Assessment of Al3+ availability in callus culture media for screening tolerant genotypes of Cynodon dactylon

• Published in: Plant cell, tissue and organ culture tissue and organ culture, 1999, Vol.56 (1), p.65-68
• Main Authors: Ramgareeb, S, Watt, M.P, Marsh, C, Cooke, J.A
• Format: Article
• Language: English
• Subjects: 2,4-D; aluminum; Biological and medical sciences; Biotechnology; callus; culture media; Cynodon dactylon; dose response; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; genetic variation; genotype; in vitro culture; Methods. Procedures. Technologies; micropropagation; mine spoil; Miscellaneous; phytotoxicity; Plant cells and fungal cells; rapid methods; regenerative ability; revegetation plants; screening; soil stabilization
• ISSN: 0167-6857; 1573-5044
• DOI: 10.1023/A:1006248916412

Aluminium-tolerant genotypes of Cynodon dactylon are potential candidates for the vegetation of gold mine tailings in South Africa. As a prerequisite to in vitro selection of tolerant genotypes, this work aimed at assessing and adapting micropropagation media to ensure Al3+ activity and toxicity. This was investigated using MINTEQA2, a chemical equilibrium speciation model. The maximum Al3+ activity achieved in any medium was 7.5 micromolar. Of the seven published media investigated, four never achieved an activity greater than 4 micromolar at 3-4 mM aluminium. The most appropriate medium was that of Yamamoto et al. (1996) (modified MS without KH2PO4 and EDTA), as it showed an increasing range of Al3+ activities from 2 to 7.5 micromolar at aluminium concentrations from 0.25-2.5 mM. An improved modified MS formulation retaining phosphate was investigated because phosphate is an important component of our medium for callus induction in C. dactylon. Using MINTEQA2, no reduction in Al3+ activity by phosphate was detected in standard MS medium at pH 4. Through further simulations a new modified MS medium was derived with 1 mM SO2(-4) and no EDTA at pH 4, which gave the maximum Al3+ activity (7.5 micromolar) at 2 mM aluminium. This medium gave the highest Al3+ activities for the 0.25-2 mM concentration range of all the tested formulations, including the seven published media. It also resulted in significantly higher callus growth rates than standard MS media and other tested media. This new medium is currently being used to screen C. dactylon for aluminium tolerance at pH 4.