1. EPSPS and Crops Developed by Dr. M. Zapiola for CSS/Hort 431

2. Glyphosate inhibits EPSPS enzyme - impeding the synthesis of aromatic amino acids EPSPS Glyphosate Herbicide EPSPS Proteins CO 2 H 2 O NH 3 aromatic amino acids Proteins CO 2 H 2 O NH 3 aromatic amino acids glyphosate

3. Commercial RR crops "Two key elements needed for the development of commercially viable glyphosate-tolerant crops are: – a resistant target enzyme – sufficient expression of that enzyme within the transgenic plant” Heck et al. 2005. Development and Characterization of a CP4 EPSPS-Based, Glyphosate-Tolerant Corn Event: Crop Sci. 45:329-339 (2005).

4. EPSPS Roundup Ready Crops When incorporated into the genome of RR- susceptible plants, the Roundup Ready TM gene (CP4 EPSPS) confers resistance to glyphosate Proteins CO 2 H 2 O NH 3 aromatic amino acids glyphosate CP4 EPSPS

5. Gene Construct The gene is introduced into the plant nuclear genome via different mechanisms The trait is then incorporated to elite varieties by backcrossing promoter(leading sequence)resistant EPSPS terminator

6. Gene Construct: RR soybean LOCUS AB209952.1 Size 2457 bp SOURCE Glycine max FEATURES – CaMV35S – Chloroplast transit peptide – CP4EPSPS – Nos 3’ terminator – Repeated fragment of cp4epsps truncate Gene bank gi|62318478|dbj|AB209952.1|[62318478]

7. PCR Primer Design Nametm Product size CaM_1F60117 CaM_1R60 CaM_2F60121 CaM_2R60 CTP_1F60150 CTP_1R60 CP4_F60286 CP4_R60 Nos_F60216 Nos_R58

8. Scanning 96 Plants