Salt stress is the most important abiotic stress limiting crop yield globally, and breeders are encouraged to test novel genotypes for resistance to this selection agent. However, getting a sufficient number of plants for the evaluation can be a lengthy and time-consuming procedure that might be shortened by applying in vitro techniques. An experiment was conducted under controlled saline circumstances using a tissue culture technique to investigate the influence of salt on several physic-biochemical parameters. Micro-shoots of Volkamariana lemon, sour orange, trifoliate orange, and Cleopatra mandarin rootstocks were re-cultured on MS media supplemented with NaCl from 25, up to 150 mM. Following the experiments, all of the tested rootstocks' morphological and biochemical characteristics, stomata behavior, and element accumulation were measured. As a result, adding NaCl from 25 to 150 mM to MS media caused a reduction in All morphogenetic parameters were measured and compared to the control, including shoot number, shoot length, leaf number, and survival percentage. Photosynthesis pigment, RWC, K, and Ca ions were decreased by increasing NaCl from 100 to 150 mM. On the contrary, proline, Cl, and Na were increased. Cleopatra mandarin and Trifoliate orange are more tolerant to salt stress than Volkamariana lemon and sour orange citrus.