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Lettuce (Lactuca sativa) is commonly produced in vertical farms. The levels of nutritionally important phytochemicals such as beta-carotene (precursor to vitamin A) are generally low in lettuce. In this study, we investigated the benefits of variable lighting strategy (i.e., varying the light quality during production) on maintaining plant growth and increasing the biosynthesis of beta-carotene and anthocyanin. We tested two variable lighting methods, using green and red romaine lettuce, namely (i) providing growth lighting (supports vegetative growth) initially (21 days) followed by a high percentage of blue light (supports biosynthesis of phytochemicals) at final stages (10 days) and (ii) providing a high percentage of blue light initially followed by growth lighting at final stages. Our results indicate that the variable lighting method with initial growth lighting and high percentage of blue at final stages can maintain vegetative growth and enhance phytochemicals such as beta-carotene in green romaine lettuce while both variable lighting methods were not effective in red romaine lettuce. In green romaine lettuce, we did not observe a significant reduction in shoot dry weight but there was an increase in beta-carotene (35.7%) in the variable compared to the fixed lighting method with growth lighting for the entire duration. The physiological bases for differences in vegetative growth and synthesis of beta-carotene and anthocyanin in the variable and fixed lighting methods are discussed.