Materials and Reagents

1. 3-4 weeks old Cucumis melo L. subsp. melo cv. Galia plants
   
2. Agrobacterium tumefaciens (A. tumefaciens) strain C58C1, or similar, transformed with the binary vector harboring the GFP-tagged viral genome
   In our case, pMNSV(Al)/GFP encoding GFP-tagged Melon necrotic spot virus (MNSV) genome under the control of the Cauliflower mosaic virus (CaMV) 35S promoter and the potato proteinase inhibitor terminator (PoPit) (Figure 1).
   
   
   Figure 1. Schematic representation of the recombinant infectious clone pMNSV(Al)/GFP used in this assay
   
   
3. Yeast extract (Difco, catalog number: 212750 )
   
4. Tryptone (Difco, catalog number: 211705 )
   
5. Sodium chloride (NaCl) (Panreac Applichem, catalog number: 131659 )
   
6. Acetosyringone (Sigma-Aldrich, catalog number: D134406 )
   
7. MES (Sigma-Aldrich, catalog number: M8250 )
   
8. Magnesium chloride (MgCl₂) (Sigma-Aldrich, catalog number: M9272 )
   
9. Antibiotics
   
10. LB medium (see Recipes)
    
11. Agrobacterium infiltration buffer (see Recipes)
    

Equipment

1. 28 °C growing chamber
   
2. 15 ml culture tubes
   
3. Swinging centrifuge rotor for 15 ml tubes
   
4. 1.5 ml tubes (standard Eppendorf tubes or similar)
   
5. BioPhotometer plus (Eppendorf, catalog number: 6132000008 )
   
6. 1 ml syringes without needle
   
7. Plant growing chamber
   
8. Leica MZ12 fluorescent stereo microscope
   

Software

1. Adobe Photoshop CS5 or higher
   
2. ImageJ, FIJI or similar software
   
3. MS Excel
   

Procedure