The objective of the present work was to evaluate in vitro growth of cassava accessions under different temperatures to improve protocols for germplasm conservation. Acclimatized chambers were tested at 18 °C, 20 °C and 22 °C, and again at 22 °C in conservation room (control) in five accessions: Mandioca João Grande (BGM1325), Fécula Branca (BGM1632), BRS Kiriris (BGM1728), S/N (121) (BGM2004) and BRS Prata (BGM2019). After 90 days of distance, the following were evaluated: shoot height (cm), number of sprouts and microcuttings, number of green and senescent leaves, fresh and dry weight of shoots and roots (g). The experiment was performed under a completely randomized design in factorial scheme 5(accessions) x 4(temperatures), with 20 replicates. All data were submitted to the F test ANOVA and evaluated by the Tukey and Scott-Knott tests at 5% probability. The control temperature promoted lower mean values for plant shoot height, with emphasis for accessions BGM1325, BGM1632 and BGM2019. In a general manner accessions BGM1325, BGM1632 and BGM2019 showed lower SH, with the first and BGM2004 outstanding with the best results for lower NSL and higher NGL.