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Annals of Botany 87: 47-52, 2001
© 2001 Annals of Botany Company

Using Confocal Laser Scanning Microscopy to Measure Apoplastic pH Change in Roots of Lupinus angustifolius L. in Response to High pH

Q. Yu, J. Kuo and C. Tang+

Centre for Legumes in Mediterranean Agriculture/Soil Science and Plant Nutrition, The University of Western Australia, Nedlands, WA 6907, Australia Centre for Microscopy and Microanalysis, The University of Western Australia, Nedlands, WA 6907, Australia College of Biotechnology, South China Agricultural University, Guangzhou, 510642, China

Received: 11 April 2000 ; Returned for revision: 19 June 2000 . Accepted: 12 September 2000


  Abstract

A confocal laser scanning microscopy method to measure apoplastic pH in situ of the tap roots of Lupinus angustifolius L. is described. Pseudo-ratiometric imaging of cell walls of lupin tap roots was achieved by loading living roots with a pH-sensitive dye, Cl-NERF-dextran, and a pH-insensitive dye, Texas Red-dextran, as a reference. In 5-d-old tap roots which had previously been grown at pH 5.4, the apoplastic pH in the elongating zone was 5.2 ± 0.06. The apoplastic pH in roots grown at pH 7.5 increased to 5.5. The method was validated by supplying different forms of nitrogen in the nutrient solution. With Formula nutrition the apoplastic pH was lowered by 0.3 units while when Formula was supplied the apoplastic pH increased by 0.2 units. Causes of the poor root growth of L. angustifolius at high pH are discussed. Copyright 2001 Annals of Botany Company

Apoplastic pH, Cl-NERF, confocal laser scanning microscopy, high pH, lupin roots


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