Chloroplasts were purified by centrifugation in Percoll gradient from low temperature (4 C), control (22 C) and high temperature (30 C) grown Malus x domestica microshoots. The proteins were digested using filter aided sample processing (FASP) and analyzed using nano-LC MS/MS. Constant scan rate of 2 Hz and range from 100 to 2,200 m/z was used for the MS spectra acquisition. Data-dependent auto-MS/MS method (Compass 1.5 acquisition and processing software, Bruker Daltonics) was used for selecting the 10 most abundant precursor ions for fragmentation. The MS/MS summation time was adjusted based on precursor intensity within the range 1-10 Hz. Active exclusion of already analyzed precursors was used with a release time of 30 s. Precursor ion isolation window and the CID collision energy were adjusted between 3-9 Th and 34-55 eV as a function of the m/z value, respectively. The data files (9 low temperature samples, 12 control samples and 9 high temperature sampes) are described in the "metadata.txt" file.