Osmolyte-related recovery of the opaque-6 lethal phenotype in Zea mays L

Endosperm growth and development is a complex phenomenon, driven by the coordinate expression of several genes. A series of endosperm mutants with altered timing and zein synthesis rate have been studied, allowing the partial unravelling of a multifarious system, integrating carbohydrate, amino acid, and storage protein me¬tabolisms, and operating during endosperm growth and development. The exact biological function of one of these loci, the Opaque-6 (O6) gene, remains to be acknowledged. The o6 locus determines a general reduction of 19- and 22 kDa zeins as well as a number of non-zein polypeptides present in the wild type endosperm. The o6 mutants present a collapsed, dull endosperm, leaf striations and early seedling death; however, o6 seedlings can survive when grown in the presence of exogenous proline. It has been suggested that, in mutant seeds and in con¬trast with the development of the normal seeds, proline does not reach the sites of protein synthesis in adequate amounts. Yet, it has been demonstrated that amino acids other than proline are also able to restore o6 seedling le¬thality, contradicting this hypothesis. In this paper, we explored the possibility that the observed proline-mediated rescue of o6 mutant seedling lethality regarded an osmolyte-mediated mitigation of aberrant protein folding rather than the restoration of a reduced proline flux needed for protein synthesis. This hypothesis was tested by means of in vitro cultivation of o6 seedlings in the presence of putative osmolytes including a series of amino acids, me¬thylamines, and polyols. Several osmolytes were identified, which were able to restore normal growth in o6 mutant seedlings. Root reestablishment required higher osmolyte concentrations than those necessary for the recovery of the aboveground plant parts. The results presented in this paper provide sufficient preliminary evidence to assume that proline-induced recovery of the o6 mutant phenotype depends on the osmolytic properties of this amino acid

Quality related traits of the maize (Zea mays L) grain: gene identification and exploitation

Developing maize plants with improved kernel quality traits involves the ability to use existing genetic variation and to identify and manipulate commercially important genes. This will open opportunities for designing novel variation in grain composition and will provide the basis for the development of the next generation of specialty maize. This paper provides an overview of current knowledge on the identification and exploitation of genes affecting the composition, development, and structure of the maize kernel with particular emphasis on pathways relevant to endosperm growth and development, differentiation of starch-filled cells, and biosynthesis of starches, storage proteins, lipids, and carotenoids. The potential that the new technologies of cell and molecular biology will provide for the creation of new variation in the future are also indicated and discussed

The b-32 ribosome inactivating protein from maize influences fumonisin accumulation in in-vitro bioassays

Fungi of the genus Fusarium are common plant pathogens mainly associated with cereal crops. In particular, Fusarium verticillioides Sacc. is the most common toxigenic fungus in maize worldwide, causing root, stalk, and ear rot. Fusarium spp. can produce a wide range of secondary metabolites, some of which can unfavourably affect human and animal health. Owing to the potential risks of fumonisins F. verticillioides secondary metabolites, new regulations for the allowable mycotoxin limit in food and feed have been put in place by most agencies worldwide. Plants act on the attack of pathogenic fungi through a complex network of active responses such as the production of proteins toxic or inhibitory to pathogens such as RIP (Ribosome-Inactivating-Protein). The RIP present in maize endosperm (termed b-32) has been widely investigated. Similarly to other RIPs, is accumulated in the seed as an inactive pro-RIP precursor, which is converted into an active form by proteolytic processing. To understand the relationships between structure and substrate specificity of the maize b-32 RIP protein, a series of recombinant b-32 sequences, by selective deleting of different domains (RIP b-32, RIP ∆N, RIP ∆C and RIP-∆C (Ala), were prepared. Recombinant sequences were expressed in Escherichia coli to obtain high levels of recombinant proteins, which were subsequently tested for their potential ability to reduce both the colonization of F. verticillioides and fumonisin accumulation

