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. 2025 Feb 20;15(4):e5215.
doi: 10.21769/BioProtoc.5215.

Micrografting Technique of Hevea brasiliensis in Vitro Plantlets

Florence Dessailly  1   2 , Pascal Montoro  1   2 , Sémi Melliti  1   2 , Julie Leclercq  1   2
Affiliations

Micrografting Technique of Hevea brasiliensis in Vitro Plantlets

Florence Dessailly et al. Bio Protoc. .

Abstract

To prepare Hevea brasiliensis plantations, selected planting material is propagated by grafting using illegitimate seedlings as rootstocks, whose paternal genotype is unknown. Recent advances in rubber tree in vitro cloning propagation open the possibility of using these techniques to supply new planting material. Micrografting is a promising technique to speed up the preparation of plant material for rootstock-scion interaction studies. This article describes the implementation of an efficient micrografting technique from Hevea in vitro plants from clone PB 260. The procedure combines several conditions to preserve the root system and the grafted scion and to prevent any breakage of rootstock buds. This technique paves the way for clonal propagation and holds potential for further development on other rubber clones for further studies on the interaction between rootstock and scion. Key features • This protocol requires rejuvenated in vitro plantlets. • Requires between 40 and 60 days before scion bud break. • Sterile working conditions. Graphical overview.

Keywords: Budding; Microcutting; Propagation; Rootstock; Rubber; Scion; Somatic embryogenesis.

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Conflict of interest statement

Competing interestsThe authors declare that they have no financial and non-financial competing interests.

Figures

Figure 1.
Figure 1.. Folding the filter paper into the glass tube.
A. Place the paper disc on the lid of a glass tube. B. Fold the paper disc over the lid. C. Pierce the center of the folded disk with a pointed object. This will allow the taproot to pass through. D. Insert the folded disc into the tube that will contain the grafted seedling.
Figure 2.
Figure 2.. Plant material preparation.
A. In vitro plantlet at the right stage of development. B. Rootstock after pruning of cotyledons and taproot. C. Grafting window preparation. D. Scion at the correct size equivalent to the diameter of the rootstock.
Figure 3.
Figure 3.. Grafting steps in optimal conditions.
A. Placement of the graft in the slit and protection with a piece of solid medium. B. Correct positioning of the grafted plant into the filter paper base.
Figure 4.
Figure 4.. Graft re-growth, acclimatization, and plant development in the greenhouse.
A. Graft re-growth. B. Grafted plantlet with fully developed leaves. C. Acclimatization step in the greenhouse. D. Development of the grafted plant after one year.
Figure 5.
Figure 5.. Effect of rootstock culture medium, the treatment at the junction, and the pruning of the rootstock buds on the development of the grafted buds.
A. Histograms showing the effectiveness of treatments on root damage, graft survival, bud breaking, and acclimatization in the greenhouse. B. Details of treatments applied and number of grafted plants used. Statistical analyses were carried out using a parametric k-proportions comparison. Values with the same letter are not significantly different at the 0.05 probability level.
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References

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