Manual removal of the main-stem growth tip is traditionally used to break the apical dominance of cotton ( Gossypium hirsutum L.). Chemical topping with plant growth regulators also effectively inhibits apical dominance. However, the effect of chemical topping on yield increases and whether plant density or ecological conditions affect its efficacy are unclear. Therefore, a three-year field experiment with a split-plot design was conducted to determine the effects of plant topping, plant density, and their interactions on cotton yield and related physiological and agronomical parameters at three sites with different ecological conditions in China. In each site, the main plots were assigned low, moderate, or high plant density and the subplots were assigned no topping, manual topping, or chemical topping. Growth, yield, yield components, earliness, and late-season leaf photosynthesis as well as labor and material inputs were examined each year. Compared with no topping, both chemical and manual topping greatly reduced plant height at all sites. Manual topping increased seed cotton yield and earliness in all tested plant densities and sites. However, plant density but not ecological condition greatly mediated the effect of chemical topping on yield. At low plant density, the yields with chemical topping were 4–6% lower than those with no topping and 5.5–10.8% lower than those with manual topping at the three sites. Although yields with chemical topping were comparable with those of manual topping at moderate and high plant densities, they were 8.6–12.8% higher at moderate density and 13.8–16.4% higher at high plant density than those with no topping across years and sites. Averaged across the sites, chemical topping reduced biological yield by 12.7% at low plant density. Although biological yield decreased slightly, chemical and manual topping increased the harvest index by 12.4% and 13.3% at moderate density and by 15.6% and 17.4% at high density, respectively. In comparison with no topping, the reduction in seed cotton yield with chemical topping at low plant density was attributed to insufficient biological yield, whereas the increase in yield at moderate and high plant densities was mainly due to greater partitioning of assimilates to reproductive tissues. Compared with manual topping, chemical topping produced 23.2% lower net returns as a result of lower seed cotton yield at low plant density but produced 8.1% and 20.9% higher net returns at moderate and high plant densities, respectively, because of savings in labor inputs and comparable seed cotton yields. In addition, chemical topping increased the earliness percentage compared with that of no topping. Overall, this study demonstrates that chemical topping is a promising alternative to traditional manual topping under moderate or high cotton plant density. • Manual topping increased cotton yield regardless of plant densities and ecological conditions relative to non-topping. • Chemical topping decreased yield at low plant density relative to non-topping because of greater biomass reduction. • Chemical topping increased cotton yield at moderate and high plant densities via greater assimilates partitioning to fruits. • Chemical topping could replace manual topping at moderate and high plant densities without ecological dependence.

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