*5.8. Plant Material and Treatments*

In the present study, the germinated seeds of *G. hirsutum* cv. Junmian 1 were grown in plastic pots containing a mixture of soil and vermiculite (3:1). The pots were then placed in an artificial growth chamber for five weeks. The growth conditions were as follows: The temperature was set to 24/16 ºC , the photoperiod was 16/8 h, and the relative humidity was 65%–70%. The two-week-old seedling was exposed to specific treatment, as follows: For heat and cold treatments, seedlings were exposed to 38 ºC and 4 ºC , respectively. For NaCl and drought stress treatments, the seedlings were cultured in a nutrient solution medium with 250 µM and 6000 PEG (*w*/*v*). All treatments were carried out in continuous time intervals of 1, 6, and 12 h, respectively. Additionally, for every specific treatment, five randomly chosen whole seedlings were pooled to form a biological replicate. After that leaf samples were quick-frozen in liquid nitrogen and stored at −80 ◦C for further use.
