The cross-laminated timber (CLT) construction system has recently emerged as an excellent alternative for civil construction. The objective of this work is to analyze the structural performance of CLT panels using plantation lumber, especially eucalyptus (Eucalyptus grandis) heartwood and pine (Pinus taeda). After visual grading, all boards were mechanically graded through the ultrasonic nondestructive testing method. Boards were organized to compose four types of three-layered CLT panels: 1) exclusively eucalyptus heartwood (EEE), 2) eucalyptus in the outer layers and pine in the central layer (EPE), 3) exclusively pine (PPP), and 4) pine in the outer layers and eucalyptus in the central layer (PEP). Three panels with graded timber were manufactured for each type, and one more panel was made out of ungraded timber, so each group had four panels altogether. Panels containing eucalyptus in the outer layers (EEE and EPE) were stiffer than the ones with pine in outer layers (PPP and PEP). However, the first two groups presented lower bending strength than the second ones. Modulus of elasticity and modulus of rupture results were compared to values observed in the literature and to the international standard that regulates CLT (American National Standards Institute/American Plywood Association PRG 320). From the four types studied, only panels containing mostly eucalyptus (EEE and EPE) could meet the PRG 320 E2 class. Panels containing mostly pine (PPP and PEP) did not reach the thresholds of any class in terms of stiffness although their resistance was much higher than that specified in the standard.