Study on Excised Leaf Tissue Inoculation of Common Leaf Spot in Alfalfa

The excised leaf tissue inoculation (ELTI) method is studied in this paper for identification of alfalfa resistance to common leaf spot. The results showed that ELTI is more efficient and dependable in resistance identification than the traditional whole plant inoculation method. Greenness of excised leaf tissues was the best maintained by adding benzimidazole into media of 0.3% water agar at 50μm/mL. The conditions required for a good inoculation of excised leaf tissue includes relative humidity about 100%, temperature of 20℃, duration of 20 hours, and complete dark. The condition required for good culture are a 12 hours interval alternation of light (9000lux) with temperature at 22℃ and dark with temperature at 18℃.