Abstract: To explore the effects of carbon sources and other main factors on the proliferation of embryogenic callus in Pinus elliottii,and provide technical methods and theoretical basis for further improving proliferation efficiency. The embryogenic callus of the elite clone PEE3-13 was used,the effects of different types and concentrations of carbon sources,pH and the organic nitrogen sources,L-glutamine and its substitutes,Lalanine-L-glutamine on its proliferation were studied. The pH and the type of carbon source had significant effects on its proliferation. The autoclaved medium pH ranged from 5.68 to 6.28,it was the most suitable for the embryogenic callus growth. The maximum proliferation rate was achieved when using 30 g·L^-1 white sugar was used as carbon source. When 450 mg·L^-1 L-glutamine was added,the embryogenic callus had the highest proliferation rate and the best activity,however,L-alanine-L-glutamine could not replace L-glutamine. The optimal medium was 30 g·L^-1 white sugar and 450 mg·L^-1 L-glutamine and autoclaved pH was ranged from 5.68 to 6.28 and was most suitable for embryogenic callus proliferation,and the maximum proliferation rate was h851.27%.