In this study, copper sulfate (CuSO4) was evaluated as affected factor on date palm plantlet in tissues culture on emission of ethyl acetate which considers the important volatile organic compounds (VOCs) of kairomone for red palm weevil, some chemical compounds and some activity of antioxidant enzymes. Plantlets were treated with different concentration with CuSO4 (0, 5, 10, 20mg/L tissue culture media). The treated plantlets with each concentration of CuSO4 were divided to healthy plantlets and mechanically injured and incubated at growth room. GC-MS analysis was used to estimation of ethyl acetate emission in plantlets samples and the high level of emesis ethyl acetate at injured plantlets with free of CuSO4. Biochemical compounds were estimated and the results referred to high level of phenol was at the treated plantlets with CuSO4 (5mg/l) with injured while high level of total indol and GA3 were at treated plantlets with CuSO4 (5mg/l) with uninjured. High level of amino acid and flavonoids were recorded at untreated plantlets with CuSO4 in uninjured plantlets in amino acid and injured in flavonoids. High level of activity antioxidants enzymes (SOD, CAT, and ADH) and the proline levels were recorded in the injured plantlets with 20mg/l of CuSO4. Chlorophyll a and b levels were the highest at treated plantlets with 20mg/L CuSO4 with uninjured samples