To investigate the genetic diversity of Nephotettix cincticeps across different regions of Hunan Province, we analyzed 10 geographic populations using inter-simple sequence repeats(ISSR) markers. The results showed that the genetic diversity among populations(Ht=0.366 8) was higher than that within the population(Hs=0.247 5), indicating the genetic heterozygosity among populations was lower than that within populations. The coefficient of genetic differentiation(Gst) was 0.325 3, indicating that 32.53% of the total genetic variation was attributed to differences among populations, while 67.47% originated from within populations. The gene flow(Nm) among different geographical populations was 1.036 8, indicating a moderate level of genetic exchange. The high intra-population genetic diversity and moderate gene flow within the Nephotettix cincticeps populations enhanced their adaptive potential, facilitating population continuity under pressures such as geographical isolation and pesticide application. Based on this result, during the control of Nephotettix cincticeps, attention should be paid to the potential risk of rapid evolution of resistance caused by its strong genetic adaptability.