Host-specific proteomic and growth analysis of maize and tomato seedlings inoculated with Azospirillum brasilense Sp7.

Azospirillum brasilense Sp7 (Sp7) is a diazotrophic, free-living plant growth-promoting rhizobacterium (PGPR) that is increasingly used for its ability to reduce stress and improve nutrient uptake by plants. To test the hypothesis that Sp7 interacts differently with the primary metabolism in C3 and C4 plants, differential proteomics were employed to study weekly protein expression in Sp7-treated maize (Zea mays cv. B73) and tomato (Solanum lycopersicum cv. Boludo) seedlings. Plant and root growth parameters were also monitored. Protein changes were most striking at the four-leaf stage (T1) for both species. Proteins related to metabolism and redox homeostasis were most abundant in tomato at T1, but later, plants experienced inhibited Calvin-Benson (CB) cycle and chloroplast development, indicating that photosynthetic proteins were damaged by reactive oxygen species (ROS). In maize, Sp7 first increased ROS-scavenging enzymes and decreased those related to metabolism, which ultimately reduced photoinhibition at later sampling times. Overall, the early interaction with maize is more complex and beneficial because the photosynthetic aparatus is protected by the C4 mechanism, thereby improving the interaction of the PGPR with maize. Better seedling emergence and vigor were observed in inoculated maize compared to tomato. This study provides an integrated perspective on the Sp7 strain-specific interactions with young C3 and C4 plants to modulate primary metabolism and photosynthesis.

Patterns of CRISPR/Cas9 activity in plants, animals and microbes.

The CRISPR/Cas9 system and related RNA-guided endonucleases can introduce double-strand breaks (DSBs) at specific sites in the genome, allowing the generation of targeted mutations in one or more genes as well as more complex genomic rearrangements. Modifications of the canonical CRISPR/Cas9 system from Streptococcus pyogenes and the introduction of related systems from other bacteria have increased the diversity of genomic sites that can be targeted, providing greater control over the resolution of DSBs, the targeting efficiency (frequency of on-target mutations), the targeting accuracy (likelihood of off-target mutations) and the type of mutations that are induced. Although much is now known about the principles of CRISPR/Cas9 genome editing, the likelihood of different outcomes is species-dependent and there have been few comparative studies looking at the basis of such diversity. Here we critically analyse the activity of CRISPR/Cas9 and related systems in different plant species and compare the outcomes in animals and microbes to draw broad conclusions about the design principles required for effective genome editing in different organisms. These principles will be important for the commercial development of crops, farm animals, animal disease models and novel microbial strains using CRISPR/Cas9 and other genome-editing tools.