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Mitochondrial diseases are a group of genetic disorders characterized by dysfunctional mitochondria. Within eukaryotic cells, mitochondria contain their own ribosomes, which synthesize small amounts of proteins, all of which are essential for the biogenesis of the oxidative phosphorylation system. The ribosome is an evolutionarily conserved macromolecular machine in nature both from a structural and functional point of view, universally responsible for the synthesis of proteins. Among the diseases afflicting humans, those of ribosomal origin - either cytoplasmic ribosomes (80S) or mitochondrial ribosomes (70S) - are relevant. These are inherited or acquired diseases most commonly caused by either ribosomal protein haploinsufficiency or defects in ribosome biogenesis. Here we review the scientific literature about the recent advances on changes in mitochondrial ribosomal structural and assembly proteins that are implicated in primary mitochondrial diseases and neurodegenerative disorders, and their possible connection with metalloid pollution and toxicity, with a focus on MRPL44, NAM9 (MNA6) and GEP3 (MTG3), whose lack or defect was associated with resistance to tellurite. Finally, we illustrate the suitability of yeast Saccharomyces cerevisiae (S. cerevisiae) and the nematode Caenorhabditis elegans (C. elegans) as model organisms for studying mitochondrial ribosome dysfunctions including those involved in human diseases.

Isolated sorghum (Sorghum bicolor) storage proteins (kafirins) have been successfully used in the production of several bio-materials including adhesives, films, micro-particles, fibers, and biological scaffold material. Comparatively little research has been conducted on the use of isolated kafirins in food products or to produce bioactive peptides via hydrolysis for nutritional uses. To support such research, the aim of this study was to compare existing methods for bulk isolation of sorghum kafirins with the goal of identifying a solvent with the least toxicity that maintained a high extraction rate from food grade sorghum flour. A secondary goal was to characterize the kafirin isolates produced from various extraction methods to provide some information on their potential use in food products to guide future research in this area. Five different extraction methods were compared including 1) aqueous ethanol containing NaOH and sodium metabisulfite, 2) glacial acetic acid, 3) aqueous ethanol with sodium metabisulfite, 4) aqueous ethanol at acidic pH, and 5) alkaline pH alone. The protein contents of the kafirin isolates obtained by the five methods ranged from 49.76% to 56.83%. Kafirin isolates were characterized using reversed phase (RP)-high performance liquid chromatography (HPLC), which revealed substantial variability in the various kafirin patterns among the extraction methods tested. However, characterization of the kafirin isolates by size exclusion chromatography (SEC) did not show a high degree of variability among the methods tested. Likewise, analysis of the samples using sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE) showed essentially the same band profiles but with different band intensities among kafirin extraction methods. Surface hydrophobicity of the kafirin isolates varied considerably with isolates extracted with glacial acetic acid and aqueous ethanol plus sodium metabisulfite the most hydrophobic as indicated by hydrophobic dye binding.

Grain sorghum [Sorghum bicolor (L.) Moench] is a gluten-free grain that is gaining attention as a food crop that can be used in the management of celiac disease. At present, sorghum is widely grown in many semiarid regions of the world. New food-grade sorghum cultivars are of particular interest in Mediterranean countries due to improved quality characteristics and gluten-free status of the grains. Until now very few studies have examined the grain yield (GYLD) and agronomic performance characteristics of food-grade sorghum hybrids in Italy. A 2 year study was conducted to evaluate the agronomic performance of eight food-grade sorghum hybrids representing different maturity classes in trials conducted in Southern Italy. The results showed wide variation in adaptation of these hybrids as measured by differences in GYLD (2.35-8.50 t ha(-1)) and other phenomorphological characteristics. Of particular interest was the fact that many of the early-flowering hybrids (e. g. SP-X303) performed better than the later-flowering hybrids (e. g. ArchX-02). These results demonstrated that flowering time of hybrid and crop cycle length are important factors to consider in selecting cultivars for production in the Mediterranean region.

