Agave REVEILLE1 regulates the onset and release of seasonal dormancy in Populus.

Deciduous woody plants like poplar (Populus spp.) have seasonal bud dormancy. It has been challenging to simultaneously delay the onset of bud dormancy in the fall and advance bud break in the spring, as bud dormancy, and bud break were thought to be controlled by different genetic factors. Here, we demonstrate that heterologous expression of the REVEILLE1 gene (named AaRVE1) from Agave (Agave americana) not only delays the onset of bud dormancy but also accelerates bud break in poplar in field trials. AaRVE1 heterologous expression increases poplar biomass yield by 166% in the greenhouse. Furthermore, we reveal that heterologous expression of AaRVE1 increases cytokinin contents, represses multiple dormancy-related genes, and up-regulates bud break-related genes, and that AaRVE1 functions as a transcriptional repressor and regulates the activity of the DORMANCY-ASSOCIATED PROTEIN 1 (DRM1) promoter. Our findings demonstrate that AaRVE1 appears to function as a regulator of bud dormancy and bud break, which has important implications for extending the growing season of deciduous trees in frost-free temperate and subtropical regions to increase crop yield.

Agrobacterium- and a single Cas9-sgRNA transcript system-mediated high efficiency gene editing in perennial ryegrass.

Genome editing technologies provide a powerful tool for genetic improvement of perennial ryegrass, an important forage and turfgrass species worldwide. The sole publication for gene editing in perennial ryegrass used gene-gun for plant transformation and a dual promoter based CRISPR/Cas9 system for editing. However, their editing efficiency was low (5.9% or only one gene-edited plant produced). To test the suitability of the maize Ubiquitin 1 (ZmUbi1) promoter in gene editing of perennial ryegrass, we produced ZmUbi1 promoter:RUBY transgenic plants. We observed that ZmUbi1 promoter was active in callus tissue prior to shoot regeneration, suggesting that the promoter is suitable for Cas9 and sgRNA expression in perennial ryegrass for high-efficiency production of bi-allelic mutant plants. We then used the ZmUbi1 promoter for controlling Cas9 and sgRNA expression in perennial ryegrass. A ribozyme cleavage target site between the Cas9 and sgRNA sequences allowed production of functional Cas9 mRNA and sgRNA after transcription. Using Agrobacterium for genetic transformation, we observed a 29% efficiency for editing the PHYTOENE DESATURASE gene in perennial ryegrass. DNA sequencing analyses revealed that most pds plants contained bi-allelic mutations. These results demonstrate that the expression of a single Cas9 and sgRNA transcript unit controlled by the ZmUbi1 promoter provides a highly efficient system for production of bi-allelic mutants of perennial ryegrass and should also be applicable in other related grass species.

Complete Chloroplast Genomes and Comparative Analyses of Three Ornamental Impatiens Species.

Impatiens L., the largest genus in the family Balsaminaceae with approximately 1,000 species, is a controversial genus. Due to the conflict of morphological features and insufficient genomic resources, the studies of systematic evolution and understanding of taxonomic identification are considered to be very limited. Hence, we have sequenced the complete chloroplast genomes of three ornamental species (Impatiens balsamina, I. hawkeri, and I. walleriana), and compared them with previously published wild species data. We performed a detailed comparison of a highly similar basic structure, size, GC content, gene number, order, and functional array among them. Similarly, most divergent genes were detected from previous work in the literature. The mutational regions containing highly variable nucleotide hotspots were identified and may be used as potential markers for species identification and taxonomy. Furthermore, using whole chloroplast genome data to analysis the phylogenetic relationship of the Balsaminaceae species, we found that they were all part of a single clade. The three phenotypically different ornamental species were clustered together, suggesting that they were very likely to be closely related. We achieved and characterized the plastid genome structure, identified the divergence hotspots, and determined the phylogenetic and taxonomic positions of the three cultivated species in the Impatiens genus. The results may show that the chloroplast genome can be used to solve phylogenetic problems in or between the Impatiens genus and also provide genomic resources for the study of the Balsaminaceae family's systematics and evolution.