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Callus, induced in the dark from leaf tissue of spinach (Spinacia oleracea L. cv. Fall Green) on Murashige and Skoog (MS) medium supplemented with (in mg·liter ^-1) 2 kinetin and 0.5 2,4-D regenerated shoots upon transfer to a medium containing 2 kinetin, 0.01 2,4-D, and 1 GA₃. Complete plants were established by stimulating rooting of the shoots with 1 mg IBA/liter and transferring them to potting soil; survival was 60%. Chemical names used: N-(2-furanylmethyl)-1H-purin-6-amine (kinetin); 2;4-dichlorophenoxy acetic acid (2,4-D); gibberellic acid (GA₃); 1H-indole-3-butanoic acid (IBA).

A preliminary study has shown that the addition of 15% (v/v) coconut water (CW) to the culture medium significantly improved callus growth, shoot-regenerative capacity, and shoot growth in leaf disk cultures of spinach (Spinacia oleracea L.). Subsequently, the influence of a range of CW concentrations, 0%, 5%, 10%, 15%, or 20% (v/v), was examined. Callus weight obtained after 5 weeks showed direct relationship to the concentration of CW. This stimulator action was observed in both cultivars tested in this study, `High Pack' and `Baker'. On CW-containing medium, shoot regeneration was expedited to 4 to 5 weeks compared with 8 to 12 weeks on a CW-free medium. Callus of `Baker' induced on a CW-free medium exhibited a significant increase in shoot regeneration frequency when transferred to a regeneration medium enriched with CW, suggesting that the addition of CW to the regeneration medium only is sufficient to achieve improved regeneration.