Grain protein has recently received attention in several countries because of a premium associated to increased protein concentration (PC) in grains. Nitrogen (N) fertilizer management with its timing of application and rates plays a crucial role in increasing PC. The objectives of the present study were to: (1) quantify the effects of different N fertilizer rates and timing of application on grain yield and grain protein of six durum wheat cultivars, and (2) identify the best cultivar in terms of net return, yield, and protein content in typical pedo-climatic condition of southern Italy. The field experiments were conducted in Foggia during three growing seasons (2002-2003, 2003-2004, and 2004-2005). Results from this study showed that N application at sowing did not influence the cultivars performance. N applied at stem elongation increased yield and protein content provided there is adequate soil water for N uptake. In absolute terms, the best combination was the cultivar Svevo managed using 104 kg N ha-1 split between tillering and stem-elongation stage. The excessive N splitting reduces yield and does not provide any improvement in terms of protein content.

Background A chemical cross-talk between plants and insects is required in order to achieve a successful co-adaptation. In response to herbivory, plants produce specific compounds, and feeding insects respond adequately7 to molecules produced by plants. Here we show the role of the gut microbial community of the mint beetle Chrysolina herbacea in the chemical cross-talk with Mentha aquatica (or watermint). Results By using two-dimensional gas chromatography–mass spectrometry we first evaluated the chemical patterns of both M. aquatica leaf and frass volatiles extracted by C. herbacea males and females feeding on plants, and observed marked differences between males and females volatiles. The sex-specific chemical pattern of the frass paralleled with sex-specific distribution of cultivable gut bacteria. Indeed, all isolated gut bacteria from females belonged to either α- or γ-Proteobacteria, whilst those from males were γ-Proteobacteria or Firmicutes. We then demonstrated that five Serratia marcescens strains from females possessed antibacterial activity against bacteria from males belonging to Firmicutes suggesting competition by production of antimicrobial compounds. By in vitro experiments, we lastly showed that the microbial communities from the two sexes were associated to specific metabolic patterns with respect to their ability to biotransform M. aquatica terpenoids, and metabolize them into an array of compounds with possible pheromone activity. Conclusions Our data suggest that cultivable gut bacteria of Chrysolina herbacea males and females influence the volatile blend of herbivory induced Mentha aquatica volatiles in a sex-specific way. Electronic supplementary material The online version of this article (doi:10.1186/s12870-017-0986-6) contains supplementary material, which is available to authorized users.

Vetiver is the only grass cultivated worldwide for the root essential oil, which is a mixture of sesquiterpene alcohols and hydrocarbons, used extensively in perfumery and cosmetics. Light and transmission electron microscopy demonstrated the presence of bacteria in the cortical parenchymatous essential oil-producing cells and in the lysigen lacunae in close association with the essential oil. This finding and the evidence that axenic Vetiver produces in vitro only trace amounts of oil with a strikingly different composition compared with the oils from in vivo Vetiver plants stimulated the hypothesis of an involvement of these bacteria in the oil metabolism. We used culture-based and culture-independent approaches to analyse the microbial community of the Vetiver root. Results demonstrate a broad phylogenetic spectrum of bacteria, including alpha-, beta- and gamma-Proteobacteria, high-G+C-content Gram-positive bacteria, and microbes belonging to the Fibrobacteres/Acidobacteria group. We isolated root-associated bacteria and showed that most of them are able to grow by using oil sesquiterpenes as a carbon source and to metabolize them releasing into the medium a large number of compounds typically found in commercial Vetiver oils. Several bacteria were also able to induce gene expression of a Vetiver sesquiterpene synthase. These results support the intriguing hypothesis that bacteria may have a role in essential oil biosynthesis opening the possibility to use them to manoeuvre the Vetiver oil molecular structure.

The transposition of the Ty mobile genetic element of Saccharomyces cerevisiae is induced by carcinogens. While the molecular background of spontaneous Ty1 transposition is well understood, the detailed mechanism of carcinogen induced Ty1 transposition is not clear. We found that mitochondrial functions participate in the Ty induced transposition induced by carcinogens. Contrary to the parental rho(+) cells rho(-) mutants (spontaneous or induced by ethidium bromide) do not increase the rate of Ty1 transposition upon treatment with carcinogens. Preliminary results strongly suggest that the absence of oxidative phosphorylation in rho(-) mutants is the reason for the inhibited Ty transposition. The lack of carcinogen induced Ty1 transposition in rho(-) cells is not specific for a particular carcinogen and represents a general feature of different carcinogenic substances inducing rho(-). It is concluded that carcinogen induced Ty1 transposition depends on the functional state of mitochondria and cannot take place in cells with compromised mitochondrial function (rho(-)).

