Cotton (Gossypium hirsutum L.) is an important crop plant that is susceptible to multiple species of parasitic nematodes including Rotylenchulus reniformis, reniform nematode (RN). There is much emphasis on breeding resistance to RN into commercially viable cotton varieties, and in 2007 LONREN-1 (LR1) and LONREN-2 (LR2), putative RN-resistant germplasm derived from Gossypium longicalyx was released. More recently Implementation of these materials in commercially useful germplasm has proven problematic. A resistant line BARBAREN-713 (B-713) has been developed with RN resistance based on Gossypium barbadense, which shows promise. To facilitate this implementation, the gene regulatory networks involved in RN-resistant and susceptible genotypes are under investigation particularly focusing on miRNAs. Initially, the role of small regulatory RNAs (srRNAs) in these regulatory networks is under investigation. In this study, LR1, LR2, and B-713 were selected as nematode resistant genotypes, and DeltaPine 90 & Suregrow 747 were susceptible lines. Small RNA libraries were prepared from greenhouse grown plants of each of these 4 genotypes either pre- or post-infection with reniform nematodes. RNA-seq was conducted on these small RNA libraries using the ABI Solid platform with single-end sequencing,