Maize grain yield is drastically reduced by heat stress (HTS) during anthesis and early grain filling. However, the mechanism of HTS in reproductive organs and kernel numbers remains poorly understood. From 2018 to 2020, two maize varieties (ND372, heat tolerant; and XY335, heat sensitive) and two temperature regimens (HTS, heat stress; and CK, natural control) were evaluated, resulting in four treatments (372CK, 372HTS, 335CK, and 335HTS). HTS was applied from the nine-leaf stage (V9) to the anthesis stage. Various morphological traits and physiological activities of the tassels, anthers, and pollen from the two varieties were evaluated to determine their correlation with kernel count. The results showed that HTS reduced the number of florets, tassel volume, and tassel length, but increased the number of tassel branches. HTS accelerates tassel degradation and reduces pollen weight, quantity, and viability. Deformation and reduction in length and volume due to HTS were observed in both the Nongda 372 (ND372) and Xianyu 335 (XY335) varieties, with the average reductions being 22.9% and 35.2%, respectively. The morphology of the anthers changed more conspicuously in XY335 maize. The number of kernels per spike was reduced in the HTS group compared with the CK group, with the ND372 and XY335 varieties showing reductions of 47.3% and 59.3%, respectively. The main factors underlying the decrease in yield caused by HTS were reductions in pollen quantity and weight, tassel rachis, and branch length. HTS had a greater effect on the anther shape, pollen viability, and phenotype of XY335 than on those of ND372. HTS had a greater impact on anther morphology, pollen viability, and the phenotype of XY335 but had no influence on the appearance or dissemination of pollen from tassel.