The maize (Zea mays) b-32 protein shows RIP activity in yeast cells

Ribosome-inactivating proteins (RIPs) are either single-chain (type 1) or two-chain (type 2) toxins. They are toxic to eukaryotic cells by cleaving a N-glycosidic bond in an extremely conserved loop located in the 28S RNA. This releases a specific adenine and inactivates the ribosome, ultimately inhibiting protein synthesis. Plant RIPs have been intensely investigated because of their projected antiviral, antifungal and insecticidal activity. RIPs also have biomedical applications as the toxic mojety of immunotoxins. Given their biotechnological potentials, it is strategic to develop platforms to rapidly evaluate the activity of recombinant RIPs. This investigation fills this need in that it reports that the yeast Saccharomyces cerevisiae is a model system to assess the impact of genetic manipulations on the functionality of a recombinant Zea mays RIP named b-32

Characterization of the maize b-32 ribosome inactivating protein and its interaction with fungal pathogen development

Plants respond to attack by pathogenic fungi with a complex network of responses, including the production and accumulation of proteins, such as the Ribosome Inactivating Proteins (RIPs) that are toxic or inhibitory to pathogens. In maize endosperm, a cytosolic albumin termed b-32 (RIP1) is synthesized in temporal and quantita¬tive coordination with the deposition of storage proteins. Research has shown that b-32 is able to i) enzymati¬cally inactive ribosomes modifying rRNA inhibiting protein synthesis in vitro, ii) inhibit the growth of Rhizoctonia solani mycelia in an in vitro and in planta assays, iii) reduce Fusarium culmorum head blight in wheat transgenic plants expressing b-32, and iv) diminish Fusarium verticillioides attack symptoms in leaf tissues assays of maize transgenic expressing ectopically b-32 protein. Similarly to other RIPs, maize b-32 is accumulated in the seed as an inactive precursor, which is converted into an active form by proteolytic processing which removes peptide segments from the N (residues 1-16 of pro-RIP) and C (residues 295-301) termini and also from the center (linker domain) of the polypeptide. In this review we will summarize evidence and advances related to the ability of the b-32 protein in contrasting pathogen attacks by considering and describing i) in vivo b-32 antifungal activity and ii) in vitro fungal development inhibition. These data provide information for assessing b-32 in developing plants with a higher capacity to contrast damages induced by pathogens

Antioxidant activity in a set of sorghum landraces and breeding lines

Sorghum (Sorghum bicolor L) is becoming an increasingly important crop in the developed world especially as a cereal grain option for patients with celiac disease, being also characterized by a high level of bioactive compounds. It is a good source of phenolic compounds, including phenolic acids, flavonoids and condensed tannins, that express antioxidant capacity and potential health benefits. A group of 210 sorghum genotypes was evaluated in terms of physical parameters and resulted to be characterized by a wide range of 1000-seeds weight (6.93 - 42.67 g) and kernel colour. A sub-set of 121 samples were selected by near infrared spectroscopy for chemical analyses, and revealed a wide range of variability for total antioxidant capacity (6.89 - 172.02 mmol TE kg-1 dm-1), phenols (0.60 - 20.73 g GAE kg-1 dm-1), condensed tannins (0 - 28,362.63 μg CE g-1 dm-1) and flavonoids (0 - 8,138.22 μg CE g-1 dm-1). A high negative correlation was observed between antioxidant compounds and the colour parameters L* and b*; on the contrary, correlation of the same parameters with a* was low and positive. The results of these preliminary analyses highlighted genotypes characterized by light-coloured grains (white or yellow), large seeds, high antioxidant properties but absence of condensed tannins, all traits which make them suitable for food industry

Spectroscopic Kernel Quality From A Symbiotic Corn Production

The management of the inoculation of a plant's roots, by means of biofertilizers (BF) containing arbuscular mycorrhizal (AM) fungi, is aimed at inducing modifications of the quality of the seeds. It is here shown that a seed-soil treatment can be elicited in the fingerprints of a symbiotic treatment using Near Infra Red (NIR)-SCiO NIR-SCiO spectra collections of single kernels: overall, a sensitivity of 73% and a specificity of 73% have been achieved, thus suggesting that it may be possible to assign the symbiotic origin of corn from just twenty kernels, provided that the dataset is adequately representative of the cultivar and AM. A global correlation study has shown a positive general trend (R2 0.45) of quality vs. quantity, in the sense that an increase in yield corresponded to an increase in the spectral differences between the symbiotic spectra and the control ones, but the inverse was also true, as a result of the parasitic behaviour of the BF treatments. The efficacy of the symbiosis can be back predicted from the NIR spectra; in fact, around 90% of the positive yield outcome results were discriminated from the negative ones. A reduction in the foliar pH (R2 0.37) and an increase in the foliar protein (R2 0.43) were observed as immediate phenotypic signs of a productive symbiosis. The commercial raw composition of the kernels appeared to only be affected slightly by the BF treatments; thus, till now uncharted secondary compounds of the maize kernels are involved, as supported by animal performances

