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This article reports on the biochemical investigation of six African cereals (fonio, teff, sorghum, African rice, finger millet, pearl millet). These cereals play an important role in food security in many African and Asian regions, despite not being internationally traded, with the exceptions of sorghum and, partially, of pearl millet. Nowadays, crop breeders and research institutions are becoming concerned with improving the productivity and the nutritional quality of these cereals for reasons connected with the problems of human population growth and climate change. However, in these species less is known about the presence and content of anti-nutritional components that may impact human health, such as polyphenols, in particular tannins, phytate and the goitrogenic C-glycosyl flavones. In this work, these compounds were quantified and identified by means of biochemical assays or HPLC and LC-MS analysis. Obtained results showed that each of the six analysed cereals contained different levels of anti-nutritional or harmful goitrogenic compounds. In particular, all three examined pearl millet lines showed the sharply highest content of C-glycosyl flavones, slightly highest content of phytic acid and a relevant average level of polyphenols. On the other side, the Fonio lines presented the lowest level of C-glycosyl flavones and condensed tannins.

A number of orphan- (understudied) cereals are staple food largely consumed in many african developing countries. These crops are often indigenous ancient species that in general are well adapted to the extreme soil and climatic conditions prevalent in many African countries. However, orphan crops have been poorly studied under nutritional and nutraceutical aspects and, due to lack of genetic improvement, produce inferior yields compared to the major cereals. In this work we focused our attention on 6 african cereals: Pennisetum glaucum (pearl millet), Eleucine coracana (finger millet), Oryza glaberrima (African rice), Digitaria exilis (fonio), three ssp. of Sorghum ssp. and three ssp. of Eragrostis teff. Seeds of the above plants were collected from a number of sources: ICRISAT, Niger (P. glaucum); dr. Njido Kane, IRSA, Senegal (O. glaberrima); dr. Moussa Daouda Sanogo, Institut d'Economie Rurale, Mali (D. exilis); local producers in Eritrea (Sorghum, teff and finger millet). The flour prepared from seeds of each sample was analysed as to nutritional, anti-nutritional and nutraceutical properties, ascribable to the presence and amount of phytic acid, anthocyanins, poliphenols and condensed tannins, as well as to the total antioxidant activity. Moreover, we investigated the occurrence of C-glycosyl flavones, goitrogenic molecules interfering with the thyroid metabolism, which are of particular relevance for consumer's health. Results obtained from this preliminary screening carried out, showed relevant differences as regards all the above cited compounds, supplying a base for planning bio-fortification and/or genetic improvement projects.

The present invention concerns the field of gene integration and durable gene expression in specific genomic sites of transgenic animals suitable for the generation of tools for biomedical research and biotechnological applications.

Pearl millet is a crop with the potential to meet real world challenges and achieve sustainable agricultural systems. Its high amount of macro and micronutrients, low vulnerability to pathogens, tolerance to drought and salinity, and short life cycle, make this crop ideal as staple crop for growing populations (Basavaraj, 2010-J SAT Agric Res 8:1). However, the nutritional quality of pearl millet is hampered by the high content of anti-nutrient molecules such as phytic acid (PA) and C-glucosylflavones (CGFs), which limit the bio-availability of minerals and cause goitrogenic effects, respectively. PA is the major source of phosphorous in seeds, but it forms mineral complexes with Fe, Zn, Ca, K and Mg on the gut, causing problems such as poor mineral bioavailability and eutrophication of waterways (Bouwman et al, 2013-PNAS 52:20882). The development of low PA (lpa) mutants of cereals crops and legumes could help to alleviate this situations. Nonetheless, lpa plants often show poor seed germination that compromise the crop yield, decreasing its agronomic value making them undesirable for farmers. (Redekar et al, 2015-BMC Gen 16:1074). With the aim to produce association studies by integrating phenotypic and GBS data, we evaluated the natural genetic variability of the levels of PA in two pearl millet populations. A total of 153 inbred lines derived from African landraces, has been screened to quantify the content of PA in flour extracts and the most contrasting lines (reduced and high PA - RPA and HPA respectively; and reduced and high CGFs - RCGF and HCGF) have been identified and further analyzed. These inbred lines were used for the evaluation of the expression profiles of genes involved in PA biosynthesis during the seed development. A total of 11 orthologous genes of the PA and one of the CGFs biosynthesis pathways were identified in the pearl millet genome and their expression was investigated through different seed developmental stages. For the expression analysis three stages of seed development were established: early, middle and late development. Total RNA was isolated and gene expression was assayed by Real Time PCR. Actually, a major MIPS (myo-inositol phosphate synthase - Pgl_GLEAN10001337) increase of expression was detected during the middle developmental stage on the HPA phenotype. A similar peak of expression was previously reported for other species as common bean, rice and soybean. Since this expression peak was not detected on the RPA inbred line, we hypothesize that the diminished MIPS expression level may be correlated with the decrease of PA content. This outcome is very interesting for our work, since MIPS catalyze the rate-limiting step in the synthesis of all inositol-containing compounds (Rasmussen et al, 2010-Biochem Soc T 38:689). For the CGFs biosynthesis, a slightly CGT (C-Glucosyl Transferase - Pgl_GLEAN10014648) increase of expression was detected on early seed development and its contribution for the observed differences in CGFs content is under evaluation. Our results contribute to increase the understanding of the glycosyl flavones biosynthesis pathway which is not yet fully clarified. This work is supported by CERES-NewPearl project, jointly funded by Fondazione Cariplo and Agropolis Fondation.