Wheat (Triticum spp. L.), rye (Secale cereal L.), and barley (Hordeum vulgare L.) seeds contain peptides toxic to celiac patients. Maize (Zea mays L.) and rice (Oryza sativa L.) are distant relatives of wheat as well as sorghum (Sorghum bicolor (L.) Moench) and are known to be safe for celiacs. Both immunochemical studies and in vitro and in vivo challenge of wheat-free sorghum food products support this conclusion, although molecular evidence is missing. The goal of the present study was to provide biochemical and genetic evidence that sorghum is safe for celiac patients. In silico analysis of the recently published sorghum genome predicts that sorghum does not contain peptides that are toxic for celiac patients. Aqueous/alcohol-soluble prolamins (kafirins) from different sorghum varieties, including pure lines and hybrids, were evaluated by SDS-PAGE and HPLC analyses as well as an established enzyme-linked immunosorbent assay (ELISA) based on the R5 antibody. These analyses provide molecular evidence for the absence of toxic gliadin-like peptides in sorghum, confirming that sorghum can be definitively considered safe for consumption by people with celiac disease.

To gain insight into essential oil metabolism we analyzed the oil produced in Vetiver ( Vetiveria zizanioides (L.) Nash) roots during early growth. Planting of Vetiver in the Campania region of southern Italy was performed in the spring using Vetiver culms with short roots and approximately 20 cm leaves. During the first two years of growth Vetiver essential oil was hydrodistilled from root samples collected at intervals of two months starting from transplantation. The production of Vetiver oil was constant during the first six months. Over the next two months there was a twofold increase in production of essential oil, which anticipated a progressive decrease during the cold months (from January to April 2003). The analysis of the Vetiver oil showed the presence of 49 constituents, mainly the tricyclic sesquiterpenes khusimol and zizanoic acid, and the bicyclic sesquiterpenes ( E )-isovalencenol, junenol, juniper camphor, nootkatone and α-vetivone. These results suggest that Vetiver essential oil production is closely related to the metabolism of plant roots, which is affected by changes in environmental temperatures.

Some nuclear genes in Saccharomyces cerevisiae (S. cerevisiae) respond to signals from the mitochondria in a process called by Butow (Cell Death Differ. 9 (2002) 1043–1045) retrograde regulation. Expression of these genes is activated in cells lacking mitochondrial function by involvement of RTG1, RTG2 and RTG3 genes whose protein products bind to “R-boxes” in the promoter region; RTG2p is a cytoplasmic protein. Since S. cerevisiae rho0 strains, lacking the entire mitochondrial genome, are resistant to lycorine, an alkaloid extracted from Amaryllis plants, it could be hypothesized that in rho0 cells the dysfunctional mitochondrial status stimulates overexpression of nuclear genes very likely involved in both nuclear and mitochondrial DNA replication. In this report we show that the resistance of rho0 cells to lycorine is affected by the deletion of RTG genes.

Under certain in vitro (salt and temperature) conditions rRNA aggregation occurs in female inflorescences but not in leaves or pollen RNA preparations from hazelnut (Corylus avellana L.), a species of economic interest. This paper describes experiments addressing an explanation of this phenomenon. The experiments demonstrate that: (i) trans-acting factors induce rRNA aggregate formation in female inflorescences RNA preparations; (ii) these factors support aggregation also of heterologous rRNA; (iii) aggregation is a function of temperature pre-treatment of rRNA and not of source 18S rRNA; (iv) the factors inducing rRNA aggregates are sensitive to RNase; (v) antisense small nucleolar RNAs (snoRNAs) participate in rRNA aggregate formation. snoRNAs are involved in pre-rRNA spacer cleavages, and are required for the two most common types of rRNA modifications: 2'-O-ribose methylation and pseudouridylation. Even though it is questionable whether rRNA aggregation really happens in female inflorescence in vivo, the phenomenon observed in vitro may reflect the abundance of snoRNAs in these reproductive structures. In fact the level of accumulation of three tested snoRNAs, R1, U14 and U3, is much higher in female inflorescence than in leaves or pollen of hazelnut. This finding opens the possibility of studying the role of snoRNAs in tissue development in plants.

