Global relative species loss due to first‐generation biofuel production for the transport sector

The global demand for biofuels in the transport sector may lead to significant biodiversity impacts via multiple human pressures. Biodiversity assessments of biofuels, however, seldom simultaneously address several impact pathways, which can lead to biased comparisons with fossil fuels. The goal of the present study was to quantify the direct influence of habitat loss, water consumption and greenhouse gas (GHG) emissions on potential global species richness loss due to the current production of first‐generation biodiesel from soybean and rapeseed and bioethanol from sugarcane and corn. We found that the global relative species loss due to biofuel production exceeded that of fossil petrol and diesel production in more than 90% of the locations considered. Habitat loss was the dominating stressor with Chinese corn, Brazilian soybean and Brazilian sugarcane having a particularly large biodiversity impact. Spatial variation within countries was high, with 90th percentiles differing by a factor of 9 to 22 between locations. We conclude that displacing fossil fuels with first‐generation biofuels will likely negatively affect global biodiversity, no matter which feedstock is used or where it is produced. Environmental policy may therefore focus on the introduction of other renewable options in the transport sector.     

Highly efficient multiplex editing: one-shot generation of 8× Nicotiana benthamiana and 12× Arabidopsis mutants

Genome editing by RNA-guided nucleases, such as SpCas9, has been used in numerous different plant species. However, to what extent multiple independent loci can be targeted simultaneously by multiplexing has not been well documented. Here, we developed a toolkit, based on a highly intron-optimized zCas9i gene, which allows assembly of nuclease constructs expressing up to 32 single guide RNAs (sgRNAs). We used this toolkit to explore the limits of multiplexing in two major model species, and report on the isolation of transgene-free octuple (8×) Nicotiana benthamiana and duodecuple (12×) Arabidopsis thaliana mutant lines in a single generation (T₁ and T₂, respectively). We developed novel counter-selection markers for N. benthamiana, most importantly Sl-FAST2, comparable to the well-established Arabidopsis seed fluorescence marker, and FCY-UPP, based on the production of toxic 5-fluorouracil in the presence of a precursor. Targeting eight genes with an array of nine different sgRNAs and relying on FCY-UPP for selection of non-transgenic T₁, we identified N. benthamiana mutant lines with astonishingly high efficiencies: All analyzed plants carried mutations in all genes (approximately 112/116 target sites edited). Furthermore, we targeted 12 genes by an array of 24 sgRNAs in A. thaliana. Efficiency was significantly lower in A. thaliana, and our results indicate Cas9 availability is the limiting factor in such higher-order multiplexing applications. We identified a duodecuple mutant line by a combination of phenotypic screening and amplicon sequencing. The resources and results presented provide new perspectives for how multiplexing can be used to generate complex genotypes or to functionally interrogate groups of candidate genes.     

DieAgavaceae HUTCHINSONs im Lichte ihrer Embryologie,ihres Gynözeum-, Staubblatt- und Blattbaues

Ohne Zusammenfassung     

Ein künstlich bestäubter Fruchtknoten vonYucca filamentosa

Ohne Zusammenfassung