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Table 1 Technical tricks for successful wedge-style inflorescence grafting in Arabidopsis

From: Inflorescence stem grafting made easy in Arabidopsis

Step

Description of method

Tips and tricks

1

Plant Growth Before Grafting

Vernalise seedlings to synchronise the initiation of floral meristem development.

  

Use young inflorescence stems of uniform age, length (<10 cm), and diameter (1.5 to 3 mm).

2

Horizontal Excision of Stem

Use flexible razor blade to cut the floral stem below the cauline node (2 cm above the shoot meristem) and immediately place scion in sterile water.

3

Preparation of Root Stock Stem

Slide 1 cm of soft and flexible silicone tubing (sizes range from 1–3 mm ID) over the root stock stem.

  

Cut vertical incision into the root stock stem.

4

Preparation of Scion Stem

Trim the scion stem using flexible razor blade into a wedge.

5

Join and Support Grafted Stems

Place scion wedge into root stock stem incision.

  

Slide the silicone tubing over the graft junction.

  

Inject sterile water on top of silicone tube to maintain turgor.

  

Pre-stretch parafilm and wrap over the graft junction supported by silicone tube so as to avoid desiccation.

6

Plant Growth After Grafting

Maintain high humidity (70%) for 1–2 weeks.

  

Slowly acclimate grafted plants to normal growth conditions.

  

Clip new rosette shoot growth to promote further development of the primary grafted floral stem.

  

Remove meristems showing reproductive siliques to promote new cauline shoot growth of new tissues.

7

Evaluation of Grafting Success

Grafting was considered successful when scions remained green and health on top of the rootstock as evident by the maintenance of apical dominance, development of new cauline leaves and branches, as well as production of reproductive siliques.

  

Use the inflorescence growth assay to select the best and most successful grafted plants for further analysis.