Auxin response factors (ARFs) are involved in auxin-mediated transcriptional regulation in plants. In this study, we performed functional characterization of SlARF6A in tomato. SlARF6A is located in the nucleus and exhibits transcriptional activator activity. Overexpression of SlARF6A increased chlorophyll contents in the fruits and leaves of tomato plants, whereas downregulation of SlARF6A decreased chlorophyll contents compared with those of wild-type (WT) plants. Analysis of chloroplasts using transmission electron microscopy indicated increased sizes of chloroplasts in SlARF6A-overexpressing plants and decreased numbers of chloroplasts in SlARF6A-downregulated plants. Overexpression of SlARF6A increased the photosynthesis rate and accumulation of starch and soluble sugars, whereas knockdown of SlARF6A resulted in opposite phenotypes in tomato leaves and fruits. RNA-sequence analysis showed that regulation of SlARF6A expression altered the expression of genes involved in chlorophyll metabolism, photosynthesis and sugar metabolism. SlARF6A directly bound to the promoters of SlGLK1, CAB, and RbcS genes and positively regulated the expression of these genes. Overexpression of SlARF6A also inhibited fruit ripening and ethylene production, whereas downregulation of SlARF6A increased fruit ripening and ethylene production. SlARF6A directly bound to the SAMS1 promoter and negatively regulated SAMS1 expression. Taken together, these results expand our understanding of ARFs with regard to photosynthesis, sugar accumulation and fruit development and provide a potential target for genetic engineering to improve fruit nutrition in horticulture crops.
SlARF10 plays an important role in the chlorophyll accumulation and photosynthesis of tomato plants, and regulation of its expression affects the starch, fructose, and sucrose content of fruit.
Tomato is a model for studying the climate for fruit development and ripening. Down-regulation of a tomato bell-like homeodomain 4 (SlBL4) resulted in a slightly darker green fruit phenotype and increased accumulation of starch, fructose and glucose. Chlorophyll content analysis and TEM observation confirmed these phenotypes, indicating that SlBL4 was involved in the chlorophyll accumulation and chloroplast formation in tomato. SlBL4-i fruits had noticeably decreased firmness and have larger intercellular spaces and thinner cell walls in the ripened fruits. RNA-Seq had identified differential expression genes involved in chlorophyll metabolism, chloroplast development, cell wall metabolism and carotenoid metabolism. ChIP-seq identified (G/A) GCCCA (A/T/C) and (C/A/T) (C/A/T) AAAAA (G/A/T) (G/A) motifs. SlBL4 directly inhibited protoporphyrinogen oxidase (SlPPO), magnesium chelatase H subunit (SlCHLD), pectinesterase (SlPE) protochlorophyllide reductase (SlPOR), chlorophyll a/b binding protein (SlCAB-3B) and homeobox protein knotted 2 (TKN2) and expressions. In addition, SlBL4 positively regulated squamosa promoter binding protein-like colorless non-ripening (LeSPL-CNR) expression. Our study indicated that SlBL4 was involved in the chlorophyll accumulation, chloroplast development, cell wall metabolism and carotenoids accumulation during tomato fruit ripening. Our data reveal novel evidence for the transcriptional regulation mechanism of BELL mediated fruit growth and ripening.
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