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Isozyme Patterns in Tobacco Plant Parts and Their Derived Calli¹

This study was initiated to determine if different tissues of the same plant maintain persistent tissue specific patterns of gene expression during in vitro culture or if in vitro cultures are characterized by identical patterns of gene expression no matter what their tissue of origin. Two-millimeter sections were excised from the root, lower leaf, upper leaf, pith, sepal, ovary, and anthers of a mature ‘Wisconsin 38’ tobacco (Nicotiana tobacum L.) plant and were grown on a common medium for callus formation. Transfer to fresh medium was made when the calli reached about 8 mm in diameter. One to five such transfers were made depending upon the growth rate of different explants. Extracts from the original explants and each callus transfer were assayed for several isozyme systems using horizontal starch-gel electrophoresis. Malate dehydrogenase and peroxidase isozymes were found to have good xesointion and to be suitable for the study. The patterns of the isozymes displayed tissue specific differences at the explant level; however, substantial differences between the isozyme patterns of the calii and that of the original explant were displayed. Initially there were differences in isozyme patterns among the calli from different origins and within the different transfers of the same explant callus. However, after the second or third callus transfer, isozyme patterns became more uniform for calli of the same explant.

Key Words: Nicotiana tabacum L. • Starchgel electrophoresis • Peroxidases • Malate dehydrogenases • Tissue specificity

² Professors.

Received for publication December 5, 1978.