Dynamics of transcription is affected by oxygen tension and developmental speed during in vitro production of bovine embryos.

This study examines the impact of oxygen tension and embryo kinetics on gene transcription dynamics in pathways crucial for embryonic preimplantation development, including lipid metabolism, carbohydrate transport and metabolism, mitochondrial function, stress response, apoptosis and transcription regulation. Bovine embryos were generated in vitro and allocated into two groups based on oxygen tension (20% or 5%) at 18 h post insemination (hpi). At 40 hpi, embryos were categorized into Fast (≥4 cells) or Slow (2 cells) groups, resulting in four experimental groups: FCL20, FCL5, SCL20 and SCL5. Embryo collection also occurred at 72 hpi (16-cell stage; groups FMO20, FMO5, SMO20 and SMO5) and at 168 hpi (expanded blastocyst (BL) stage; groups FBL20, FBL5, SBL20 and SBL5). Pools of three embryos per group were analysed in four replicates using inventoried TaqMan assays specific for Bos taurus, targeting 93 genes. Gene expression patterns were analysed using the K-means algorithm, revealing three main clusters: genes with low relative abundance at the cleavage (CL) and 16-cell morula (MO) stages but increased at the BL stage (cluster 1); genes with higher abundances at CL but decreasing at MO and BL (cluster 2); and genes with low levels at CL, higher levels at MO and decreased levels at BL (cluster 3). Within each cluster, genes related to epigenetic mechanisms, cell differentiation events and glucose metabolism were particularly influenced by differences in developmental kinetics and oxygen tension. Fast-developing embryos, particularly those cultured under low oxygen tension, exhibited transcript dynamics more closely resembling that reported in vivo-produced embryos.

Streptomyces brasiliscabiei, a new species causing potato scab in south Brazil.

This study aimed to characterize six Streptomyces strains associated with potato scab in south Brazil through polyphasic taxonomy involving morphology, pathogenicity and genetic features. These strains were compared with other potato-scab Streptomyces species mainly S. europaeiscabiei, S. scabiei and S. stelliscabiei. South-Brazilian Streptomyces strains were morphologically distinct from the type strains of S. scabiei (CFBP 4517T) and their genomospecies S. europaeiscabiei (CFBP 4497 T) and S. stelliscabiei (CFBP 4521T), producing a brown substrate mycelium with red borders and cream-grey brown aerial spores. Red-brown diffusible pigment on YME was also observed. The carbon sources L-Arabinose, D-Fructose, D-Glucose, D-Mannitol, meso-Inositol, Raffinose, Rhamnose, Sucrose, D-Xylose were tested for these strains. All strains were pathogenic causing symptoms of necrosis on radish and several potato cultivars commonly used in potato growing areas in Brazil. In greenhouse conditions, the strains caused scab disease and produced deep-pitted lesions covering large areas of the tuber. These results were correlated with presence of pathogenicity marker genes (txtAB, tomA or nec1) detected by PCR amplifications. In both phylogenetic analyses, 16S rRNA and MLSA, Streptomyces sp. Brazilian strains were closely related to S. europaeiscabiei, S. scabiei and S. stelliscabiei species, but they were allocated in separated branches supported by high bootstrap values and/or with low sequence similarity values. Sequencing of whole genome showed an 10,846,379 bp linear chromosome with high GC content (71.3%) consisting of 9179 putative genes, 3 rRNAs, 89 tRNAs and 1 CRISPRS. The molecular data, including genomic features, associated with morphological, biochemical and pathogenic characteristics warrant that the six Streptomyces Brazilian strains represent a new species associated with potato scab in Brazil, which would be named Streptomyces brasiliscabiei with IBSBF 2867T as the type strain.

The potential of genome-wide RAD sequences for resolving rapid radiations: a case study in Cactaceae.

The reconstruction of relationships within recently radiated groups is challenging even when massive amounts of sequencing data are available. The use of restriction site-associated DNA sequencing (RAD-Seq) to this end is promising. Here, we assessed the performance of RAD-Seq to infer the species-level phylogeny of the rapidly radiating genus Cereus (Cactaceae). To examine how the amount of genomic data affects resolution in this group, we used datasets and implemented different analyses. We sampled 52 individuals of Cereus, representing 18 of the 25 species currently recognized, plus members of the closely allied genera Cipocereus and Praecereus, and other 11 Cactaceae genera as outgroups. Three scenarios of permissiveness to missing data were carried out in iPyRAD, assembling datasets with 30% (333 loci), 45% (1440 loci), and 70% (6141 loci) of missing data. For each dataset, Maximum Likelihood (ML) trees were generated using two supermatrices, i.e., only SNPs and SNPs plus invariant sites. Accuracy and resolution were improved when the dataset with the highest number of loci was used (6141 loci), despite the high percentage of missing data included (70%). Coalescent trees estimated using SVDQuartets and ASTRAL are similar to those obtained by the ML reconstructions. Overall, we reconstruct a well-supported phylogeny of Cereus, which is resolved as monophyletic and composed of four main clades with high support in their internal relationships. Our findings also provide insights into the impact of missing data for phylogeny reconstruction using RAD loci.