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Grain size and shape are important factors in determining the grain yield. In this study, OsNF-YC10, a member of the NF-Y transcription factor family encoding a putative histone transcription factor, was cloned and characterized. qRT-PCR and mRNA in situ hybridization analysis revealed that OsNF-YC10 was highly expressed in endosperm and spikelet hull at late developmental stages. The results showed that OsNF-YC10 was a nuclear protein showing transcription activation activity. The osnf-yc10 lines, produced using CRISPR/Cas9 technology, showed narrow, thin and light grains. Cytological experiments revealed significantly reduced cell number of spikelet hull in osnf-yc10 lines compared with that in WT. Narrow, thin, and light grains were found consistently in OsNF-YC10 RNAi transgenic lines. Moreover, the number of cells decreased in the grain-width direction than WT. These results indicated that OsNF-YC10 plays an important role in determining grain size and shape. OsNF-YC10 was further revealed to influence the expression of GW8 (a positive regulator of grain width), GW7 (a negative regulator of grain width) and cell cycle-regulated genes CYCD4, CYCA2.1, CYCB2.1, CYCB2.2, E2F2. Taken together, it is suggested that OsNF-YC10 regulates the grains size and shape by influencing the cell proliferation of spikelet hulls.

High temperature often leads to the failure of grain filling in rice (Oryza sativa) to cause yield loss, while the mechanism is not well elucidated yet. Here, we report that two seed-specific NAM/ATAF/CUC domain transcription factors, ONAC127 and ONAC129, are responsive to heat stress and involved in the grain filling process of rice. ONAC127 and ONAC129 are dominantly expressed in the pericarp and can form a heterodimer during rice grain filling. CRISPR/Cas9 induced mutants and overexpression lines were then generated to investigate the functions of these two transcription factors. Interestingly, both knock-out and overexpression plants showed incomplete grain filling and shrunken grains, which became more severe under heat stress. Transcriptome analysis revealed that ONAC127 and ONAC129 mainly regulate stimulus response and nutrient transport. ChIP-seq analysis identified that the direct targets of ONAC127 and ONAC129 in developing rice seeds include monosaccharide transporter OsMST6, sugar transporter OsSWEET4, calmodulin-like protein OsMSR2 and AP2/ERF factor OsEATB. These results suggest that ONAC127 and ONAC129 may regulate grain filling through affecting sugar transportation and abiotic stress responses. Overall, this study demonstrates a transcriptional regulatory network involving ONAC127 and ONAC129 and coordinating multiple pathways to modulate seed development and heat stress response at rice reproductive stage.HighlightA NAC transcription factor heterodimer plays vital roles in heat stress response and sugar transportation at rice grain filling stage.

Nuclear factor Y (NF-Y) is a ubiquitous transcription factor that regulates important physiological and developmental processes. In this study, we identified 34 OsNF-Y genes in rice, including 6 newly identified genes. Expression profile analysis covering the whole life cycle revealed that transcripts of OsNF-Y differentially accumulated in a tissue-specific, preferential or constitutive manner. In addition, gene duplication studies and expression analyses were performed to determine the evolutionary origins of the OsNF-Y gene family. Nine OsNF-Y genes were differentially expressed after treatment of seedlings with one or more abiotic stresses such as drought, salt and cold. Analysis of expression correlation and Gene Ontology annotation suggested that OsNF-Y genes were co-expressed with genes that participated in stress, accumulation of seed storage reserves, and plant development. Co-expression analysis also revealed that OsNF-Y genes might interact with each other, suggesting that NF-Y subunits formed complexes that take part in transcriptional regulation. These results provide useful information for further elucidating the function of the NF-Y family and their regulatory pathways.