With the advent of next-generation Roche 454 pyrosequencing technologies, transcriptome level sequence collections are arising as prominent resources for the discovery of gene-based molecular markers. In a previous study more than 1450 simple sequence repeats (SSRs) in expressed sequence tag (EST) sequences resulting from 454 pyrosequencing of Laodelphax striatellus cDNA were identified. From these we developed PCR primers for 40 di- or tri-candidate SSRs (minimum repeats > 8) from the combined EST library. After extensive optimization, 13 pairs were end labeled with a fluorescent dye. Here, we also tested these 13 SSRs for cross-species amplification in two other planthoppers, Nilaparvata lugens and Sogatella furcifera. The marker transferability was considerably high in N. lugens (92.31%) and in S. furcifera (61.54%). All of the 13 fluorescent SSRs were polymorphic, with allele numbers ranging from two to seven for 24 male and 24 female L. striatellus individuals. Observed heterozygosities ranged from 0.016 to 0.621. Development of SSR markers from ESTs will be a valuable tool to improve our understanding of population structure in this important rice pest.