Improving micropropagation of some grape cultivars via boron, calcium and phosphate

Abstract

Background: An efficient regeneration protocol is a priority for the successful application of plant biotechnology. Grape nodal explants were used to develop a micropropagation protocol for Thompson Seedless and Taify cvs. Explants were cultured on MS medium supplemented with Kinetin or benzylaminopurine (BA) and indolebutyric acid (IBA).

Results: For both cultivars, axillary buds were grown, only, on a medium enriched with kinetin, moreover, shoot tip necrosis and callus formation were observed on Thompson Seedless cv. cultures grown on a medium with BA. Supplementing the growth medium with 100 mM (boron) B and 2.5 mM (calcium) Ca successfully help overcome these phenomena. The highest regenerated shoot numbers (14 and 6.2 explant⁻¹) for Taify and Thompson Seedless cvs., respectively, were on media supplemented with 13.2 µM BA + 4.9 µM IBA and BA 13.2 µM + 5.8 µM IBA, respectively. Moreover, these media supported the developing shoots to have the heaviest dry weights (1.46 and 0.72 mg explant⁻¹) for Taify and Thompson Seedless cvs., respectively. Thompson Seedless cv. regenerated shoot numbers and their dry weights were significantly increased by increasing the MS medium PO₄⁻ concentration. However, these two parameters were significantly decreased for Taify cv. Developing shoots were elongated and rooted on MS medium enriched with 4.9 µM, IBA 100 mM B and 2.5 mM Ca. Plantlets were acclimatized and successfully transferred to the greenhouse conditions.

Conclusions: A novel promising protocol for Thomson Seedless and Taify cvs. micropropagation using single nodes has been developed.