Productivité des aigrettes et hérons nicheurs en six sites asiatiques. — La production, la taille des oeufs et les conditions corporelles des poussins ont été étudiées chez le Héron garde-boeufs (Bubulcus ibis), l’Aigrette intermédiaire (Egretta intermedia), l’Aigrette garzette (Egretta garzetta), le Crabier chinois (Ardeola bacchus) et le Bihoreau gris (Nycticorax nycticorax) en deux sites de Chine centrale et dans quatre autres au Pakistan. La production en Chine s’est avérée très forte, plus importante qu’au Pakistan, probablement en relation avec une grande disponibilité de zones humides pour l’alimentation. La forme sombre de l’Aigrette garzette de la côte du Pakistan présentait une productivité significativement plus faible que celle des formes blanches., Fasola Mauro,Canova Luca,Boncompagni Eleonora,Muhammad Ashiq,Najam Rahat,Wei Liu,Dong Yuanhua,Zhang Yingmei. Breeding productivity of egrets and herons at six sites in Asia. In: Revue d'Écologie (La Terre et La Vie), tome 62, n°4, 2007. pp. 369-374.

Pearl millet is the sixth world most important cereal grain. It is is produced as a rain-fed grain and fodder crop in the hottest, driest regions of sub-Saharan Africa and the Indian sub-continent, where it ensures food security by providing both calories as well as essential micronutrients more reliably than any other cereal. However, the nutritional quality of pearl millet is serioulsy hampered by its high content of phytic acid (IP6) and C-glucosylflavones (CGFs), both well-known anti-nutritional compounds that limit mineral bioavailability and cause goitrogenic effects, respectively. High-iron content advanced breeding lines and germplams of pearl millet have been recently identified, however, their high content in IP6 and polyphenols inhibits iron absorption (Tako et al., 2015, Nutr J, 14:11). Iodine is another essential micronutrient. Its deficiency is also prevalent in developing countries and is frequently the casue of goitre formation. Pearl millet consumption is considered one of the factors responsible for the high incidence of goitre in rural populations. C-glucosylflavones related to vitexin, glucosyl vitexin and glucosyl orientin are supposed to be responsible of the goitrogen effects (Gaitan et al., 1995, J Clin Endocrinol Metabol 80:1144-7). The finding that yellow coloured pearl millet is less goitrogenic than brown or grey millet is a good indication that a strategy based on the reduction or suppression of the accumulation of C-glucosylflavone(s) could be effective to reduce the goitrogenic effects associated to pearl millet consumption. The aim of the NewPearl project is to provide tools and knowledge to accelerate the breeding of new pearl millet varieties with increased nutritional qualities, as well as with better adaptation to environmental stresses. With the aim to make association studies by integreting phenotypic and GBS data, we will evaluate the natural genetic variability of the levels of IP6, CGFs and essential minerals (such as Fe and Zn) in pearl millet seeds. A total of 153 inbred lines derived from Western and Central African landraces, provided by ICRISAT, is currently under screening to quantify the content of such compounds in flour extracts. Data on the genetic variability of these traits will be presented. A further objective of this project is the identification in the pearl millet genome of relevant genes involved in IP6 and CGFs synthesis. Their expression will be investigated in the more contrasting lines at different seed developmental stages. Moreover, as future forecasted activity, a pearl millet TILLING population will be screened to search for low phytic acid (lpa) mutants, a strategy to reduce the antinutrient effects of IP6 which showed successful in other crops. This work is supported by CERES-NewPearl project, jointly funded by Fondazione Cariplo and Agropolis Fondation.