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104 reviewsAbstract22 Carbohydrate partitioning from photosynthetic sources to non-photosynthetic sinks is 23 essential for plant development and crop yield. Using a maize-teosinte BC2S324 population, we identify Chlorotic Leaf Spot1 (CLS1), a fumarylacetoacetate hydrolase 25 (FAH) in the tyrosine degradation pathway that plays an essential role in carbohydrate 26 partitioning in maize. CLS1 localizes to the plasma membrane, cytoplasm, and 27 nucleus. Allelic tests and sequence analysis reveal that the teosinte parent 28 CIMMYT8759 carries a weak allele of CLS1, likely due to rare amino acid 29 substitutions at residues 175 and 355. Loss-of-function mutants of CLS1 develop30 chlorotic leaf spots accompanied by carbohydrate hyperaccumulation, reduced 31 photosynthetic efficiency, chloroplast damage, and impaired transient starch 32 conversion. Critically, cls1 mutants exhibit ectopic callose accumulation and aberrant 33 plasmodesmata ultrastructure at the mesophyll-bundle sheath and bundle 34 sheath-vascular parenchyma interfaces. This defect causes starch granule and soluble 35 sugar accumulation in chlorotic leaf tissues, indicating a disruption of the symplastic 36 transport pathway. Collectively, our results uncover an important role for FAH in 37 plant development and identify CLS1 as a key regulator of symplastic carbohydrate 38 partitioning.39 Keywords: Maize; Fumarylacetoacetate hydrolase; Carbohydrate partitioning; 40 Plasmodesmata; Tyrosine degradation 41