The phenotype identification， microscopic observation of endosperm cells， determining the content of storage substance in endosperm， and map-based cloning of a defective maize kernel mutant named as empty pericarp35 （emp35） were conducted to study its physiological function in the development of maize kernel and map the Emp35 gene. The results showed that the development of emp35 kernels was slow and significantly smaller than that of the wild-type kernels during the same period of development， resulted in the shrunken and empty pericarp kernels. The development of embryo and endosperm in the emp35 was severely delayed， and the mitochondrial structure in endosperm cells was abnormal. The accumulation of starch and protein in the emp35 was reduced. The seeds in the F₂ population segregated in a ratio of 3 wild type to 1 empty pericarp， indicating that the defective kernel phenotype is controlled by a single recessive nuclear gene. The Emp35 gene was mapped at the interval between 127.90 Mb and 163.36 Mb on the chromosome 8 of maize with a bulked segregant analysis （BSA）. Four InDel markers were developed within this interval， and Emp35 gene was finely mapped at the interval between 139 571 117-146 176 858.