Declining temperature and low light often appear together to affect cotton (Gossypium hirsutum L.) growth and development. To investigate the interaction on fibre elongation, two cultivars were grown in fields in 2010 and 2011 and in pots in 2011 under three shading levels for three planting dates, and the differences of environmental conditions between different planting dates were primarily on temperature. Fibre length in the late planting date 25 May was the longest instead of the normal planting date. Late planting prolonged fibre elongation period and the effect of late planting on fibre length formation was greater than low light. In the normal planting date, shading increased fibre length through delaying the peak of b-1,3-glucanase gene expression and bringing the peak of b-1,3-glucan synthase gene expression forward, leading to a longer duration of plasmodesmata(PD) closure to increase fibre length, instead of changing sucrose contents or relate enzyme activities. However, in the late planting dates, the difference of the duration of PD closure between shading treatments was not obvious, but low light had a negative impact on sucrose contents, sucrose synthase (SuSy) and vacuolar invertase(VIN) activities during fibre rapid elongation period, leading to the decline of fibre length. Due to late planting and low light, the decreased extent of fibre length of Sumian 15 was larger than Kemian 1. Under the combined condition, Sumian 15 had a shorter gene expression of Expansin, and more sensitive sucrose content, VIN and SuSy activity during fibre rapid elongation period. This resulted in the length formation of Sumian 15 which was more sensitive than Kemian 1, when the cotton suffered the combined effects.