Tocopherols are a class of four natural compounds that can provide nutrition and function as antioxidant in both plants and animals.Maize kernels have low α-tocopherol content,the compound with the highest vitamin E activity,thus,raising the risk of vitamin E deficiency in human populations relying on maize as their primary vitamin E source.In this study,two insertion/deletions (InDels) within a gene encoding γ-tocopherol methyltransferase,Zea mays VTE4 (ZmVTE4),and a single nucleotide polymorphism (SNP) located ~85 kb upstream of ZmVTE4 were identified to be significantly associated with α-tocopherol levels in maize kernels by conducting an association study with a panel of~500 diverse inbred lines.Linkage analysis in three populations that segregated at either one of these three polymorphisms but not at the other two suggested that the three polymorphisms can affect α-tocopherol content independently.Furthermore,we found that haplotypes of thetwo InDels could explain ~33% of α-tocopherol variation in the association panel,suggesting ZmVTE4 is a major gene involved in natural phenotypic variation of α-tocopherol.One of the two InDels is located within the promoter region and associates with ZmVTE4 transcript level.This information can not only help in understanding the underlying mechanism of natural tocopherol variations in maize kernels,but also provide valuable markers for marker-assisted breeding of α-tocopherol content in maize kernels,which will then facilitate the improvement of maize as a better source of daily vitamin E nutrition.