Sperm competition commonly occurs whenever females mate multiply, leading to variation in male paternity success. This can be due to variation in the various traits that might affect sperm competitive ability, which itself depends on both genetic and environmental factors, as well as on genotype-by-environment interactions (GEI). Seminal fluid is a major component of the male ejaculate that is often expected to mediate sperm competition, where different genotypes can differ in their seminal fluid expression as a response to different level of sperm competition (i.e., exhibit GEI). We therefore here focussed on testing for GEI in expression of two recently identified seminal fluid transcripts, suckless-1 and suckless-2, which potentially modulate sperm competitive ability in the simultaneously hermaphroditic flatworm Macrostomum lignano via their effects on manipulating post-mating partner behaviour and ultimately the fate of transferred ejaculates. In addition, we sought to test for GEI in sperm competitive ability in a standardised sperm competition (P₁ & P₂) assay, to investigate the relationship between natural variation in the expression of these seminal fluid transcripts generated through GEI and relative paternity success. We found GEI for the expression level of suckless-1 and suckless-2, as well as for sperm competitive ability. Moreover, we found a positive relation between the expression of suckless-1 and relative paternity success (P₁). This suggests that natural variation in the expression of this seminal fluid transcript indeed can influence sperm competition outcomes in M. lignano.