The present research investigated copper and cadmium stress-induced differentially expressed genes (DEGs) using annealing control primers (ACP) with the differential display reverse transcription polymerase chain reaction technique in alfalfa (Medicago sativa L. cv. Vernal) leaves. Alfalfa leaves were subjected to 250 μM of copper and cadmium treatment for a period of 6 h. A total of 120 ACPs was used. During copper and cadmium treatment, 6 DEGs were found to be up or down regulated. During copper stress treatment, 1 DEG was up-regulated, and 3 novel genes were discovered. Similarly, during cadmium stress treatment, 1 DEG was up-regulated and 5 novel genes were identified. Among all 6 DEGs, DEG-4 was identified as the gene for trans-2,3-enoyl-CoA reductase, DEG-5 was identified as the gene for senescence-associated protein DIN1 and DEG-6 was identified for caffeic acid O-methyltransferase. All the up-regulated genes may play a role in copper and cadmium stress tolerance in alfalfa.

Ⅰ. INTRODUCTION
Ⅱ. MATERIALS AND METHODS
Ⅲ . RESULTS AND DISCUSSION
Ⅳ. CONCLUSION
Ⅴ. REFERENCES

• Ki-Won Lee(Grassland & Forages Division, National Institute of Animal Science, Rural Development Administration)
• Atikur Rahman(Grassland & Forages Division, National Institute of Animal Science, Rural Development Administration)
• Muhammad Zada(Grassland & Forages Division, National Institute of Animal Science, Rural Development Administration)
• Dong-Gi Lee(Division of Life Sciences, Korea Basic Science Institute)
• Ki-Yong Kim(Grassland & Forages Division, National Institute of Animal Science, Rural Development Administration)
• Tae Young Hwang(Grassland & Forages Division, National Institute of Animal Science, Rural Development Administration)
• Hee Jung Ji(Grassland & Forages Division, National Institute of Animal Science, Rural Development Administration)
• Sang-Hoon Lee(Grassland & Forages Division, National Institute of Animal Science, Rural Development Administration) Corresponding author