A field experiment was conducted in north Gondar to investigate the response of malt barley varieties to different inter-row spacing under contrasted environment. The experiment was laid out using randomized complete block design with three replications. The treatments were three different inter-row spacing (i.e., 20, 25, and 30 cm), six malt barley varieties (i.e., Holker, HB1963, Sabine, Ibone174/03, EH1847, and Freygebse), and two locations (Miligebsa and Kino). The result sowed that days to 50% heading, days to 90% physiological maturity, plant height, spike length, and number of kernels per spike were significantly (
p
<
0.01
) affected by the main effect of location and varieties. Similarly, effective tiller number, biomass yield, grain yield, and straw yield were significantly (
p
<
0.01
) influenced by the main effect of variety, inter-row spacing, and location. Important interaction was also observed between varieties × inter-row spacing × location on days to 50% heading, spike length, biomass yield, grain yield, and straw yield. Location × variety interaction significantly influences the days to 90% physiological maturity and effective tiller number. Similar significant (
p
<
0.05
) trend was also observed between varieties with inter-row spacing on effective tiller number and thousand kernel weight even at
p
<
0.01
level of significance. It was found that optimization of row spacing was found to be influenced by location and varietal difference. Hence, on the basis of the result, it can be inferred that 25 cm row spacing combined with the varieties “HB1963,” “Hollker,” and “Sabine” was optimum for maximum yielding potential under potential environment (Miligebsa).
