Leaf development, especially in
leafy greens, is crucial for
crop productivity. In various plants, DA1, a pivotal
ubiquitin receptor that negatively regulates leaf growth, and EOD1 (
ENHANCER1 OF DA1), an E3
ubiquitin ligases that interacts with DA1 through its RING domain, influence organ size. In this disclosure, in lettuce (
Lactuca sativa), homologs genes were identified for both DA1 and EOD1, and da1 single mutants, da1 / eod1 double mutants, and DA1 overexpressing lines (DA1ox) were created. The da1 / eod1
double mutant exhibited a striking
phenotype with larger organ size, accelerated growth, and increased
biomass, resulting in higher yields and rapid growth traits pivotal for Controlled Environment
Agriculture (CEA). Additionally, the da1 / eod1
double mutant displayed a pronounced ‘stay-green’
phenotype, characterized by elevated
chlorophyll b levels, which implies extended
shelf life, and enhanced
postharvest quality, both traits indispensable for
leafy vegetables in CEA settings. In contrast, the DA1ox line exhibited early
senescence and diminished organ size. Based on these findings that spotlight the nuanced roles of the DA1-EOD1 module in modulating
plant growth phases and its potential to revolutionize breeding strategies for improved yield and
postharvest longevity in CEA-cultivated
leafy greens, this disclosure provides guide RNAs for
gene editing of DA1 and EOD1, and transformed lettuce having improved organ size and
postharvest quality.