We previously previously reported that G protein alpha subunit 1 (GPA1) is essential for sexual reproduction in the homothallic ascomycete fungus Gibberella zeae. In this study we performed microarray analyses on a GPA1 deletion mutant of G. zeae (Delta gpa1) to identify genes involved in the sexual reproduction of this fungus. In the Delta gpa1 strain, 645 genes were down-regulated and 550 genes were up-regulated during sexual reproduction when compared to the wild-type strain. One hundred of the down-regulated genes were selected for further investigation based on orthologous group clusters and differences in transcript levels. Quantitative real time-PCR was used to determine transcriptional profiles of these genes at various sexual and vegetative stages. We observed that transcript levels of 78 of these genes were dramatically increased in the wild-type strain during sexual reproduction compared to levels observed during vegetative growth, and were down-regulated in Delta gpa1 compared to the wild-type strain. We deleted 57 of these genes and found that four of the deletion mutants lost self-fertility and five produced fewer perithecia compared to the wild-type strain. Two mutants produced wild-type numbers of perithecia, but maturation of perithecia and ascospores was delayed. In all we identified 11 genes that are involved in sexual reproduction of G. zeae and present evidence that some of these genes function at distinct stages during sexual reproduction in the fungus. (C) 2010 Elsevier Inc. All rights reserved.