Immunogold localization of trehalose-6-phosphate synthase in leaf segments of wild-type and transgenic tobacco plants expressing the AtTPS1 gene from Arabidopsis thaliana.
Following the establishment of a transgenic line of tobacco (B5H) expressing the trehalose-6-phosphate synthase (TPS) gene from Arabidopsis thaliana, a preliminary immunolocalization study was conducted using leaves of adequately watered B5H and wild-type plants. Immunocytochemical staining, followed by electron microscopy showed that the enzyme could be detected in both B5H and wild-type plants at two different levels. Quantification showed the signal to be two to three times higher in transgenic plants than in the wild type. This enzyme was markedly present in the vacuoles and the cell wall, and to a lesser extent in the cytosol. Moreover, a high profusion of gold particles was detected in adjacent cells and in the sieve elements. Occasional spots were also detected in chloroplasts and the nucleus, especially in the transgenic B5H line. No labeling signal was detected in mitochondria. Protein localization seems to confirm the important role of TPS in sugar metabolism and transport through the plant, which could explain its role in plant stress tolerance. Finally, it can be expected that TPS from tobacco has a relatively high similarity to the TPS of Arabidopsis thaliana.
Almeida
A M
AM
Laboratório de Biotecnologia de Células Vegetais, Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal. amalmeid@itqb.unl.pt
Santos
M
M
Villalobos
E
E
Araújo
S S
SS
van Dijck
P
P
Leyman
B
B
Cardoso
L A
LA
Santos
D
D
Fevereiro
P S
PS
Torné
J M
JM
D017360:Arabidopsis / Q000235:genetics*; D015152:Blotting, Northern; D015153:Blotting, Western; D015870:Gene Expression; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism*; D007150:Immunohistochemistry; D008954:Models, Biological; D018515:Plant Leaves / Q000378:metabolism*; D030821:Plants, Genetically Modified; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism
D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase