Position of water in vitrified plants visualised by NMR imaging

Abstract

Vitrification of plants in vitro is a physiological abnormality of tissue-cultured plants which causes significant losses in the micropropagation industry. Vitrified plants are waterlogged but the position of water within plants has not been identified. Nuclear magnetic resonance (NMR) imaging of normal tissue-cultured, vitrified tissue-cultured, and glasshouse-grown leaves ofGypsophila paniculata showed the distribution of water within the leaves. Normal tissue-cultured and glasshouse-grown leaves had a high concentration of water within leaf vascular bundles and lower concentrations elsewhere. In contrast, vitrified leaves had a relatively even distribution of high water concentration throughout the leaves. When imaging parameters were changed, so that only water associated with cell membranes was shown, the images of normal tissue-cultured and glasshouse-grown leaves did not change. However, the image of the vitrified leaves showed a general lowering of intensity across the whole of the leaf. The appearance of the NMR images, together with those obtained by light microscopy, suggest that the excess water associated with vitrified plants is located in the intercellular air spaces. The blockage of these spaces may lead to a cycle of perturbations in the plant's physiology culminating in the development of vitrification.