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This study was conducted to compare and estimate the genetic variability among durum wheat lines in response to drought according to their grain yields and physiological traits. The use of fluorescence parameters as drought selection criteria for durum wheat was investigated in a population of 249 recombinant inbred lines (RILs) derived from a cross between two cultivars of durum wheat: ‘Svevo’ and ‘Kofa’. The durum wheat RILs considered were analyzed to determine the relationship between their grain yields and chlorophyll fluorescence parameters at the grain filling stage under drought stress (rainfed) and supplemental irrigation (irrigated) conditions during the 2004 and 2005 seasons at the ICARDA center. The results obtained show that the durum wheat line performance was influenced by drought stress, indicating significant differences between the grain yields and fluorescence parameters recorded under drought rainfed (RF) and irrigation (IR) conditions. Significant differences between the experimental groups of durum wheat RILs (P<0.05) were only recorded under drought conditions, with the exception of leaf water potential (Lwp). Under such conditions, nearly all the parameters examined were significantly increased in the high-yielding group (with the exception of the Que parameter), thus revealing the genetic variability of the durum wheat lines considered in response to drought stress. The potential quantum efficiency of photosystem II (Fv/Fm) was found to be positively associated with the grain yield parameter. The mean values of Fv/Fm in both the high- and low-yielding groups significantly dropped under drought stress (0.71 and 0.68, respectively) compared to the Fv/Fm values recorded under irrigated conditions (0.80 and 0.81). Under drought conditions, slopes were highly significant (P<0.001) nearly for all the fluorescence parameters examined (with the exception of CHLSPAD) compared to those recorded under irrigation conditions. It was concluded that chlorophyll content (SPAD), F0, Fm, Fv, Fv/Fm, Lwp, and Que could be used as additional indicators in screening wheat germplasm for drought tolerance.