An efficient method has been developed to improve preparation of phage particles by ammonium sulfate precipitation and to yield high quality DNA. The method, that has been used to screen plant DNA libraries constructed in λvectors, is inexpensive, does not require purification of phage particles, and can be used from either plate stocks or liquid lysates. Up to 1100 μg DNA was produced from 5 ml lysate obtained from agar plates.

In this paper we report the isolation and preliminary characterisation of nuclear mutants with increased mitochondrial mutability in fission yeast. Screening of about 2000 clones after nitrosoguanidine mutagenesis led to the isolation of ten mutator mutants. For one of them (mut-1) we show that the mutation is chromosomally encoded. The activity of the mutator is restricted to the mitochondrial genome, since it increases the mutation rate to mitochondrially encoded drug resistance considerably, whereas the mutability of nuclear genes is not altered.

Expression of serogroup B meningococcal capsular polysaccharide undergoes frequent phase variation involving reversible frameshift mutations within a homopolymeric repeat in the siaD gene. A high rate of phase variation is the consequence of a biochemical defect in methyl-directed mismatch repair. The mutator phenotype is associated to the absence of DNA adenine methyltransferase (Dam) activity in all pathogenic isolates and in 50% of commensal strains. Analysis of the meningococcal dam gene region revealed that in all Dam- strains a gene encoding a putative restriction endonuclease (drg) that cleaves only the methylated DNA sequence 5'-GmeATC-3' replaced the dam gene. Insertional inactivation of the dam and/or drg genes indicated that high rates of phase variation and hypermutator phenotype are caused by absence of a functional dam gene.

Using as a template genomic DNA extracted from the cultivar Ofanto of durum wheat (Triticum turgidum var. durum Desf), and as DNA primers two oligonucleotides spanning Lipoxygenase-1 (LOX-1) gene region, evolutionary conserved in different plants, we have amplified in a PCR reaction a 1200 bp fragment. This DNA fragment has been subcloned and used to prepare probes in Slot-blot experiments. Our aim was to detect the expression LOX gene(s), at level of RNA, in different cultivars of Durum wheat. Preliminary results indicates that there are significant variations in the expression of LOX gene(s) in the tested samples, and a good negative correlation of LOX-1 mRNA levels with both semolina p-carotene content and semolina yellow index of durm wheat varieties utilized. The physiological, significance of this relation will be discussed.

Encapsulating peritoneal sclerosis (EPS) represents a critical complication of peritoneal dialysis (PD). EPS is characterized by abdominal discomfort, often leading to fatal outcomes with limited pharmaceutical and surgical options. Herein is described a case of EPS with a favorable outcome in an African male treated with PD for 15 years. Repeated courses of prednisone and tamoxifen significantly attenuated the abdominal symptoms and the peritoneal membrane thickening. This case suggests a time dependent effect of medical treatment encouraging clinical efforts to maintain a mild immunosuppressant regimen and tamoxifen in the presence of EPS also on the long run. Future and ad hoc studies should test this hypothesis.

Vetiver is the only grass cultivated worldwide for the root essential oil, which is a mixture of sesquiterpene alcohols and hydrocarbons, used extensively in perfumery and cosmetics. Light and transmission electron microscopy demonstrated the presence of bacteria in the cortical parenchymatous essential oil-producing cells and in the lysigen lacunae in close association with the essential oil. This finding and the evidence that axenic Vetiver produces in vitro only trace amounts of oil with a strikingly different composition compared with the oils from in vivo Vetiver plants stimulated the hypothesis of an involvement of these bacteria in the oil metabolism. We used culture-based and culture-independent approaches to analyse the microbial community of the Vetiver root. Results demonstrate a broad phylogenetic spectrum of bacteria, including alpha-, beta- and gamma-Proteobacteria, high-G+C-content Gram-positive bacteria, and microbes belonging to the Fibrobacteres/Acidobacteria group. We isolated root-associated bacteria and showed that most of them are able to grow by using oil sesquiterpenes as a carbon source and to metabolize them releasing into the medium a large number of compounds typically found in commercial Vetiver oils. Several bacteria were also able to induce gene expression of a Vetiver sesquiterpene synthase. These results support the intriguing hypothesis that bacteria may have a role in essential oil biosynthesis opening the possibility to use them to manoeuvre the Vetiver oil molecular structure.

