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Sorghum (Sorghum bicolor L.) is a gluten-free cereal with many varieties containing greater amounts of bioactive compounds than other grains. In this work, the phenolic composition and antioxidant capacity were compared in three sorghum varieties grown in the Mediterranean, consisting of different pericarp colors: white, red, and black. The qualitative and quantitative profiles of acidified aqueous methanol extracts of whole grain sorghum flours were elucidated through ultrahigh performance liquid chromatography (UHPLC) coupled to photodiode array (PDA) and ion trap time-of-flight (IT-TOF) mass spectrometry. The investigated sorghum varieties contained polyamines, such as spermidines, and polyphenolic compounds belonging to different classes: hydroxycinnamic acids, flavanols, flavones, and flavanones. Pyrano-flavanone-flavanols were detected only in the black sorghum. Precolumn and postcolumn 2,2′-diphenyl-1-picrylhydrazyl (DPPH)· radical scavenging assays to determine the main antioxidant compounds isolated from sorghum were developed. These results will further help develop sorghum varieties containing polyphenols relevant to the potential prevention of human diseases.

A considerable body of evidence links together mitochondrial dysfunctions, toxic action of metalloid oxyanions, and system and neurodegenerative disorders. In this study we have used the model yeast Saccharomyces cerevisiae to investigate the genetic determinants associated with tellurite resistance/sensitivity. Nitrosoguanidine-induced K2TeO3-resistant mutants were isolated, and one of these mutants, named Sc57-Te5R, was characterized. Both random spore analysis and tetrad analysis and growth of heterozygous (TeS/Te5R) diploid from Sc57-Te5R mutant revealed that nuclear and recessive mutation(s) was responsible for the resistance. To get insight into the mechanisms responsible for K2TeO3-resistance, RNA microarray analyses were performed with K2TeO3-treated and untreated Sc57-Te5R cells. A total of 372 differentially expressed loci were identified corresponding to 6.37% of the S. cerevisiae transcriptome. Of these, 288 transcripts were up-regulated upon K2TeO3 treatment. About half of up-regulated transcripts were associated with the following molecular functions: oxidoreductase activity, structural constituent of cell wall, transporter activity. Comparative whole-genome sequencing allowed us to identify nucleotide variants distinguishing Sc57-Te5R from parental strain Sc57. We detected 15 CDS-inactivating mutations, and found that 3 of them affected genes coding mitochondrial ribosomal proteins (MRPL44 and NAM9) and mitochondrial ribosomal biogenesis (GEP3) pointing out to alteration of mitochondrial ribosome as main determinant of tellurite resistance.

A white food-grade waxy sorghum Tw variety, grown in two Mediterranean sites (named Tw1M and Tw1S) was evaluated for nutrient composition and fatty acid- and mineral concentrations in order to determine the suitability of producing waxy sorghum for human uses in southern Italy. The nutritional values of the grains of the Tw inbred line grown in the two trial fields were substantially the same, except for slight differences in ash level and accordingly slight variation in mineral composition. In samples from both locations, a higher percentage of K was observed among the nutritionally essential macro-elements, and higher percentages of Zn, Fe, Mn, Cu, Al among the nutritionally essential micro-element along with a strong difference in Cd content was among trace elements. Across both sites linoleic, oleic and palmitic were the most abundant fatty acids, while very slight variations in the content of minerals were found among the two samples examined. These results demonstrate the importance of developing agronomically productive waxy sorghum varieties suitable for growth in non-traditional sorghum producing regions both as a food and feed crop (i.e. with good nutritional quality) and for utilization in new products at the industrial level.

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.

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.

Sorghum has been grown as a food crop for many centuries in Africa and Asia and it is a cereal option for celiac patients. Due to its properties as a wheat-free food, interest is increasing in cultivating sorghum in Mediterranean countries. To evaluate the agronomic characteristics of growth in the Mediterranean area, six hybrids of sorghum (Sorghum bicolor L. Moench), representing different maturity classes, were selected and tested in experimental trials for 2 years in San Bartolomeo in Galdo (BN) hill, South Italy. The results showed moderate variation in adaptation of these hybrids as measured by differences in grain yields (4.60 to 7.30 t ha-1) and other pheno-morphological traits. In particular, the hybrid SW6129W from Bolivia and the hybrid SASG05W from Argentina had grain yields higher than others tested in these trials. Our results demonstrated that selected food-grade hybrids from South America, have agronomic characteristics well-suited for cultivation in Mediterranean countries.

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.

Mitochondria are eukaryotic organelles which contain the own genetic material and evolved from free-living Eubacteria, namely hydrogen-producing Alphaproteobacteria. Since 1965, biologists provided, by research at molecular level, evidence for the prokaryotic origins of mitochondria. However, determining the precise origins of mitochondria is challenging due to inherent difficulties in phylogenetically reconstructing ancient evolutionary events. The use of new tools to evidence the prokaryotic origin of mitochondria could be useful to gain an insight into the bacterial endosymbiotic event that resulted in the permanent acquisition of bacteria, from the ancestral cell, that through time were transformed into mitochondria. Electron microscopy has shown that both proteobacterial and yeast cells during their growth in the presence of increasing amount of tellurite resulted in dose-dependent blackening of the culture due to elemental tellurium (Te 0 ) that formed large deposits either along the proteobacterial membrane or along the yeast cell wall and mitochondria. Since the mitochondrial inner membrane composition is similar to that of proteobacterial membrane, in the present work we evidenced the black tellurium deposits on both, cell wall and mitochondria of ρ + and respiratory deficient ρ − mutants of yeast. A possible role of tellurite in studying the evolutionary origins of mitochondria will be discussed.

