A revisit the original Plant Physiology classic and trace how one mutant phenotype reshaped modern understanding of plant carbon partitioning, transcriptional regulation, and metabolic engineering.
Abstract
Abstract Plant seeds accumulate triacylglycerol (TAG) as a major storage reserve that supports postgermination growth and seedling establishment. Vegetable oils are also essential for human nutrition and provide renewable feedstocks for industrial and biotechnological applications. In 1998, Focks and Benning published a landmark study in Plant Physiology describing the Arabidopsis WRINKLED1 mutants (wri1), which display a distinctive wrinkled seed phenotype and a dramatic reduction in seed oil accumulation. The conceptual importance of this discovery was not simply the identification of a low-oil mutant, but the demonstration that seed oil accumulation depends on developmental control of carbon flux from carbohydrates into fatty acid precursors. Cloning of the Arabidopsis WRI1 (AtWRI1) gene in 2004 transformed this physiological phenotype into a molecular framework by identifying WRI1 as a member of the APETALA2 (AP2) family of transcription factors that activates late glycolytic and fatty acid biosynthetic genes. Subsequent work uncovered the AW-box cis-element, upstream seed-maturation regulators, WRI1-interacting partners, post-transcriptional and post-translational modification mechanisms controlling WRI1 stability and activity, and the structural basis of WRI1-DNA recognition. These discoveries established WRI1 as a central regulatory node linking seed development, carbohydrate metabolism, and seed oil accumulation. More recent studies have broadened WRI1 biology beyond canonical seed oil biosynthesis to include non-seed oil-storing tissues, hormone and nutrient-associated processes, environmental responses, and structure-guided crop engineering. Here, we revisit the original Plant Physiology classic and trace how one mutant phenotype reshaped modern understanding of plant carbon partitioning, transcriptional regulation, and metabolic engineering
Introduction Triacylglycerols (TAGs) are the main storage lipids in oilseeds, providing energy and carbon during seed germination, and serving as valuable feedstocks for food, feed, and industrial applications. In plants, the terminal acylation of diacylglycerol (DAG) to form TAG is catalyzed by diacylglycerol acyltransferases (DGATs) or phospholipid:diacylglycerol acyltransferase (PDAT). Although sunflower (Helianthus annuus) is one of the world´s leading oilseed crops, the contributions of individual DGAT isoforms to seed TAG biosynthesis remain poorly characterized. Methods We identified six DGAT genes in sunflower —three DGAT1, two DGAT2, and one DGAT3—and examined their genomic organization, phylogenetic relationships, predicted structural features, expression profiles, subcellular localization, in planta functional behaviour and TAG profiles in the TAG deficient yeast strain H1246. Results Gene expression analysis revealed that HaDGAT2A and HaDGAT2D transcripts predominated in developing seeds during the main oil accumulation phase, whereas HaDGAT1A and HaDGAT1B showed lower but detectable expression. Confocal microscopy confirmed the endoplasmic reticulum localization of representative HaDGAT1A and HaDGAT2A isoforms, consistent with their potential involvement in ER-associated TAG assembly. Transient expression in Nicotiana benthamiana leaves revealed functional differences between sunflower DGAT isoforms, HaDGAT1 isoforms were associated with TAG profiles enriched in less unsaturated molecular species, whereas HaDGAT2 isoforms, particularly HaDGAT2D, were associated with increased TAG accumulation and a greater representation of polyunsaturated TAG species under the heterologous conditions tested. Functional complementation in yeast further showed that the analyzed sunflower DGAT isoforms restored TAG synthesis in the H1246 mutant and produced distinct TAG molecular profiles, supporting functional diversification among DGAT1 and DGAT2 isoforms. Discussion These results indicate that sunflower DGAT1 and DGAT2 paralogs contribute differently to TAG accumulation and composition in heterologous systems and identify HaDGAT2A and HaDGAT2D as relevant candidate isoforms for future studies on sunflower seed oil biosynthesis. Direct biochemical assays will be required to confirm the intrinsic substrate specificity of each isoform.
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