The effects of an animal RNase fused to the late cotton pollen-specific promoter G9 in a plant system were investigated. Expression of the chimeric genes G9-uidA and G9-RNase in tobacco plants showed that the 1.2-kb promoter fragment of the G9 gene was sufficient to maintain tissue and temporal specificity in a heterologous system. GUS (beta-glucuronidase) expression was detected only in pollen from anther stage 6 through anthesis, with maximal GUS activity in pollen from stage 10 anthers. Investigating the effects of the rat RNase on pollen viability at stage 10, we found that pollen viability was reduced from 79 to 8% and from 89 to 40%, in pollen germination and fluoresceine diacetate assays, respectively, in one G9-RNase transgenic line, suggesting a lethal effect of the RNase gene. This indicates that the rat RNase produces deleterious effects in this plant system and may be useful for engineering male sterility.
