In this study, the principal objective was to investigate the effect of light quality and vessel ventilation on the growth and development, physiology, activities of antioxidant enzymes, and contents of mineral nutrients of carnation (Dianthus caryophyllus L.) ‘Marble Beauty’. Single node cuttings stuck into the plant growth regulator (PGR)-free MS medium in containers covered with caps with or without a ventilation filter were cultured first four weeks under white and then additional four weeks under either white (control), blue, red, or red + blue light emitting diodes (LEDs) for 56 days. Interestingly, a ventilated culture condition not only reduced the percentage of the hyperhydricity, but also increased the total chlorophyll content (Chl a + Chl b) of the plantlets as compared to the non-ventilated condition. In addition, blue LEDs produced plantlets with the greatest number of shoots and red LEDs produced plantlets with the greatest shoot length. The quality of plantlets was improved under a ventilation condition. Besides, under a ventilated condition, red + blue LEDs raised APX activity, and blue LEDs not only raised the activity of the CAT, but also increased tissue contents of such elements as K, Ca, Mg, Zn, Mn and Fe. The red LEDs increased contents of B and Si under a ventilated condition, and Na accumulation under a non-ventilated condition. Thus, including blue or red LEDs as the light source in a ventilated culture condition will produce plantlets of carnation ‘Marble Beauty’ in vitro with improved quality.