Table 1.
Media recipes for Agrobacterium growth and induction and pathogen growth
Reagent | Amount per liter |
---|---|
523 Medium (1 L)a | |
Sucrose | 10 g |
Casein enzymatic hydrolysate | 8 g |
Yeast extract | 4 g |
K2HPO4 anhydrous | 2 g |
MgSO4 anhydrous | 0.15 g |
Induction medium (recipe per 30 mL volume)b | |
Liquid ED (recipe described in [30]) | 30 mL |
0.1 M acetosyringone (Sigma Cat. # D134406) | 30 μL |
l-Proline | 0.00465 g |
20 % V8 Media (1 L)c | |
Bacto Agar | 15 g |
Calcium carbonate (CaCO3) | 3 g |
Campbell’s V8 Vegetable Juice | 200 mL |
aAdd distilled water to 1 L. Adjust pH to 7.1 and autoclave
bAdjust pH to 5.25–5.3 using 0.1 M KOH. Discard if pH exceeds 5.32. Do not adjust pH using HCl. Prepare induction medium on morning of leaf infiltration experiment. Use liquid ED less than 30 days old
cAdd distilled water to 1 L. Adjust pH to 7.1 and autoclave. Shake frequently while pouring into petri dishes to maintain homogeneity of media. Pour about 20 mL of media into each plate to ensure that agar plugs and do not fall during leaf assay