E. colona is a C4 annual summer grass which is troublesome to major summer annual and perennial crops. Due to recent complaints by the farmers, the objectives of the present study were to evaluate the efficacy of penoxsulam, profoxydim, cycloxydim, cyhalofop-butyl, florpyrauxifen-benzyl and glyphosate against six E. colona accessions, and also to evaluate the response of these accessions to different rates of glyphosate in a dose-response experiment. In the first experiment, herbicides were applied at their maximum recommended label rates, while in the dose-response experiment, glyphosate was applied at six doses corresponding to 0, 1/4X, 1/2X, X, 2X, and 4X of the recommended rate. The dry weight of the biotypes TH8 and TH7 treated with profoxydim was 66% and 68% of the untreated control, respectively. The efficacy of cyhalofop-butyl against three accessions was lower than 30%, while two accessions were susceptible to this herbicide. The efficacy of penoxsulam against the biotypes ET2 and ET4 was lower than 10%, while dry weight of FT5 and TH8 was only reduced by 23%–28% as compared to the control. Cycloxydim application provided control higher than 75% at 21 days after treatment (DAT) of three accessions, while the majority of E. colona accessions was adequately controlled by the application of florpyrauxifen-benzyl. The response of the different accessions to glyphosate varied. The results of the glyphosate dose-response experiment revealed that the GR50 values of the resistant E. colona accessions ET2 and ET4 were up to 1098 and 1220 g a.e. ha−1 of glyphosate, respectively, whereas the GR50 value of the susceptible accession (FT5) was only 98 g a.e. ha−1. The resistance indices of ET2 and ET4 were 12.4 and 11.2, respectively, indicating that they have already developed resistance to glyphosate. Three more accessions could be also of developing resistant to glyphosate. This is the first report of glyphosate resistance from E. colona accessions in Greece, with indications of multiple resistance also present. Further research is needed in order to evaluate the efficacy of several herbicides under different soil and climatic conditions, conduct baseline sensitivity studies, reveal the evolvement of resistance patterns to glyphosate from accessions of Echinochloa spp., and search for alternative options of weed management in annual and perennial crops due to the clear indications of multiple resistance situations.