The 45-km-long, E-W to NW-SE-striking Manisa Fault Zone, which constitutes the western section of the Gediz Graben, is characterized by a pure normal sense of motion with a minor strike-slip component. Even though there are numerous historical earthquakes have been listed in Western Anatolia Graben System, a few studies have been addressed on seismic sources of the earthquakes so far. According to existing literature, the western segment of the Manisa Fault Zone is well known characterized; however, the eastern segment of it has rarely been addressed. To decipher the Holocene seismotectonic behavior of the eastern segment of Manisa Fault Zone, trench-based paleoseismological analyses for the first time were performed along with it. To constrain the timing and frequency of past earthquakes, and elapse time from the last activation using the optically stimulated luminescence (OSL) and radiocarbon (14C) dating, we collected twenty-six colluvial and paleosol samples from the trenches. The obtained paleoseismic data show that (i) Manisa Fault Zone is responsible for six surface rupturing earthquakes since late Pleistocene-Holocene, which is occurred at 30.6 ± 8.8 ka (E1), 15.0 ± 5.0 ka (E2), 6.6 ± 1.3 ka (E3), 2.9 ± 1.3 ka (E4), 0.8 ± 0.4 ka (E5), and 0.1 ± 0.1 ka (E6), (ii) the recurrences interval of destructive earthquakes on Manisa Fault Zone was found between 0.95 kyr to 3.8 kyr for Holocene, and (iii) the elapsed time since the most recent surface ruptured earthquake on the Manisa Fault Zone is 159 year.