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.

The effects of potassium tellurite on growth and survival of rho + and rho0 Saccharomyces cerevisiae strains were investigated. Both rho+ and rho0 strains grew on a fermentable carbon source with up to 1.2 mM K2TeO3, while rho+ yeast cells grown on a non-fermentable carbon source were inhibited at tellurite levels as low as 50 μM suggesting that this metalloid specifically inhibited mitochondrial functions. Growth of rho+ yeast cells in the presence of increasing amount of tellurite resulted in dose-dependent blackening of the culture, a phenomenon not observed with rho0 cultures. Transmission electron microscopy of S. cerevisiae rho+ cells grown in the presence of tellurite showed that blackening was likely due to elemental tellurium (Te0) that formed large deposits along the cell wall and small precipitates in both the cytoplasm and mitochondria. © 2009 Springer Science+Business Media, LLC.

Although sorghum has been used for centuries as a food-crop in Africa and India, researchers in the United States and Europe have only relatively recently become interested in the potential of this unique cereal. Much of this interest focuses on the potential use of sorghum in food product development for individuals with allergies to foods containing wheat-based flours. Because it lacks gluten, sorghum is considered safe for people diagnosed with celiac disease, a condition marked by intolerance to gluten. Recent studies have shown that certain sorghum varieties, tan-plant sorghums, can be used to produce high-quality food and beverage products including cookies, waffles, flour, bread, noodles and beer. The intention is to promote the use and marketing of these sorghums in the Italian Campania Region for sorghum flours suitable as food for celiac patients.

BACKGROUND & AIMS: Celiac disease is a condition in which genetically predisposed people have an autoimmune reaction to gluten proteins found in all wheat types and closely related cereals such as barley and rye. This reaction causes the formation of autoantibodies and the destruction of the villi in the small intestine, which results in malabsorption of nutrientsand other gluten-induced autoimmune diseases. Sorghum is a cereal grain with potential to be developed into an important crop for human food products. The flour produced from white sorghum hybrids is light in color and has a bland, neutral taste that does not impart unusual colors or flavors to food products. These attributes make it desirable for use in wheat-free food products. While sorghum is considered as a safe food for celiac patients, primarily due to its relationship to maize, no direct testing has been conducted on its safety for gluten intolerance. Therefore studies are needed to assess its safety and tolerability in celiac patients. Thus the aim of the present study was to assess safety and tolerability of sorghum flour products in adult celiac disease patients, utilizing an in vitro and in vivo challenge. RESULTS: Sorghum protein digests did not elicit any morphometric or immunomediated alteration of duodenal explants from celiac patients. Patients fed daily for 5 days with sorghum-derived food product did not experience gastrointestinal or non-gastrointestinal symptoms and the level of anti-transglutaminase antibodies was unmodified at the end of the 5-days challenge. CONCLUSIONS: Sorghum-derived products did not show toxicity for celiac patients in both in vitro and in vivo challenge. Therefore sorghum can be considered safe for people with celiac disease.

The Italian cigar manufacturing process includes a fermentation step that leads to accumulation of nitrite and tobacco-specific nitrosamines (TSNA), undesirable by-products due to their negative impact on health. In this study, growth and biochemical properties of Debaryomyces hansenii TOB-Y7, a yeast strain that predominates during the early phase of fermentation, have been investigated. With respect to other D. hansenii collection strains (Y7426, J26, and CBS 1796), TOB-Y7 was characterized by the ability to tolerate very high nitrite levels and to utilize nitrite, but not nitrate, as a sole nitrogen source in a chemically defined medium, a property that was enhanced in microaerophilic environment. The ability to assimilate nitrite was associated to the presence of YNI1, the gene encoding the assimilatory NAD(P)H:nitrite reductase (NiR), absent in Y7426, J26, and CBS 1796 by Southern blot data. YNI1 from TOB-Y7 was entirely sequenced, and its expression was analyzed in different media by Northern blot and reverse transcriptase polymerase chain reaction. The evidence that, in D. hansenii TOB-Y7, YNI1 was transcriptional active also in the presence of high ammonia concentration typical of tobacco fermentation, stimulated the development of an improved process that, on a laboratory scale, was proved to be effective in minimizing nitrite and TSNA accumulation.

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.

The open reading frame of the first intron of the mitochondrial cox1 gene (cox1I1) was expressed in Escherichia coli. The putative intron-encoded protein stimulated the formation of intra-chromosomal lac +-recombinants about threefold. No stimulation was found when the reading frame was inserted in the opposite direction, or when it was interrupted by a deletion. The intronic open reading frame did not complement recA − or recB − mutants of E. coli. In S. pombe, elimination of this intron did not abolish homologous recombination in mitochondria. A possible role of the recombinase activity in yeast mitochondria will be discussed.

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.