Abstract: In order to investigate the variation and influencing factors of phytolith morphology in culm sheath of Phyllostachys aureosulcata from different cultivation sites, phytoliths collected in Huzhou of Zhejiang, Chengdu of Sichuan, and Chishui of Guizhou were extracted, and differences in phytolith content, concentration, area, and proportion were analyzed. The relationship between phytolith morphology and annual average temperature, precipitation, light, and soil was analyzed. The results showed that the phytolith morphotypes in Ph. aureosulcata culm sheaths from 3 cultivation sites were consistent, with a total of 28 morphotypes observed. The highest proportion of phytolith morphotypes was rondels, saddle, linear, and acute, while the number of other phytolith morphotypes was relatively small. Phytolith content in Ph. aureosulcata culm sheath was increased in the order of Chishui of Guizhou > Chengdu of Sichuan > Huzhou of Zhejiang. In contrast, phytolith concentration and area exhibited the opposite trend: Huzhou of Zhejiang> Chengdu of Sichuan> Chishui of Guizhou. The area and concentration of phytoliths were significantly positively correlated with the annual average annual temperature and light, and negatively correlated with precipitation. Phytolith content was significantly positively correlated with annual precipitation, and negatively correlated with average yearly temperature and light. The contents of total potassium, total magnesium, and total silicon in soil were significantly positively correlated with phytolith content in culm sheath. This study provided a theoretical basis for enriching the phytolith morphology of gramineous bamboos and explored the relationship between phytoliths and environmental factors.