Production of the Phanerochaete flavido-alba laccase in Aspergillus niger for synthetic dyes decolorization and biotransformation

We investigated the expression of Phanerochaete flavido-alba laccase gene in Aspergillus niger and the physical and biochemical properties of the recombinant enzyme (rLac-LPFA) in order to test it for synthetic dye biotransformation. A. niger was able to produce high levels of active recombinant enzyme (30 mgL^?1), whose identity was further confirmed by immunodetection using Western blot analysis and N-terminal sequencing. Interestingly, rLac-LPFA exhibited an improved stability at pH (2–9) and organic solvents tested. Furthermore, the percentage of decoloration and biotransformation of synthetic textile dyes, Remazol Brilliant Blue R (RBBR) and Acid Red 299 (NY1), was higher than for the native enzyme. Its high production, simple purification, high activity, stability and ability to transform textile dyes make rLac-LPFA a good candidate for industrial applications.     

Identifying Cell Types from Spatially Referenced Single-Cell Expression Datasets

Complex tissues, such as the brain, are composed of multiple different cell types, each of which have distinct and important roles, for example in neural function. Moreover, it has recently been appreciated that the cells that make up these sub-cell types themselves harbour significant cell-to-cell heterogeneity, in particular at the level of gene expression. The ability to study this heterogeneity has been revolutionised by advances in experimental technology, such as Wholemount in Situ Hybridizations (WiSH) and single-cell RNA-sequencing. Consequently, it is now possible to study gene expression levels in thousands of cells from the same tissue type. After generating such data one of the key goals is to cluster the cells into groups that correspond to both known and putatively novel cell types. Whilst many clustering algorithms exist, they are typically unable to incorporate information about the spatial dependence between cells within the tissue under study. When such information exists it provides important insights that should be directly included in the clustering scheme. To this end we have developed a clustering method that uses a Hidden Markov Random Field (HMRF) model to exploit both quantitative measures of expression and spatial information. To accurately reflect the underlying biology, we extend current HMRF approaches by allowing the degree of spatial coherency to differ between clusters. We demonstrate the utility of our method using simulated data before applying it to cluster single cell gene expression data generated by applying WiSH to study expression patterns in the brain of the marine annelid Platynereis dumereilii. Our approach allows known cell types to be identified as well as revealing new, previously unexplored cell types within the brain of this important model system.     

Is nitrogen accumulation in grain legumes responsive to growth or ontogeny?

Nitrogen (N) accumulation in legumes is one of the main determinants of crop yield. Although N accumulation from symbiotic nitrogen fixation or N absorption from the soil has been widely investigated, there is no clear consensus on timing of the beginning of N accumulation and the termination of N accumulation and the physiological events that may be associated with these two events. The analyses conducted in this study aimed at identifying the determinant of N accumulation in two grain legume species. Nitrogen accumulation dynamics and mass accumulation and development stages were recorded in the field for several genotypes of common bean (Phaseolus vulgaris) and faba bean (Vicia faba) under different growing conditions. This study showed that during the vegetative stages, N accumulation rate was correlated with mass accumulation rate. However, the maximum accumulation of N did not correspond to the time of the maximum mass accumulation. In fact, for both species, N accumulation was found to persist in seed growth. This challenges a common hypothesis that seed growth causes a decrease in N accumulation because of a shift of the photosynthate supply to support the seed growth. Even more surprising was the shift of the active accumulation of N in faba bean to late in the growing season as compared with common bean. N accumulation by faba bean only was initiated at high rates very late in vegetative growth and persisted at high rates well into seed fill.