Molecular genetic diversity of ICARDA’s worldwide barley landrace collection

The use of barley grain as a whole grain food or food ingredient through the development of various products from improved varieties has increased sharply in developed countries and is linked to health claims. Barley landraces are preferred for making traditional dishes and are conserved in genebanks worldwide. International barley landraces from the collection at ICARDA were assessed for food quality traits and the genetic diversity was investigated using twenty genic- and genomic SSR markers. SNP in several candidate genes for these traits were also assessed. This molecular analysis of food quality traits should allow accelerated use of the germplasm in barley landraces in the development of superior barley varieties for human food use

Two-Dimensional Isoelectric Focusing OFFGEL, Micro-Fluidic Lab-on-Chip Electrophoresis and FTIR for Assessment of Long-Term Stability of rhG-CSF Formulation

Recombinant human granulocyte colony-stimulating factor (rhG-CSF) has been increasingly recognized from among one of the most abundant families of biosimilars. Upon long-term storage, the rhG-CSF is subject to subtle chemical modifications that rapidly occur and, in particular, produce deaminated variants with divergent charge. Indeed, changes in charge from glutamine deamination may alter the way rhG-SCF will refold and the structure of resulting molecule. To assess this charge heterogeneity, 2-D gel electrophoresis has limited application. Recent micro-fluidic-based technical advances offer a great alternative method to better control liquid volumes on a minute scale. Here, we used IEF OFFGEL-lab-on-chip electrophoresis for 2-D separation of the rhG-CSF peptides according to their isoelectric point (pI) and molecular weight (kDa). We used an rhG-CSF commercial therapeutic formulation, kept refrigerated 24 months after expiry. The samples were analyzed for particulate matter and charge variants. Subsequently, the secondary structure was assessed by FTIR spectroscopy and residual biological activity was recorded. Interestingly, we showed an additional band in the acidic gel area above and below the most intense protein band (fractions 10, 11, and 12 at 22.84s). This observation reveals the presence of the rhG-CSF variant charges without any additional high molecular weight impurity or biological activity decrease. We conclude that after two years of storage, the rhG-CSF solution maintained its native secondary structure with little beta-sheet deviation, as reflected in the 1622 cm(-1) and 1695 cm(-1). These data demonstrated that a combined strategy is a more suitable and accurate analytical assessment of the rhG-CSF and recombinant protein-based biosimilars

Genetic diversity of ICARDA's worldwide barley landrace collection

Twenty genic- and genomic SSR markers were used to study genetic diversity and geographical differentiation of barley from 29 countries through analysis of a worldwide collection of 304 ICARDA's barley landraces. Of these, 19 loci were highly polymorphic in the material studied. Based on Nei-distance matrix, Principal Component Analysis (PCoA) and cluster analysis using UPGMA associated with AMOVA the data revealed countries' grouping within regions. Three distinct germplasm pools were identified in the landraces. The first of these was from Eastern Africa (Eritrea and Ethiopia) and South America (Ecuador, Peru and Chile) suggesting that barley introduced to South America might have originated specifically from East Africa or that they share a common genetic basis for adaptation. The second was the Caucasus (Armenia and Georgia) and the third included the remaining regions of Central Asia, Near East, Northern Africa and Eastern Asia. Genetic diversity of barley subspecies (Six-rowed barley, Two-rowed barley, H. spontaneum C. Koch and H. agriocrithon A...berg) also discriminates them into three groups: cultivated barleys (Six-rowed barley and Two-rowed barley), wild barley H. spontaneum and subspecies H. agriocrithon. These results associated with parsimony analysis demonstrate that H. agriocrithon and H. spontaneum might be distinct and do not support a hybrid origin for H. agriocrithon suggesting further investigation of the basis of more intense sampling of the two subspecies H. spontaneum and H. agriocrithon