This investigation was planned to select and develop date palm plants from selected resistant callus variants to wilt disease caused by Botryodiplodia theobromae. The chemical analysis of selected date palm resistant plants was also investigated. Virulent strains of B. theobromae fungus was tested as a selective agent for use in vitro selection for resistance by measuring the response of explants (callus) of two date palm genotypes (Bartamoda and Samani). It was obviously dear that the higher concentrations of culture filtrate (CF) B. theobromae showed destructive effect on the callus growth. Bartamoda cv. recorded the highest survival percentage of callus growth (74.99%) comparing with Samani cv. (41.66%). Cain culturing on culture filtrate of causal fungus increased reducing sugars content with Samani cv. and reduced those sugars with Bartamoda cv. While pathogen culture filtrate mostly decreased non-reducing sugars into treated callus of both Samani and Bartamoda compared with control. Increasable rates of 8.theobromae culture filtrate than 5 % decreased total sugars into treated callus of both cultivars. Increasable rates of pathogenic fungus culture filtrate increased amino acids content and total phenol contents. Different concentrations of culture filtrate decreased indoles content in treated callus of the two cultivars. Use of descending rates of 8.theobromae culture filtrate treatments resulted in reasonable resistance at the rates of 5 and 7.5% v/v. Higher rate (10%) highly decreased growing individuals. Whereas lowest rate (2.5 %) resulted in complete growth of treated callus.