• Applied Microscopy
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• v.36 no.3
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• pp.217-226
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• 2006

Image processing by ultra high voltage electron microscopy (UHVEM) and tomography has offered major contributions to research in the field of cellular ultrastructure. Furthermore, such advancements also have enabled the improved analysis of three-dimensional cellular structures in botany. In the present study. using UHVEM and tomography, we attempted to reconstruct the three-dimensional images of plastid inclusions that probably differentiate during photosynthesis. The foliar tissues were studied Primarily with the TEM and further examined with UHVEM. The spatial relationship between tubular elements and the thylakoidal membrane and/or starch grains within plastids mainly have been investigated in CAM-performing Sedum as well as in $C_4$ Salsola species. The inclusion bodies were found to occur only in early development in the former, while they were found only in mesophyll cells in the latter. The specimens were tilted every two degrees to obtain two-dimensional images with UHVEM and subsequently comparison has been made between the two types. Digital image processing was performed on the elements of the inclusion body using tilting, tomography, and IMOD program to generate and reconstruct three-dimensional images on the cellular level. In Sedum plastids, the inclusion bodies consisted of tubular elements exhibiting about 20 nm distance between elements. However, in Salsola, plastid inclusion bodies demonstrated quite different element structure, displaying pattern, and origin relative to those of the Sedum. The inclusion bodies had an integrative relationship with the starch grains in both species.