WRKY transcription factors play key roles in stress responses, growth, and development. We previously reported on the evolution of WRKYs from unicellular green algae to land plants. To address recent evolution events, we studied three domesticated and eight wild species in the genus Oryza, an ideal model due to its long history of domestication, economic importance, and central role as a model system. We have identified prevalence of Group III WRKYs despite differences in breeding of cultivated and wild species. Same groups of WRKY genes tend to cluster together, suggesting recent, multiple duplication events. Duplications followed by divergence may result in neofunctionalizations of co-expressed WRKY genes that finely tune the regulation of target genes in a same metabolic or response pathway. WRKY genes have undergone recent rearrangements to form novel genes. Group Ib WRKYs, unique to AA genome type Oryza species, are derived from Group III genes dated back to 6.76 million years ago. Gene tree reconciliation analysis with the species tree revealed details of duplication and loss events in the 11 genomes. Selection analysis on single copy orthologs reveals the highly conserved nature of the WRKY domain and clusters of fast evolving sites under strong positive selection pressure. Also, the numbers of single copy orthologs under positive or negative selection almost evenly split. Our results provide valuable insights into the preservation and diversification of an important gene family under strong selective pressure for biotechnological improvements of the world’s most valued food crop.
Rice is the primary staple food for half the world’s population. Climate change challenges and food insecurity supports the need for rice with agronomically advantageous traits. We report on a transposon insertional rice mutant with enhanced germination rates. This trait is advantageous for rice growth in limited water regions and to reduce yield constraints caused by weed and bird competition. Evaluations of vital nutritional components, compositional analysis, and comparative metabolomics on threshed grain samples are performed, as these assays are those used to assess the safety of foods from genetically modified crops. Compared with the wild type (cv. Nipponbare), oswrky71 mutant grains have a similar size, shape, amount of crude fiber, crude fat, and ash content but higher crude protein. Mineral analyses reveal higher contents of phosphorus and zinc but lower calcium, potassium, sodium, and manganese in the mutant. Analysis of B vitamins reveals significantly higher riboflavin concentrations but lower choline chloride, calcium pantothenate, and thiamine. In addition, untargeted metabolomics analyses identify approximately 50 metabolites whose levels differed between the mutant and its wild type. Physical traits and compositional parameters analyzed are mostly similar and within the range or very close to being considered safe for consumption by the International Life Sciences Institute Crop Composition Database. Further agronomic evaluation and cooked rice sensory properties assessment are needed before positioning this mutant for human consumption.
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