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Natural variation in the ORL4 promoter regulates rice leaf rolling to modulate leaf architecture and yield
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DOI：10.1093/plcell/koag086.png)
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Abstract
En 中文
Rice (Oryza sativa L.) leaf rolling enhances canopy architecture and photosynthetic efficiency, thereby improving yield potential. Through genome-wide association studies (GWAS) of 802 Xian and Geng rice accessions, we identified 32 loci associated with the leaf rolling index (LRI) and characterized OUTWARD ROLLED LEAF 4 (ORL4) as a regulatory gene from natural germplasm. CRISPR/Cas9-mediated knockout of ORL4 resulted in abaxially rolled leaves, driven by excessive bulliform cell proliferation and disrupted vascular patterning. A promoter variant (Chr04_19562737-C/T) modulates ORL4 expression by altering the binding affinity of the transcription factor RICE OUTERMOST CELL-SPECIFIC 8 (ROC8). Carriers of the ORL4C allele exhibit stronger ROC8 binding, higher ORL4 expression, and adaxially rolled or flat leaves; ORL4T carriers show the opposite phenotype. ORL4 forms a ternary complex with ROC8 and TOPLESS-RELATED PROTEIN 2(TPL2) to suppress ABAXIALLY CURLED LEAF 1(ACL1), a positive regulator of bulliform cell differentiation. Evolutionary analyses indicated that the ORL4C and ORL4T alleles originated from Oryza nivara and Oryza rufipogon, with ORL4C favored in Geng and ORL4T in Xian during domestication. Promoter editing of ORL4 in the elite cultivar Shennong 89-366 generated lines with enhanced LRI, achieving a significant yield increase in field trials. This study elucidates the genetic and molecular basis of leaf rolling and provides a promising strategy for rice yield improvement via precise promoter editing. ORL4, a genome-wide association studies-identified regulator of bulliform cells and leaf rolling, is fine-tuned by a promoter single nucleotide polymorphism that alters ROC8 binding and forms a complex with ROC8 and TPL2 to suppress ACL1 expression.
Keywords:
TRANSCRIPTION FACTOR
GENE-EXPRESSION
ORYZA-SATIVA
PROTEIN
OVEREXPRESSION
TRANSFORMATION
AGROBACTERIUM
ENCODES
VARIANT
CELLS
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