Materials and Reagents

1. Arabidopsis thaliana Col-0 ecotype seeds from Arabidopsis Information Service Collection (www.arabidopsis.info)
   
2. Trichoderma harzianum CECT 2413 (Spanish Type Culture Collection, Valencia, Spain) [referred to as T34 along the paper is the strain used in this work. T. harzianum T34 is grown on Potato Dextrose Agar (PDA) and spores are maintained at -80 °C in a 30% glycerol solution]
   
3. Murashige & Skoog medium (MS), including B5 vitamins (Duchefa Biochemia, catalog number: M0255.0050 )
   
4. Sucrose (Applichem Panreac, catalog number: 141621.1211 )
   
5. 85% potassium hydroxide pellets (KOH) (Applichem Panreac, catalog number: 141515.1210 )
   
6. 0.15% agarose (Conda Pronadisa, catalog number: 8016 )
   
7. 0.39% potato dextrose agar (PDA) (Sigma-Aldrich, catalog number: P2182 )
   
8. 0.24% potato dextrose broth (PDB) (Sigma-Aldrich, catalog number: P6685 )
   
9. Potato dextrose agar (PDA) (Conda, catalog number: 1022.00 )
   
10. Potato dextrose broth (PDB) (Difco, catalog number: 254920 )
    
11. Glass wool washed QP (Panreac, catalog number: 211376.1208 )
    
12. Resma filter paper (420 x 500 mm) (Auxilab S.L, catalog number: 80250452 )
    
13. Liquid nitrogen (Air Liquide)
    
14. Ethanol (Panreac, catalog number: 161086 )
    
15. Triton X-100 (Sigma-Aldrich, catalog number: T8787 )
    
16. Sodium hypochlorite
    
17. Sterilization solution (see Recipes)
    
18. MS Medium (see Recipes)
    
19. Washing solution (see Recipes)
    

Equipment

1. Sterile distilled water purification system (EMD Millipore, model: ELIX35 )
2. Brand cotton roving (Sigma-Aldrich, catalog number: BR28205 )
3. Sterile stainless steel screen (Alunet, catalog number: 174562 )
4. Surgical Micropore tape (3M, catalog number: 1530-0 )
   
5. 1.5 ml microtubes (Deltalab, catalog number: 200400P )
   
6. Cold chamber
   
7. Phytatray II boxes [114 mm, 86 mm, 102 mm (W x D x H)] (Sigma-Aldrich, catalog number: P5929 )
   
8. Laminar flow cabinet (Telstar, model: AV-100 )
   
9. Plant Growth Chamber AGP-1400-HR (Radiber SA)
   
10. Shaker Certomat^® R (B. Braun, model: 986302/4 )
    
11. Petri dishes (90 x 14 mm) (Deltalab, catalog number: 200209 )
    
12. Surgeon carbon steel (surgical blade sterile) (Jai Surgicals, catalog number: 0835147 )
    
13. 1.5 ml microtubes with glass wool (homemade)
    
14. 15 ml Tubes (Deltalab, catalog number: 401402 )
    
15. Thoma cell counting chamber (BRAND, catalog number: 7180 05 )
    
16. Coverslip EUROTUBO (22 x 22 mm) (Deltalab, catalog number: D102222 )
    
17. Optical microscope (Leica Microsystems AG, model: DC300F; catalog number: 10447115 )
    
18. Erlenmeyer flask (Thermo Fischer Scientific, catalog number: 11972233 )
    
19. Kühner shaker (Thermo Fisher Scientific, model: ISF-1-W )
    
20. Vacuum/pressure pump PALL (Life Sciences, model: DOA-P730-BN )
    
21. Safety glass (vacuum flask, 10 ml pipette and gums) (homemade)
    
22. X5 Graduated pipette (type1, class B, ISO 835) (Thermo Fisher Scientific, catalog number: 11912178 )
    
23. Medical grade silicone tubing (1x 1.5, internal diameter x external diameter) (Deltalab, catalog number: 3500115 )
    
24. Rubber plugs (VWR International, catalog number: 217-9463 )
    
25. Forceps (stainless, L= 105 mm) (Thermo Fisher Scientific, catalog number: 10458242 )
    
26. Vacuum flask pirex (1,000 ml) (Thermo Fisher Scientific, catalog number: 12693182 )
    
27. Magnetic filter funnel (VWR International, catalog number: 516-7590 )
    
28. Scissors stainless 170 mm (Thermo Fisher Scientific, catalog number: 12693182)
    
29. Lyophilizer Virtis Advantage (SP Scientific)
    

Procedure