridisation with chromosome-specific BAC clones were performed on Brachypodium distachyon mitotic metaphase chromosomes to determine specific DNA methylation patterns of each chromosome in the complement. In the majority of cells examined, chromosomes Bd4 and Bd5, which bear the loci of 5S and 35S ribosomal DNA, respectively, had characteristic 5-MeC patterns. In contrast, the distribution of 5-MeC along the metacentric chromosome pairs Bd1, Bd2 and Bd3 was more variable. There were numerous differences in distribution of methylated sites between homologous chromosomes as well as between chromosome arms. Some chromosome sites, such as pericentromeric regions, were highly methylated in all chromosomes. Additionally, the influence of a hypomethylating agent, 5-azacytidine, on B. distachyon chromosome methylation patterns was confirmed. It was found that some chromosome pairs underwent demethylation more easily than others, but there was no apparent regularity in demethylation of particular chromosome segments

The Arabidopsis thaliana knockout mutant for phytochelatin synthase1 (cad1-3) Is defective in callose deposition, bacterial pathogen defense and auxin content, but shows an increased stem lignification

The enzyme phytochelatin synthase (PCS) has long been studied with regard to its role in metal(loid) detoxification in several organisms, i.e., plants, yeasts, and nematodes. It is in fact widely recognized that PCS detoxifies a number of heavy metals by catalyzing the formation of thiol-rich oligomers, namely phytochelatins, from glutathione and related peptides. However, recent investigations have highlighted other possible roles played by the PCS enzyme in the plant cell, e.g., the control of pathogen-triggered callose deposition. In order to examine novel aspects of Arabidopsis thaliana PCS1 (AtPCS1) functions and to elucidate its possible roles in the secondary metabolism, metabolomic data of A. thaliana wild-type and cad1-3 mutant were compared, the latter lacking AtPCS1. HPLC-ESI-MS analysis showed differences in the relative levels of metabolites from the glucosinolate and phenylpropanoid pathways between cad1-3 and wild-type plants. Specifically, in control (Cd-untreated) plants, higher levels of 4-methoxy-indol 3-ylmethylglucosinolate were found in cad1-3 plants vs. wild-type. Moreover, the cad1-3 mutant showed to be impaired in the deposit of callose after Cd exposure, suggesting that AtPCS1 protects the plant against the toxicity of heavy metals not only by synthesizing PCs, but also by contributing to callose deposition. In line with the contribution of callose in counteracting Cd toxicity, we found that another callose-defective mutant, pen2-1, was more sensitive to high concentrations of Cd than wild-type plants. Moreover, cad1-3 plants were more susceptible than wild-type to the hemibiotrophic bacterial pathogen Pseudomonas syringae. The metabolome also revealed differences in the relative levels of hydroxycinnamic acids and flavonols,with consequences on cell wall properties and auxin content, respectively. First, increased lignification in the cad1-3 stems was found, probably aimed at counteracting the entry of Cd into the inner tissues. Second, in cad1-3 shoots, increased relative levels of kaempferol 3,7 dirhamnoside and quercetin hexoside rhamnoside were detected. These flavonols are endogenous inhibitors of auxin transport in planta; auxin levels in both roots and shoots of the cad1-3 mutant were in fact lower than those of the wild-type. Overall, our data highlight novel aspects of AtPCS1 functions in A. thaliana