Evaluation of ear rot (Fusarium verticillioides) resistance and fumonisin accumulation in Italian maize inbred lines

Mycotoxin contamination of maize (Zea mays L.) grain is a global threat to the safety of both human food and animal feed. Hence, the development of maize genotypes with reduced mycotoxin accumulation in grain is of major importance. In order to find maize germplasm sources of resistance to Fusarium ear rot, 34 Italian and six public inbred lines were evaluated by means of artificial inoculation in field experiments during 2009 and 2010. Relationships between ear rot and fumonisin concentration in the ears were investigated. Primary ears were challenged with a mixture of two Fusarium verticillioides isolates from Northern Italy, through kernel inoculation, and ear rot severity was assessed.The average number of visibly infected kernels per ear, after inoculation, ranged from 2 to 68 in 2009 and from 0 to 120 in 2010. Fumonisin concentrations in the inoculated ears were greater than in the experimental controls for both years. Variability was found between the inbred lines: fumonisin accumulation ranged from 0.56 to 240.83 mg kg-1 in 2009 and from 1.09 to 190.60 mg kg-1 in 2010. In both years, six inbred lines showed high fumonisin content (≥100 mg kg-1), while the other genotypes were almost equally split into two groups, low (≤10 mg kg-1) and medium (from 11 to 100 mg kg-1) fumonisin content. The number of infected kernels after artificial inoculation correlated with fumonisin concentration both in 2009 (r = 0.94; P≤0.01) and 2010 (r = 0.67; P≤0.01). Additionally, the percentage of internally infected kernels correlated positively with fumonisin concentration (r = 0.37; P≤0.01) and with the number of infected kernels (r = 0.29; P≤0.05). This research has demonstrated that Italian maize germplasm is a valid source of resistance to Fusarium ear rot. Furthermore, there is a strong association of visible Fusarium symptoms with fumonisin concentration, suggesting that selection in maize for reduced visible moulds should reduce the risk of mycotoxin contamination

Traditional Foods From Maize (Zea mays L.) in Europe

Maize (Zea mays L.) is one of the major crops of the world for feed, food, and industrial uses. It was originated in Central America and introduced into Europe and other continents after Columbus trips at the end of the 15th century. Due to the large adaptability of maize, farmers have originated a wide variability of genetic resources with wide diversity of adaptation, characteristics, and uses. Nowadays, in Europe, maize is mainly used for feed, but several food specialties were originated during these five centuries of maize history and became traditional food specialties. This review summarizes the state of the art of traditional foodstuffs made with maize in Southern, South-Western and South-Eastern Europe, from an historic evolution to the last research activities that focus on improving sustainability, quality and safety of food production

Genetic and phenotypic evaluation of european maize landraces as a tool for conservation and valorization of agrobiodiversity

The ECPGR European Evaluation Network (EVA) for Maize involves genebanks, research institutions, and private breeding companies from nine countries focusing on the valorization of maize genetic resources across Europe. This study describes a diverse collection of 626 local landraces and traditional varieties of maize (Zea mays L.) from nine European genebanks, including criteria for selection of the collection and its genetic and phenotypic diversity. High-throughput pool genotyping grouped the landraces into nine genetic groups with a threshold of 0.6 admixture, while 277 accessions were designated admixed and likely to have resulted from previous breeding activities. The grouping correlated well with the geographic origins of the collection, also reflecting the various pathways of introduction of maize to Europe. Phenotypic evaluations of 588 accessions for flowering time and plant architecture in multilocation trials over three years confirmed the great diversity within the collection, although phenotypic clusters only partially correlated with the genetic grouping. The EVA approach promotes conservation of genetic resources and opens an opportunity to increase genetic variability for developing improved varieties and populations for farmers, with better adaptation to specific environments and greater tolerance to various stresses. As such, the EVA maize collection provides valuable sources of diversity for facing climate change due to the varieties’ local adaptation