In a previous work temporal succession of two sheathed, filamentous methanotrophic γ-Proteobacteria, Crenothrix polyspora and Clonothrix fusca, was observed by analyzing the microbial community of an artesian well. Although a correlation between physical-chemical environmental parameters and growth of these microorganisms was suggested, the link with the microbial physiology remained elusive. In this study we demonstrate that phosphatidylethanolamine and cardiolipin, generally abundant in most γ-Proteobacteria, could not be detected in C. fusca membranes by thin-layer chromatography. The absence of cardiolipin was supported by results of enzymatic assay of cardiolipin synthase activity and by Southern blot analysis that failed to detect cls homologous sequences in the genomic DNA of this microorganism. The absence of phosphatidylethanolamine and cardiolipin might account for requirement of millimolar concentrations of magnesium and calcium for growth, and for massive over-growth of this microorganism during the winter season when the water levels of magnesium and calcium increased in the artesian well. © 2007 Taylor & Francis.

In the yeast Saccharomyces cerevisiae, mutants are viable with large deletions (rho-), or even complete loss of the mitochondrial genome (rho0). One class of rho-mutants, which is called hypersuppressive, is characterised by a high transmission of the mutated mitochondrial genome to the diploid progeny when mated to a wild-type (rho+) haploid. The nuclear gene CCE1 encodes a cruciform cutting endonuclease, which is located in the mitochondrion and is responsible for the highly biased transmission of the hypersuppressive rho-genome. CCE1 is a Holliday junction specific endonuclease that resolves recombination intermediates in mitochondrial DNA. The cleavage activity shows a strong preference for cutting after a 5'-CT dinucleotide. In the absence of the CCE1 gene product, the mitochondrial genomes remain interconnected and have difficulty segregating to the daughter cells. As a consequence, there is an increase in the fraction of daughter cells that are rho0. In this paper we demonstrate the usefulness of lycorine, together with staining by 4',6-diamidino-2-phenylindole (DAPI), to assay for the mitotic stability of a variety of mitochondrial genomes. We have found that rho+and rho-strains that contain CT sequences produce a large fraction of rho0progeny in the absence of CCE1 activity. Only those rho-mitochondrial genomes lacking the CT recognition sequence are unaffected by the cce1 allele.Key words: yeast, mitochondria, hypersuppressive, Saccharomyces cerevisiae, lycorine.

Summary Thirty-nine nitrosoguanidine-induced respiratory-deficient mutants of Kluyveromyces lactis strain CBS 2359 were isolated. These mutants do not grow on glycerol and have a lower respiration rate associated with a reduced capacity to react with tetrazolium salts. Mitochondria isolated from these mutants are altered in the oxidation of several substrates of the respiratory chain. From the spectroscopic analysis of the cytochromes it is possible to distinguish four groups of mutants in which: first group) all cytochromes are missing; second group) only cytochrome a is missing; third group) cytochromes a , b , c 1 are missing; fourth group) no cytochrome is absent. Tetrade analyses show that the respiratory-deficient mutants are of chromosomal type. The existence of these mutants demonstrates that the failure in obtaining cytoplasmic respiratory-deficient mutants in Kluyveromyces lactis does not depend on the lethality of the alteration of the respiratory chain.

It is now well documented that mitochondrial (mt) genomes of all eukaryotes contain genes encoding elements of mt protein synthesis (rRNAs and tRNAs) as well as a few proteins for respiration and energy conservation. Genes encoding apo- cytochrome b (cob) or subunit I of cytochrome c oxidase (coxl) can be found, with no exception, in every mt genome. These genes are sufficiently similar in sequence, that comparison of DNA sequences allows to assign their functions in the different organisms. This set of genes is referred to as the “basic set” of mt genetic information (Kotylak et al., 1985). Lower eukaryotes possess additional genetic information in their mt genomes, which has been termed by Slonimski and collaborators (Kotylak et al., 1985) the “extra set” of mt genetic information. This information is deposited as introns of the genes forming the “basic set”. It is, however, important to note, that not all mt introns contain the genetic information for a protein.