Transposable element-mediated DNA methylation of the NAC20 and NAC26 promoters led to a maternal effect on grain filling.

Parent-of-origin effects are usually caused by selective expression of maternal or paternal alleles. Although genome-wide studies suggest that imprinted gene expression occurs primarily in the endosperm in plants, detailed studies of allele-specific gene expression and its associations with parent-of-origin phenotypes are scarce. NAC20 and NAC26 (NAC20/26 hereafter), a pair of tightly linked NAC-family transcription factors, redundantly regulate grain filling and albumin accumulation in rice endosperm. Here, we show that NAC20/26 exhibited allele-specific maternal expression, and the floury endosperm phenotype of the nac20/26 double mutant was inherited with a maternal effect. Further studies showed that the imprinted NAC20/26 expression and floury endosperm phenotype with a maternal effect are associated with insertions of two TEs in NAC20/26 of two Japonica rice varieties, but not in two Indica ones examined. The maternal NAC20/26 expression was associated with elevated DNA methylation in their paternal DMRs, and deletions of those TEs by gene editing led to decreased methylation in these DMRs, and biallelic NAC20/26 expression. Geographical analyses showed that Japonica varieties with high-latitude origins examined carried these TEs. These results establish that TE-mediated DNA methylation lead to grain filling with a maternal effect in high-latitude Japonica rice varieties, which may associate with northward expansion of rice during domestication.